TCF7L2 gene polymorphism as a risk for type 2 diabetes mellitus and diabetic microvascular complications.

BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is a chronic metabolic condition with various genetics and environmental influences that affects the capacity of the body to produce or use insulin resulting in hyperglycemia, which may lead to variable complications. It is one of the world's rising health problems. There is emerging evidence that some genetic polymorphisms can impact the risk of evolving T2DM. We try to determine the relationship of (rs7903146) variant of the Transcription factor 7-like 2 (TCF7L2) gene with T2DM and its microvascular complications. METHODS AND RESULTS: This case-control study included 180 subjects: 60 diabetic patients without complications, 60 diabetic patients with microvascular complications and 60 matched healthy controls. Genotypes of rs7903146 (C/T) SNP in the TCF7L2 gene were evaluated by real-time polymerase chain reaction via TaqMan allelic discrimination. Logistic regression was used to detect the most independent factor for development of diabetes and diabetic microvascular complications. Variant homozygous TT and heterozygous CT genotypes were significantly increased in diabetic without complications and diabetic with complications groups than controls (p = 0.003, 0.001) respectively. The T allele was more represented in both patient groups than controls with no significant difference between patient groups. TT genotype as well as T allele was significantly associated with increased T2DM risk. CONCLUSION: The T allele of rs7903146 polymorphism of TCF7L2 confers susceptibility to development of T2DM. However, no significant association was found for diabetic complications.

Study the Association of Tumor Necrosis Factor Promoter Polymorphism with Type 2 Diabetic Nephropathy.

Type 2 Diabetes Mellitus (T2DM) is well known to include an inflammatory component that has been considered to be related to diabetic complications. Diabetic nephropathy (DN) is one of the significant complications as it constitutes the most frequent cause of end-stage renal disease. Tumor Necrosis Factor-α (TNF-α) is known as a multifunctional proinflammatory cytokine which is associated with some pathological processes such as immunoregulation, proliferation, inflammation, and apoptosis. The aim was to explore the association between the TNF-α promoter -1031T/C single nucleotide polymorphism (SNP) and the serum TNF-α level in addition to nephropathy among type 2 diabetic patients. The study included 38 T2DM subjects without nephropathy (DM group), 40 subjects with DN, and 20 controls. Identification of TNF-α promoter gene polymorphism -1031T/C was done by PCR-RFLP, and genotyping was confirmed by direct sequencing. The serum TNF-α level was assessed by ELISA. Correlations were tested by Pearson's correlation analysis. Logistic regression was used to detect the most independent factor for development of DN. The serum level of TNF-α in the DM group was significantly higher than controls (p < 0.001); also, the DN group was considerably higher than controls and DM without nephropathy (p < 0.001). Also, there was a significant positive correlation between serum levels of TNF-α with FBG (fasting blood glucose), creatinine, total cholesterol, LDL-C, HbA1c, and microalbumin/creatinine ratio (ACR) among the DN group (p = 0.042, <0.001, <0.001, <0.001, 0.027, and 0.043, respectively). Mutant homozygous CC and heterozygous TC genotypes were higher in DN than in DM and controls. C allele was more represented in DN than in DM and controls (p = 0.003) while T allele was higher in controls than in DM and DN patients. The levels of TNF-α were higher in subjects who had mutant CC than the wild TT genotype among DN (p < 0.001). C allele was more risky for DN than T allele between DN and controls by 5.4-fold (CI: 1.75-16.68) as well as between DN and DM by 2.25-fold (CI: 1.1-4.59). Conclusion. Serum levels of TNF-α were higher in individuals with mutant CC genotype of -1031T/C TNF-α gene, and C allele could be associated with increased risk for nephropathy among patients with T2DM.

Telomere length and 8-hydroxy-2-deoxyguanosine as markers for early prediction of Alzheimer disease.

BACKGROUND: Becoming shorter by each cell division, telomere length (TL) is regarded as a marker of cellular aging. Relative TL (T/S) depends on the quantitation of telomere hexamer repeat copy number normalized to autosomal single-copy gene copy number. TL is influenced by several factors, including oxidative stress (OS) and inflammation. This study aimed to investigate the possible role of TL and OS as markers for Alzheimer's disease (AD). MATERIALS AND METHODS: One hundred and eighty participants were categorized into three groups. Group 1: Included 60 patients with AD. Group II: included 60 age-matched nondemented subjects. Group III (pregeriatric group): included 60 healthy controls with their ages ranging between 30 and 60 years. TL was determined by the quantitative Real time-PCR method, plasma levels of 8-OHdG by enzyme-linked immunosorbent assay, and total antioxidant capacity (TAC) by colorimetery. RESULTS: In comparison to the other two groups, patients with AD showed shortened TL, increased plasma 8-OHdG concentration, and decreased TAC. The sensitivity of T/S ratio to predict AD was 86.67%, whereas the specificity was 96.67%. The sensitivity of 8-OHdG to predict AD was 96.67%, whereas the specificity was 86.67%. CONCLUSION: AD is associated with shortened TL and increased OS as manifested by decreased TAC and increased serum 8-OHdG. T/S and 8-OHdG could be used as early predictors for AD.

T-Regulatory Cell Subsets in Children with Type 1 Diabetes Mellitus: Relation to Control of the Disease.

BACKGROUND: Type 1 diabetes mellitus is described as a chronic metabolic disorder characterized by aggressive immune &#946;-cell destruction. There are a number of varied immune mechanisms for sustaining self-tolerance in opposition to the autoimmune disorders. A recessive tolerance is accomplished by thymic gland via a negative assortment of different clones, while a dominant tolerance is accomplished by the regulatory T cells (Treg) in the periphery. Treg (CD4+ CD25+FOXP3+) are subsets of T cells which have an essential role in maintaining tolerance. OBJECTIVE: To evaluate peripheral Treg (CD4+; CD25+; FOXP3+) in children cohort with T1DM. METHODS: This study included 64 children diagnosed with T1DM and 35 age- and sex-matched healthy children as controls. All children were clinically evaluated and subjected to assessment of complete blood count (CBC), glycated hemoglobin, surface and cytoplasmic detection of Treg by flow cytometry. RESULTS: This study showed that the frequency of Treg (CD4+; CD25+; FOXP3+) was significantly lower in diabetic children than with normal controls (P<0.001). There was a significant (P <0.001) reduction in the Treg (CD4+; CD25+; FOXP3+) in T1DM children with uncontrolled (Hemoglobin A1c>7%) as compared to those with controlled (Hemoglobin A1c<7%) disease. CONCLUSION: Diminished Treg in T1DM proved that auto-reactivation of T-cell as a result of the breakdown of immune tolerance takes part in the elaboration of autoimmune disorders as T1DM. Treg may be used in immunotherapy, thus preventing T1DM development due to its pivotal role in immune tolerance.

Serum PAI-1 and PAI-1 4G/5G Polymorphism in Hepatitis C Virus-Induced Cirrhosis and Hepatitis C Virus-Induced Hepatocellular Carcinoma Patients.

Association between variable agent-induced hepatocellular carcinoma (HCC) and both PAI-1 4G/5G polymorphism and plasminogen activator inhibitor (PAI-1) levels compared to healthy controls have been reported in earlier studies. We aimed to assess serum PAI-1 and PAI-1 4G/5G polymorphism in hepatitis C virus (HCV)-induced HCC, HCV-induced liver cirrhosis, and viral infection-free apparently healthy control subjects. Forty nine HCC, 52 cirrhosis, and 105 controls were genotyped for PAI-1 4G/5G using an allele-specific polymerase chain reaction analysis. In addition, for 31 HCC, 24 cirrhosis, and 28 controls, serum PAI-1 level was measured by enzyme-linked immunosorbent assay (ELISA). There was no significant difference in PAI-1 4G/5G genotype distribution between cirrhosis and controls (p = 0.33, p = 0.15, and p = 0.38 for the codominant, dominant, and recessive models, respectively) or between HCC and cirrhosis (p = 0.5, p = 0.24, and p = 0.69 for the codominant, dominant, and recessive models, respectively). Serum PAI-1 was significantly higher in cirrhosis than controls and significantly lower in HCC than cirrhosis (p < 0.001 for both). Serum PAI-1 did not differ significantly among the three PAI-1 4G/5G genotypes in controls, cirrhosis, and HCC (p = 0.29, p = 0.28, and p = 0.73 respectively). We documented higher serum PAI-1 in HCV-induced HCC than viral infection-free controls, but interestingly, lower than HCV-induced liver cirrhosis patients. This was not genotype related. Further studies will be needed to clearly elucidate the underlying mechanism.

Mean Platelet Volume as a Fibrosis Marker in Patients with Chronic Hepatitis C

Background and study aim: Liver biopsy limitations push us to search for new non invasive methods to detect liver fibrosis such as serum markers. The aim of this study is to evaluate mean platelet volume (MPV) as a fibrosis marker in patient with chronic hepatitis C. Patients and methods: 150 patients diagnosed with chronic hepatitis C infection refereed to Tanta Fever Hospital in period from May 2013 to January 2014 and 20 healthy volunteers as a control were included. All of them were tested for Mean Platelet Volume (MPV) in comparison with who done liver biopsy as standard. Results: Statistically significant differences in MPV and Platelet Count were seen in patients with chronic hepatitis C (CHC) compared to healthy controls (MPV: 8.95 ± 1.39fL vs. 7.57 ± 0.68 fl, P-value = 0.043; PC 226.03 ±68.36 vs. 188.9±46.49, P-value = 0.02) Multi-variate Logistic regression analysis shows only 5 variables remained as independent risk factors for fibrosis progression: (MPV, Schistosomiasis, ALT, AST and Prothrombin time). AST (OR 1.11, 95% CI 1.02 to 1.21), ALT (OR 0.92, 95% CI 0.86 to 0.99), PT (OR 2.11, 95% CI 1.15 to 3.88), and MPV (OR 2.28, 95% CI 1.22 to 4.25). Cut-off values were calculated for diagnostic performance, and the cut-off value for MPV was 9.22 fl., sensitivity 75.5%, specificity 62%, PPV 40.3%, NPPV 93.4% and Accuracy rate 61.8%. Conclusion: We suggest that high MPV levels (especially those over 9.22 fl) may help to predict advanced fibrosis in patients with CHC. However, it should not be forgotten that MPV is not a specific marker for fibrosis, and the negative predictive rate seems more valuable to exclude a high fibrosis ratio in patients with CHC

Bone mineral density and vitamin D receptor polymorphism in beta-thalassemia major.

Osteoporosis is the most prevalent bone complication in beta-thalassemic patients despite regular transfusions and iron chelation therapy. Although its etiology is multi-factorial, genetic factors play an important role in pathogenesis. These factors have not yet been clearly defined, however, osteoporosis may be related to vitamin D receptor gene BsmI polymorphism. In this study, BsmI vitamin D receptor gene polymorphism was analyzed using polymerase chain reaction and BsmI restriction fragment length polymorphism in 42 regularly treated-beta-thalassemic patients of different ages. Bone mineral density was measured by peripheral quantitative ultrasound at the heel of the foot. Serum levels of alkaline phosphatase, calcium, phosphorus, ferritin and 25-hydroxyvitamin D3 were determined. Patients were divided into two groups according to pubertal signs: group I (22 children), and group II (20 adolescents and adults). The Z-scores of bone mineral density in both groups were -1.32 +/- -0.9 and -2.30 +/- -1.02 respectively, with a significant difference between the two groups. The height standard deviation and 25-hydroxyvitamin D3 were significantly decreased in group II compared to group I. Moreover, significantly lower bone mineral density and height standard deviation were detected among patients with BB vitamin D receptor genotype. Therefore, this genotype may be considered as a risk factor for osteoporosis in beta-thalassemic patients.

Experimental diabetic nephropathy can be prevented by propolis: Effect on metabolic disturbances and renal oxidative parameters.

Oxidative stress may play a key role in the pathogenesis of diabetic nephropathy. Propolis and its extract have antioxidant properties. The effect of ethanolic extract of propolis against experimental diabetes mellitus-associated changes was examined. Diabetes was induced experimentally in rats by i.p. injection of streptozotocin (STZ) in a dose of 60 mg/kg bwt for 3 successive days. Blood urea nitrogen (BNU), creatinine, glucose, lipid profile, malondialdehyde (MDA) and urinary albumin were measured. Superoxide dimutase (SOD), glutathione (GSH), catalase (CAT) and MDA were measured in the renal tissue. The results showed decreased body weight and increased kidney weight in diabetic animals. Compared to the control normal rats, diabetic rats had higher blood glucose, BNU, creatinine, total cholesterol, triglycerides, low-density lipoprotein-cholesterol (LDL-C), MDA and urinary albumin and lower high-density lipoprotein-cholesterol (HDL-C) levels. Moreover, renal tissue MDA was markedly increased while SOD, GSH and CAT were significantly decreased. Oral administration of propolis extract in doses of 100,200 & 300 mg/kg bwt improved the body and kidney weights, serum glucose, lipid profile, MDA and renal function tests. Renal GSH, SOD and CAT were significantly increased while MDA was markedly reduced. These results may suggest a strong antioxidant effect of propolis which can ameliorate oxidative stress and delay the occurrence of diabetic nephropathy in diabetes mellitus.

Autoantibodies in chronic liver disease.

Some hepatotropic viruses (HBV and HCV) are capable of triggering autoimmune phenomena and manifest the features of autoimmune hepatitis (AIH) in the course of the disease. Careful attention is required to differentiate between AIH and chronic viral hepatitis (CVH) before the selection of treatment. This study was performed to assess the prevalence of rheumatoid factor (RF), antinuclear antibodies (ANA), anti-smooth muscle antibodies (ASMA), anti-mitochondrial antibodies (AMA), anti-parietal cell antibodies (APCA), anti-liver/kidney microsomal antibodies type I (ALKMA1) and anti-neutrophil cytoplasmic antibodies (ANCA) among patients with chronic liver diseases (CLD), and to assess the diagnostic value of these autoantibodies and their relation to HCV viral load and genotype and treatment with interferon-alpha (IFN-alpha). Five groups of patients with CLD (HCV, HBV, HCC, AIH and schistosomal hepatic fibrosis {SHF}) as well as a group of age- and gender-matched healthy controls were enrolled in the study. All the studied persons were subjected to full clinical assessment and laboratory investigations, including liver function tests, hepatitis markers, and HCV RNA by PCR. Detection of ANA, ASMA, APCA, AMA and ALKMA-1 was done by indirect immunofluorescence technique, while ANCA and RF were detected by EIA and latex agglutination test respectively. Results showed a significantly higher prevalence of RF, ASMA and ANCA among patients with CHC, RF and ASMA among HCC patients and ASMA and ALKMA1, among AIH patients as compared to the control group. Patients with HBV and those with SHF had a non-significantly higher prevalence of RF, ASMA and ANCA compared to controls. However, AMA was not detected in this study, and APCA showed no significant difference between the studied groups. The occurrence of these autoantibodies was not significantly related to HCV viral load, HCV genotype or treatment with IFN-alpha. There was a significant association between the occurrence of RF, ANA, ASMA, and ALKMA1 and high ALT levels, and between the occurrence of ANA, ASMA and ALKMA-1 and high AST and ALP levels. In conclusion, autoantibodies are commonly found among patients with HCV infection. The co-existence of HCV infection and autoimmune hepatitis should be considered in patients who are positive for both viral markers and autoantibodies and thorough evaluation of patients must be performed before selection of treatment. Testing for RF, ASMA and ANCA may have a good diagnostic value, however, AMA is the least useful in diagnosis.