Oxidative Stress, Ocular Disease and Diabetes Retinopathy.

BACKGROUND: Oxidative stress is caused by free radicals or oxidant productions, including lipid peroxidation, protein modification, DNA damage and apoptosis or cell death and results in cellular degeneration and neurodegeneration from damage to macromolecules. RESULTS: Accumulation of the DNA damage (8HOdG) products and the end products of LPO (including aldehyde, diene, triene conjugates and Schiff's bases) were noted in the research studies. Significantly higher levels of these products in comparison with the controls were observed. Oxidative stress induced changes to ocular cells and tissues. Typical changes include ECM accumulation, cell dysfunction, cell death, advanced senescence, disarrangement or rearrangement of the cytoskeleton and released inflammatory cytokines. It is involved in ocular diseases, including keratoconus, Fuchs endothelial corneal dystrophy, and granular corneal dystrophy type 2, cataract, age-related macular degeneration, primary open-angle glaucoma, retinal light damage, and retinopathy of prematurity. These ocular diseases are the cause of irreversible blindness worldwide. CONCLUSIONS: Oxidative stress, inflammation and autophagy are implicated in biochemical and morphological changes in these ocular tissues. The development of therapy is a major target for the management care of these ocular diseases.

Using Red Cell Indices and Reticulocyte Parameters for Carrier Screening of Various Thalassemia Syndromes.

Primary screening for thalassemia carriers usually involves an accurate blood count using an automated blood cell analyzer. We analyzed the red cell and reticulocyte parameters from 200 samples of various types of thalassemias and identified the discrimination criteria for differential diagnosis. These were separated into four groups based on genotypes. These groups included α-thalassemia, ß-thalassemia, ß-thalassemia with Hb E and Hb E trait, which are the important target of thalassemia screening. To compare the effectiveness of the screening strategies, seven selected screening tools were compared, including MCV alone (cutoff <80 fL); MRV alone (cutoff <100 fL); SD-C-NR alone (cutoff >28.5); a combined MCV and MRV; a combined MCV and SD-C-NR; a combined MRV and SD-C-NR; and a combined MCV, MRV and SD-C-NR. The combination of MCV, MRV and SD-C-NR has highest sensitivity for discrimination in all thalassemia, ß-thalassemia, α-thalassemia, Homozygous Hb E and Hb E trait groups as 99.5, 100, 98, 100 and 100 %, respectively. The effectiveness of the combination of MCV, MRV and SD-C-NR parameters in the present study was determined as the faster and higher sensitivity screening tool than the other methods, including simple, reliable, cost-effective, and using only one automated hematology analyzer, saving labor costs.

Association of Elevated Serum Lipoprotein(a), Inflammation, Oxidative Stress and Chronic Kidney Disease with Hypertension in Non-diabetes Hypertensive Patients.

Hypertension is the most common cardiovascular risk factor. Lipoprotein(a) [Lp(a)], inflammation, oxidative stress and chronic kidney disease (CKD) exacerbate the response to tissue injury and acts as markers of the vascular disease, especially in glomerulosclerosis. We compared the clinical characteristics of 138 non-diabetes hypertensive women (ndHT) patients with 417 non-diabetes normotensive subjects and tested the association of hypertension with Lp(a), inflammation, CKD and oxidative stress by using multiple logistic regression. BP, BMI, waist circumference, creatinine, Lp(a), inflammation and malondialdehyde levels were significantly higher and CKD state in the ndHT patients (p < 0.05). Multiple logistic regression showed hypertension associated with increased Lp(a), inflammation, ORs and 95 % CIs were 2.52 (1.33, 4.80), 2.75 (1.44, 5.27) after adjusting for their covariates. Elevated serum Lp(a) and inflammation levels concomitants with increased oxidative stress and CKD were the major risk factors associated with hypertension and implications for the increased risk of HT and vascular disease.

Tumor Necrosis Factor-Α, Interleukin-6, C-Reactive Protein Levels and Insulin Resistance Associated with Type 2 Diabetes in Abdominal Obesity Women.

We aim to investigate the association between elevated tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and high sensitivity-C-reactive protein (hs-CRP) with type 2 diabetes mellitus (T2DM) in abdominal obesity (AO) women subjects. A total of 428 AO subjects (age 48.4 ± 10.2 years), and 107 non-AO women subjects (age 48.8 ± 11.8 years) were enrolled for the all biochemistry testing, inflammatory cytokines, fasting insulin and Homeostasis Model Assessment of insulin resistance (HOMA-IR). Body mass index, waist circumference (WC), blood pressure (BP), plasma glucose (Glu), triglyceride (TG), insulin, HOMA-IR and inflammatory cytokines were significantly higher and lower total antioxidant capacity, HDL-C in AO subjects (p < 0.05). WC was significantly correlated with BP, Glu, TG, LDL-C, insulin, HOMA-IR, TNF-α, IL-6 and negative correlation with HDL-C in AO subjects. Elevation of TNF-α, IL-6, hs-CRP and insulin resistance were significantly associated with T2DM in AO subjects, after adjusting with insulin resistance, increased oxidative stress, elevated TG and reduced HDL-C by using multiple logistic regression analysis. In conclusions, elevation of inflammatory cytokines, oxidative stress and insulin resistance were associated with T2DM in AO women subjects. These inflammatory cytokines are positively associated with T2DM and may have a causal relation with an increased oxidative stress and insulin resistance in these AO women subjects.

Effect of cinnamon supplementation on glucose, lipids levels, glomerular filtration rate, and blood pressure of subjects with type 2 diabetes mellitus.

A randomized, double-blind, placebo-controlled trial was performed to investigate the effect of cinnamon supplementation on glucose, triglyceride (TG), and high density lipoprotein-cholesterol (HDL-C) levels, TG/HDL-C ratio, blood pressure (BP), and estimated glomerular filtration rate (eGFR) for ninety-nine type 2 diabetes mellitus (T2DM) patients. Forty nine (49) patients with T2DM were in the cinnamon group whereas 50 were in the placebo group. All participants received either a cinnamon or placebo capsule daily for the 60-day study period. At the end of the study, median glucose, TG, TG/HDL-C ratio, and BP were significantly decreased (p < 0.005) whereas HDL-C and eGFR levels were significantly increased (p < 0.005) in the cinnamon supplementation group. Interestingly, the study also revealed reduction of HbA1c levels after the 60-day study period. In the placebo group, glucose, TG levels tended to increase and HDL-C levels tended to decrease whereas systolic and diastolic BP, HbA1c, and BUN CT were significantly increased and eGFR was significantly reduced (p < 0.005). Cinnamon supplementation reduced plasma glucose, HbA1c, triglyceride, TG/HDL-C ratio, and BP and increased HDL-C levels and eGFR in subjects with T2DM. Cinnamon supplementation may be beneficial for those with T2DM to prevent and control diabetic complications.

Abdominal Obesity Associated with Elevated Serum Butyrylcholinesterase Activity, Insulin Resistance and Reduced High Density Lipoprotein-Cholesterol Levels.

Abdominal obesity (AO) has a strong correlation with cardiovascular disease and has been linked to Alzheimer's disease and type 2 diabetes. We investigated the association between AO and elevated serum butyrylcholinesterase (BChE) activity, insulin resistance and the serum lipid profile, including triglyceride (TG), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C) levels in AO and non-AO women subjects. A total of 500 AO subjects (age 49.1 ± 10.5 years), and 142 non-AO women subjects (age 49.9 ± 11.9 years) were enrolled for the general biochemistry tests, serum BChE, fasting insulin and homeostasis model assessment of insulin resistance (HOMA-IR). Body mass index, waist circumference, Blood pressure (BP), plasma glucose (Glu), triglyceride (TG), BChE, insulin, HOMA-IR were significantly higher and HDL-C levels were significantly lower in AO subjects (p < 0.05). Waist circumference was significantly correlated with BP, Glu, TG, BChE, insulin and HOMA-IR in AO subjects. Multiple logistic regression demonstrated that AO was associated with elevated BChE, HOMA-IR, hypertension and reduced HDL-C after adjusting for these variables. AO is associated with elevated BChE, insulin resistance, HT and reduced HDL-C. These may predict the development of type 2 diabetes mellitus and may be associated with cognitive disorder in the future, both are mediated through insulin resistance.

Association of Elevated Serum Uric Acid with the Components of Metabolic Syndrome and Oxidative Stress in Abdominal Obesity Subjects.

Abdominal obesity (AO) and metabolic syndrome (MetS) are associated with the cardiovascular disease and type 2 diabetes. Serum uric acid (SUA) is often elevated in subjects with the AO. We aimed to investigate the association of elevated SUA with the components of MetS, oxidative stress and TG/HDL-C ratio in AO subjects. This cross-sectional study used data from a Health Survey for Prevention of Hypertension and Type 2 Diabetes Mellitus in residents of two districts in Phitsanulok province, including 443 subjects. Anthropometric, blood pressure (BP) and biochemical variables were measured. We categorized the participants to two-group as 248 AO subjects (median age = 58, interquartile range 50.0-65.0 years) and 195 non-AO subjects (median age = 53, interquartile range 47.0-62.0 years). Waist circumference was significantly correlated with SystBP, DiastBP, Glu and SUA (P < 0.05) and SUA was significantly correlated with Glu, TG, HDL-C and TG/HDL-C ratio (P < 0.05). By using multiple logistic regression, we found the association of elevated SUA with abdominal obesity, hyperglycemia, hypertriglyceridemia, reduced HDL-C, elevated TG/HDL-C ratio, MetS and increased oxidative stress after adjusting for their covariates. Our study demonstrated that circulating UA is a major antioxidant and might help protect against free-radical oxidative damage. However, elevated SUA concentrations associated with oxidative stress, MetS, insulin resistance, and components of MetS. Then, SUA may be a marker of increased oxidative stress, insulin resistance and MetS, implying an increased risk of vascular disease and T2DM.

Alloimmunization to Red Cells and the Association of Alloantibodies Formation with Splenectomy Among Transfusion-Dependent ß-Thalassemia Major/HbE Patients.

Severe hemolytic anemia in ß-thalassemia major and ß-thalassemias/HbE (ß-TM) patients requires giving blood transfusions. Chronic blood transfusions lead to iron overload consequence with organs damage and risk of alloantibody-formation. This study evaluates the prevalence of red cell alloimmunization and estimates the risk of alloantibody-formation in chronic transfusion-dependent ß-TM patients. This cross sectional study was conducted on 143 ß-TM patients receiving regular transfusions. We tried to determine the frequency, types and factors influencing red cell alloimmunization in these transfusion-dependent ß-TM patients. Median age of 25 (17.5 %) alloantibody-formation ß-TM patients was 19.0 years (inter quartile 15.5-24.0 years). The alloantibodies were Anti-Rh (E) (13.1 %), Anti-Rh (D) (0.7 %). Thirty-four patients (23.8 %) of the sample had splenectomies of which 10 (29.4 %) had alloantibody-formation. The interval from first transfusion to antibody development varied from 1.5 to 14 years. Alloantibody-formation correlated with splenectomy and splenectomy correlated with number of transfusion (p < 0.005). In multiple logistic regression used to estimate the risk of alloantibodies formation with splenectomy; OR and 95 % CI were 2.88 (1.07-7.80), p = 0.037 after adjusting for other co-variates. The rate of red cell alloimmunization was 17.5 % and splenectomy associated with increased alloantibody-formation in these transfusion-dependent ß-TM patients.

Oxidative stress, insulin resistance, dyslipidemia and type 2 diabetes mellitus.

Oxidative stress is increased in metabolic syndrome and type 2 diabetes mellitus (T2DM) and this appears to underlie the development of cardiovascular disease, T2DM and diabetic complications. Increased oxidative stress appears to be a deleterious factor leading to insulin resistance, dyslipidemia, ß-cell dysfunction, impaired glucose tolerance and ultimately leading to T2DM. Chronic oxidative stress, hyperglycemia and dyslipidemia are particularly dangerous for ß-cells from lowest levels of antioxidant, have high oxidative energy requirements, decrease the gene expression of key ß-cell genes and induce cell death. If ß-cell functioning is impaired, it results in an under production of insulin, impairs glucose stimulated insulin secretion, fasting hyperglycemia and eventually the development of T2DM.

The plasma DNA concentration as a potential breast cancer screening marker.

Increased levels of plasma DNA have frequently been noticed in the blood plasma of cancer patients. The possibility of using plasma DNA level as the indicator of tumor stage in breast cancer was investigated in plasma samples obtained from 100 breast cancer patients and 100 healthy women who were included as controls. Circulatory plasma free DNA was extracted from plasma samples and quantified by fluorometer. The median concentration of plasma DNA in the plasma samples from breast cancer patients classified by TNM staging system as stage I, II, III, IV and breast surgical patients were 0.5, 235, 422, 1,280 and 0.5 ng/ml, respectively. The level of plasma DNA in the stage II- IV group was significantly higher than those in the surgical group with breast cancer and control group (P value < 0.001). The plasma DNA concentration in stage II, III and IV of breast cancer were higher when compared with healthy group. These tumor size, TNM stage and metastasis were significantly correlated with plasma DNA. The cut point of 120 ng/ml was early screening and treatment follow up breast cancer.

Association of Iron Overload with Oxidative Stress, Hepatic Damage and Dyslipidemia in Transfusion-Dependent ß-Thalassemia/HbE Patients.

Blood transfusion can be a life-saving therapy for ß-thalassemia major and ß-thalassemia/HbE (ß-TM) patients with chronic anemia, major caused severe iron overload particularly in ß-TM patients received only blood transfusion therapy. We aim to evaluate the association of iron overload with oxidative stress, liver damage, and elevated very low density lipoprotein cholesterol (VLDL-C) in transfusion-dependent ß-TM patients. Serum ferritin, malondialdehyde (MDA), liver profiles, triglycerides levels, and VLDL-C were significantly higher while total cholesterol, low-density lipoprotein cholesterol, high density lipoprotein cholesterol and total antioxidant capacity were lower in ß-TM than controls. Serum ferritin was significantly correlated with MDA, liver enzymes and lipid profiles (p < 0.05). Multiple forward stepwise linear regression analyses of the significant variables showed that in these ß-TM patients, independent predictors of iron overload were MDA (ß = 0.410, r (2) = 0.671, p < 0.001), ALT (ß = 0.493, r (2) = 0.578, p < 0.001), and VLDL-C (ß = 0.253, r (2) = 0.711, p < 0.001). In conclusion, iron overload associated with increased oxidative stress, lipid peroxidation, liver damage, decreased TC, LDL-C, HDL-C and over production of VLDL-C, is significantly problem in transfusion-dependent ß-TM patients. These appeared the major cause of future morbidity and mortality in ß-TM patients.

Association of iron overload and oxidative stress with insulin resistance in transfusion-dependent beta-thalassemia major and beta-thalassemia/HbE patients.

BACKGROUND: Beta-thalassemia causes a severe hemolytic anemia in patients necessitating frequent transfusions leading to iron overload and endocrine complications, especially diabetes mellitus. We tried to determine the change or effect on carbohydrate physiology and oxidation markers and the association of these markers in chronic transfusion-dependent beta-thalassemia major and beta-thalassemia/HbE (beta-TM) patients. METHODS: Prospective study on 60 beta-TM patients, receiving only regular blood transfusions, and 20 healthy controls were enrolled for oral glucose tolerance test, fasting insulin, Homeostasis Model Assessment of insulin resistance (HOMA-IR), liver function test, iron overload, oxidative stress, and lipid profile at baseline. The same tests were repeated after 6 months. RESULTS: One beta-TM patient had impaired glucose tolerance. Fasting glucose, insulin, ferritin, MDA, TG concentrations, HOMA-IR, and liver profiles were significantly higher while Hb, Hct, TC, HDL-C, LDL-C concentrations and TAC were significantly lower in beta-TM patients than controls (p < 0.001). Fasting glucose, HOMA-IR and beta-cell function were significantly correlated with MDA and glucose, AST, ALT, MDA were significantly correlated with ferritin (p < 0.05). Multiple forward stepwise linear regression analyses of the significant variables showed that in these beta-TM patients, independent predictors of HOMA-IR were fasting glucose (beta = 0.634, r2 = 0.374, p < 0.001), HDL-C (beta = 0.249, r2 = 0.418, p = 0.043) and MDA (beta = 0.225, r2 = 0.466, p = 0.029). CONCLUSIONS: Progression of iron overload, oxidative stress and hyperinsulinemia were substantially and persistently higher in beta-TM patients. We observed a positive association between oxidative stress, iron overload and insulin resistance in these beta-TM patients.

Association of elevated urinary cadmium with urinary stone, hypercalciuria and renal tubular dysfunction in the population of cadmium-contaminated area.

Excessive urinary calcium is the major risk of renal tubular dysfunction and urinary stone formation. We examined the association of elevated urinary cadmium with urinary stones and chronic kidney disease (CKD) in 1,085 study residents of 13 cadmium-contaminated villages. Elevated urinary cadmium was significantly correlated with urinary stone and CKD. Elevated urinary cadmium appeared to increase risk of urinary stone and CKD; ORs and 95 % CIs were 2.73 (1.16, 6.42) and 3.73 (2.50, 5.57) after adjusting for other co-variables. This study revealed that elevated urinary stone and CKD induced by cadmium might increase the risk of urinary stone and CKD.

Estimated Creatinine Clearance, Homocysteine and High Sensitivity-C-Reactive Protein Levels Determination for Early Prediction of Nephropathy and Atherosclerosis Risk In Type 2 Diabetic Patients.

Type 2 diabetes mellitus (T2D) patients are increased risk for cardiovascular disease (CVD) and chronic kidney disease (CKD). Many studies had demonstrated that CKD is significantly associated with CVD. We aim to indicate the using estimated creatinine clearance (eCrCl), homocysteine (tHcy), and high sensitivity-C-reactive protein (hs-CRP) levels, may have an impact on the interpretation risk for nephropathy and CVD. eCrCl was using the Cockroft-Gault formula, eCrCl levels were stratified according to the Kidney Disease Outcome Quality Initiative definition. We measured serum tHcy, hs-CRP, and the other biochemical variables in 54 T2D patients compared with 40 age matched healthy controls (NDM). T2D patients were significantly lower eCrCl than NDM (P < 0.05). T2D patients also showed significantly higher in tHcy, hs-CRP, and MDA levels than NDM subjects (P < 0.05). The eCrCl was significantly correlated with tHcy and hs-CRP levels in T2D patients (r = -0.504, P < 0.001; r = -0.282, P = 0.047). eCrCl had an impact on interpretation for CKD, especially in T2D patients. Decrease eCrCl concomitant with increased in tHcy, hs-CRP, and MDA levels may present a higher risk for future diabetic nephropathy and CVD.

Association of serum lipoprotein ratios with insulin resistance in type 2 diabetes mellitus.

BACKGROUND: The fasting serum total cholesterol (TC), triglycerides (TG), high density lipoprotein (HDL-C) and low density lipoprotein (LDL-C) levels are used to calculate following lipid ratios: TC/ HDL-C, TG/HDL-C, and LDL-C/HDL-C. Cholesterol retention fraction (CRF), non-HDL-C, LDL-C, TG and waist circumference (WC) one considered as markers for the identification of individuals with an increased risk for cardiovascular diseases (CVD). These individuals frequently show insulin resistance as well. We analyzed the association of lipoprotein ratios with the homeostasis model assessment of insulin resistance (HOMA-IR). METHODS: Type 2 diabetes mellitus (T2D) patients (92) and 40 age match healthy controls were randomized from the Tapho Primary Health Care Unit and the area in the same district. The HOMA-IR was used to calculate for insulin resistance. The areas under the curves (AUC) of the receiver operating characteristic curves (ROC) were used to compare the power of these serum lipoprotein ratios markers. RESULTS: All lipoprotein ratios, lipid profile, blood pressure, and WC were significantly higher in T2D patients as compared to healthy controls (P<0.05). TC/HDL-C ratio, TG/HDL-C ratio, non-HDL-C, WC, TG, and TC were significantly correlated with HOMA-IR (P<0.05) as obtained by Spearman correlation analysis. The largest AUC of the ROC curve was obtained with the TC/HDL-C ratio as one parameter. CONCLUSION: TC/HDL-C ratio, TG/HDL-C, ratio, Non-HDL-C, WC, TG, and TC can be used as the markers of insulin resistance and CVD risk in T2D patients.

An effective method for hemoglobin E detection: DEAE sepharose microcolumn.

Hemoglobin E (Hb E) is a variant hemoglobin that can lead to considerable morbidity when it occurs in a compound heterozygous state with beta-thalassemia. Therefore, its detection is important because it permits antenatal counseling. Hb E is prevalent in economically weaker regions of the world. Thus, its recognition is often hindered by the high costs of sophisticated Hb E-detection assays. We developed a simple visual test based on identifying Hb E in blood samples with DEAE Sepharose microcolumn chromatography. The diagnostic accuracy of our new Hb E microcolumn assay was 100% for sensitivity, specificity, and positive and negative predictive values. Furthermore, the Hb E microcolumn assay is rapid, reproducible, and economical; hence, it offers affordable screening for Hb E in less well-equipped laboratories.

Whatman 3 MM dried blood spots for identifying alpha-thalassemia-1.

The present study was undertaken to assess the feasibility of whole dried blood spots to identify alpha-thalassemia-1 by PCR technique. Logistical constraints associated with sample collection and processing make the measurement of alpha-thalassemia-1 at community laboratories difficult. Dried blood spot samples provide a relatively convenient and minimally invasive means for collecting and transporting samples. We present Whatman 3 MM paper for spotting blood samples used for the first time for alpha-thalassemia-1 detection. Measurements of fresh EDTA whole blood and Whatman 3 MM dried blood spots prepared from venous blood were performed in 60 subjects. Whatman 3 MM dried blood spots results were precise and reliable. There was no actual difference in comparison to fresh EDTA whole blood samples. Whatman 3 MM dried blood spots for alpha-thalassemia-1 detection using the PCR technique showed no changes for 6 months.

Antimutagenic activity of Sesbania javanica Miq. flower DMSO extract and its major flavonoid glycoside.

The antimutagenic activity of Sesbania javanica Miq. or Sano, an edible vegetable flower DMSO extract against aflatoxin B1 (AFB1) benzo (a) pyrene [B(a)P], was evaluated by means of the Ames' test. The Sesbania javanica Miq. flower DMSO extract showed a strong inhibitory effect against AFB1 and B(a)P mutagens. A search to isolate the major flavonoid in Sesbania javanica Miq. flower extract found the flavonol glycoside, Quercetin 3-2(G)-rhamnosylrutinoside, which was confirmed by its physicochemical properties as a major constituent of the flower. Quercetin 3-2(G)-rhamnosylrutinoside (207 microg/plate) also showeda strong inhibitory effect against AFB1 and B(a)P with a more than 70% inhibition rate.