Helio Vannucchi

DNA oxidative damage in patients with dialysis treatment

Background/Aims. Chronic renal patients on hemodialysis (HD) and peritoneal dialysis (PD) treatment are exposed to oxidative stress and DNA damage. The objective of this study was to assess the oxidative damage to DNA in end-stage chronic renal failure, before and after vitamin E supplementation. Methods. Patients on HD (n = 29) and PD (n = 22) received oral supplementation with 300 mg vitamin E three times a week for 4 weeks. A blood sample was collected at the beginning and at the end of the supplementation cycle for the determination of vitamin E levels (high-performance liquid chromatography), carbonyl groups, and DNA damage (8-hydroxy 2′-deoxyguanosine [8-OHdG] and comet assay). Results. After supplementation, vitamin E concentration was increased by about 50%. Protein oxidation was initially observed in both groups, with a reduction after supplementation. DNA damage detected by the comet assay and by 8-OHdG analysis was significantly reduced (p< 0.05) after supplementation in both groups. Conclusions. Vitamin E supplementation reduced oxidative DNA damage in both HD and PD patients. Treatments such as HD and PD induce oxidative stress and consequent DNA damage, and increased plasma vitamin E levels significantly contribute to the normalization of these events.

Effects of creatine supplementation on homocysteine levels and lipid peroxidation in rats.

Hyperhomocysteinaemia is an independent risk factor for CVD. Recent data show a relationship between homocysteine (Hcy) and free radical formation. Since creatine synthesis is responsible for most of the methyl group transfers that result in Hcy formation, creatine supplementation might inhibit Hcy production and reduce free radical formation. The present study investigated the effects of creatine supplementation on Hcy levels and lipid peroxidation biomarkers. Thirty rats were divided into three groups: control group; diet with creatine group (DCr; 2 % creatine in the diet for 28 d); creatine overload plus diet with creatine group (CrO + D; 5 g creatine/kg by oral administration for 5 d+2 % in the diet for 23 d). Plasma Hcy was significantly lower (P < 0.05) in DCr (7.5 (sd 1.2) micromol/l) and CrO + D (7.2 (sd 1.7) micromol/l) groups compared with the control group (12.4 (sd 2.2) micromol/l). Both plasma thiobarbituric acid-reactive species (TBARS) (control, 10 (sd 3.4); DCr, 4.9 (sd 0.7); CrO + D, 2.4 (sd 1) micromol/l) and plasma total glutathione (control, 4.3 (sd 1.9); DCr, 2.5 (sd 0.8); CrO + D, 1.8 (sd 0.5) micromol/l) were lower in the groups that received creatine (P < 0.05). In addition, Hcy showed significant negative correlation (P < 0.05) with plasma creatine (r - 0.61) and positive correlation with plasma TBARS (r 0.74). Plasma creatine was negatively correlated with plasma TBARS (r - 0.75) and total peroxide (r - 0.40). We conclude that creatine supplementation reduces plasma Hcy levels and lipid peroxidation biomarkers, suggesting a protective role against oxidative damage. Modulating Hcy formation may, however, influence glutathione synthesis and thereby affect the redox state of the cells.

Maternal risk for Down syndrome is modulated by genes involved in folate metabolism

Studies have shown that the maternal risk for Down syndrome (DS) may be modulated by alterations in folate metabolism. The aim of this study was to evaluate the influence of 12 genetic polymorphisms involved in folate metabolism on maternal risk for DS. In addition, we evaluated the impact of these polymorphisms on serum folate and plasma methylmalonic acid (MMA, an indicator of vitamin B12 status) concentrations. The polymorphisms transcobalamin II (TCN2) c.776C>G, betaine-homocysteine S-methyltransferase (BHMT) c.742A>G, methylenetetrahydrofolate reductase (NAD(P)H) (MTHFR) c.677 C>T and the MTHFR 677C-1298A-1317T haplotype modulate DS risk. The polymorphisms MTHFR c.677C>T and solute carrier family 19 (folate transporter), member 1 (SLC19A1) c.80 A>G modulate folate concentrations, whereas the 5-methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR) c.66A>G polymorphism affects the MMA concentration. These results are consistent with the modulation of the maternal risk for DS by these polymorphisms.

Bioavailability of iron added to the diet by cooking food in an iron pot

Abstract The iron bioavailability of food cooked in an iron pot was studied. Iron deficiency was produced in 42 21-day old Wistar rats over a period of 5 weeks, while 18 rats receiving a complete diet throughout the experiment were used as a control group (CG). After the period of iron depletion, the animals were subdivided into 3 subgroups: one continued to receive the basal diet (BB), the second was fed a complete diet (BC), and the third received the basal diet cooked in an iron pot (BI). After 4 weeks, the BC and BI group did not differ significantly (p>0.05) from CG in hemoglobin, hematocrit, protoporphyrin, serum iron and transferrin saturation and that it promoted the correction of the parameters studied within four weeks.

Antioxidant Effect of Thiamine on Acutely Alcoholized Rats and Lack of Efficacy Using Thiamine or Glucose to Reduce Blood Alcohol Content

Although there is no consensus about the use of glucose and thiamine for the treatment of acute ethanol intoxication, this is a routine practice in many countries. Our objective was to determine the efficacy of this treatment and the changes it causes in the antioxidant status of the liver. Male Wistar rats were intoxicated with an ethanol dose of 5 g/kg and divided into three groups: ethanol (EtOH; untreated), EtOH+G (treated with glucose), and EtOH+B1 (treated with thiamine). Blood and urinary ethanol as well as hepatic malondialdehyde, reduced glutathione and vitamin E were determined in all animals. Blood alcohol levels did not differ between groups, although urinary excretion was about four times higher in the group treated with thiamine (EtOH+B1). The malondialdehyde, reduced glutathione and vitamin E values used here as parameters of the antioxidant system of the liver showed improvement for the thiamine-treated group (EtOH+B1). Treatment with glucose or thiamine was ineffective in reducing blood alcohol levels in rats with acute ethanol intoxication. However, the beneficial effect of thiamine as an antioxidant for ethanol metabolism was demonstrated. Further investigations are necessary to clarify the urinary excretion of ethanol reported here for the first time and the possibility of using thiamine as an antioxidant in situations of chronic alcohol use.

Chemical composition of the fruit mesocarp of three peach palm (Bactris gasipaes) populations grown in central Amazonia, Brazil.

The percent composition, soluble and insoluble food fibers, oil fatty acids and minerals were determined in the mesocarp of fruits of three peach palm (Bactris gasipaes Kunth) populations grown in Central Amazonia, Brazil. Amino acids were also determined in one of the populations. The mean protein levels ranged from 1.8 to 2.7%, lipid levels ranged from 3.5 to 11.1%, the nitrogen free fraction ranged from 24.3 to 35%, food fiber ranged from 5.2% to 8.7%, and energy ranged from 179.1 to 207.4 kcal%. All essential, as well as non-essential, amino acids were present, with tryptophan and methionine presenting the lowest mean concentrations. The mono-unsaturated oleic acid predominated in the oil, ranging from 42.8 to 60.8%, and palmitic acid was the most abundant saturated fatty acid, ranging from 24.1 to 42.3%. Among the essential fatty acids, linoleic acid was the most abundant, with a maximum of 5.4% in Pampa-8. The most important mineral elements were potassium, selenium and chromium, respectively corresponding to 12%, 9% and 9% of daily recommended allowances. Considering the nutritional potential of the fruit, we suggest its more frequent incorporation into the diet of the Amazonian population.

Lipid peroxidation in nicotinamide-deficient and nicotinamide-supplemented rats with streptozotocin-induced diabetes.

Abstract Reactive oxygen species have been related to the pathogenesis of various diseases, including diabetes mellitus. Nicotinamide has been used for the prevention of the diabetogenic effects of streptozotocin (STZ) in animals. In the present study we assessed the effect of diets with deficient, normal or 17-fold supplemented nicotinamide concentrations on the rate of lipopoeroxidation in animals with STZ-induced diabetes. Male Wistar rats were divided into three groups kept on one of the diets for six weeks: DD, diabetic rats on a nicotinamide-deficient diet; DN, diabetic rats on a normal nicotinamide diet; and DS, diabetic rats on a nicotinamide-supplemented diet. During the fourth week of the experiment all animals were fasted for 24 hours and injected into the tail vein with a single STZ dose (40 mg/kg weight). Eight animals from each of the six groups were then sacrificed 24 hours, 1 week and 2 weeks after STZ injection. Mean pancreatic thiobarbituric acid reactive substances (TBARS) (nmol/mg tissue) were significantly lower in the DS group (p < 0.05) compared to the DN and DD groups at 24 hours and during the first week. Hepatic TBARS concentrations (nmol/mg protein) did not differ between groups. Mean hepatic reduced glutathione (GSH) levels were significantly higher (46.76 ± 12.33 nmol/mg protein) in the DS group compared to the DD (32.90 ± 6.70) and DN (24.55 ± 6.41) groups, but only after the 24-hour period. Hepatic vitamin E consumption (Μg/g tissue) was considerable in the groups not supplemented with nicotinamide, whereas vitamin E levels were unchanged in the supplemented group. In contrast, plasma vitamin E levels were decreased in the normal and supplemented groups after 1 and 2 weeks. A higher N-methylnicotinamide excretion (μg/ 24 hours) occurred in the supplemented group. We conclude that, after induction of diabetes with STZ, nicotinamide supplementation protected from the damage caused by the toxic action of STZ, promoting lower lipid peroxidation.

Vitamin A deficiency in children aged 6 to 24 months in S~ao Paulo State, Brazil

Abstract Vitamin A deficiency (VAD), mainly the subclinical form, is endemic in several areas of the Brazilian Northeast. However, studies concerning its prevalence and etiology (risk factors) in urban areas of Sao Paulo State are lacking. The objective of the present study was to identify VAD and the risk factors among children attending a Child Care outpatient clinic in Ribeirao Preto city, Sao Paulo State, Brazil. A total of 103 children aged 6 to 24 months without any diarrhea or fever illness were selected from a pediatric outpatient clinic. A careful clinical history and physical examination provided information about breast-feeding, parental education, family income, family size, birth weight and anthropometric data. The children also underwent ophthalmologic examination to check for signs of xerophthalmia and were submitted to blood tests in order to determine hemoglobin and serum iron, zinc and retinol levels. Serum retinol levels £ 0.70 μmol/l are considered to be deficient by the World Health Organization. Retinol levels, determined by high performance liquid chromatography (HPLC), were £? 0.70 μ?mol/l in 22 children (21.4%). No child had xerophthalmia. The overall mean serum zinc level was 108.9 μg% (2SD ± 43.1 μg%) and the values for children with and without VAD were 105.1 μg% (2SD ± 44.1 μg%) and 110.0 μg% (2SD ± 43.2 μg%) respectively, with no child presenting serum zinc levels below the normal range. None of them showed a

Choline Supplementation Protects against Liver Damage by Normalizing Cholesterol Metabolism in Pemt/Ldlr Knockout Mice Fed a High-Fat Diet

Dietary choline is required for proper structure and dynamics of cell membranes, lipoprotein synthesis, and methyl-group metabolism. In mammals, choline is synthesized via phosphatidylethanolamine N-methyltransferase (Pemt), which converts phosphatidylethanolamine to phosphatidylcholine. Pemt(-/-) mice have impaired VLDL secretion and developed fatty liver when fed a high-fat (HF) diet. Because of the reduction in plasma lipids, Pemt(-/-)/low-density lipoprotein receptor knockout (Ldlr(-/-)) mice are protected from atherosclerosis. The goal of this study was to investigate the importance of dietary choline in the metabolic phenotype of Pemt(-/-)/Ldlr(-/-) male mice. At 10-12 wk of age, Pemt(+/+)/Ldlr(-/-) (HF(+/+)) and half of the Pemt(-/-)/Ldlr(-/-) (HF(-/-)) mice were fed an HF diet with normal (1.3 g/kg) choline. The remaining Pemt(-/-)/Ldlr(-/-) mice were fed an HF diet supplemented (5 g/kg) with choline (HFCS(-/-) mice). The HF diet contained 60% of calories from fat and 1% cholesterol, and the mice were fed for 16 d. HF(-/-) mice lost weight and developed hepatomegaly, steatohepatitis, and liver damage. Hepatic concentrations of free cholesterol, cholesterol-esters, and triglyceride (TG) were elevated by 30%, 1.1-fold and 3.1-fold, respectively, in HF(-/-) compared with HF(+/+) mice. Choline supplementation normalized hepatic cholesterol, but not TG, and dramatically improved liver function. The expression of genes involved in cholesterol transport and esterification increased by 50% to 5.6-fold in HF(-/-) mice when compared with HF(+/+) mice. Markers of macrophages, oxidative stress, and fibrosis were elevated in the HF(-/-) mice. Choline supplementation normalized the expression of these genes. In conclusion, HF(-/-) mice develop liver failure associated with altered cholesterol metabolism when fed an HF/normal choline diet. Choline supplementation normalized cholesterol metabolism, which was sufficient to prevent nonalcoholic steatohepatitis development and improve liver function. Our data suggest that choline can promote liver health by maintaining cholesterol homeostasis.

Peroxisome proliferator-activated receptors alpha and gamma2 polymorphisms in nonalcoholic fatty liver disease: a study in Brazilian patients.

BACKGROUND Non-alcoholic fatty liver disease (NAFLD) refers to the accumulation of hepatic steatosis in the absence of excess alcohol consumption. The pathogenesis of fatty liver disease and steatohepatitis (NASH) is not fully elucidated, but the common association with visceral obesity, hyperlipidemia, hypertension and type 2 diabetes mellitus (T2DM) suggests that it is the hepatic manifestation of metabolic syndrome. Peroxisome proliferator-activated receptor PPARα and PPARγ are members of a family of nuclear receptors involved in the metabolism of lipids and carbohydrates, adipogenesis and sensitivity to insulin. The objective of this study was to analyze the polymorphisms Leu162Val of PPARα and Pro12Ala of PPARγ as genetic risk factors for the development and progression of NAFLD. METHODS One hundred and three NAFLD patients (89 NASH, 14 pure steatosis) and 103 healthy volunteers were included. Single nucleotide polymorphisms (SNPs) Leu162Val and Pro12Ala were analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). RESULTS NASH patients presented higher BMI, AST and prevalence of T2DM than patients with pure steatosis. A higher prevalence of 12Ala allele was observed in the NASH Subgroup when compared to Control Group. When we grouped NASH and Steatosis Subgroups (NAFLD), we found lower serum glucose and more advanced fibrosis in the Leu162Val SNP. On the other hand, there was no statistical difference in clinical, laboratorial and histological parameters according to the Pro12Ala SNP. CONCLUSIONS We documented a lower prevalence of 12Ala allele of gene PPARγ in the NASH Subgroup when compared to Control Group. In NAFLD patients, there were no associations among the occurrence of Pro12Ala SNP with clinical, laboratorial and histological parameters. We also documented more advanced fibrosis in the Leu162Val SNP. The obtained data suggest that Pro12Ala SNP may result in protection against liver injury and that Leu162Val SNP may be involved in the progression of NAFLD.