Railson Henneberg

The FXR agonist 6ECDCA reduces hepatic steatosis and oxidative stress induced by ethanol and low-protein diet in mice

BACKGROUND AND AIM Excessive ethanol consumption can lead to development of hepatic steatosis. Since the FXR receptor regulates adipose cell function and liver lipid metabolism, the aim of this work was to examine the effects of the FXR agonist 6ECDCA on alcoholic liver steatosis development and on oxidative stress induced by ethanol consumption. METHODS Swiss mice (n=24) received a low-protein diet (6%) and a liquid diet containing 10% ethanol or water for 6weeks. In the last 15days mice received oral treatment with 6ECDCA (3mgkg(-1)) or 1% tween (vehicle). The experimental groups (n=6) were: water+tween, water+6ECDCA, ethanol+tween and ethanol+6ECDCA. Moreover, as a diet control, we used a basal group (n=6), fed by a normal-proteic diet (23%) and water. After the treatment period, the animals were anesthetized for sample collection to perform plasma biochemistry assays, hepatic oxidative stress assays, hepatic cholesterol and triglycerides measurements, liver histology and hepatic gene expression. RESULTS Ethanol associated with low-protein diet induced hepatic oxidative stress, increased plasma transaminases and induced hepatic lipid accumulation. Many of these parameters were reversed by the administration of 6ECDCA, including amelioration of lipid accumulation and lipoperoxidation, and reduction of reactive oxygen species. These effects were possibly mediated by regulation of Srebpf1 and FAS gene expression, both reduced by the FXR agonist. CONCLUSIONS Our data demonstrated that 6ECDCA reverses the accumulation of lipids in the liver and decreases the oxidative stress induced by ethanol and low-protein diet. This FXR agonist is promising as a potential therapy for alcoholic liver steatosis.

Erythrocyte oxidative stress markers in children with sickle cell disease

Objective To determine eight parameters of oxidative stress markers in erythrocytes from children with sickle cell disease and compare with the same parameters in erythrocytes from healthy children, since oxidative stress plays an important role in the pathophysiology of sickle cell disease and because this disease is a serious public health problem in many countries.OBJECTIVE To determine eight parameters of oxidative stress markers in erythrocytes from children with sickle cell disease and compare with the same parameters in erythrocytes from healthy children, since oxidative stress plays an important role in the pathophysiology of sickle cell disease and because this disease is a serious public health problem in many countries. METHODS Blood samples were obtained from 45 children with sickle cell disease (21 males and 24 females with a mean age of 9 years; range: 3-13 years) and 280 blood samples were obtained from children without hemoglobinopathies (137 males and 143 females with a mean age of 10 years; range: 8-11 years), as a control group. All blood samples were analyzed for methemoglobin, reduced glutathione, thiobarbituric acid reactive substances, percentage of hemolysis, reactive oxygen species, and activity of the enzymes glucose 6-phosphate dehydrogenase, superoxide dismutase, and catalase. Data were analyzed using Students t-test and were expressed as the mean±standard deviation. A p-value of <0.05 was considered significant. RESULTS Significant differences were observed between children with sickle cell disease and the control group for the parameters methemoglobin, thiobarbituric acid reactive substances, hemolysis, glucose 6-phosphate dehydrogenase activity, and reactive oxygen species, with higher levels in the patients than in the controls. CONCLUSIONS Oxidative stress parameters in childrens erythrocytes were determined using simple laboratory methods with small volumes of blood; these biomarkers can be useful to evaluate disease progression and outcomes in patients.

Reference values for methemoglobin concentrations in children

Objective The aim of this work was to establish reference values for methemoglobin levels in 6 to 10-year-old children. Methods Methemoglobin concentrations were studied in clinically healthy children. The method for methemoglobin measurement used, neither uses highly toxic chemical compounds nor expensive enzymatic methods, thus it is feasible in the laboratory routine. Results The results showed higher reference values for clinically healthy children (from 3.61 to 6.44%) than for adults (from 1.9 to 3.8%). Conclusion The higher concentrations of methemoglobin in children may be explained by smaller amounts of soluble cofactor cytochrome b5 and reduced activity of the cytochrome b5 reductase enzyme in red blood cells which make children particularly susceptible to the development of methemoglobinemia. Methemoglobin concentrations in children are higher than in normal adult subjects thus, adult reference values cannot be used to interpret infant methemoglobinemia.

Ginkgo biloba extract (EGb 761) attenuates oxidative stress induction in erythrocytes of sickle cell disease patients

A doenca falciforme promove anemia hemolitica e oclusao dos pequenos vasos, causados pela presenca de altas concentracoes de hemoglobina S, cujas consequencias incluem a producao aumentada de especies reativas de oxigenio e diminuicao da capacidade de defesa antioxidante. O objetivo desse estudo foi avaliar a acao protetora de um extrato padronizado de Ginkgo biloba (EGb 761), selecionado devido ao seu alto conteudo de flavonoides e terpenoides, em eritrocitos de pacientes com anemia falciforme (HbSS, eritrocitos SS) submetidos ao estresse oxidativo usando terc-butil-hidroperoxido e 2,2-azobis-(amidinopropano)-diidrocloreto, in vitro. Indices de hemolise, glutationa reduzida, concentracao de metemoglobina, peroxidacao lipidica e especies reativas de oxigenio foram determinados. Eritrocitos de pacientes com anemia falciforme apresentaram taxas aumentadas de oxidacao da hemoglobina e peroxidacao lipidica e a concentracao de EGb 761 necessaria para atingir o mesmo efeito antioxidante foi pelo menos duas vezes maior em relacao aos eritrocitos normais (HbAA, eritrocitos AA), inibindo a formacao de especies reativas de oxigenio (IC50 de 13.6 µg/mL), prevenindo parcialmente a peroxidacao lipidica (IC50 de 242.5 µg/mL) e prevenindo a hemolise (IC50 de 10.5 µg/mL). Portanto, EGb 761 possui um efeito benefico no estado oxidativo dos eritrocitos SS. Entretanto, o EGb 761 nao preveniu a oxidacao da hemoglobina e da glutationa reduzida, nas concentracoes examinadas.

Original articleErythrocyte oxidative stress markers in children with sickle cell diseaseMarcadores de estresse oxidativo em eritrócitos de crianças com doença falciforme

Objective To determine eight parameters of oxidative stress markers in erythrocytes from children with sickle cell disease and compare with the same parameters in erythrocytes from healthy children, since oxidative stress plays an important role in the pathophysiology of sickle cell disease and because this disease is a serious public health problem in many countries.OBJECTIVE To determine eight parameters of oxidative stress markers in erythrocytes from children with sickle cell disease and compare with the same parameters in erythrocytes from healthy children, since oxidative stress plays an important role in the pathophysiology of sickle cell disease and because this disease is a serious public health problem in many countries. METHODS Blood samples were obtained from 45 children with sickle cell disease (21 males and 24 females with a mean age of 9 years; range: 3-13 years) and 280 blood samples were obtained from children without hemoglobinopathies (137 males and 143 females with a mean age of 10 years; range: 8-11 years), as a control group. All blood samples were analyzed for methemoglobin, reduced glutathione, thiobarbituric acid reactive substances, percentage of hemolysis, reactive oxygen species, and activity of the enzymes glucose 6-phosphate dehydrogenase, superoxide dismutase, and catalase. Data were analyzed using Students t-test and were expressed as the mean±standard deviation. A p-value of <0.05 was considered significant. RESULTS Significant differences were observed between children with sickle cell disease and the control group for the parameters methemoglobin, thiobarbituric acid reactive substances, hemolysis, glucose 6-phosphate dehydrogenase activity, and reactive oxygen species, with higher levels in the patients than in the controls. CONCLUSIONS Oxidative stress parameters in childrens erythrocytes were determined using simple laboratory methods with small volumes of blood; these biomarkers can be useful to evaluate disease progression and outcomes in patients.

Data for serum 1,5 anhydroglucitol concentration in different populations

1,5 anhydroglucitol (1,5-AG), is a nonmetabolized 1-deoxy form of glucose, originate mainly from the diet. 1,5-AG is a biomarker to detect and magnify hyperglycemic excursions (postprandial hyperglycemia) in diabetic patients. Concentrations of 1,5-AG has been applied as supporting biomarker to diagnosis of the major forms of diabetes (type 1, type 2, and gestational). The serum 1,5-AG reference interval is relevant to the appropriate clinical application of this biomarker. This article contains data regards to serum concentration of the biomarker primarily for healthy subjects, capture from the literature, in different populations. Correlation analysis between 1,5-AG and markers associated with diabetes and its complication were presented. The data was complementary to the study “Reference intervals for serum 1,5-anhydroglucitol in children, adolescents, adults, and pregnant women” (Welter et al., 2018). The data present in this article improve the comparisons for 1,5-AG in different conditions and methodologies.

Reference intervals for serum 1,5-anhydroglucitol in children, adolescents, adults, and pregnant women

BACKGROUND 1,5-anhydroglucitol (1,5-AG) is a validated marker of short-term glycemic control. We determined the reference intervals of 1,5-AG in different age groups and during pregnancy. METHODS Blood samples were collected from 2303 Euro-Brazilian healthy subjects: 580 children, 496 adolescents, 922 adults matched by age and sex, and 305 pregnant women in four gestational periods. Serum 1,5-AG was measured using an enzymatic reagent in an automated system. RESULTS The calculated reference intervals (nonparametric, 2.5th-97.5th) for males and females were, respectively: children, 96-302 and 89-277 μmol/l; adolescents, 84-311 and 79-277 μmol/l; and adults, 80-260 and 62-241 μmol/l. Males consistently showed significantly higher concentrations than females. 1,5-AG reference intervals in pregnant women were 56-298 μmol/l at <23 weeks gestation (n = 110), 37-166 μmol/l at 24-28 weeks gestation (n = 106), 34-155 μmol/l at 29-32 weeks gestation (n = 52), and 33-246 μmol/l at >32 weeks gestation (n = 37). No significant differences in 1,5-AG concentration were observed between non-pregnant and pregnant women at <23 weeks of gestation. A negative correlation (r = -0.287; p < .001) between 1,5-AG concentration and age was observed. CONCLUSIONS The reference intervals for 1,5-AG were affected by sex and age.

Von Willebrand Disease Lab Diagnosis.

AbstractThe hemorrhagic diseases are characterized by bleeding which can vary considerably according to their severity. The von Willebrand disease (VWD) is the most frequent hereditary hemorrhagic disease and the prevalence of clinically significant disease is probably closer to 1:1000, being an extremely heterogeneous and complex disorder that is related to the deficiency in concentration, structure or function of von Willebrand factor (VWF). The VWD is divided into type 1, with partial deficiency of the VWF, type 2, with qualitative defects in the molecule with four subdivisions, and type 3, with very low or undetectable levels of plasma and platelet VWF and ristocetin cofactor activity. The laboratory diagnosis of VWD is complex. Specific tests that assess the functionality and concentrations of the VWF and FVIII are needed. The routine tests are the bleeding time, the activated partial thromboplastin time and the platelet count, however, singly, they may not suggest the diagnosis of VWD, requiring further specific tests, such as VWF function evaluation through its ristocetin cofactor assay (VWF:RCo), VWF protein concentration immunoassay (VWF:Ag), the factor VIII coagulation assay (FVIII:C), VWF binding to immobilized collagen (VWF:CB), ristocetin-induced platelet aggregation (RIPA), VWF multimers patterns, factor VIII binding of immobilized VWF (VWF:FVIIIB), among others. From the moment the diagnosis is confirmed, the appropriate treatment for each patient is sought, with the purpose of increasing plasma concentrations of the deficient protein, both in bleeding episodes, as for invasive procedures. Although diagnosis facilitates treatment other approach in the present scenario is prenatal diagnosis which, is the need of the hour.

Hematological reference ranges among healthy adults of Curitiba, PR, Brazil

The development of this work focused on the establishment of new reference values for the complete blood count (CBC) in the city of Curitiba as this was last studied back in the 1980s.(1,2) With the improvement in cell counting and analysis technologies, the need to update hematologic reference values is evident with the inclusion of the new parameters provided by the most up-to-date equipment. Besides the methodological aspect, the updating of hematologic indices is also important in respect to the new socioeconomic conditions of the Brazilian population, as well as the new lifestyle in our society. Venous blood samples were obtained from 1000 individuals (500 women and 500 men) for routine hematological examinations. Blood collection was made in several healthcare clinics throughout the city and transported under controlled conditions to the Laboratorio Municipal de Curitiba where all analyzes were carried out. Hematology analyses were performed in an ABX Pentra 120 - Horiba cell counter. Participants for this study were randomly selected between June and October 2007 from the data base according to the following criteria: (i) age between 12 to 60 years old of both genders; (ii) exams identified as solely routine; (iii) cross file examinations to discard suspected pathologies.(3-5) Reference ranges were obtained from 2.5 and 97.5 accumulated percentiles in normal distributions. When normality could not be achieved, the 95% reference interval was obtained with the help of non-parametric ordinal descriptive statistics. Analyses were carried out using an Excel spreadsheet (Microsoft) and the statistical package Statistica 8.0 (StatSoft). Statistical significance was set for a p-value < 0.05. There were statistically significant differences (p-value < 0.05) between men and women for most hematology parameters. Men had higher red blood cell, hemoglobin, eosinophil, basophil and monocyte counts and higher hematocrit, mean cell hemoglobin, mean cell hemoglobin concentration and red cell distribution width values compared to women. Women had higher neutrophil and platelet counts than men. The red cell distribution width for both men and women was higher than the values commonly found in the literature.(6,7) The maximum limits of 16% found in our work are generally above the limit cited by the manufacturers of automated cell counters 14.5%). These differences may be explained by the different types of equipment on the market today where the cell measurement methods may be different. Regarding erythrogram and leukogram parameters, the values found for both women and men did not show significant differences compared to values reported in the literature.(1,2,8) Reference values, stratified by gender, are described in Table 1. Table 1 Mean and reference ranges for hematology laboratory values in the Municipal Laboratory of Curitiba, PR An important factor to be noted is the characteristic of the population involved in the current study. The Municipal Laboratory receives blood samples from about 100 government healthcare clinics scattered around the city of Curitiba that attend the citys most needy populations. Therefore, our results must be analyzed cautiously to avoid erroneous comparisons. In summary, our results showed that despite the improved technology of cell counting and analysis, the hematological parameters of the adult population of the city of Curitiba suffered no major changes compared to studies performed in the 1980s. However, our work incorporates one new parameter (red cell distribution width), and provides values that better reflect the current conditions of the majority of the adult population of the city of Curitiba, thereby allowing greater accuracy in the interpretation of the data provided by the complete blood count.

Prevalence of anemia in schools of the metropolitan region of Curitiba, Brazil

Background Anemia during childhood is one of the biggest public health problems worldwide, including Brazil. Insufficient or abnormal production of hemoglobin, loss of iron and excessive destruction of red blood cells are the most common causes of anemia. Among the reasons of anemia, iron deficiency accounts for 50% of anemia cases in developing countries. Affected individuals present a wide range of clinical problems, including delayed neuropsychomotor progression, impaired cellular immunity and reduction of intellectual capacity. This study aimed to evaluate the prevalence of anemia in children attending public schools in the metropolitan region of Curitiba, Paraná, Brazil. Method A retrospective study was conducted of 409 children aged 8–12 years old included in an extension project of the Universidade Federal do Paraná. The results of complete blood count and hemoglobin electrophoresis of all children were evaluated. Anemia was considered when the hemoglobin levels were <11.5 g/dL. Results The prevalence of anemia was found to be 2.2% of the population studied, with hypochromic microcytic anemia being the most common type. Seven children had sickle cell trait and one had β-thalassemia. Conclusion The prevalence of anemia in this study was considered normal according the World Health Organization classification, which is different from the data found in other Brazilian regions.