Guanosine enhances glutamate uptake and oxidation, preventing oxidative stress in mouse hippocampal slices submitted to high glutamate levels.

Glutamate (Glu) is the main mammalian brain neurotransmitter. Concerning the glutamatergic neurotransmission, excessive levels of glutamate in the synaptic cleft are extremally harmful. This phenomenon, named as excitotoxicity is involved in various acute and chronic brain diseases. Guanosine (GUO), an endogenous guanine nucleoside, possesses neuroprotective effects in several experimental models of glutamatergic excitotoxicity, an effect accompanied by an increase in astrocytic glutamate uptake. Therefore, the objective of this study was to investigate the involvement of an additional putative parameter, glutamate oxidation to CO2, involved in ex-vivo GUO neuroprotective effects in mouse hippocampal slices submitted to glutamatergic excitotoxicity. Mice were sacrificed by decapitation, the hippocampi were removed and sliced. The slices were incubated for various times and concentrations of Glu and GUO. First, the concentration of Glu that produced an increase in L-[14C(U)]-Glu oxidation to CO2 without cell injury was determined at different time points (between 0 and 90 min); 1000 µM Glu increased Glu oxidation between 30 and 60 min of incubation without cell injury. Under these conditions (Glu concentration and incubation time), 100 µM GUO increased Glu oxidation (35%). Additionally, 100 µM GUO increased L-[3,4-3H]-glutamate uptake (45%) in slices incubated with 1000 µM Glu (0-30 min). Furthermore, 1000  µM Glu increased reactive species levels, SOD activity, and decreased GPx activity, and GSH content after 30 and 60 min; 100 µM GUO prevented these effects. This is the first study demonstrating that GUO simultaneously promoted an increase in the uptake and utilization of Glu in excitotoxicity-like conditions preventing redox imbalance.

Behavioral, Neurochemical and Brain Oscillation Abnormalities in an Experimental Model of Acute Liver Failure.

Hepatic encephalopathy (HE) represents a brain dysfunction caused by both acute and chronic liver failures, and its severity deeply affects the prognosis of patients with impaired liver function. In its pathophysiology, ammonia levels and glutamatergic system hyperactivity seem to play a pivotal role in the disruption of brain homeostasis. Here, we investigate important outcomes involved in behavioral performance, electroencephalographic patterns, and neurochemical parameters to better characterize the well-accepted animal model of acute liver failure (ALF) induced by subtotal hepatectomy (92% removal of tissue) that produces ALF. This study was divided into three cohorts: (1) rats clinically monitored after hepatectomy every 6 h for 96 h or until death; (2) rats tested in an open-field task (OFT) before and after surgery and had blood, cerebrospinal fluid, and brain tissue collected after the last OFT; and (3) rats that had continuous EEGs recorded before and after surgery for 3 days. The hepatectomized rats presented significant motor behavioral changes accompanied by important abnormalities in classical blood laboratory parameters of ALF, and EEG features suggestive of HE and deep disturbances in the brain glutamatergic system. Using an animal model of ALF induced via subtotal hepatectomy, this work provides a comprehensive and reliable experimental model that increases the opportunity for studying the effects of new treatment strategies to be explored in an unprecedented way. It also presents insights into the pathophysiology of HE in a reproducible model of ALF, which correlates important neurochemical and EEG aspects of the syndrome.

Guanosine Exerts Neuroprotective Effect in an Experimental Model of Acute Ammonia Intoxication.

The nucleoside guanosine (GUO) increases glutamate uptake by astrocytes and acts as antioxidant, thereby providing neuroprotection against glutamatergic excitotoxicity, as we have recently demonstrated in an animal model of chronic hepatic encephalopathy. Here, we investigated the neuroprotective effect of GUO in an acute ammonia intoxication model. Adult male Wistar rats received an intraperitoneal (i.p.) injection of vehicle or GUO 60 mg/kg, followed 20 min later by an i.p. injection of vehicle or 550 mg/kg of ammonium acetate. Afterwards, animals were observed for 45 min, being evaluated as normal, coma (i.e., absence of corneal reflex), or death status. In a second cohort of rats, video-electroencephalogram (EEG) recordings were performed. In a third cohort of rats, the following were measured: (i) plasma levels of glucose, transaminases, and urea; (ii) cerebrospinal fluid (CSF) levels of ammonia, glutamine, glutamate, and alanine; (iii) glutamate uptake in brain slices; and (iv) brain redox status and glutamine synthetase activity in cerebral cortex. GUO drastically reduced the lethality rate and the duration of coma. Animals treated with GUO had improved EEG traces, decreased CSF levels of glutamate and alanine, lowered oxidative stress in the cerebral cortex, and increased glutamate uptake by astrocytes in brain slices compared with animals that received vehicle prior to ammonium acetate administration. This study provides new evidence on mechanisms of guanine-derived purines in their potential modulation of glutamatergic system, contributing to GUO neuroprotective effects in a rodent model of by acute ammonia intoxication.

Obesity and vitamin D deficiency: a systematic review and meta-analysis.

Over the past decade, there have been an increasing number of studies on the association between vitamin D deficiency and anthropometric state. However, we did not identify any meta-analyses of the relationship between obesity and vitamin D deficiency in different age groups. Thus, we evaluated the association between obesity and vitamin D deficiency. We searched for observational studies published up to April 2014 in PubMed/Medline, Web of Science and Scopus databases. We performed a meta-analysis in accordance with the random-effects model to obtain the summary measurement (prevalence ratio, PR). Among the 29,882 articles identified, 23 met the inclusion criteria. The prevalence of vitamin D deficiency was 35% higher in obese subjects compared to the eutrophic group (PR: 1.35; 95% CI: 1.21-1.50) and 24% higher than in the overweight group (PR: 1.24; 95% CI: 1.14-1.34). These results indicate that the prevalence of vitamin D deficiency was more elevated in obese subjects. The vitamin D deficiency was associated with obesity irrespective of age, latitude, cut-offs to define vitamin D deficiency and the Human Development Index of the study location.

Neuroprotective effects of guanosine administration on behavioral, brain activity, neurochemical and redox parameters in a rat model of chronic hepatic encephalopathy.

It is well known that glutamatergic excitotoxicity and oxidative stress are implicated in the pathogenesis of hepatic encephalopathy (HE). The nucleoside guanosine exerts neuroprotective effects through the antagonism against glutamate neurotoxicity and antioxidant properties. In this study, we evaluated the neuroprotective effect of guanosine in an animal model of chronic HE. Rats underwent bile duct ligation (BDL) and 2 weeks later they were treated with i.p. injection of guanosine 7.5 mg/kg once a day for 1-week. We evaluated the effects of guanosine in HE studying several aspects: a) animal behavior using open field and Y-maze tasks; b) brain rhythm changes in electroencephalogram (EEG) recordings; c) purines and glutamate levels in the cerebral spinal fluid (CSF); and d) oxidative stress parameters in the brain. BDL rats presented increased levels of glutamate, purines and metabolites in the CSF, as well as increased oxidative damage. Guanosine was able not only to prevent these effects but also to attenuate the behavioral and EEG impairment induced by BDL. Our study shows the neuroprotective effects of systemic administration of guanosine in a rat model of HE and highlights the involvement of purinergic system in the physiopathology of this disease.

Chronic sulforaphane oral treatment accentuates blood glucose impairment and may affect GLUT3 expression in the cerebral cortex and hypothalamus of rats fed with a highly palatable diet.

Obesity and insulin resistance are the key factors underlying the etiology of major health problems such as hypertension, diabetes and stroke. These important health issues lead researchers to investigate new approaches to prevent and treat obesity and insulin resistance. Good candidates are the phytochemical compounds that have been extensively studied in the field. Therefore, the aim of this study was to test whether sulforaphane (SFN, 1 mg kg⁻¹, 4 months treatment), a potent inducer of antioxidant enzymes present in cruciferous vegetables, had some beneficial effects on obesity and insulin resistance induced by a highly palatable (HP) diet in male Wistar rats. Glucose tolerance, serum and hepatic lipid levels, lipid profile, ALT, AST, urea and creatinine, GLUT1 and GLUT3 levels in the cerebral cortex, hippocampus and hypothalamus were analyzed. Glucose tolerance was lower in the HP diet groups, especially in the HP group treated with SFN. Except for the liver triacylglycerols, no differences were found in serum lipids, hepatic and kidney markers of the HP diet groups. Although expression of GLUT1 was similar between groups for all three brain structures analyzed, expression of GLUT3 in the cortex and hypothalamus had a tendency to decrease in the HP diet group treated with SFN. In conclusion, SFN at the specific dose was able to accentuate glucose intolerance and may affect GLUT3 expression in the cerebral cortex and hypothalamus.

Evidences that maternal swimming exercise improves antioxidant defenses and induces mitochondrial biogenesis in the brain of young Wistar rats.

Physical exercise during pregnancy has been considered beneficial to mother and child. Recent studies showed that maternal swimming improves memory in the offspring, increases hippocampal neurogenesis and levels of neurotrophic factors. The objective of this work was to investigate the effect of maternal swimming during pregnancy on redox status and mitochondrial parameters in brain structures from the offspring. Adult female Wistar rats were submitted to five swimming sessions (30 min/day) prior to mating with adult male Wistar rats, and then trained during the pregnancy (five sessions of 30-min swimming/week). The litter was sacrificed when 7 days old, when cerebellum, parietal cortex, hippocampus, and striatum were dissected. We evaluated the production of reactive species and antioxidant status, measuring the activities of superoxide-dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GPx), as well as non-enzymatic antioxidants. We also investigated a potential mitochondrial biogenesis regarding mitochondrion mass and membrane potential, through cytometric approaches. Our results showed that maternal swimming exercise promoted an increase in reactive species levels in cerebellum, parietal cortex, and hippocampus, demonstrated by an increase in dichlorofluorescein oxidation. Mitochondrial superoxide was reduced in cerebellum and parietal cortex, while nitrite levels were increased in cerebellum, parietal cortex, hippocampus, and striatum. Antioxidant status was improved in cerebellum, parietal cortex, and hippocampus. SOD activity was increased in parietal cortex, and was not altered in the remaining brain structures. CAT and GPx activities, as well as non-enzymatic antioxidant potential, were increased in cerebellum, parietal cortex, and hippocampus of rats whose mothers were exercised. Finally, we observed an increased mitochondrial mass and membrane potential, suggesting mitochondriogenesis, in cerebellum and parietal cortex of pups subjected to maternal swimming. In conclusion, maternal swimming exercise induced neurometabolic programing in the offspring that could be of benefit to the rats against future cerebral insults.

Validation of a food frequency questionnaire for children and adolescents aged 4 to 11 years living in Salvador, Bahia.

OBJECTIVE: To assess the validity of a food frequency questionnaire (FFQ) by applying it to children and adolescents living in Salvador, Bahia. METHODS: The validity of this FFQ with 98 food items was investigated among 108 children and adolescents who were selected from a sample of 1445 that had been planned for a study on the risk factors for asthma and other allergic diseases. The adults responsible for these children and adolescents gave responses for a 24-hour recall (R24h) and an FFQ. The average energy and nutrient values from the FFQ were compared with those from the R24h by means of the paired t test and Pearson correlation coefficients. The concordance was evaluated using the Bland-Altman method and kappa statistics. RESULTS: The energy and nutrient intake estimated using the FFQ was significantly higher than what was obtained using the R24h. The correlation coefficients adjusted for energy were statistically significant for protein, fat, vitamin C and zinc. The weighted kappa values ranged from 0.06 for vitamin A (p = 0.24) to 0.34 for energy (p < 0.00). The results from the Bland-Altman plots for lipid, protein and zinc showed the most significant validity parameters, and zinc was found to show the best concordance. CONCLUSION: The results suggest that the FFQ showed satisfactory validity for use in studies involving children and adolescents.

Individual and contextual determinants of the duration of diarrhoeal episodes in preschool children: a longitudinal study in an urban setting.

This study investigated individual and contextual factors associated with the duration of diarrhoeal episodes in 693 young children living in a large Brazilian city who were followed-up for at least 3 months. The outcome is analysed as a continuous variable, by means of a hierarchical conceptual model organizing the factors in meaningful blocks. A total of 2397 episodes were recorded (median duration 2 days, interquartile range 1-3 days). Low percentage of households connected to the sewerage system in the neighbourhood, low family purchasing power, high agglomeration, mother aged <19 years, low zinc content in child's diet, and episode severity were significantly associated with longer duration (0·26-0·69 days more). Purchasing power effect was largely mediated by environmental conditions, characteristics of the child, and hygienic behaviour. Environmental conditions acted as a possible effect modifier, enhancing the effect on duration of diarrhoea of the child not having being vaccinated against measles or breastfed for >6 months.

The effects of hypercaloric diets on glucose homeostasis in the rat: influence of saturated and monounsaturated dietary lipids.

Consumption of energy-dense/high-fat diets is strongly and positively associated with overweight and obesity, which are associated with increase in the prevalence of certain chronic diseases. We evaluated the effect of hypercaloric/fat or normocaloric diets on some biochemical parameters in rats. Seventy-two rats were divided into four groups that were fed for 16 weeks with diets: normocaloric [9.12% soy oil, normocaloric soy oil (NSO)], hypercaloric olive oil [43.8% olive oil, hypercaloric olive oil (HOO)], hypercaloric saturated fat [43.8% saturated fat, hypercaloric saturated fat (HSF)] and normocaloric saturated fat [43.8% saturated fat, normocaloric saturated fat (NSF)]. HSF rats consumed more calories daily than the others and gained more retroperitoneal fat, although HSF and HOO rats had higher body weight. In liver, glycogen synthesis and concentration were higher in rats HSF and NSF. In plasma, total cholesterol (TC) levels were higher in HSF rats than in the others, and triacylglycerol (TAG) levels were lower in HOO and higher in HSF rats in relation to the others. In liver, TC and TAG were elevated in HSF, NSF and HOO rats. Paraoxonase 1 activity, which is related to high-density lipoprotein cholesterol and has anti-atherogenic role was lower in rats HSF. In HOO rats, glucose tolerance test was altered, but insulin tolerance test was normal. These results suggest that consumption of energy-dense/high-fat diets, both saturated or monounsaturated, causes damaging effects. However, more studies are necessary to understand the mechanisms by which these diets cause the metabolic alterations observed.

Is overweight a risk factor for wheezing in pre-school children? A study in 14 Brazilian communities.

OBJECTIVE: To evaluate the association between overweight and wheezing in pre-school children in 14 small Brazilian communities. METHODS: Cross-sectional epidemiological study, conducted between 2001 and 2002. A sample of 3453 children under 5 years of age was taken from nine communities in the state of Bahia and five in the state of São Paulo. Data on housing, family and children were obtained by applying structured questionnaires in loco. Weight and height for each child were also measured. The association between wheezing and overweight was assessed by unconditional logistic multivariate regression models. RESULTS: Overweight children had a greater frequency of wheezing and an odds ratio of 2.57 (95% confidence interval 1.51-4.37) was estimated after controlling for several potential confounding variables. The magnitude of the risk was not affected by several different model specifications. CONCLUSION: Excess weight is associated with increased risk for wheezing in this population of children below 5 years of age.

Three novel mutations in the activin receptor-like kinase 1 (ALK-1) gene in hereditary hemorrhagic telangiectasia type 2 in Brazilian patients.

Hereditary hemorrhagic telangiectasia (HHT) or Osler-Rendu-Weber disease is a systemic fibrovascular dysplasia with an autosomal dominant inheritance pattern. Mutations in two genes, endoglin and ALK-1, are known to cause HHT, both of which mediate signaling by transforming growth factor beta ligands in vascular endothelial cells. Ten patients were analyzed. Diagnosis of HHT was carried out by means of family history, recurrent bleeding, and the presence of multiple telangiectases lesions. Conformation-sensitive gel electrophoresis analyses with consistent abnormal migration patterns were cloned and sequenced using the MegaBace 1000 DNA automated analyzer. Three novel mutations were identified in the coding sequence of the ALK-1 gene in five patients and their families, which demonstrated clinical manifestations of HHT type 2. These mutations included a G insertion and a T deletion of single base pairs in exons 3 and 7, as well as missense mutations in exons 7 and 8 of the ALK-1 gene. These data indicate that loss-of-function mutations in a single allele of the ALK1 locus are sufficient to contribute to defects in maintaining endothelial integrity. We suggest the high rate of mutation detection and the small size of the ALK-1 gene make genomic sequencing a viable diagnostic test for HHT2.

Growth faltering in childhood related to diarrhea: a longitudinal community based study.

OBJECTIVE: This study aimed to evaluate the association of diarrhea and acute lower respiratory tract infections (ALRI) with growth of preschool children. DESIGN: A longitudinal community-based study over a 12-month period. Children were followed up with thrice-weekly household visits for collection of morbidity data. Every 4 months (round) clinical and anthropometric examinations were performed. At baseline a questionnaire was used to collect socioeconomic family data and environmental household variables. Generalized estimating equation was used in the statistical analysis. The variations in weight-for-age or height-for-age Z-scores in each round were the dependent variables, while the main independent variables were the number of days with diarrhea and ALRI. SETTING: Serrinha, located in Northeast Brazil. SUBJECTS: In total, 487 children, aged 6-48 months at baseline, with 1-y complete follow-up. RESULTS: The number of sick days with diarrhea or ALRI was not associated with mean changes in weight-for-age Z-scores. However, the mean of height-for-age Z-scores was found to decrease in those children with 7 or more days of diarrhea (beta=-0.0472; P=0.016) but not with 1 or more days of ALRI (beta=0.0022; P=0.406) in all rounds of the follow-up period. CONCLUSION: Results of the study reinforce the concept of diarrhea burden as a major determinant of poor growth in children under 5 y of age. Actions targeted to decrease the risk factors for the occurrence of diarrhea may represent an important component of interventions aimed to ensure satisfactory child growth.

Asymptomatic giardiasis and growth in young children; a longitudinal study in Salvador, Brazil.

This study sought to assess the effect of giardiasis on growth of young children. In Salvador, northeast Brazil, 597 children initially aged 6 to 45 months were followed for a year in 1998/9, measured anthropometrically thrice, every 6 months, and monitored for diarrhoea prevalence twice weekly. Stool samples were collected and examined during the second round of anthropometry, and infected children were treated 39 days later, on average (S.D. 20 days). For each 6-month interval, the gains in z-scores of infected and uninfected children were compared, after adjustment for potential confounding factors, including longitudinal prevalence of diarrhoea. No significant difference was found for the first interval but in the second, the gain in adjusted height-for-age z-score was 0.09 less in infected than uninfected children, equivalent to a difference in height gain of 0.5 cm. The shortfall in growth was greater in children who remained free of diarrhoea, and was significantly correlated with the proportion of the second interval during which the child had remained untreated. We conclude that Giardia can impede child growth even when asymptomatic, presumably through malabsorption. This finding challenges the view that young children found to have asymptomatic giardiasis in developing countries should not be treated.

Consenso sobre a transdisciplinaridade na medicina periodontal 2005 / Consensus on periodontal medicine transdisciplinarity 2005

Objetivo : estabelecer diretrizes para as investigações sobre a transdisciplinaridade na Medicina Periodontal. Métodos: representantes das diferentes especialidades que estudam e pesquisam diversoa aspectos da Medicina Periodontal participaram do II Seminário de Periodontia em Saúde Coletiva - Uma Visão Transdisciplinar na Medicina Periodontal (Feira de Santana-Bahia, 2005) e prepararam este consenso, no qual estabeleceram as diretrizes para as investigações sobre a transdisciplinaridade na Medicina Periodontal, particularmente na questão da possível associação entre a doença periodontal e a prematuridade e baixo peso ao nascer. Resultados: Este consenso foi produzido após a apresentação de cada questão e extensa discussão pelos participantes do evento

The WHO dose pole for the administration of praziquantel is also accurate in non-African populations.

In 2001, WHO developed a pole for the administration of praziquantel without the use of weighing scales, with encouraging results in African populations. In the present study, the pole was tested on height/weight data from 9354 individuals from 11 non-African countries. In more than 98% of the individuals (95% CI 97.8-98.4) the pole estimated an acceptable dosage (30-60 mg/kg), a performance statistically similar to that observed in African populations. Reproducing the present pole in the form of a strip of paper and including it in each container of praziquantel would greatly facilitate the administration of the drug in large-scale interventions.

Childhood stunting in Northeast Brazil: the role of Schistosoma mansoni infection and inadequate dietary intake.

OBJECTIVE: To evaluate the relationship between stunting, Schistosoma mansoni infection and dietary intake in schoolchildren. DESIGN: This is a cross-sectional study. Two stool samples were obtained from each child and examined quantitatively for the presence of S. mansoni, Ascaris lumbricoides and Trichuris trichiuria eggs. Information on dietary intake, and demographic, biologic and socioeconomic variables was elicited during the in-home survey. Logistic regression was used to evaluate the association between stunting (height for age < -2s.d.), parasitic infection and food consumption. SETTING: The study was carried out in the city of Nazaré, located in the Recôncavo region of the State of Bahia, Northeastern Brazil. SUBJECTS: The sample consisted of 461 children 7-14 y old, 228 boys and 233 girls, recruited from public schools. RESULTS: Of the children studied, 55.1% presented with S. mansoni infection and 22.1% were stunted. The median protein, lipid and carbohydrates intake were 47.8, 36.0 and 248.2 g/day, respectively. The median caloric consumption was 1527.0 kcal (6388.97 kJ/day). The analysis indicated that children heavily infected (> or = 400 eggs/g of stool) with S. mansoni had a 2.74-fold (95% CI: 1.32-5.67) higher risk of stunting than uninfected children, and those with inadequate intake of lipid (< 36 g/day) had a 1.83-fold (95% CI: 1.05-3.20) increased risk of stunting compared to those with adequate diets. CONCLUSION: Heavy S. mansoni infection and inadequate dietary intake of fat in schoolchildren play a significant and independent role in the development of stunting. This meaning that nutritional interventions in this age group in S. mansoni endemic areas must include the diagnosis and treatment of the infection associated with dietary measures. SPONSORSHIP: This study was supported by the Thrasher Foundation. Sandra Maria Conceição Pinheiro is a National Council on Technological Development Scholarship Awardee (CNPq), #302228/81-0.

The contribution of 700,000 ORF sequence tags to the definition of the human transcriptome.

Open reading frame expressed sequences tags (ORESTES) differ from conventional ESTs by providing sequence data from the central protein coding portion of transcripts. We generated a total of 696,745 ORESTES sequences from 24 human tissues and used a subset of the data that correspond to a set of 15,095 full-length mRNAs as a means of assessing the efficiency of the strategy and its potential contribution to the definition of the human transcriptome. We estimate that ORESTES sampled over 80% of all highly and moderately expressed, and between 40% and 50% of rarely expressed, human genes. In our most thoroughly sequenced tissue, the breast, the 130,000 ORESTES generated are derived from transcripts from an estimated 70% of all genes expressed in that tissue, with an equally efficient representation of both highly and poorly expressed genes. In this respect, we find that the capacity of the ORESTES strategy both for gene discovery and shotgun transcript sequence generation significantly exceeds that of conventional ESTs. The distribution of ORESTES is such that many human transcripts are now represented by a scaffold of partial sequences distributed along the length of each gene product. The experimental joining of the scaffold components, by reverse transcription-PCR, represents a direct route to transcript finishing that may represent a useful alternative to full-length cDNA cloning.