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No / Glutathione depletion is a consistent feature of the progression of mild to severe acute pancreatitis. In this study, we examined the temporal relationship between cysteine, homocysteine, and cysteinyl-glycine levels; total reduced erythrocyte glutathione; gamma-glutamyl transpeptidase activity; and disease severity. Initially, cysteine concentration was low, at levels similar to those of healthy controls. However, glutathione was reduced whilst cysteinyl glycine and gamma-glutamyl transpeptidase activity were increased in both mild and severe attacks. As the disease progressed, glutathione and cysteinyl glycine were further increased in mild attacks and cysteine levels correlated with homocysteine (r = 0.8, P < 0.001) and gamma-glutamyl transpeptidase activity (r = 0.75, P < 0.001). The progress of severe attacks was associated with glutathione depletion, reduced gamma-glutamyl transpeptidase activity, and increased cysteinyl glycine that correlated with glutathione depletion (r = 0.99, P = 0.01). These results show that glutathione depletion associated with severe acute pancreatitis occurs despite an adequate cysteine supply and could be attributed to heightened oxidative stress coupled to impaired downstream biosynthesis.

M. Tech. (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal University of Technology. / Background: Increased serum homocysteine is well known as an independent cardiovascular risk factor. Hyperhomocysteinemia may be due to several factors such as nutritional deficiencies and genetics. The MTHFR C677T polymorphism is associated with increased serum homocysteine. Folate and vitamin B12 play essential roles in lowering homocysteine levels. Limitations have been identified using serum vitamin B12 as a marker for vitamin B12 status due to lack an efficient of test. Holotranscobalamin has been reported as a more accurate marker for vitamin B12 status. Cardiovascular risk due to hyperhomocysteinemia has been confirmed among the elderly in Sharpeville. Knowledge of the prevalence of MTHFR C677T polymorphism among Black elderlies in South Africa is limited. Objectives: The main aim of the study was to evaluate the prevalence of MTHFR C677T polymorphism as a cardiovascular risk in an elderly black population in Sharpeville. Correlations between the presence of MTHFR C677T polymorphism and homocysteine metabolic markers were evaluated. Holotranscobalamin as a diagnostic test for vitamin B12 status was also assessed in this study. Materials and methods: This study was ethically approved by the Vaal University of Technology ethics committee (20140827-1ms). It was an observational, experimental study conducted in 102 elderly (≥60 years) attending the day-care centre in Sharpeville. Real-Time PCR was used to determine MTHFR genotypes. Folate and vitamin B12 were measured with AIA-PACK. Homocysteine levels were determined with an automated Konelab™ 20i and holotranscobalamin by ELISA. STATA 12 software was used for analysis of descriptive and inferential statistics. Results: The prevalence of MTHFR C677T polymorphism in this sample population was 19%. Heterozygous CT single nucleotide polymorphism was 17% and mutant homozygous TT was 2%. The majority (81%) of the subjects had wild type homozygous CC genotypes. No associations were found between MTHFR C677T genotypes and homocysteine and folate levels. Hyperhomocysteinemia was high (54%) and low (5%) folate deficiency found. No vitamin B12 deficiency was found however 7% were on the category of likely to be deficient. Sensitivity and specificity of holotranscobalamin were 100% and 95% respectively. Conclusion: The conclusions drawn from the study is that the prevalence of MTHFR C677T polymorphism is low within elderly in Sharpeville. There is a high risk of cardiovascular disease as a result of high prevalence of hyperhomocysteinemia. An intervention to lower homocysteine concentration of elderlies residing in Sharpeville is needed. Other genetic predisposing factors of increased homocysteine levels should be investigated.

Homocysteine (Hcy) has been identified as a primary risk factor for atherosclerosis as it induces endothelial cell (EC) activation/dysfunction and thus potentially initiating atherosclerotic plaque formation. There is accumulating evidence indicating a key role for oxidative stress in mediating Hcy atherogenic effects. The aim of this study was to evaluate the effects of chronic treatment with Hcy on EC activation and to explore the role of oxidative stress in these effects. Human umbilical vein endothelial cells (HUVEC) were cultured and treated chronically with DL-Hcy for 5-9 days. An in vitro flow system was also used to characterize the different types of interactions between DL-Hcy-treated HUVEC and neutrophils under physiological flow conditions. EC activation was studied by characterizing the activation of the JNK pathway and the up-regulation of different cell adhesion molecules (CAM) and cytokines, using different techniques including western blot, immunohistochemical staining, enzyme-linked immunosorbent assay and polymerase chain reaction. The role of oxidative stress was investigated by measuring the production of ROS and evaluating the efficiency of antioxidants. Furthermore, the role of nitric oxide and nitric oxide synthase in modulating Hcy effects was investigated. Chronic treatment with DL-Hcy did not kill the EC however, it inhibited cell proliferation. Furthermore, this treatment induced EC activation/dysfunction which was characterized by sustained activation of the JNK pathway, which in turn mediated up-regulation of E-selectin, ICAM-1 and to lesser extent P-selectin. Furthermore, DL-Hcy induced production of IL-8 protein. These CAM and chemokines collectively mediated different interactions between DL-Hcy-treated HUVEC and neutrophils under flow conditions including tethering, rolling, adherence and transmigration. DL-Hcy was also shown to induce significant ROS generation which mediated activation of the JNK pathway. Antioxidants restored DL-Hcy-induced interactions under flow to the basal level. DL-Hcy was shown to induce eNOS uncoupling which mediated, at least in part, the DL-Hcy-induced ROS production. Furthermore, short term treatment with NO inhibited DL-Hcy-induced HUVEC:neutrophil interactions in a cGMP-independent manner. In summary, this research showed that DL-Hcy has several proatherogenic effects, mediated at least in part by the JNK pathway, and induces EC activation/dysfunction priming for atherosclerosis initiation. The data supports that oxidative stress mediates the majority of Hcy atherosclerotic effects. Antioxidants tested, JNK inhibitors and NO showed promising results in reversing all DL-Hcy effects and restoring EC normal status.

Background: In the past 37 years, increased efforts have been directed toward a better understanding of the importance of Hcy in disease and it has now become clear that hyperhomocysteinemia is a major independent risk factor for CVD. Extensive research on the influence of vitamin supplementation leading to reductions in Hcy levels and improvements in cardiovascular function has been done. The importance of exercise in the lowering of cardiovascular risk factors, as well as its favourable influence on cardiovascular function has also been indicated in several studies, however, the limited number of studies investigating the effect of exercise on Hcy concentrations revealed contradicting results. Furthermore, a relationship between Hcy concentration and cardiovascular function with the intervention of an exercise training and a vitamin supplementation programme respectively has also not been investigated. Objective: The objective of this study was to examine the effect of a 12-week exercise training and a 12-week vitamin supplementation intervention respectively on tHcy concentrations and cardiovascular function, and whether the change in tHcy concentration within the different interventions correlated with the change in cardiovascular function. Methods: In a randomised controlled cross-over intervention study, 52 men matched for age, cardiorespiratory fitness levels and cardiovascular risk factors were randomly assigned to one of 3 groups (Group A = exercise training programme, 20-30min. at 70-80% of HRmax; Group B = 400 g folic acid and 25 g vitamin B12 supplement; Group C = control). Group A and B were crossed over for phase 11, and Group C remained the control. The questionnaires were completed, and the body composition variables (BMI, WHR and body fat percentage), cardiovascular function (Finometer), tHcy concentrations and VO2max, were measured before and after each 12-week intervention period. A 6-week washout period separated the crossovers. Results: The ANCOVA, adjusted for age and BMI, showed that the percentage change from baseline to end, corrected for baseline of the tHcy concentration increased significantly (p ≤ .05) by 9.7% with the exercise training intervention and decreased significantly (p ≤ .05) by 12.9%, with the vitamin supplementation intervention. The ANCOVA of the percentage change from baseline to end in cardiovascular function showed that the vitamin supplementation intervention resulted in improvements in cardiovascular function (decreased resting MAP, TPR and increased resting SV, CO, Cw) in comparison to the impairment in cardiovascular function with the exercise training intervention (increased resting DBP, MAP and TPR). The relationship between the tHcy concentration and cardiovascular function at baseline and within each of the different interventions were assessed by partial correlations adjusted for age, BMI and VO2max. Significant (p ≤ .05) relationships only occurred within the vitamin supplementation intervention, where decreased percentage change in tHcy concentration significantly correlated with increased percentage change of resting SV and CO and decreased percentage change of resting TPR. Conclusion: The general conclusion that can be drawn is that a 12-week vitamin supplementation intervention showed increased health related results, e.g. a significant reduction in tHcy concentration, an improvement in cardiovascular function and a significant positive relationship between these b o factors, in comparison to the 12-week exercise training intervention that significantly increased the tHcy concentration and did not show increased health related results. Due to inadequate compliance to the exercise training intervention, no conclusion can be drawn with regard to the effect of exercise training on tHcy concentrations and cardiovascular function. / Thesis (M.Sc. (Human Movement Science))--North-West University, Potchefstroom Campus, 2007

Au cours du développement, les modifications du métabolisme des monocarbones liées à une malnutrition peuvent être délétères autant pour la mère que pour le nouveau-né. De plus, les conséquences à long terme d'une carence en période gestationnelle et périnatale sont mal connues, notamment en ce qui concerne les pathologies cardiaques et hépatiques. Nous avons mis en oeuvre un modèle nutritionnel de rates adultes carencées en donneurs de groupements méthyles (vitamines B12, folates et choline) avant la gestation. Ces micronutriments participent à la régulation de différentes enzymes impliquées dans le métabolisme de l'homocystéine. Afin de se placer dans un contexte de physiopathologie, proche de la situation clinique évaluée, nous avons choisi d'alimenter les rates avec un régime carencé un mois avant la mise en accouplement et de poursuivre ce régime pendant la période d'allaitement. Nous avons évalué les répercussions métaboliques et fonctionnelles du régime sur les tissus myocardique et hépatique, chez le nouveau né à 21 jours. Nous avons étudié l'effet de cette carence en groupements méthyles sur le métabolisme énergétique lipidique et sur la carnitine. Conséquences de la carence au niveau myocardique : Le régime carencé en donneurs de méthyles induit une hypertrophie cardiaque avec une augmentation de l'épaisseur du myocarde et un agrandissement des cardiomyocytes. L'étude protéomique du myocarde et l'analyse des données par bioinformatique identifient PGC-1[alpha], PPAR[alpha] et ERR[alpha] comme principaux déterminants des variations d'expression des protéines du métabolisme oxydatif mitochondrial. Nous avons observé une diminution d'expression de PPAR[alpha] et ERR[alpha] et une inactivation de PGC1[alpha] par hypométhylation et hyperacétylation, en lien avec une diminution d'expression de PRMT1 et de SIRT1 et une augmentation de SAH. Conséquences de la carence au niveau du foie : La carence s'accompagne de l'apparition d'une stéatose hépatique microvésiculaire, avec une élévation des taux tissulaires de lipides et de triglycérides. De plus, nous avons observé une augmentation des marqueurs pro inflammatoires sans augmentation des marqueurs de fibrose. A cet égard, nos résultats ont montré qu'un déficit de synthèse de carnitine, impliquée dans la bêta-oxydation et le stockage des acides gras, jouerait un rôle déterminant dans la pathogenèse de la stéatose chez le nouveau-né. Il existe également une dérégulation du métabolisme oxydatif des acides gras, avec diminution d'activité des complexes I et II de la chaîne respiratoire, qui résulte d'une hypométhylation de PGC1 et d'une diminution d'expression de PPAR[alpha], ER[alpha] et ERR[alpha]. En conclusion, nos résultats montrent que la carence maternelle en donneurs de méthyles, induit des modifications sur la fonction de PGC-1[alpha]. Ces modifications sont associées à des altérations de l'oxydation des acides gras et sur la fonction mitochondriale pendant la période néonatale, ce qui entraîne l'accumulation de lipides dans les tissus myocardique et hépatique. Le lien entre la carence en donneurs de méthyles et l'altération de la méthylation de PGC-1[alpha] modifie les activités des enzymes impliquées dans la méthylation et l'acétylation de PGC-1. Ces enzymes sont aussi liées à des modifications épigénomiques qui modulent la fonction et l'expression des protéines. Nos résultats sont en accord avec les études de population de Barker et al, qui suggèrent que la nutrition maternelle pendant les étapes précoces de la vie est corrélée avec le risque de maladies cardio-vasculaires dans la vie adulte indépendamment des autres facteurs de risque / During development, changes in carbon metabolism related to malnutrition may be deleterious for both the mother and the newborn. In addition, long-term consequences of a methyl deficiency gestational and prenatal are poorly understood. We are particularly interested in studying these effects on the heart and liver. We have used a nutritional model of adult rats deficient in methyl donors (vitamin B12, folate and choline) before pregnancy. To be placed in a context of pathophysiology, close to the clinical situation, we chose to feed the rats with a methyl deficient diet one month before mating and continue this diet during the suckling period. We evaluated the metabolic and functional effects of this diet on myocardial and hepatic tissues, in the newborn pups in 21 days old. We studied the effects of methyl deficient diet on lipid and energy metabolism and carnitine. Consequences of the deficiency at the myocardial: The diet deficient in methyl donors induces cardiac hypertrophy with an increase in myocardial thickness and enlargement of cardiomyocytes. The proteomics analysis of the bioinformatics data identifies PGC-1[alpha], ERR[alpha] and PPAR[alpha] as major determinants of changes in protein expression of mitochondrial oxidative metabolism. We observed a decreased expression of PPAR[alpha] and ERR[alpha] and an inactivation of PGC1[alpha] by hypomethylation and hyperacetylation, in conjunction with a decrease in the expression of PRMT1 and SIRT1 and increased the level of SAH. Consequences of the deficiency at liver: Deficiency is accompanied by the appearance of microvesicular hepatic steatosis, with elevated tissue levels of lipids and triglycerides. Increase of inflammation was observed in this model with no changes in fibrosis score. In this respect, our results showed a deficiency of carnitine synthesis, involved in the beta-oxidation and storage of fatty acids, play a role in the pathogenesis of hepatic steatosis in the newborn. There is also a deregulation of the oxidative metabolism of fatty acids, with decreased activity of complex I and II of the respiratory chain, resulting in hypomethylation of PGC1 and decreased expression of PPAR[alpha], ER[alpha] and ERR[alpha]. In conclusion, our results show that maternal deprivation in methyl donors impaired the function of PGC-1[alpha]. These changes are associated with alterations in fatty acid oxidation and mitochondrial function during the neonatal period, with lipid accumulation in myocardial tissue and liver. The link between the methyl donor deficiency and impaired methylation of PGC-1[alpha] alters the activities of enzymes involved in methylation and acetylation of PGC-1[alpha]. These enzymes are also associated with epigenetic changes and the function and gene expression. Our results are consistent with population studies by Barker and colleagues, who suggest that maternal nutrition during early stages of life is correlated with the risk of cardiovascular disease in adult, independent of other risk factors

Lúpus eritematoso sistêmico (LES) é uma doença multisistêmica crônica e autoimune de etiologia desconhecida. O LES é considerado um fator de risco independente para eventos cardiovasculares em qualquer faixa etária. A coexistência de fatores de risco tradicionais e não tradicionais (alto nível plasmático de homocisteína) são as causas para o risco do desenvolvimento de doenças cardiovasculares (DCV) em portadores de LES. A hiperhomocisteinemia é considerada um fator de risco independente para DCV e cerebrovasculares e tem como causas fatores nutricionais, genéticos e fisiológicos. Proteína C reativa de alta sensibilidade (hs-PCR), fator de necrose tumoral-? (TNF- ?), interferon ?, MCP-1 (Monocyte Chemoattractant Protein- 1) e leptina tem sido descritos como importantes biomarcadores para DCV. Por outro lado, adiponectina e grelina tem sido retratadas como potentes protetores contra o desenvolvimento da aterosclerose. Parâmetros antropométricos aumentados também podem estar associados ao risco de DCV como aumento da espessura da camada íntima da carótida. O processo de inflamação crônica existente no LES está diretamente associado a alterações no perfil lipídico e no metabolismo de lipoproteínas. Análises em larga escala dos perfis de expressão de proteínas estão se tornando uma importante ferramenta na investigação de várias patologias associadas a resposta autoimune. Objetivos: Descrever e comparar medidas antropométricas, ingestão alimentar e proteoma plasmático em pacientes com lúpus eritematoso sistêmico juvenil (LESJ) e controles saudáveis. Comparar metabólitos e antropometria entre dois clusters metabólicos de pacientes com LESJ (Lúpus Melhor Perfil Metabólico - LMPM e Lúpus Pior Perfil Metabólico - LPPM) e um cluster com controles saudáveis (Controle Melhor Perfil Metabólico) e, também, em dois clusters de pacientes com LESJ definidos pela dose diária de corticoide administrada. Métodos: Foram recrutadas 19 adolescentes portadoras de LESJ e 39 controles saudáveis. Foram mensurados índice de massa corporal (IMC), peso, estatura, circunferência da cintura (CC), ingestão alimentar (Recordatório de 24 horas), SLEDAI (Systemic Lupus Erythematosus Disease Activity Index), SLICC (Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index), níveis plasmáticos de homocisteína, vitamina B12, 6 folato, TNF-?, hs-PCR, MCP-1, adiponectina, leptina, grelina, perfil lipídico e proteoma plasmático pela técnica Shotgun proteomics com Isobaric Tag for Relative and Absolute Quantitation. Os grupos foram comparados por ANCOVA. k-cluster foi usado para separar LESJ e controles em dois clusters extremos de melhor e pior perfil metabólico de acordo com os níveis plasmáticos de homocisteína, TNF-?, hsPCR e folato para a análise da proteômica. Resultados: Pacientes com LESJ apresentaram maior IMC, CC, homocisteína, triglicérides, TNF-?, hsPCR e menor folato plasmático quando comparados ao grupo controle. Foram encontradas 10 proteínas com expressão significativamente diferente entre os clusters: Cluster Lúpus Melhor Perfil Metabólico (LMPM), Cluster Lúpus Pior Perfil Metabólico (LPPM) e Cluster Controle Melhor Perfil Metabólico (CMPM) (? -2-macroglobulina, ? -1-antitripsina, apoliproteína AI, apoliproteína E, ceruloplasmina, complemento C3, fibrinogênio de cadeia ?, haptoglobina, hemopexina e sorotransferrina). Oito proteínas foram mais expressas no LMPM e menos expressas no LPPM comparados com CMPM. As proteínas menos expressas no LPPM foram negativamente correlacionadas com maior risco para DCV. Conclusão: O presente estudo mostrou que as adolescentes com LESJ apresentaram maior IMC, circunferência da cintura, concentrações séricas de homocisteína, triglicérides, TNF-?, hs-PCR e, menor estatura e concentração de folato sérico quando comparadas com adolescentes saudáveis. Além disso, o cluster LPPM apresentou uma expressão diminuída das proteínas apolipoproteína AI, apolipoproteína E, alfa-2- macroglobulina, alfa-1-antitripsina, ceruloplasmina, complemento C3, hemopexina e sorotransferrina quando comparado aos clusters CMPM e LMPM. Sendo assim, é possível concluir que o presente estudo sinaliza possíveis complicações cardiovasculares futuras em pacientes com LESJ e, sugerem a necessidade de novos estudos, a fim de elucidar a interação do estado nutricional e das proteínas encontradas pela proteômica no contexto de sistemas biológicos em pacientes com LESJ. / Systemic lupus erythematosus (SLE) is a, autoimmune, chronic and inflammatory disease of unknown etiology. SLE is considered an independent risk factor for cardiovascular diseases at any age. Reasons for the increased risk of CVD in SLE include the co-existence of traditional cardiac risk factors and nontraditional risk factors such as high concentrations of homocysteine. Elevated homocysteine (Hcy) levels is considered an independent risk factor for CVD. Hiperhomocysteine can indicate undernourishment due to a lack of vitamins or genetic alterations. Inflammatory biomarkers have been consistently associated with the presence of CVD in multiple studies from different populations, including SLE, such as high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor - ? (TNF - ?), type I interferon (IFN), monocyte chemoattractant protein-1 (MCP-1) and leptin. On the other hand, some cytokines and hormones are related to atherosclerosis prevention, such as adiponectin and ghrelin. The existing chronic inflammation process in SLE is directly associated with changes in lipid and lipoprotein metabolism. Some comprehensive studies have been conducted at multiple biological levels including DNA (or genomics), mRNA (or transcriptomics), protein (or proteomics) and metabolites (or metabolomics). The \'omics\' platforms allow us to re-examine SLE at a greater degree of molecular resolution. Objectives: To describe and compare anthropometric measurements, food intake, metabolites and plasma proteomic analysis in child-hood onset systemic lupus erythematosus (c-SLE) and healthy controls. To compare metabolites and anthropometric parameters between two clusters with c-SLE (lupus cluster with the best - LCBMP and the worst metabolic profile - LCWMP) and one cluster with controls (healthy cluster with the best metabolic profile - HCBMP) and to compare metabolites and anthropometric parameters between two clusters of c-SLE patients defined by daily corticosteroid doses intake. Methods: 19 c-SLE and 39 healthy volunteers were recruited. We evaluated BMI, weight, height, waist circumference, food intake through a 24-hours recalls, SLEDAI (Systemic Lupus Erythematosus Disease Activity Index), SLICC (Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index), SLICC, serum levels of homocysteine, vitamin B12, 8 folate, TNF-?, hs-C reactive protein, monocyte chemoattractant protein-1, adiponectin, leptin, ghrelin, lipid profile and plasma proteomic by Shotgun proteomics with Isobaric Tag for Relative and Absolute Quantitation. The groups were compared by ANCOVA. k-cluster were used to separate SLE and control groups into two different clusters with the best and the worst metabolic profile according to homocysteine, TNF-?, hsCRP and folate plasma levels for proteomic assessment. Results: SLE patients presented higher BMI, WC, homocysteine, triglycerides, TNF-?, hsCRP levels and lower plasma folate when compared to controls. We found 10 proteins with significantly different expression between healthy cluster with the best metabolic profile (HCBMP) and lupus cluster with the best (LCBMP) and the worst metabolic profile (LCWMP) (alpha-2- macroglobulin; alpha-1-antitripsin, apoliprotein AI, apoliprotein E, ceruloplasmin, complement C3, fibrinogen ? chain, haptoglobin, hemopexin and serum transferrin). Eight proteins were higher expressed in LCBMP and lower expressed in LCWMP compared with HCBMP. Proteins less expressed in LCWMP were negatively correlated with a higher risk for cardiovascular disease. Conclusion: Our results described some previously cardiovascular risk factors in c-SLE patients and possible associations between nutritional status and cardiovascular disease risk factors. Proteomic results showed acute phase proteins and pro-inflammatory proteins more expressed in c-SLE patients compared to controls. These results might allow us to treat c-SLE with personalized diets to avoid cardiovascular complications in future. The small sample size and the cross-sectional design were the limitations of the study, but it is a rare disease in the pediatric field and, despite this fact, the present study was able to characterize two distinct nutritional and metabolic groups with uncommon proteins expressed. These results deserve further investigations to better elucidate the whole of these proteins in the context of systems biology interactions in SLE pediatric patients.

Made available in DSpace on 2016-01-26T12:51:12Z (GMT). No. of bitstreams: 1 mariapaulaalvarenga_tese_parte 1.pdf: 552363 bytes, checksum: a74c19218fe01b85e1a7c0331018df0a (MD5) Previous issue date: 2006-12-15 / The chronic allograft nephropathy (CAN) accounts for approximately 60% of late renal allograft loss. High homocysteine level (Hcy) and genetic variability of renin-angiotensin system are the possibly risk factors. Objectives: To investigate the frequency of Angiotensin Conversion Enzyme (ACE I/D) gene deletion, and Metilenetetrahydrofolate reductase (MTHFR C677T and A1298C) variants, as well as to quantify the plasma homocysteine concentrations in 300 patients submitted to renal transplantation to evaluate these factors participation in CAN development. Furthermore the ingestion of micronutrients and the frequency of MTHFR T1317C polymorphism have been investigated. Material and Method: The molecular study was performed by polymerase chain reaction (PCR) and RFPL (Restriction fragment length polymorphism)techniques for polymorphism investigation. In accordance with clinical criteria established, the patients had been subdivided in patients with CAN and patients with normal renal function (NRF). The automatic sequencing was performed for MTHFR polymorphism A1298C confirmation and identification of MTHFR T1317C polymorphism. Plasma Hcy concentration was measured by liquid chromatography tandem mass spectrometry (LS-MS/MS) technique. Dietary intakes were evaluated by a validated dietary questionnaire. Results: There was no correlation between the ingestion of micronutrients and CAN. The 1317C polymorphism frequency was 7% in Brazilian recipients. The presence, at least, of one MTHFR (677T/1298C)variant was significantly more frequent in CAN (p=0.049; OR = 1.7; 95% IC: 1.0 3.1), and a higher risk for disease was observed in the presence of the polymorphic $&( (D) variant (677T/1298AC/ACED) (p=0.009; OR = 2.2; 95% IC: 1.2 4.2). The hyperhomocysteinemia was observed in 82.1% of the CAN group patients, and 68.2% from MRF group. and plasmatic medium values of Hcy have presented statistical significant difference between the groups (p=0.005 and p<0.0005, respectively). High medium level of Hcy were associated with 677TT genotype and 677TT1298AA combined genotype in CAN group (p=0.002 and p=0.018, respectively) and with the 1298A allele from NRF group (p=0.009). The individuals of NRF group with 1298AA genotype have presented higher medium levels than 1298AC (p=0.033) genotype, suggesting a protector factor for 1298A allele. In relation to the distribution of Hcy levels, the CAN group has presented greater number of patients classified with severe and intermediate yperhomocysteinemia (>30&#956;mol/L) (p=0.0005), in relation to NRF group. The 1298C allele presence, as well as the combination, at least, one 07+)5 (677T/1298C) polymorphic allele in patients with hyperhomocysteinemia, were more frequent in CAN group (p=0.007 and p=0.002). A risk for CAN was observed in this combination (MTHFR 677T/1298C) in hyperhomocysteinemia patients (OR = 2.8; 95% IC: 1.4 6.0). This risk is increased in the presence of the polymorphic $&( (D) variant (OR = 3.4; 95% IC: 1.5 8.1). This suggests that the combination of, at least, one polymorphic 07+)5 allele and ECA(in those patients with hyperhomocysteinemia can predispose recipients to CAN development. / A disfunção crônica do transplante (DCTx) constitui aproximadamente 60% das causas de perda do transplante. Entre os possíveis fatores envolvidos estão o aumento do nível da homocisteína (Hcy) no plasma e a variabilidade genética no sistema angiotensina renina. Objetivos: investigar as freqüências do polimorfismo de deleção do gene ECA (enzima conversora da angiotensina) e das variantes 677 e 1298 do gene MTHFR (metilenotetrahidrofolato redutase), bem como dosar a concentração plasmática de Hcy em pacientes submetidos a transplante renal, visando avaliar a participação destes fatores no desenvolvimento da DCTx. Também foram investigadas a ingestão de micronutrientes e a freqüência do polimorfismo MTHFR T13317C. Casuística e Métodos: Foram investigados 300 pacientes submetidos a transplante renal há no mínimo 12 meses. De acordo com critérios clínicos estabelecidos os pacientes foram subdivididos em pacientes com DCTx e pacientes com função renal normal (FN). O estudo molecular utilizou as técnicas de reação em cadeia da polimerase seguida de digestão enzimática para a investigação dos polimorfismos. O seqüenciamento automático foi realizado para confirmação do polimorfismo MTHFR A1298C e para identificação do polimorfismo MTHFR T1317C. A concentração plasmática de Hcy foi dosada por meio da técnica de cromatografia líquida/espectrometria de massas seqüencial. A ingestão de micronutrientes foi avaliada por meio de questionário validado cientificamente. Resultados: Não foi encontrada associação entre a ingestão de micronutrientes e a DCTx. A freqüência do polimorfismo 1317C foi de 7% na população de indivíduos transplantados brasileiros. A presença de pelo menos uma variante MTHFR (677T/1298C), foi significantemente mais freqüente em DCTx (p=0,049; OR = 1,7; 95% IC: 1,0 3,1) e um risco aumentado para doença foi observado na presença da variante polimórfica ECA (D) (677T/1298AC/ECAD) (p=0,009; OR = 2,2; 95% IC: 1,2 4,2). A hiper-homocisteinemia foi observada em 82,1% dos pacientes do grupo com DCTx e 68,2% do grupo de FN e os valores médios de Hcy plasmática apresentaram diferença estatisticamente significante entre os grupos (p=0,005 e p<0,0005, respectivamente). Nível médio elevado de Hcy foi associado com o genótipo 677TT e com o genótipo combinado 677TT/1298AA no grupo DCTx (p=0,002 e p=0,018, respectivamente) e ao alelo 1298A do grupo FN (p=0,009). Os indivíduos FN com o genótipo 1298AA apresentaram níveis médios mais elevados que àqueles com o genótipo 1298AC (p=0,033), sugerindo um fator protetor para o alelo 1298A. Em relação à distribuição dos níveis de Hcy, o grupo DCTx apresentou maior número de pacientes classificados com hiper-homocisteinemia intermediária e grave (>30&#956;mol/L) (p=0,0005), em relação ao grupo FN. A presença do alelo 1298C, bem como a combinação de pelo menos um alelo polimórfico MTHFR (677T/1298C) em pacientes com hiper-homocisteinemia foi mais freqüente no grupo DCTx (p=0,007 e p=0,002, respectivamente). Um risco para DCTx foi observado para essa combinação MTHFR 677T/1298C) (OR = 2,8; 95% IC: 1,4 6,0) e é aumentado na presença da variante polimórfica ECA (D) (OR = 3,4; 95% IC: 1,5 8,1). Conclusão: A combinação de pelo menos um alelo polimórfico MTHFR e ECA naqueles pacientes que apresentam hiper-homocisteinemia parece predispor o paciente transplantado a DCTx.

A homocisteína é um aminoácido sulfurado derivado do metabolismo da metionina. Quando os níveis plasmáticos de homocisteína ultrapassam 10-15 μM, tem-se uma condição conhecida como hiperhomocisteinemia, a qual pode ser classificada em leve (>10 μM), moderada (>30 μM) ou severa (>100 μM). A hiperhomocisteinemia leve não tem origem genética, sendo considerada um fator de risco para o desenvolvimento de doenças neurodegenerativas e vasculares, incluindo isquemia cerebral e cardíaca. Nosso grupo de pesquisa desenvolveu um modelo químico induzido de hiperhomocisteinemia leve crônica em ratos adultos jovens e, usando esse modelo foi mostrado que existe associação entre essa condição e alterações em parâmetros de inflamação e estresse oxidativo/nitrativo em tecido cerebral. O objetivo do presente estudo foi avaliar se o ácido acetilsalicílico tem papel neuroprotetor no efeito da homocisteína sobre os níveis de interleucinas IL-1β e IL-6, atividade e imunoconteúdo da acetilcolinesterase, biodisponibilidade de óxido nítrico, atividade e imunoconteúdo das enzimas antioxidantes superóxido dismutase e catalase, conteúdo de sulfidrilas e índice de dano ao DNA. Também realizamos análise morfológica por microscopia eletrônica de transmissão em córtex cerebral de ratos submetidos ao modelo. Ratos Wistar receberam homocisteína (0,03 μmol/g de peso corporal) por injeções subcutâneas duas vezes ao dia e ácido acetilsalicílico (25 mg/Kg de peso corporal) por injeções intraperitoneais uma vez ao dia, do dia 30 ao 60º dia pós-parto Ratos controles receberam o mesmo volume da solução veículo. Doze horas após a última injeção, alguns animais foram decapitados para posteriores análises bioquímicas, e outros animais foram perfundidos para posterior análise morfológica. Os resultados mostraram que os ratos submetidos à hiperhomocisteinemia leve apresentaram aumento significativo dos níveis de IL-1β, IL-6 e da atividade da acetilcolinesterase, bem como níveis reduzidos de nitritos. A homocisteína também diminuiu as atividades da superóxido dismutase e catalase, bem como o imunoconteúdo da catalase. Danos às proteínas e DNA, assim como alterações ultraestruturais também foram observadas no córtex cerebral dos animais hiperhomocisteinêmicos. O ácido acetilsalicílico preveniu totalmente o efeito da homocisteína sobre a atividade da acetilcolinesterase, atividade e imunoconteúdo da catalase, e alterações ultraestruturais. As alterações nos níveis de IL-1β, atividade de superóxido dismutase, conteúdo de sulfidrilas e dano ao DNA foram parcialmente prevenidas pelo ácido acetilsalicílico. Nossos achados mostraram que o modelo induzido quimicamente de hiperhomocisteinemia leve alterou alguns parâmetros inflamatórios, oxidativos/nitrativos e morfológicos. Nossos resultados também sugerem que o ácido acetilsalicílico desempenha um papel neuroprotetor nas condições apresentadas, pois preveniu a maior parte dessas alterações. Porém, a administração crônica do ácido acetilsalícico também apresentou efeito per se significativo de dano ao DNA, o qual deve ser melhor elucidado em estudos posteriores. / Homocysteine is a sulfur amino acid derived from methionine metabolism. When plasma homocysteine levels exceed 10-15 μM, there is a condition known as hyperhomocysteinemia, which can be classified as mild (>10 μM), moderate (>30 μM), or severe (>100 μM). Mild hyperhomocysteinemia does not have genetic origin and it is considered a risk factor for the development of neurodegenerative and vascular diseases, including cerebral and cardiac ischemia. Our research group has developed an induced chemical model of chronic mild hyperhomocysteinemia in young adult rats and using this model, it has been shown an association between this condition and changes in parameters of inflammation and oxidative/nitrative stress in brain tissue. The objective of the present study was to evaluate if acetylsalicylic acid has a neuroprotective role in the effect of homocysteine on IL-1β and IL-6 interleukin levels, acetylcholinesterase activity and immunocontent, nitric oxide bioavailability, activity and immunocontent of antioxidant enzymes superoxide dismutase and catalase, sulfhydryl content and DNA damage index. We also performed morphological analysis by transmission electron microscopy in the cerebral cortex of rats submitted to the model. Wistar male rats received homocysteine (0.03 μmol/g of body weight) by subcutaneous injections twice a day and acetylsalicylic acid (25 mg/Kg of body weight) by intraperitoneal injections once a day from the 30th to the 60th postpartum day Control rats received the same volume of vehicle solution. Twelve hours after the last injection, some animals were decapitated for subsequent biochemical analyzes, and other animals were perfused for subsequent morphological analysis. The results showed that rats submitted to mild hyperhomocysteinemia significantly increased levels of IL-1β, IL-6, acetylcholinesterase activity and reduced nitrite levels. Homocysteine also decreased the activities of superoxide dismutase and catalase, as well as catalase's immunocontent. Damage to proteins and DNA as well as ultrastructural changes were also observed in the cerebral cortex of hyperhomocysteinemic animals. Acetylsalicylic acid totally prevented the effect of homocysteine on acetylcholinesterase activity, catalase activity and immunocontent, and ultrastructural changes. Alterations in IL-1β levels, superoxide dismutase activity, sulfhydryl content and DNA damage were partially prevented by acetylsalicylic acid. Our findings showed that the chemically induced model of mild hyperhomocysteinemia altered some inflammatory, oxidative/nitrative and morphological parameters. Our results also suggest that acetylsalicylic acid plays a neuroprotective role in the conditions presented, as it prevented most of these changes. However, chronic administration of acetylsalicylic acid also had a significant effect of DNA damage, which should be better elucidated in later studies.

Introdução: O café é uma das bebidas mais apreciadas e consumidas no mundo, por suas características organolépticas e efeito estimulante. Pelos potenciais efeitos na saúde causados por esta bebida surgiu, desde cedo, o interesse da comunidade científica. Como ainda ha controvérsias a respeito do café quanto aos seus reais efeitos, justifica-se este estudo em voluntários saudáveis e consumidores habituais de café. Objetivo: Comparar o consumo de dois diferentes graus de torras de café (torra média e escura) em 3 tipos de café: 100% arábica, blend e blend descafeinado e seus efeitos sobre o perfil metabólico em indivíduos saudáveis. Métodos: Em estudo prospectivo, foram avaliados 70 indivíduos sem qualquer doença associada, sendo 50 mulheres, com idade média de 47 ± 12 anos. Durante o período de seguimento no estudo, os voluntários que preencheram os critérios de inclusão, assinaram o Termo de Consentimento Livre e Esclarecido e iniciaram no estudo, que teve um total de 77 dias. Cada voluntário realizou quatro visitas (T0, T1, T2 e T3). Primeiramente, os participantes foram randomizados para cada tipo de café: 100% arábica, blend ou blend descafeinado. Depois uma nova randomização foi feita para estabelecer a ordem de consumo dos dois graus de torra (média e escura), num estudo do tipo crossover. Na visita T0, o participante foi orientado a parar a ingestão do café ou qualquer alimento fonte de cafeína por 21 dias. Na visita T1 foram randomizados para iniciar o consumo de café filtrado primeiro com um tipo de torra (torra média ou torra escura) por 4 semanas e então com \"crossover\" para o outro tipo (visita T2), com um período total de 8 semanas de consumo de café. O café foi fornecido aos pacientes, sendo do mesmo tipo, do mesmo produtor e com a forma de preparo padronizada e consumo diário de café de 450-600 ml/dia. Após período de \"washout\" (basal) e após cada período de tomada de café por tipo de torra, os pacientes foram submetidos aos exames laboratoriais (Colesterol total e frações, triglicérides, glicemia de jejum, PCR, Lp(a), homocisteína e acido fólico) e avaliação antropométrica (peso e IMC). Resultados: Foi observado aumento do colesterol total, do LDL e do HDL no grupo que consumiu café do tipo Blend, para os dois tipos de torra quando comparados ao basal. Houve aumento significativo da homocisteína no grupo que consumiu café 100% arábica e Blend. No grupo que consumiu café 100% arábica houve aumento significativo da homocisteína somente na torra média quando comparado ao basal. Já no grupo que consumiu o café tipo blend a diferença significativa foi apenas entre as duas torras escura e média, na qual a escura apresentou valor menor que a torra média. Conclusão: Café promove aumento discreto nas dosagens de homocisteína quando consumido o café blend e 100% arábica. Aumentou também o colesterol total, LDL, e ao mesmo tempo o HDL, quando consumido café tipo Blend. Apesar de significativas, as alterações no perfil metabólico foram muito discretas. Seria muito difícil determinar a influência que o café tem no metabolismo lipídico dos indivíduos, mas talvez esse discreto aumento juntamente com algum efeito antioxidante pode contribuir para a redução de morte por doenças cardiovasculares como já foi observado em estudos epidemiológicos / Introduction: Coffee is one of the most popular beverages in the world. Given its high consumption, potential health effects caused by this beverage brought the interest of the scientific community. Considering the current knowledge and controversies about drinking coffee daily, justifies this study in healthy volunteers and habitual coffee drinkers. Objectives: To compare the consumption of two different coffee roasts degrees (medium and dark roast) on 3 types of coffee: 100% arabic, blend and decaffeinated blend coffee and its effects on metabolic profile in healthy subjects. Results: 70 healthy subjects, age 47 ± 12 years old participated in the trial. Most subjects were female (71,4%). This randomized crossover clinical trial lasted 77 days. All the volunteers performed four visits (T0 - T1 - T2 -T3). In the visits, the participants had blood samples taken, held clinical examinations and nutritional evaluation, aside from receiving orientation. Randomly, participants should drink 100% Arabic, blend coffee or decaffeinated coffee. Then, in a second stage, again randomly, volunteers were draw to see which roast they would start drinking. In T0, subjects were oriented to stop consuming all the foods and beverages, which contained coffee or caffeine for 21 days. In T1 subjects start consuming medium or dark roast paper-filtered coffee for 4 weeks. In T2 they start the other roast for another additional 4 weeks. In T3, subjects stopped the coffee consumption, thus the study was completed. Participants received the coffee powder according to randomization criteria previously described. It is the same kind of coffee from the same producer and way to prepare. The coffee daily consumption were between 450 -600 ml. After \"washout\" period and after each period drinking coffee, all the subjects had sample bloods taken, to assess total cholesterol and fractions, triglycerides, glycaemia, homocisteine, acid folic and anthropometric (weight and BMI - body mass index). After analyzing the results, we observed an increase in total cholesterol, LDL and HDL levels after blend coffee intake. A significative increase in homocisteine levels after 100% Arabic and blend coffee intake. The significant difference in the group that consumed 100% arabic coffee were in between basal and medium roast. But in in group that consumed blend coffee this significant difference in homocisteine were in between the two roasts, higher during medium roast. Conclusions: Those results demonstrate a slight increase in homocisteine, when consumed blend coffee and 100% Arabic coffee. Also increased total cholesterol and LDL but at the same time increased the HDL, when they drank blend coffee. Despite significant, those metabolic alterations were discrete. It is difficult to determine the coffee influence on lipid metabolism, but maybe this slight increase tin HDL together with some antioxidant effect can contribute to the reduction of death from cardiovascular disease as already noted in epidemiologic studies