Diversity of Apolipoprotein E genetic polymorphism significance on cardiovascular risk is determined by the presence of Metabolic Syndrome among hypertensive patients

Background: Hypertension has a significant relevance as a cardiovascular risk factor. A consistent increase on world's Metabolic Syndrome (MetS) incidence has been associated with an epidemic cardiovascular risk in different populations. Dislipidemia plays a major role determining the epidemic CV burden attributed to MetS. Apolipoprotein E (ApoE) is involved on cholesterol and triglycerides metabolism regulation. Once ApoE polymorphism may influence lipid metabolism, it is possible that it brings on individual susceptibility consequences for the development of MetS and cardiovascular risk. the objective of the study is to measure the discriminatory power of ApoE polymorphism in determining cardiovascular risk stratification based on the presence MetS in a cohort of hypertensive patients.Methods: It was enrolled 383 patients, divided in two groups, classified by MetS presence (IDF criteria): Group 1: 266 patients with MetS (MetS +) and Group 2: 117 patients without Mets (MetS -). Patient's data were collected by clinical evaluation, physical exam, file reviews and laboratory testing. Polymorphic ApoE analysis was performed by PCR amplification. Groups were compared on clinical and laboratory characteristics as well as allele and genotype distribution towards ApoE polymorphism. Mets CVD prevalence was analysed according to E4 allele prevalence.Results: the results evidenced 184 men (48%), 63,7% whites, 45,1% diabetics and 11,7% of patients were smokers. Mean age was 64,0 +/- 12,0 years. When genotypic distribution was analyzed, E3/3 genotype and E3 allele frequencies were more prevalent. Among patients with MetS, we observed an independent association between CVD prevalence and E4 allele frequency (OR 2.42 (1.17-5.0, p < 0,05)). On the opposite direction, in those without MetS, there was lesser CVD burden in E4 allele carriers (OR 0,14 (0,02-0,75)). These associations remained significant even after confounding factor corrections.Conclusions: the results presented demonstrate that the association between ApoE gene and CVD may be modulated by the presence of MetS, with an increased CV burden observed among E4 allele carriers with the syndrome. On the opposite way, E4 allele carriers without visceral obesity had lesser prevalence of CVD.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Div Nephrol, BR-04039032 São Paulo, BrazilTufts Univ, New England Med Ctr, Div Nephrol, Boston, MA 02111 USAHosp Israelita Albert Einstein, Res & Educ Inst, São Paulo, BrazilUniversidade Federal de São Paulo, Div Nephrol, BR-04039032 São Paulo, BrazilWeb of Scienc

Apolipoprotein E polymorphism modulation of asymmetric dimethylarginine in hypertensive patients is determined by renal function

Background: Endothelial dysfunction is considered an early step of atherosclerotic vascular disease. Asymmetric dimethylarginine (ADMA), the main endogenous inhibitor of nitric oxide synthase (NOS), plays a critical role in the process of atherosclerosis in a uremic environment. Increased plasma ADMA not only works as a cardiovascular morbidity biomarker but it is also involved in the genesis of atherosclerosis in renal disease. Considering the relationships of apolipoprotein E(ApoE) polymorphism with LDL cholesterol (LDL-C) levels and coronary risk, it is possible that it brings on susceptibility to endothelial dysfunction and atherogenesis seen on uremia. Methods: Six hundred twenty patients were stratified according to glomerular filtration rate (GFR) estimated by Chronic Kidney Disease Epidemiology Collaboration (CKDEPI) formula: group I > 60 mL/min, group II 15 mL/min, and group III <= 15 mL/min or in hemodialysis. Polymorphic ApoE analysis was performed by polymerase chain reaction amplification (PCR). Plasma ADMA levels were measured by high performance liquid chromatography (HPLC). Groups were compared on clinical and laboratory characteristics as well as allele and genotype distribution towards. Results: The epsilon 2 allele of ApoE was present in 62 (10.3 %) patients, epsilon 3 allele in 581 (96.2 %), and epsilon 4 allele in 114 (18.9 %). Their distribution among the 3 groups was uniform. Such uniformity was not observed when we considered endothelial function measured by asymmetric dimethylarginine. In group III, the frequency of epsilon 4 allele was significantly lower in the third tertile compared with the first tertile (14.7 versus 53.3 %, P = 0.000; Pearson chisquare). In groups I and II, there was no difference in allele frequency according to ADMA levels. This association remained significant even after confouding factors corrections (OR 0.329, 95 % CI 0.155 - 0.699, P = 0.004). Conclusions: The results of this study shows that the frequency of epsilon 4 allele of ApoE is significantly lower among hypertensive patients on hemodialysis with the highest levels of ADMA. Uremia is capable of determining lower plasma ADMA levels in hypertensive epsilon 4 allele carriers.Fundo de Amparo a Pesquisa do Estado de Sao Paulo/FAPESPUniv Fed Sao Paulo, Nephrol Div, R Pedro de Toledo 781 14 Andar, BR-04039032 Sao Paulo, BrazilTufts Univ, New England Med Ctr, Nephrol Div, Boston, MA 02111 USAHosp Israelita Albert Einstein, Res & Educ Inst, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Nephrol Div, R Pedro de Toledo 781 14 Andar, BR-04039032 Sao Paulo, BrazilWeb of Scienc

Effects of simvastatin on cytokines secretion from mononuclear cells from critically ill patients with acute kidney injury

Purpose: To assess the in vitro effects of simvastatin on IL-10 and TNF-alpha secretion from peripheral blood mononuclear cells (PBMC) of critically ill patients with and without acute kidney injury (AKI).Methods: PBMC were collected from 63 patients admitted to the intensive care unit (ICU) and from 20 healthy controls. Patients were divided in 3 subgroups: with AKI, with sepsis and without AKI and with AKI and sepsis. After isolation by ficoll-gradient centrifugation cells were incubated in vitro with LPS 1 ng/mL, simvastatin (10(-8)M) and with LPS plus simvastatin for 24 h. TNF-alpha and IL-10 concentrations on cells surnatant were determined by ELISA.Results: Cells isolated from critically ill patients showed a decreased spontaneous production of TNF-alpha and IL-10 compared to healthy controls (6.7(0.2-12) vs 103(64-257) pg/mL and (20 (13-58) vs 315(105-510) pg/mL, respectively, p < 0.05). Under LPS-stimulus, IL-10 production remains lower in patients compared to healthy control (451 (176-850) vs 1150(874-1521) pg/mL,p < 0.05) but TNF-alpha production was higher (641 (609-841) vs 406 (201-841) pg/mL, p < 0.05). the simultaneous incubation with LPS and simvastatin caused decreased IL-10 production in cells from patients compared to control (337 (135-626) vs 540 (345-871) pg/mL, p < 0.05) and increased TNF-alpha release (711 (619-832) vs 324 (155-355) pg/mL, p < 0.05). Comparison between subgroups showed that the results observed in TNF-alpha and IL-10 production by PBMC from critically ill patients was independent of AKI occurrence.Conclusions: the PBMC treatment with simvastatin resulted in attenuation on pro-inflammatory cytokine spontaneous production that was no longer observed when these cells were submitted to a second inflammatory stimulus. Our study shows an imbalance between pro and anti-inflammatory cytokine production in PBMC from critically ill patients regardless the presence of AKI. (C) 2011 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Instituto de Ensino e Pesquisa do Hospital Israelita Albert EinsteinUniversidade Federal de São Paulo, Div Nephrol, Dept Med, São Paulo, BrazilIAEH IEP Hosp Israelita Albert Einstein Inst Ensi, São Paulo, BrazilUniversidade Federal de São Paulo, Div Nephrol, Dept Med, São Paulo, BrazilWeb of Scienc

Serum soluble-Fas is a predictor of red blood cell transfusion in critically ill patients

OBJECTIVE: To investigate the relation between the need for red blood cell transfusion and serum levels of soluble-Fas, erythropoietin and inflammatory cytokines in critically ill patients with and without acute kidney injury. METHODS: We studied critically ill patients with acute kidney injury (n=30) and without acute kidney injury (n=13), end-stage renal disease patients on hemodialysis (n=25) and healthy subjects (n=21). Serum levels of soluble-Fas, erythropoietin, interleukin 6, interleukin 10, iron status, hemoglobin and hematocrit concentration were analyzed in all groups. The association between these variables in critically ill patients was investigated. RESULTS: Critically ill patients (acute kidney injury and non-acute kidney injury patients) had higher serum levels of erythropoietin than the other groups. Hemoglobin concentration was lower in the acute kidney injury patients than in other groups. Serum soluble-Fas levels were higher in acute kidney injury and end-stage renal disease patients. Critically ill patients requiring red blood cell transfusions had higher serum levels of soluble-Fas (5,906±2,047 and 1,920±1,060; p<0.001), interleukin 6 (518±537 and 255+502; p=0.02) and interleukin 10 (35.8±30.7 and 18.5±10.9; p=0.02), better iron status and higher mortality rates in the first 28 days in intensive care unit. Serum soluble-Fas levels were independently associated with the number of red blood cell units transfused (p=0.02). Serum soluble-Fas behaved as an independent predictor of the need for red blood cell transfusion in critically ill patients (p=0.01). CONCLUSIONS: Serum soluble-Fas level is an independent predictor of the need for red blood cell transfusion in critically ill patients with or without acute kidney injury. Further studies are warranted to reconfirm this finding.OBJETIVO: Investigar a relação entre a transfusão de hemácias e os níveis séricos de Fas solúvel, eritropoietina e citocinas inflamatórias em pacientes gravemente enfermos, com e sem insuficiência renal aguda. MÉTODOS: Os seguintes grupos foram estudados: pacientes gravemente enfermos com insuficiência renal aguda (n=30) e sem insuficiência renal aguda (n=13), pacientes portadores de doença renal crônica terminal em hemodiálise (n=25) e indivíduos saudáveis (n=21). Os níveis séricos de Fas solúvel, eritropoietina, interleucina 6, interleucina 10 e ferro, além da concentração de hemoglobina e de hematócrito, foram analisados em todos os grupos. A associação entre tais variáveis foram estudadas nos pacientes gravemente enfermos. RESULTADOS: Os níveis séricos de eritropoietina mostraram-se mais elevados nos pacientes gravemente enfermos do que nos dos demais grupos. Concentrações mais baixas de hemoglobina foram documentadas nos pacientes com insuficiência renal aguda em relação aos demais. Níveis séricos mais elevados de Fas solúvel foram observados nos pacientes com insuficiência renal aguda e doença renal crônica terminal. Pacientes gravemente enfermos transfundidos apresentaram níveis séricos mais elevados de Fas solúvel (5.906±2.047 e 1.920±1.060; p<0,001), interleucina 6 (518±537 e 255±502; p=0,02), interleucina 10 (35,8±30,7 e 18,5±10,9; p=0,02) e ferro, além de maior mortalidade em 28 dias. Os níveis séricos de Fas solúvel mostraram-se independentemente associados ao número de transfusões (p=0,02). O nível sérico de Fas solúvel foi um preditor independente da necessidade de transfusão de hemácias em pacientes gravemente enfermos (p=0,01). CONCLUSÃO: O nível sérico de Fas solúvel é um preditor independente da necessidade de transfusão de hemácias em pacientes gravemente enfermos, com ou sem insuficiência renal aguda. Mais estudos clínicos e laboratoriais são necessários para confirmar tal resultado.Universidade Federal de São Paulo (UNIFESP)Hospital Israelita Albert EinsteinUNIFESPSciEL

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure &lt; 100 mmHg (n = 1127), estimated glomerular filtration rate &lt; 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Identification of a new site of angiotensin-I converting enzyme (ACE) production in IMCD cells and study of interation between the bradykinin receptors and ACE using ACE inhibitors

Enzima conversora de angiotensina I (ECA) ou kininase II é uma dipeptidil- carboxipepitidase (EC 3.4.15.1) que tem um importante papel na regulação de pressão sangüínea, cliva o dipeptídeo C-terminal da angiotensina I(AI)i produzindo o potente peptídeo vasopressor angiotensina II (AII) e inativa bradicinina (BK), um potente vasodilatador. Baseados no estudo de Casarini e cols. (1997) no qual um novo sítio produção de ECA foi localizado no ducto coletor, o objetivo deste trabalho foi purificar e caracterizar na cultura de células mIMCD-3 a presença e/ou síntese da ECA, enfatizando desta forma um papel fisiológico importante para esta enzima nestas células, uma vez que todo sistema calicreína-cininas encontra-se presente nesta região. Outro objetivo foi estudar a interação da ECA com os receptores B2 da BK utilizando inibidores da ECA. Inicialmente, isolamos a atividade da ECA presente nas células mIMCD no meio de cultura, por gel filtração em coluna empacotada com resina AcA-34 previamente calibrada. Cada isoforma foi recromatografada em coluna afinidade Lisinopril-sepharose. As enzimas isoladas do meio foram denomina M1 e M2, e as ECAs extraídas das células, C1 e C2. As massas moleculares determinadas por eletroforese em gel de poliacrilamida foram: M1, 60 kDa e M2 117 kDa (isoformas secretadas) e C1, 63 kDa e C2, 130 kDa (isoformas intracelulares). A expressão protéica foi verificada por Western Blotting utilizando anticorpo monoclonal 9B9, nas isoformas purificadas e nas frações do citoplasma da membrana e do núcleo…(au).BV UNIFESP: Teses e dissertaçõe

Study of kininases, purified from human urine and from fresh urine using internally quenched fluorescents substrates

Cininase e uma denominacao geral para enzimas proteoliticas capazes de inativar cininas como bradicinina (BK), lisil-bradicinina (LBK) e metionil-lisil-bradicinina (MLBK). Em estudos anteriores realizados em nosso laboratorio, foram purificadas de urina de individuos normais, atraves de cromatografia em DEAE-celulose, 8 enzimas com atividade cininasica: duas atividades do tipo carboxipeptidase, duas enzimas conversaras de angiotensina I, duas serino endopeptidase (Hl e H2), uma prolil endopeptidase (H) e uma endopeptidase neutra (NEP-Iike). Neste trabalho, com o objetivo de estabelecer um metodo extremamente sensivel para a quantificacao de endopeptidases, purificamos e caracterizamos 4 enzimas, a partir de urina humana, que foram classificadas com base nos estudos de inibicao e ligacao hidrolisada na molecula de BK, como serino tiol endopeptidase (H2), serino endopeptidase (Hl), prolil endopeptidase (H) e endopeptidase neutra (NEP-Iike). Para tanto utilizamos o substrato com apagamento intramolecular da fluorescencia AbzOBKQOEDDnp, bem como testamos diversos substratos fluorogenicos para estudar a especificidade das referidas enzimas. A prolil endopeptidase e uma tiol metalo endopeptidase sendo inibida por EDTA, 2-ME e POHMB. A serino endopeptidase Hl foi inibida por PMSF e a serino tio[ endopeptidase H2 alem de ser inibida por PMSF foi tambem inibida por E64 e POHMB. A NEP-Iike foi inibida por fosforamidom, tiorfam e OPA. Os substratos fluorogenicos Abz-FPQ-EDDnp, Abz-FGQ-EDDnp, AbzFRQ-EDDnp e Abz-FDQ-EDDnp foram determinados especificos para as enzimas H, Hl, H2 e NEP-Iike respectivamente. O Km encontrado foi da ordem de mM para estes substratos, bem como para Abz-BKQ-EDDnp. Utilizando o substrato Abz-BKQ-EDDnp a atividade maxima encontrada para a prolil endopeptidase foi em 9,0; para a serino endopeptidase Hl foi em 7,0 e 8,5, e para a NEP-Iike foi em 7,0 e 8,0. Quando utilizamos os substratos especificos para cada enzima encontramos atividade maxima nos seguintes pHs: 6,5 e 8,0 para a prolil endopeptidase, utilizando o substrato Abz-FPQ-EDDnp; 5,5 e 8,0 para a serino Hl utilizando o substrato Abz-FRQ-EDDnp; e 6,0 e 7,0 para a NEP-/ike utilizando o substrato Abz-FDQ-EDDnp. Para a endopeptidase H2 nao foi possivel determinar o pH otimo de atividade. Os pesos moleculares determinados para a prolil endopeptidase, 45 kDa; para a serino endopeptidase Hl, 49 kDa...(au)BV UNIFESP: Teses e dissertaçõe

Multiple angiotensin I-converting enzyme (ACE) isoforms from WKY and SHR rats.

UNIFESP, Dept Med, Div Nephrol, Sao Paulo, BrazilUNIFESP, Dept Med, Div Nephrol, Sao Paulo, BrazilWeb of Scienc