Valores de referência para o teste de caminhada com carga progressiva em indivíduos saudáveis: da distância percorrida às respostas fisiológicas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytokines and acute-phase serum proteins as markers of inflammatory regression during pulmonary tuberculosis treatment

Tuberculosis is still increasing and was declared a worldwide sanitary emergency by the World Health Organization (WHO) in 1995. Its control is difficult due to long treatment duration and lack of markers of treatment success or failure. Cytokines such as IFN-gamma and TNF-alpha, a central factor in immune response against Mycobacterium tuberculosis, are responsible for the interaction between T lymphocytes and the infected macrophage and are also produced during this interaction. As proinflammatory cytokines have a close relationship with mycobacteria clearance, in fact even preceding it, they could be used as markers for inflammatory activity and response to treatment. Proinflammatory cytokines act in the liver and stimulate a strong local and systemic acute-phase response as a result of homeostatic and physiological responses also induced by them. Acute-phase proteins produced by cytokine activity are useful diagnostic markers that could also be used to monitor treatment response as they can be serially quantified. The objective of this study was to evaluate IFN-gamma, TNF-alpha, IL-10 and TGF-beta production in supernatant of peripheral blood mononuclear cell (PBMC) and monocyte (MO) cultures, as well as serum acute-phase response through total protein, albumin, globulin, C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), and erythrocyte sedimentation rate (ESR) as regression markers of inflammatory response during pulmonary tuberculosis treatment. Twenty blood donors (G1) from the Blood Bank at Botucatu School of Medicine's University Hospital (BSM-UH) were evaluated once and 28 pulmonary tuberculosis patients (G2): 13 from BSM-UH and 15 from the Bauru State Health Secretariat. Patients were evaluated at three moments of treatment: before (M1), at three months (M2), and at the end (M3). Cytokines were determined in 20ml of peripheral blood (ELISA), with or without activation: lipopolysaccharide (LPS) for MO culture and phytohemagglutinin (PHA) for PBMC culture. Acute-phase protein behavior in G2 throughout treatment was: Globulins: M1> M2, M1> M3 (rho < 0.001); CRP: M1> M2> M3 (.< 0.001); AGP for men: M1> M2, M1> M3 (rho < 0.001); ESR for men: M1> M2, M1> M3 (rho < 0.0016) and for women: M1> M2 (.< 0.025). Comparison between cytokine levels found in supernatant of MO and PBMC cultures, with and without stimulus, in G1 and G2 during treatment showed: TNF-alpha (with/ without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); (without LPS) at M3: G2> G1 (rho < 0.001), (with LPS) at M3: G2> G1 (rho < 0.028); IFN-. (with and without PHA) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); IL-10 (with and without LPS) at M1: G2> G1; at M2: G2> G1; at M3: G2> G1 (rho < 0.001); TGF-beta (with and without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001), (without LPS) at M3: G2> G1 (rho < 0.001). In G2, all cytokines in supernatant of MO and PBMC cultures, with and without stimulus, showed: M1> M2> M3 (rho < 0.01). Levels of globulins, CRP, AGP, and ESR in patients with pulmonary tuberculosis before treatment (M1) were significantly higher than reference values, suggesting their use as diagnostic markers and indicators of treatment. The CRP decreasing values along treatment could be taken as a marker of the regression of inflammatory process and of response to treatment in patients with pulmonary tuberculosis.Regarding cytokines, there was significant increase in TNF-alpha, IFN-gamma, IL-10, and TGF-alpha levels before and at three months treatment, with and without stimulus; in TNF-a and IL-10 lvels, with and without stimulus, as well as in TGF-alpha levels without stimulus at six months. Patients had higher levels of all studied cytokines than controls before treatment, and these values decreased along treatment. In this study, pulmonary tuberculosis patients showed a Th0 cytokine profile before treatment, with the production of both Th1 (IFN-gamma) and Th2 (IL-10) cytokines, in addition to TNF-alpha inflammatory and TGF-alpha regulatory and fibrosis-inducer cytokines. At the end of treatment, all had evolved to Th2 profile, probably in an attempt to reduce the harmful effects of the proinflammatory activity of the Th1 cytokine profile and of the still above-normal levels of TNF-alpha. The high levels of TGF-alpha, also found in these patients, are related to its important role in the extracellular matrix deposition and fibrosis induction that characterize tuberculosis healing process. IFN-gamma was the only cytokine reaching normal levels at the end of treatment, which suggests its use as a marker of response to treatment.

Changes in apoptosis and Bcl-2 expression in human hyperglycemic, term placental trophoblast

Apoptosis and its associated regulatory mechanisms are physiological events crucial to the maintenance of placental homeostasis; imbalance of these processes, however, such as occurs under various pathological conditions, may compromise placenta function and, consequently, pregnancy success. Increased apoptosis occurs in the placentas of pregnant women with several developmental disabilities, while increased Bcl-2 expression is generally associated with pregnancy-associated tumors. Herein, we tested the hypothesis that apoptosis-associated disturbs might be involved in the placental physiopathology subjected to different maternal hyperglycemic conditions.Thus, in the present study we investigated and compared the incidence of apoptosis using TUNEL reaction and Bcl-2 expression, in term-placentas of normoglycemic, diabetic and daily hyperglycemic patients. Tissue samples were collected from 37 placentas, being 15 from healthy mothers with normally delivered healthy babies, and 22 from mothers with glucose disturbances. From these latter 22 patients, 10 showed maternal daily hyperglycemia and 12 were clinically diabetics. Both Bcl-2 expression and apoptotic DNA fragmentation were established and quantified in the trophoblasts of healthy mothers. Compared to these reference values, a higher apoptosis index and lower Bcl-2 expression were disclosed in the placentas of the diabetic women, while in the daily hyperglycemic group, values were intermediate between the diabetic and normoglycemic patients. The TUNEL/Bcl-2 index ratio in the placentas varied from 0.02 to 0.09 for pregnant normoglycemic and diabetic women, respectively, revealing a predominance of apoptosis in the diabetic group. Our findings suggest that hyperglycemia may be a key factor evoking apoptosis in the placental trophoblast, and therefore, is relevant to diabetic placenta function. (c) 2006 Elsevier B.V.. All rights reserved.

Avaliação da posição do supercílio por medidas angulares

OBJETIVO: Avaliar a posição do supercílio em diferentes idades, utilizando medidas angulares. MÉTODOS: Foram avaliados indivíduos com idade de 4 a 6 anos (Grupo de crianças) e igual ou superior a 50 anos (Grupo de idosos), separados em faixas etárias, avaliando-se a posição do supercílio por meio de imagens digitais, utilizando medidas angulares. As imagens foram tomadas em posição primária do olhar, utilizando filmadora Sony Lithium, e posteriormente transferidas para computador MacIntosh G4 e processadas pelo programa NIH 1,58. Os parâmetros analisados foram: ângulo interno, externo e vertical da cauda do supercílio. As comparações foram entre sexos, faixas etárias e lateralidade. Os resultados obtidos foram submetidos à análise estatística. RESULTADOS: A comparação das medidas angulares mostrou que houve diferença significativa na posição da cauda do supercílio entre os grupos estudados quando comparados dentro do grupo com faixa etária semelhante. Porém, comparando-se crianças e adultos, houve diferença em todos os tipos de ângulos estudados. CONCLUSÕES: A posição do supercílio avaliada por medidas angulares mostrou diferenças entre crianças e idosos, revelando associação positiva com a idade.

The HUman MicroNucleus project on eXfoLiated buccal cells (HUMNXL): The role of life-style, host factors, occupational exposures, health status, and assay protocol

The human buccal micronucleus cytome assay (BMCyt) is one of the most widely used techniques to measure genetic damage in human population studies. Reducing protocol variability, assessing the role of confounders, and estimating a range of reference values are research priorities that will be addressed by the HUMNXL, collaborative study. The HUMNXL, project evaluates the impact of host factors, occupation, life-style, disease status, and protocol features on the occurrence of MN in exfoliated buccal cells. In addition, the study will provide a range of reference values for all cytome endpoints. A database of 5424 subjects with buccal MN values obtained from 30 laboratories worldwide was compiled and analyzed to investigate the influence of several conditions affecting MN frequency. Random effects models were mostly used to investigate MN predictors. The estimated spontaneous MN frequency was 0.74 parts per thousand (95% CI 0.52-1.05). Only staining among technical features influenced MN frequency, with an abnormal increase for non-DNA-specific stains. No effect of gender was evident, while the trend for age was highly significant (p < 0.001). Most occupational exposures and a diagnosis of cancer significantly increased MN and other endpoints frequencies. MN frequency increased in heavy smoking (>= 40 cig/day. FR = 1.37:95% CI 1.03-.82) and decreased with daily fruit consumption (FR = 0.68; 95% CI 0.50-0.91). The results of the HUMNXL, project identified priorities for validation studies, increased the basic knowledge of the assay, and contributed to the creation of a laboratory network which in perspective may allow the evaluation of disease risk associated with MN frequency. (C) 2011 Elsevier B.V. All rights reserved.

Salivary cortisol to assess the hypothalamic-pituItary-adrenal axis in healthy children under 3 years old

Objective: To establish reference concentration intervals for salivary cortisol in healthy children, in the morning and in the afternoon, investigating factors that interfere with the concentration measured and the possibility that circadian rhythms are present.Methods: A controlled observational study was carried out with 91 children aged 45 days to 36 months, selected at random and living in Santo Andre, state of São Paulo, Brazil. Inclusion criteria were: healthy, well-nourished, free from fever and corticoid use, subdivided by age group (five subsets) at 6-month intervals. Saliva was collected during home visits in the morning and afternoon. Cortisol was radioimmunoassayed with cortisol 3-oxime-bovine albumin antiserum.Results: the five subsets exhibited higher cortisol concentration during the morning than in the afternoon (p < 0.001), and this difference passed 30% from 1 year of age onwards. Mean concentrations, in nmol/L, were 557.86 (morning) and 346.36 (afternoon). A negative linear correlation was observed between morning concentrations and hours' sleep and frequency of meals (p < 0,05), and in the afternoon with anthropometric measurements (p < 0.05).Conclusions: Reference values for normal salivary cortisol in healthy children were established. At:45 days it was possible to observe circadian rhythms, which reached maturity at 12 months of life. Sleep and food deprivation increased morning cortisol levels.

Curvas de percentis de valores normais de medidas ecocardiográficas em crianças eutróficas procedentes da região centro-sul do Estado de São Paulo

OBJETIVO: Avaliar os valores de medidas ecocardiográficas em crianças eutróficas sem cardiopatia, relacionando-os com a superfície corporal (SC, m²), e construir curvas de percentis que relacionem as variáveis ecocardiográficas estudadas com a SC. MÉTODOS: Foram analisadas medidas ecocardiográficas unidimensionais de crianças entre 1 e 144 meses de idade. Avaliaram-se: diâmetros diastólicos dos ventrículos direito (VDd, mm) e esquerdo (VEd, mm), sistólico do VE (VEs, mm), da via de saída do VD (VSVD, mm), da aorta (DAo, mm) e do átrio esquerdo (DAE, mm); fração de ejeção do VE (FEVE, %); porcentagem da variação do diâmetro ventricular esquerdo (deltaVE, %); espessura diastólica do septo interventricular (ESIV, mm) e da parede posterior do VE (EPPVE, mm); massa (MVE, g) e índice de massa muscular do VE (IMVE, g/m²). RESULTADOS: Ao final do estudo, 595 crianças (326 do sexo masculino) foram avaliadas. Os valores das medidas ecocardiográficas apresentaram boa correlação com a SC e possibilitaram a construção de curvas de percentis (3%, 25%, 50%, 75% e 97%). Diferenças estatisticamente significantes, entre os sexos, foram evidenciadas para as variáveis VEs, VEd, VSVD, DAo, MVE e IMVE, sendo os maiores valores observados em crianças do sexo masculino. CONCLUSÃO: As curvas de percentis dos valores obtidas podem ser utilizadas como referência para a avaliação de crianças com suspeita de cardiopatia ou para o acompanhamento daquelas já diagnosticadas como cardiopatas ou em tratamento com agentes potencialmente cardiotóxicos.

Características eletrocardiográficas em eqüinos clinicamente normais da raça Puro Sangue Inglês

Determinaram-se as características eletrocardiográficas de amplitude, duração, ritmo e eixo cardíaco no plano frontal de potros, sobreanos e éguas prenhes clinicamente normais, da raça Puro Sangue Inglês. Estudaram-se 50 potros e 50 sobreanos, machos e fêmeas, e 11 éguas prenhes. Concluiu-se que a freqüência cardíaca diminui com a idade e que nos potros e sobreanos as características apresentaram valores intermediários entre os valores padrão de neonatos e adultos. Nos sobreanos eles estão mais próximos dos de referência para adultos. Nas éguas em gestação a única diferença observada com o padrão foi o leve desvio para a direita do eixo cardíaco.

Níveis séricos de enzimas de função hepática em frangos de corte de criação industrial clinicamente saudáveis

The values for the main hepatic enzymes included in the profiles of screen clinical biochemistry, alanine-aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (FA), lactate desidrogenase (LDH) and gamaglutamiltransferase (GGT), in samples of serum of broiler chickens in industrial system, clinically healthy, starting from the seventh day of life, until the slaughter (42 days) in weekly intervals were determined. Significant variations were not observed in the analyses in relation to the age of the birds for none of the appraised enzymes.

Valores séricos de cálcio, fósforo, sódio, potássio e proteínas totais em caprinos fêmeas da raça Parda Alpina

Por ser de grande importância o conhecimento de valores séricos referenciais do mineralograma e do proteinograma, a sua utilização na medicina veterinária, e a escassez de trabalhos brasileiros relacionados à espécie caprina, este estudo objetivou determinar e comparar os níveis séricos de cálcio, fósforo, sódio, potássio e proteínas (proteína total, albumina, globulina e a relação albumina/globulina) em caprinos fêmeas da raça Parda Alpina, em três idades diferentes. Foram utilizadas 30 fêmeas caprinas da raça Parda Alpina criadas em regime intensivo, sendo separadas em três grupos: Grupo 1 - 10 fêmeas jovens com idade de quatro meses; Grupo 2 - 10 fêmeas adultas jovens com idade de 18 meses; Grupo 3 - 10 fêmeas adultas lactantes com idade de 24 a 60 meses. Os animais foram avaliados por meio de exames clínico e coproparasitológico. Foram encontrados os seguintes valores médios de cálcio, fósforo, sódio e potássio no grupo G1: 10,10± 1,70, 10,13± 2,04, 151± 3,30 e 5,16± 0,41; no grupo G2: 10,36± 0,94, 7,49± 1,34, 145,9± 3,35 e 4,56± 0,48; e para o grupo G3: 9,47± 1,45, 5,90± 1,21, 153,3± 5,17 e 4,20± 0,55. Para as variáveis proteína total sérica, albumina, globulina e relação albumina/globulina foram encontrados os seguintes valores médios, para o grupo G1: 6,29± 0,94, 3,5± 10,41, 2,78± 0,95 e 1,40± 0,48; para o grupo G2: 7,27± 0,73, 4,92± 1,06, 3,06± 0,56, 1,43± 0,44; e para o grupo G3: 7,14± 0,84, 3,70± 0,20, 3,44± 0,79 e 1,14± 0,30. A análise estatística dos dados demonstra que existe influência da faixa etária nas variáveis fósforo, sódio, potássio, proteína total e albumina.

Efeito da idade e sexo sobre a concentração sérica de eritropoietina em equinos da raça Árabe

A EPO é um fator de crescimento glicoprotéico sintetizado pelas células adjacentes aos túbulos proximais renais regulada via mecanismo de feed back envolvendo a tensão de oxigênio tissular. Na baixa tensão de oxigênio arterial, a produção de EPO aumenta causando uma maior produção de eritrócitos na medula óssea. Devido ao pouco conhecimento da concentração de EPO sérica em equinos e a ausência de trabalhos sobre o efeito da idade e sexo sobre a sua concentração o trabalho teve como objetivo comparar a concentração sérica de eritropoietina em equinos da raça Árabe de sexos e idades diferentes. Foram utilizados 31 equinos da raça Árabe, com idades de seis a 12 meses (jovens) e acima de 24 meses (adultos), sendo 13 machos (seis jovens e sete adultos) e 18 fêmeas (oito jovens e 10 adultas), clinicamente sadios. As amostras de sangue foram colhidas por venipunção jugular e o soro armazenado até o momento do processamento. A concentração sérica de eritropoetina foi determinada pelo método de radioimunoensaio (RIA) utilizando kit comercial (EPO Trac TM125I RIA, Diagnostic Systems Laboratories, Webster, Texas, USA). Para análise estatística dos dados utilizou-se o Teste t de Student ao nível de 5% de significância (P<0,05). Não foram observadas diferenças significativas (P<0,05) quando se compararam os animais divididos entre sexos e as idades de seis a 12 meses (jovens) e acima de 24 meses (adultos). Conclui-se que a concentração de eritropoietina em equinos da raça Árabe independe do sexo e idade, e pode ser utilizada como valores de referência, porém ressalta-se a necessidade da obtenção de valores de referência para cada laboratório.

Radiographic measurement of tibial joint angles in sheep

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Perfil bioquímico das galinhas poedeiras na região de Araçatuba-SP

The establishment of reference values is extremely important for successful diagnosis and treatament. Considering that in most species the serum chemistry profile is influenced by race, climate and management, we decided to determine the values of aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid, creatinine, creatine kinase (CK), phosphatase alkaline (ALP), gamma-glutamyltransferase (GGT), total protein (TP) and albumin of Dekalb hens in the region of Aracatuba - SP. All samples were processed soon after harvesting in an automatic biochemical analyzer calibrated and monitored with control serum levels I and II. The following confidence intervals were obtained: 44-65,5 U / L (AST); 18,4-21,2 U / L (ALT), 2.1-2.5 mg / dL (uric acid); 1.7 to 5.7 U / L (CK); CI 1.2-2.2 mg / dL (creatinine), 1276-1506 U / L (FA); 18-23,4 U / L (GGT); 27.12 to 29 g / L (PT), from 11.4 to 12.16 g / L (albumin).

Placental Volumes Measured by 3-Dimensional Ultrasonography in Normal Pregnancies From 12 to 40 Weeks' Gestation

Objective. The purpose of this study was to construct nomograms of placental volumes according to gestational age and estimated fetal weight. Methods. From March to November 2007, placental volumes were prospectively measured by ultrasonography in 295 normal pregnancies from 12 to 40 weeks' gestation and correlated with gestational age and estimated fetal weight. Inclusion criteria were healthy women, singleton pregnancies with normal fetal morphologic characteristics on ultrasonography, and confirmed gestational age by first-trimester ultrasonography. Results. The mean placental volume ranged from 83 cm(3) at 12 weeks to 427.7 cm(3) at 40 weeks. Linear regression yielded the following formula for the expected placental volumes (ePV) according to gestational age (GA): ePV` (cm(3)) = -64.68 + 12.31 x GA (r = 0.572; P < .001). Placental volumes also varied according to estimated fetal weight (EFW), and the following mathematical equation was also obtained by linear regression: ePV = 94.19 + 0.09 x EFW (r = 0.505; P < 0.001). Conclusions. Nomograms of placental volumes according to gestational age and estimated fetal weight were constructed, generating reference values.

Quantitative estimation of Br, Cl, K and Na in sample blood by NAA

Neutron activation analysis, using Au as flux monitor, was applied to determine the concentrations of Br, Cl, K and Na in blood of healthy male and female blood donors, selected from blood banks and hematological laboratories from different regions of Brazil. The aims of this study were to collect more reference values of the Brazilian population as well as to perform hematological investigations. The advantages as well as the limitations of using this nuclear procedure are discussed.