Evolution of potential biomarkers of acute muscle injury after physical exercise

Skeletal muscle injury is a frequent event and diagnosis using the classical blood markers sometimes produces unsatisfactory results. Therefore, objective of the study was to detect new biomarkers in plasma, saliva and urine in response to acute muscle damage induced by physical exercise. A cross-sectional study was conducted with 27 American football players. Before the physical exercises (T0), 60 minutes (T1) and 24 hours (T2) after physical exercise, was determined the clinical, biochemical and molecular parameters, including ADA, TBARS, leukocytes, lymphocytes and comet assay. The serum ADA was significantly higher in T1 and T2, in the urine there was a significant increase in T1, in the saliva there was no significant differences. There was an increase in serum TBARS in T2, saliva and urine in T1. The leukocytes increased in T1 and decreased in T2. Through the comet assay was observed significant DNA damage in T1 and T2. Serum and urinary ADA activity, serum, urinary and salivary TBARS are robust and promising biomarkers of acute muscle injury and that the comet assay allows a quick and effective evaluation of DNA lesions induced by physical exercise and could be used to monitor athletes avoiding injuries that are more serious.

Tuberculosis abdominal: patología infrecuente en un paciente joven: reporte de un caso / Abdominal tuberculosis: an unusual disease in a young patient: a case report

Resumen Introducción: La tuberculosis abdominal es un problema reemergente, y es una de las enfermedades transmisibles más importante en todo el mundo. A pesar de las expectativas acerca de su erradicación en países en desarrollo, ha sido recientemente declarada de nuevo como una patología de emergencia mundial. Con el aumento de su incidencia y prevalencia, su forma abdominal es una de las presentaciones de afectación extrapulmonar más comunes. Objetivo: Dado que la tuberculosis puede afectar diversos órganos, tiene una amplia gama y gran espectro de signos y síntomas que dificultan su diagnóstico y retrasan el tratamiento. Por esto, se realiza esta revisión de tema, concentrándonos en que el alto índice de sospecha debe ser un factor importante en el diagnóstico precoz, para que una vez establecido, se pueda iniciar el tratamiento ayudando a prevenir y disminuir las altas tasas de morbilidad y mortalidad evidenciadas en la actualidad. Caso Clínico: Paciente joven con presencia de ascitis secundaria a tuberculosis abdominal confirmada por una biopsia y el aumento de la adenosin deaminasa en el líquido peritoneal. Se describen los principales hallazgos clínicos, paraclínicos, estudios imagenológicos y tratamiento.

Introduction: Abdominal tuberculosis is a reemerging problem and is one of the most important communicable diseases in the world. Despite expectations about the eradication in developing countries, it has recently been re-declared as a global emergency pathology. The increased incidence and prevalence shows an abdominal shape as one of the most common extrapulmonary involvement presentations. Objective: Since tuberculosis can affect various organs, it has a wide range and spectrum of signs and symptoms that make diagnosis difficult and delay treatment. Therefore, this review of the topic is done, concentrating on the fact that the high suspicion index should be an important factor in the early diagnosis. Treatment can be initiated helping to prevent and reduce high morbidity and mortality rates. Case Report: We present a case of a young patient with ascites secondary to abdominal tuberculosis confirmed by biopsy and increased adenosine deaminase in the peritoneal fluid. The main clinical findings, paraclinic, imaging studies and treatment are described.

Effects of purine nucleotide administration on purine nucleotide metabolism in brains of heroin-dependent rats

ABSTRACT Heroin is known to enhance catabolism and inhibit anabolism of purine nucleotides, leading to purine nucleotide deficiencies in rat brains. Here, we determined the effect of exogenous purine nucleotide administration on purine nucleotide metabolism in the brains of heroin-dependent rats. Heroin was administrated in increasing doses for 9 consecutive days to induce addiction, and the biochemical changes associated with heroin and purine nucleotide administration were compared among the treated groups. HPLC was performed to detect the absolute concentrations of purine nucleotides in the rat brain cortices. The enzymatic activities of adenosine deaminase (ADA) and xanthine oxidase (XO) in the treated rat cortices were analyzed, and qRT-PCR was performed to determine the relative expression of ADA, XO, adenine phosphoribosyl transferase (APRT), hypoxanthine-guaninephosphoribosyl transferase (HGPRT), and adenosine kinase (AK). Heroin increased the enzymatic activity of ADA and XO, and up-regulated the transcription of ADA and XO. Alternatively, heroin decreased the transcription of AK, APRT, and HGPRT in the rat cortices. Furthermore, purine nucleotide administration alleviated the effect of heroin on purine nucleotide content, activity of essential purine nucleotide metabolic enzymes, and transcript levels of these genes. Our findings therefore represent a novel, putative approach to the treatment of heroin addiction.

Pleural effusion adenosine deaminase: a candidate biomarker to discriminate between Gram-negative and Gram-positive bacterial infections of the pleural space

OBJECTIVES: Delay in the treatment of pleural infection may contribute to its high mortality. In this retrospective study, we aimed to evaluate the diagnostic accuracy of pleural adenosine deaminase in discrimination between Gram-negative and Gram-positive bacterial infections of the pleural space prior to selecting antibiotics. METHODS: A total of 76 patients were enrolled and grouped into subgroups according to Gram staining: 1) patients with Gram-negative bacterial infections, aged 53.2±18.6 years old, of whom 44.7% had empyemas and 2) patients with Gram-positive bacterial infections, aged 53.5±21.5 years old, of whom 63.1% had empyemas. The pleural effusion was sampled by thoracocentesis and then sent for adenosine deaminase testing, biochemical testing and microbiological culture. The Mann-Whitney U test was used to examine the differences in adenosine deaminase levels between the groups. Correlations between adenosine deaminase and specified variables were also quantified using Spearman’s correlation coefficient. Moreover, receiver operator characteristic analysis was performed to evaluate the diagnostic accuracy of pleural effusion adenosine deaminase. RESULTS: Mean pleural adenosine deaminase levels differed significantly between Gram-negative and Gram-positive bacterial infections of the pleural space (191.8±32.1 U/L vs 81.0±16.9 U/L, p<0.01). The area under the receiver operator characteristic curve was 0.689 (95% confidence interval: 0.570, 0.792, p<0.01) at the cutoff value of 86 U/L. Additionally, pleural adenosine deaminase had a sensitivity of 63.2% (46.0-78.2%); a specificity of 73.7% (56.9-86.6%); positive and negative likelihood ratios of 2.18 and 0.50, respectively; and positive and negative predictive values of 70.6% and 66.7%, respectively. CONCLUSIONS: Pleural effusion adenosine deaminase is a helpful alternative biomarker for early and quick discrimination of Gram-negative from Gram-positive bacterial infections of the pleural space, which is useful for the selection of antibiotics.

Enfoque diagnóstico en el paciente con derrame pleural / Diagnostic approach in pleural effusion

En el estudio diagnóstico del paciente con derrame pleural se deben considerar la historia clínica y el análisis de las imágenes para acotar el diagnóstico diferencial. El uso adecuado de las técnicas de imágenes contribuye a realizar procedimientos en forma segura. Se debe realizar una toracocentesis diagnóstica y/o evacuadora y se debe analizar completamente el líquido pleural. A veces es necesario realizar biopsia pleural para lo cual existen diversas técnicas disponibles. En los pacientes con pleuritis crónica inespecífica se debe hacer seguimiento por dos años para evaluar el desarrollo de malignidad.

The diagnostic approach in patients with pleural effusion must begin considering clinical aspects and image interpretation. Different imaging techniques can safely guide invasive procedures. Diagnostic or therapeutic thoracentesis must be performed and pleural fluid must be completely analyzed. Some patient will require pleural biopsy, and different techniques are available. Patients with chronic unspecific pleuritis histological diagnosis after pleural biopsy, must be followed for two years long to be sure no malignancy is developed.

Teste de dosagem de adenosina deaminase (ADA) no diagnóstico precoce de tuberculose extrapulmonar / Adenosine deaminase (ADA) dosage test for the early diagnosis of extrapulmonary tuberculosis

Tecnologia: Teste de Dosagem de Adenosina Deaminase (ADA). Indicação: Diagnóstico de tuberculose pleural. Demandante: Secretaria de Saúde do Estado de São Paulo e Secretarias de Atenção à Saúde e de Vigilância em Saúde do Ministério da Saúde. Contexto: A tuberculose (TB) continua sendo mundialmente um importante problema de saúde, exigindo o desenvolvimento de estratégias para o seu controle. As apresentações extrapulmonares da TB têm seus sinais e sintomas dependentes dos órgãos e/ou sistemas acometidos. Sua ocorrência aumenta entre pacientes com aids, especialmente entre aqueles com imunocomprometimento grave. A tuberculose pleural é a mais comum forma de TB extrapulmonar em indivíduos HIV soronegativos, ocorre mais em jovens, cursa com dor torácica do tipo pleurítica. A tuberculose pleural é causa freqüente de derrame pleural, do tipo exsudato, e deve entrar no diagnóstico diferencial de exsudatos pleurais. Na maioria das vezes, a tuberculose pleural implica numa reação de hipersensibilidade da pleura ao bacilo da tuberculose, o que explica a baixíssima positividade em outros métodos diagnósticos. O Manual de recomendações para o controle da tuberculose no Brasil elaborado pelo Ministério da Saúde indica o teste ADA como método diagnóstico complementar a ser utilizado nos casos de suspeita de TB pleural, porém este teste não está disponível no SUS. O teste é colorimétrico, de fácil execução e é rápido, não necessitando de outras tecnologias para sua incorporação e nem treinamento especializado. Pergunta: O teste de dosagem de ADA é acurado para o diagnóstico de tuberculose em derrame pleural? Evidências científicas: foi elaborado parecer técnico-científico (PTC) pelo DECIT/SCTIE/MS sobre o Teste de Dosagem de ADA, para Diagnóstico Precoce da Tuberculose Pleural, que atualizou outro PTC emitido pelo Hospital João de Barros Barreto da Universidade Federal do Pará. A análise de todos os estudos mostrou heterogeneidade entre eles, porém foi possível recomendar a utilização do teste ADA para diagnóstico precoce de tuberculose pleural. Entretanto, quando o teste ADA for positivo, deverá haver a tentativa de isolamento do bacilo por cultura. Avaliação de Impacto Orçamentário: A Secretaria de Vigilância em Saúde levantou os custos para a realização do teste de ADA em 6 laboratórios públicos e privados de referência. Discussão: De acordo com as evidências apresentadas no PTC, a ADA quando dosada no líquido pleural é capaz de auxiliar no diagnóstico de tuberculose pleural precocemente, principalmente no que se refere ao seu valor preditivo negativo (em torno de 0,98), o que favorece a exclusão de pacientes suspeitos que não tenham a TB pleural. O teste ADA se trata de método colorimétrico, com técnica de dosagem fácil, rápida, reprodutível e de baixo custo. Sua inclusão no protocolo do SUS possibilitaria a redução do número de exames desnecessários e invasivos, reduzindo gastos e a exposição do paciente ao risco de exames invasivos, visto que um resultado negativo praticamente afasta a possibilidade de tuberculose pleural. No caso de valores considerados positivos devem ser associados a outras evidências ou testes para se confirmar o diagnóstico, como o isolamento do bacilo por cultura. Deste modo, a incorporação da tecnologia deve ser realizada mediante integração da mesma ao protocolo de diagnóstico da tuberculose do Ministério da Saúde, e, além disso, os resultados sobre acurácia do mesmo na prática clínica devem ser monitorados em centros de referência para fins de avaliação do impacto da introdução do método para a saúde da população. Deliberação final: o s membros da CONITEC presentes na reunião do plenário do dia 1º de agosto de 2013, por unanimidade, ratificaram a deliberação por recomendar a incorporação do Teste de Dosagem de Adenosina Deaminase (ADA) no Diagnóstico Precoce de Tuberculose Extrapulmonar.

Role of interleukin-6, gamma interferon and adenosine deaminase markers in management of pleural effusion patients / Papel de la interleucina-6, el interferón gamma, y la adenosina desaminasa como marcadores en el tratamiento de pacientes con derrame pleural

OBJECTIVE: Pleural effusion is a common diagnostic and clinical problem. Neoplasms and tuberculosis are the most frequent diagnostic causes of such effusions. Conventional laboratory methods for diagnosis of such effusion are inefficient because tubercle bacilli are rarely seen in direct examinations of pleural fluid. The present study evaluates interleukin-6 (IL-6), gamma interferon (IFN-γ) and adenosine deaminase (ADA) as diagnostic tools in pleural effusion. METHODS: Interleukin-6, IFN-γ and ADA were measured in pleural fluid from the patients, with exudative pleural effusion from tuberculous, malignant and postpneumonic origin and transudative pleural effusion ofsystemic origin in order to evaluate the diagnostic utility ofthese. RESULTS: The three markers were detectable in all effusions with significantly high levels in exudative as compared to transudative effusions. There was a statically significant difference noticed in tuberculous as compared to malignant andpostpneumonic origin and transudative pleural effusion. CONCLUSION: We concluded that IL-6, IFN-γ and ADA levels in pleural effusion are sensitive parameters to differentiate an exudate from a transudate and they can also differentiate exudates of different aetiology. Finally, the results suggest that there is a remarkable difference in production of these three markers in exudative pleural effusions as compared to transudative pleural effusions.

OBJETIVO: El derrame pleural es un problema diagnóstico y clínico común. Las neoplasias y la tuberculosis son las causas más frecuentes en los diagnósticos de tales derrames. Los métodos de laboratorio convencionales para el diagnóstico de tales derrames son ineficientes, porque los bacilos de la tuberculosis raramente se ven en los exámenes directos del líquido pleural. El presente estudio evalúa la interleucina-6, el interferón gamma (IFN-γ) y la adenosina desaminasa (ADA) como herramientas de diagnóstico en el derrame pleural. MÉTODOS: La interleucina-6, el IFN-γ, y la ADA fueron medidos en el líquido pleural de los pacientes con derrame pleural exudativo de origen tuberculoso, maligno y post-pneumónico, y el derrame pleural trasudativo de origen sistémico, con el fin de evaluar la utilidad diagnóstica de éstos. RESULTADOS: Los tres marcadores eran observables en todos los derrames, con niveles significativamente altos en los exudativos en comparación con los trasudativos. Se notó una diferencia estadísticamente significativa en los derrames de origen tuberculoso en comparación con los de origen maligno y postpneumónico, y los derrames pleurales trasudativos. CONCLUSIÓN: Llegamos a la conclusión de que los niveles de IL-6, IFN-Correspondence: Dr M Marie, College of Applied Medical Sciences, Clinical Laboratory Department, King Saud University, PO Box 10219, Riyadh 11433, Kingdom of Saudi Arabia. E-mail: drmmarie.2000@ gmail.com *Equally contributed to the manuscript YADA en el derrame pleural, son parámetros sensibles para diferenciar un derrame pleural exudado de uno trasudado, pudiendo por otra parte ayudar también a distinguir exudados de diferentes etiologías. Finalmente, los resultados sugieren que existe una diferencia notable en la forma en que se producen estos tres marcadores en los derrames efusiones pleurales exudativos en comparación con los derrames pleurales trasudativos.

Biomarcadores diagnósticos de tuberculose em pleurologia / Diagnostic biomarkers of tuberculosis in pleurologia

Atualmente o diagnóstico de tuberculose pleural pode ser realizado com a dosagem de biomarcadores diagnósticos no líquido pleural, especificamente com a dosagem da enzima adenosina desaminase. Os quadros clínico, laboratorial, imagem e citopatologia sugestivos sempre devem ser valorizados no conjunto do diagnóstico. Tal abordagem elege somente o procedimento de toracocentese como necessário para início do diagnóstico. Na maioria das apresentações clínicas, procedimentos cirúrgicos mais invasivos (biopsias pleurais), com complicações potencialmente fatais, não precisam ser realizados para exame histopatológico

Currently the diagnosis of pleural tuberculosis can be performed with the dosage of diagnostic biomarkers in pleural fluid, specifically the enzyme adenosine deaminase. The clinical, imaging and cytology suggestive should always be valued in the set of diagnosis together laboratory measurements. This approach selects only a thoracentesis procedure for early diagnosis. In most clinical presentations, more invasive surgical procedures (pleural biopsies) with life-threatening complications for histopathological examination

Association between Elevated Pleural Interleukin-33 Levels and Tuberculous Pleurisy

BACKGROUND: Interferon-gamma (IFN-gamma) plays a crucial role in Mycobacterium tuberculosis induced pleural responses. Interleukin (IL)-33 up-regulates the production of IFN-gamma. We aimed to identify whether an association between pleural IL-33 levels and tuberculous pleurisy exists and determine its diagnostic value. METHODS: Pleural IL-33, ST2 (a receptor of IL-33), adenosine deaminase (ADA), and IFN-gamma, as well as serum IL-33 and ST2 were measured in 220 patients with pleural effusions (PEs). Patients with malignant (MPEs), parapneumonic (PPEs), tuberculous (TPEs), and cardiogenic (CPEs) pleural effusions were included. RESULTS: Pleural and serum IL-33 levels were highest or tended to be higher in patients with TPEs than in those with other types of PEs. The median pleural fluid-to-serum IL-33 ratio was higher in TPE cases (> or = 0.91) than in other PE cases (< or = 0.56). Pleural IL-33 levels correlated with those of pleural ADA and IFN-gamma. However, the diagnostic accuracies of pleural IL-33 (0.74) and pleural fluid-to-serum IL-33 ratio (0.75) were lower than that of ADA (0.95) or IFN-gamma (0.97). Pleural ST2 levels in patients with MPEs were higher than in patients with TPEs. Serum ST2 levels did not differ among the groups. CONCLUSIONS: We identified an association between elevated pleural IL-33 levels and tuberculous pleurisy. However, we recommend conventional pleural markers (ADA or IFN-gamma) as diagnostic markers of TPE.

Pleural tuberculosis: is radiological evidence of pulmonary-associated disease related to the exacerbation of the inflammatory response?

OBJECTIVE: Pleural tuberculosis is the most frequently occurring form of extra pulmonary disease in adults. In up to 40% of cases, the lung parenchyma is concomitantly involved, which can have an epidemiological impact. This study aims to evaluate the pleural and systemic inflammatory response of patients with pleural or pleuropulmonary tuberculosis. METHODS: A prospective study of 39 patients with confirmed pleural tuberculosis. After thoracentesis, a high resolution chest tomography was performed to evaluate the pulmonary involvement. Of the 39 patients, 20 exhibited only pleural effusion, and high resolution chest tomography revealed active associated-pulmonary disease in 19 patients. The total protein, lactic dehydrogenase, adenosine deaminase, vascular endothelial growth factor, interleukin-8, tumor necrosis factor-α, and transforming growth factor-β1 levels were quantified in the patient serum and pleural fluid. RESULTS: All of the effusions were exudates with high levels of adenosine deaminase. The levels of vascular endothelial growth factor and transforming growth factor-β1 were increased in the blood and pleural fluid of all of the patients with pleural tuberculosis, with no differences between the two forms of tuberculosis. The tumor necrosis factor-α levels were significantly higher in the pleural fluid of the patients with the pleuropulmonary form of tuberculosis. The interleukin-8 levels were high in the pleural fluid of all of the patients, without any differences between the forms of tuberculosis. CONCLUSION: Tumor necrosis factor-α was the single cytokine that significantly increased in the pleural fluid of the patients with pulmonary involvement. However, an overlap in the results does not permit us to suggest that cytokine is a biological marker of concomitant parenchymal involvement. Although high resolution chest tomography can be useful in identifying these patients, the investigation of fast acid bacilli and cultures for M. tuberculosis in the sputum is recommended for all patients who are diagnosed with pleural tuberculosis.

Tuberculose / Tuberculosis

O diagnóstico laboratorial da tuberculose é de fundamental importância para o correto tratamento e controle da disseminação da doença. Dentre os principais métodos diagnósticos, os testes baseados em biologia molecular vêm ocupando cada vez mais um papel de destaque; entretanto, ainda não substituem por completo os métodos tradicionais, como a cultura e a pesquisa direta. A dosagem da adenosina deaminase (ADA) tem utilidade no diagnóstico de doença extrapulmonar e o teste de liberação de interferon gama linfocitário (IGRA) é de utilidade no diagnóstico de tuberculose latente. A utilização conjunta dos diferentes métodos disponíveis tem trazido grandes vantagens clínicas

The laboratory diagnosis of tuberculosis is of fundamental importance for the correct treatment and control the spread of the disease. Among the main diagnostic methods, the tests based on molecular biology are occupying an increasingly prominent role, however, has not yet completely replace traditional methods such as culture and direct search. The determination of adenosine deaminase (ADA) has useful in diagnosis of extrapulmonary disease and the interferon gamma release assay (IGRA) is useful in the diagnosis of latent tuberculosis. The combination use of different methods available has brought great clinical advantages

Characteristics of ascitic fluid from patients with suspected spontaneous bacterial peritonitis in emergency units at a tertiary hospital / Características do líquido ascítico de pacientes com suspeita de peritonite bacteriana espontânea nas unidades de emergência de um hospital terciário

CONTEXT AND OBJECTIVE: Spontaneous bacterial peritonitis (SBP) is a complication of ascites, especially in cirrhosis. Ascitic fluid with 250 or more neutrophils/mm³ is an acceptable criterion for diagnosis, even when bacterial fluid cultures are negative. The aims here were to estimate SBP frequency among emergency room patients based on cellular criteria and evaluate the biochemical profile of these fluids. DESIGN AND SETTING: Retrospective study at a public tertiary hospital. METHODS: Laboratory records of patients with ascites attended in emergency rooms between November 2001 and November 2006, from whom ascitic fluid samples were sent to the laboratory due to suspected SBP, were evaluated. The 691 samples included were divided into group A (presumed SBP: > 250 neutrophils/mm³; n = 219; 31.7 percent) and group B (no presumed SBP: < 250 neutrophils/mm3; n = 472; 68.3 percent). Patients' sex and age; ascitic fluid characteristics (numbers of neutrophils, leukocytes and nucleated cells); bacteriological characteristics; and protein, lactate dehydrogenase, adenosine deaminase and glucose concentrations were evaluated. RESULTS: Among group A cultured samples, 63 (33.8 percent) had positive bacterial cultures with growth of pathogens commonly associated with SBP. In total, the group A samples showed higher lactate dehydrogenase levels than seen in the group B samples. The latter presented predominance of lymphocytes and macrophages. CONCLUSION: Among the ascitic fluid samples with clinically suspected SBP, 31.7 percent fulfilled the cellular diagnostic criteria. Positive bacterial isolation was found in 33.8 percent of the cultured samples from the presumed SBP group.

CONTEXTO E OBJETIVO: Peritonite bacteriana espontânea (PBE) é uma complicação da ascite, especialmente na cirrose. Líquido ascítico com 250 ou mais neutrófilos/mm³ é um critério aceitável para o diagnóstico, mesmo com cultura bacteriana negativa. Os objetivos foram estimar a frequência de PBE em pacientes atendidos na sala de emergência, baseando-se no critério celular e avaliar o perfil bioquímico desses líquidos peritoneais. TIPO DE ESTUDO E LOCAL: Estudo retrospectivo em hospital público terciário. MÉTODOS: Foram avaliados registros laboratoriais de pacientes com ascite atendidos no setor de emergência entre novembro de 2001 e novembro de 2006, cujas amostras de líquido ascítico foram encaminhadas ao laboratório por suspeita de PBE. As 691 amostras incluídas foram divididas em grupo A (PBE presumida: > 250 neutrófilos/mm³; n = 219; 31.7 por cento) e grupo B (Ausência de PBE presumida: < 250 neutrófilos/mm3; n = 472; 68.3 por cento). Também foram avaliados sexo e idade dos pacientes além de características dos líquidos ascíticos: número de neutrófilos, leucócitos e células nucleadas; bacteriologia; e concentrações de proteínas, desidrogenase láctica, adenosina deaminase e glicose. RESULTADOS: Das amostras cultivadas do grupo A, 63 (33,8 por cento) tiveram cultura bacteriana positiva com crescimento de patógenos comumente associados à PBE. O total de amostras do grupo A exibiu maiores níveis de desidrogenase lática que as do grupo B. Este último demonstrou predomínio de linfócitos e macrófagos. CONCLUSÃO: Dos líquidos ascíticos com suspeita clínica de PBE, 31.7 por cento preencheram o critério diagnóstico celular. O isolamento bacteriano foi positivo em 33.8 por cento das amostras cultivadas no grupo PBE presumida.

Ciertas enzimas se relacionan con mecanismos de defensa en los desequilibrios nutricionales / Certain enzymes are related to defence mechanisms in cases of eating disorders

Algunos investigadores señalan la estrecha interrelación entre el desarrollo y funcionamiento de los linfocitos T con la actividad de la adenosina desaminasa (ADA). Trabajos previos de nuestro grupo, en modelos experimentales, demostraron que el estrés nutricional causado por la distorsión de nutrientes en la dieta provoca incremento en la actividad de la ADA en timo de rata. Se demostró aumento en la actividad de esta enzima en suero de pacientes con enfermedades que comprometen los mecanismos de defensa. Se analizó si la actividad de ADA sérica podría considerarse parámetro bioquímico funcional en el seguimiento de poblaciones en riesgo nutricional. Para ello se la estudió en suero de individuos, con compromiso del estado nutricional evaluado a través de diferentes indicadores, en diferentes situaciones fisiopatológicas. Se estudiaron mujeres con anorexia nerviosa; escolares obesos y niños con fibrosis quística. Los valores de cada grupo fueron comparados con los obtenidos en individuos sanos de igual edad (C). Los resultados muestran un incremento en la actividad de ADA estadísticamente significativo con respecto a C. El análisis integral refuerza la hipótesis de proponer la determinación en la actividad sérica de ADA como un indicador funcional relacionado con los mecanismos de defensa en los estudios de nutrición.

Validação laboratorial de um método automatizado de dosagem da atividade de adenosina desaminase em líquido pleural e em líquido cefalorraquidiano / Laboratorial validation of an automated assay for the determination of adenosine deaminase activity in pleural fluid and cerebrospinal fluid

OBJETIVO: A incidência global de tuberculose reforça a necessidade de melhores ensaios para o diagnóstico desta doença, principalmente da tuberculose extrapulmonar. O objetivo do trabalho foi validar o desempenho de um método automatizado para a determinação da atividade de adenosina desaminase (ADA) no líquido pleural (LP) e no líquido cefalorraquidiano (LCR), comparando-o com um método convencional (Giusti modificado). MÉTODOS: Selecionaram-se 134 amostras da rotina laboratorial: 94 de LP e 40 de LCR. Foram realizadas as determinações da atividade de ADA através dos dois métodos. Calculou-se a precisão inter- e intra-ensaios, análise de regressão linear, testes de concordância simples e médias das diferenças. RESULTADOS: Os coeficientes de correlação para as amostras de LP e LCR foram, respectivamente, 0,96 e 0,95. A precisão interensaio foi determinada pela média de 21 amostras replicadas em ensaios diferentes para 3 níveis de atividade: baixa, média e alta. Os coeficientes de variação em porcentagem ( por centoCV) foram, respectivamente, 5,9, 8,1 e 5,8 para amostras de LP; e 21,9, 18,6 e 13,8 para amostras de LCR, respectivamente. A precisão intra-ensaio em por centoCV foi, respectivamente, 1,3 e 11,7 por cento para amostras de LP e LCR. A concordância entre os dois métodos em amostras de LP e LCR foi, respectivamente, 96,8 por cento e 100 por cento, considerando-se como valores de referência para o diagnóstico de TB 40 U/L (convencional) e 30 U/L (automatizado) em amostras de LP, e 9 U/L em amostras de LCR para os dois métodos. CONCLUSÕES: Os resultados validaram o método automatizado de determinação da atividade de ADA para o uso em amostras de LP e LCR como alternativa ao método convencional.

OBJECTIVE: The incidence of tuberculosis worldwide has emphasized the need for better assays designed to diagnose the disease, principally the extrapulmonary form. The objective of the present study was to validate the performance of an automated method for the determination of adenosine deaminase (ADA) activity in pleural fluid (PF) and cerebrospinal fluid (CSF), comparing it with a conventional method (the modified Giusti method). METHODS: In total, 134 samples were selected from among those tested in our laboratory: 94 PF samples and 40 CSF samples. The ADA activity was determined using the two methods. Inter- and intra-assay precision was determined, linear regression analysis was performed, simple concordance tests were conducted, and the means of the differences were calculated. RESULTS: The correlation coefficients for PF and CSF samples were, respectively, 0.96 and 0.95. Inter-assay precision was determined using 21 replicates at 3 different activity levels: low, medium and high. The percentage coefficient of variation ( percentCV) was, respectively, 5.9, 8.1 and 5.8 for PF samples, compared with 21.9, 18.6 and 13.8 for CSF samples. Intra-assay precision in percentCV was 1.3 and 11.7, respectively, for PF and CSF samples. The concordance between the methods in PF and CRF samples was, respectively, 96.8 percent and 100 percent, considering the reference values for the diagnosis of TB to be 40 U/L (conventional) and 30 U/L (automated) in PF samples, versus 9 U/L (for both methods) in CSF samples. CONCLUSIONS: The results validate the use of the automated method of determining ADA activity in PF and CSF samples as an alternative to the conventional method.

Atividade da adenosinadeaminase no líquido cefalorraquiano / Adenosine deaminase activity in cerebrospinal fluid

Introdução. A adenosinadeaminase (ADA) é uma enzima que participa no metabolismo das purinas e, quando aumentada, foi tida por alguns autores como útil e até mesmo patognomônica da meningoencefalite tuberculosa (MTB). O objetivo deste trabalho é determinar o nível da atividade da adenosinadeaminase no líquido cefalorraquiano (LCR) em diversas condições patológicas, com especial referência a MTB. Métodos. A atividade da ADA foi estudada em 321 amostras de LCR de pacientes com meningite de diversas etiologias e outras alterações do LCR. Resultados. Amostras obtidas de pacientes portadores de MTB apresentaram um aumento estatisticamente significante entre a média da atividade da ADA e a média do grupo controle, e também em relação à média dos outros grupos: meningite séptica, meningite linfocitária e amostras em que se adicionou sangue humano fresco. Quando comparado ao grupo controle, amostras provenientes de pacientes com meningites de outras etiologias que não a tuberculosa também apresentavam aumento na atividade da ADA. Discussão. Verificou-se que há correlação direta entre a atividade da ADA e o número de linfócitos presentes na amostra de LCR. Assim sendo, o aumento da atividade desta enzima não é patognomônico de MTB, aumentando inclusive com a mistura de sangue acidental no LCR. Entretanto, verificou-se que a atividade da ADA acima de 24,1U/L em amostra de LCR é altamente sugestiva do diagnóstico de MTB.

Introduction. Adenosine deaminase (ADA) is an enzyme that participates in the metabolism of purines and, when its levels are high, some authors considered that it is useful or even pathognomonic of tuberculous meningitis (TBM). The objective of this study is to evaluate the levels of ADA activity in cerebrospinal fluid (CSF) of several pathologic conditions, especially TBM. Methods. Adenosine deaminase activity was studied in 321 samples of CSF from patients with meningitis from several etiologies and with other CSF alterations. Results. Samples from patients with TBM presented high ADA activity when compared with controls and when compared with the other groups. When compared to the controls, samples of CSF from patients with meningitis other than TBM also presented high ADA activity. Discussion. There is a direct correlation between ADA activity and the number of CSF lymphocytes, showing that a high ADA activity is not pathognomonic of TBM, increasing even with accidental blood mixture. However, ADA levels above 24.1U/L in a CSF sample are highly suggestive of TBM.

Avaliação da adenosina desaminase no diagnóstico da tuberculose pleural: uma metanálise brasileira / Evaluation of adenosine deaminase in the diagnosis of pleural tuberculosis: a Brazilian meta-analysis

OBJETIVO: Avaliar trabalhos brasileiros resumindo a acurácia da adenosina desaminase no diagnóstico da tuberculose pleural, com o intuito de contribuir para a concretização do exame como rotina na investigação dos derrames pleurais. MÉTODOS: Depois de realizada uma busca por trabalhos brasileiros referentes à dosagem da adenosina desaminase no líquido pleural, estes foram avaliados e incluídos no estudo. A análise dos dados foi feita por meio da curva summary receiver operating characteristic (SROC) que possibilitou a reunião dos estudos quanto a acurácia para o diagnóstico. Com os valores globais de sensibilidade e especificidade foi aplicado o teorema de Bayes para calcular as probabilidades pós-teste em diferentes prevalências da doença. RESULTADOS: Entre 1987 e 2005 foram encontrados 25 estudos contendo informações suficientes que poderiam ser utilizadas na metanálise. Após avaliação, foram incluídos nove estudos, totalizando 1.674 pacientes. De acordo com a curva SROC, foi encontrada uma sensibilidade de 91,8 por cento (IC95 por cento: 89,8-93,6 por cento) e uma especificidade de 88,4 por cento (IC95 por cento: 86,0-90,5 por cento), com uma área abaixo da curva de 0,969. O odds ratio global foi de 112,0 (IC95 por cento: 51,6-243,2). Considerando uma prevalência da tuberculose pleural de 50 por cento (considerada neutra), a probabilidade do diagnóstico pós-teste positivo é de 88,7 por cento e sua exclusão após resultado negativo de 91,5 por cento. CONCLUSÕES: Apesar das diferenças encontradas entre os estudos, é possível concluir que a adenosina desaminase possui alta acurácia no diagnóstico da tuberculose pleural, devendo ser utilizada de rotina em sua investigação.

OBJECTIVE: To evaluate Brazilian studies by summarizing the accuracy of adenosine deaminase in the diagnosis of pleural tuberculosis, with the objective of lending support to the movement to make the test part of the routine investigation of pleural effusions. METHODS: A search for Brazilian studies related to the determination of adenosine deaminase levels in the pleural liquid was carried out. These studies were evaluated and included in this study. The data were analyzed using summary receiver operating characteristic (SROC) curves, which enabled the studies to be collected and evaluated regarding the accuracy of the diagnosis. As for the global values of sensitivity and specificity, the Bayes' theorem was applied to calculate the post-test probabilities in different prevalences of the disease. RESULTS: Twenty-five studies dating from 1987 to 2005 and including enough information to be used in the meta-analysis were identified. After evaluation, nine studies were included, totaling 1674 patients. According to the SROC curve, a sensitivity of 91.8 percent (95 percent CI: 89.8-93.6 percent) and a specificity of 88.4 percent (95 percent CI: 86.0-90.5 percent) were found, with an area of 0.969 below the curve. The overall odds ratio was 112.0 (95 percent CI: 51.6-243.2). Considering a prevalence of tuberculosis of 50 percent (considered neutral), the post-test probability was 88.7 percent for a positive test and 91.5 percent for a negative test. CONCLUSIONS: Despite the differences found among studies, it is possible to conclude that the determination of adenosine deaminase levels has high accuracy in the diagnosis of the pleural tuberculosis and should be used as a routine test in its investigation.

The usefulness of adenosine deaminase in the diagnosis of tuberculous pericarditis

The objective of this study was to evaluate the adenosine deaminase (ADA) activity usefulness in the diagnosis of tuberculous pericarditis (TP), comparing its value with pericardial effusions (PE) caused by other pericardial diseases. A retrospective case-control study was conducted with nine cases of TP and 39 other than TP diseases (12 neoplastic, 11 septic and 16 unknown origin). Every patient included in this study had PE samples submitted to ADA activity measures and microbiological analysis, and then had pericardial tissue samples submitted to microbiological and histopathological examination. Considering the value of 40 U/L as the cut-off for the diagnosis of TP, the specificity and sensitivity were respectively of 72 percent and 89 percent. The specificity of ADA activity for the TP was best applied in the differential diagnosis from PE of unknown origin. The present study demonstrates the clinical value of the measurement of ADA activity in PE in the diagnosis of TP.

O objetivo deste estudo foi avaliar a atividade da adenosina deaminase (ADA) como auxiliar no diagnóstico da tuberculose pericárdica (TP), comparando o seu valor no derrame pericárdico com outras doenças pericárdicas. Um estudo retrospectivo tipo caso-controle foi conduzido com nove casos de TP e 39 pacientes com outras doenças pericárdicas (12 neoplasias, 11 pericardites bacterianas e 16 pericardites de etiologia indeterminada). Cada paciente incluído no estudo teve sua amostra de tecido pericárdico encaminhada para estudo microbiológico e histopatológico. Considerando o valor de 40 U/L como corte para o diagnóstico de TP, a especificidade e sensibilidade foram respectivamente 72 e 89 por cento. A especificidade da atividade de ADA para a TP foi melhor aplicada no diagnóstico diferencial entre derrame pericárdico de origem indeterminada. O presente estudo demonstrou o valor clínico da mensuração da atividade de ADA no diagnóstico de TP.

Diagnostic utility of adenosine deaminase (ADA) activity in pleural fluid and serum of tuberculous and non-tuberculous respiratory disease patients.

Adenosine deaminase activity (ADA) was assayed in pleural fluid and serum of 42 subjects with pleural effusion. Twenty-nine of them had TB pleural effusion and the remaining 13 had pleural effusion due to non-TB respiratory diseases. Serum adenosine deaminase activity were also measured in 32 pulmonary tuberculosis patients without pleural effusion and equal numbers of healthy controls without systemic diseases for comparative analysis. The patients attending the medicine out-patient department (MOPD) of the B. P. Koirala Institute of Health Sciences, Dharan, Nepal were taken as study subjects. Serum and pleural fluid ADA activities were assayed spectrophotometrically by the method of Guisti and Gallanti. The mean serum ADA activity was significantly increased in patients with tubercular pleural effusion (34.53 +/- 10.27 IU/l) compared to pulmonary tuberculosis patients without pleural effusion (26.54 +/- 4.76 IU/l), (p = 0.004), those with non-TB respiratory disease (16.71 +/- 5.16 IU/l), (p = 0.0001) and healthy controls (15.53 +/- 4.4 IU/l) (p = 0.0001). The mean ADA in the pleural fluid of tubercular pleural effusion patients (90.29 +/- 54.80 IU/l) was significantly higher compared to those with non-TB respiratory disease (24.43 +/- 9.28 IU/l) (p = 0.0001). Using the lowest cutoff value for enzyme activity in the serum of patients with TB pleural effusion (25 IU/l), a test sensitivity of 72.41% and specificity of 81.53% were obtained. Using the lowest cutoff value for enzyme activity in pleural fluid of patients with TB pleural effusion (45 IU/l) the sensitivity and specificity for diagnosis were 76.10% and 100%, respectively. Therefore, the measurement of ADA in tubercular pleural effusion has a utility in the diagnosis of tuberculosis when other clinical and laboratory tests are negative.

Predictive model for the diagnosis of tuberculous pleural effusion

This study developed a predictive model to identify pleural tuberculosis. A consecutive cases study of patients investigating the cause of pleural effusion, in an area of high prevalence of tuberculosis (Rio de Janeiro, Brazil). Clinical and laboratory variables were compared among patients with tuberculosis (TB) and without tuberculosis (NTB), individually and using logistic regression. The performance was described as diagnostic accuracy, compared to a gold standard in a masked way. We have studied 104 TB patients, 41 with malignant, 29 transudates, 28 parapneumonic, 13 with miscellaneous diseases. After identification of individual discrimination power aided by clinical, radiological and laboratory variables, the following ones were included in a multivariate analysis: ADA, total leukocytes, percentile of lymphocytes, protein, lactate dehydrogenase, duration of disease, age and gender. A logistic regression model to predict pleural tuberculosis including the five first variables showed the best performance. A receiver operating characteristic curve identified the best cutoff at 0.7, resulting in a sensitivity and specificity of more then 95 percent. The predictive model improved the specificity of ADA alone, keeping its sensitivity. This model seems helpful when a microbiological or histological diagnosis of pleural tuberculosis could not be established. External validation of these results is necessary before recommendation for routine application.

Etiologic and Laboratory Analyses of Ascites in Patients who Underwent Diagnostic Paracentesis / 대한간학회지

BACKGROUND/AIMS: Liver cirrhosis and malignant tumors are two major causes of ascites according to the reports from Western countries, 80% and 10% respectively. Assuming that there might be regional differences in etiologies and changes in their frequency over time, we investigated causes of ascites and the diagnostic usefulness of various laboratory tests. METHODS: Medical records of 366 patients, who underwent diagnostic paracentesis in the mid-1990s (1996 and 1997) and early 2000s (2001 and 2002), were retrospectively reviewed. The etiology was confirmed by histology, imaging studies, and ascites analyses. RESULTS: The frequency of cirrhotic ascites was 59.6%, cancer-related 25.7%, tuberculous peritonitis 6.6%, and others 8.1%. Among cirrhotics, the frequency of cases related to hepatitis B decreased significantly from 72% to 55% over time, and alcoholic cirrhosis increased from 18% to 34%. Among cancer-related ascites, peritoneal carcinomatosis type was 75.5% (primary sites: stomach 24.5%, pancreas 15.9%, colon 15.9%, lung 7.4%, etc), metastatic liver cancers 8.5%, hepatocellular carcinoma without cirrhosis 6.4%, etc. The sensitivity of serum-ascites albumin gradient for the diagnosis of cirrhotic ascites was 91.4%, and total protein in ascites also revealed a comparable diagnostic sensitivity, 90%. The diagnostic sensitivity of adenosine deaminase for tuberculous peritonitis was 94.2%, and its positive predictive value was 75%. CONCLUSIONS: Liver cirrhosis is the leading cause of ascites, especially alcoholic cirrhosis has significantly increased. The next common etiology is cancer-related, and its frequency in Korea is higher than in western countries. Tuberculous peritonitis is still prevalent, and adenosine deaminase could precisely differentiate it from other causes.