ADAR1-mediated RNA-editing of 3'UTRs in breast cancer

BACKGROUND: Whole transcriptome RNA variant analyses have shown that adenosine deaminases acting on RNA ( ADAR ) enzymes modify a large proportion of cellular RNAs, contributing to transcriptome diversity and cancer evolution. Despite the advances in the understanding of ADAR function in breast cancer, ADAR RNA editing functional consequences are not fully addressed. RESULTS: We characterized A to G(I) mRNA editing in 81 breast cell lines, showing increased editing at 3'UTR and exonic regions in breast cancer cells compared to immortalized non-malignant cell lines. In addition, tumors from the BRCA TCGA cohort show a 24% increase in editing over normal breast samples when looking at 571 well-characterized UTRs targeted by ADAR1. Basal-like subtype breast cancer patients with high level of ADAR1 mRNA expression shows a worse clinical outcome and increased editing in their 3'UTRs. Interestingly, editing was particularly increased in the 3'UTRs of ATM, GINS4 and POLH transcripts in tumors, which correlated with their mRNA expression. We confirmed the role of ADAR1 in this regulation using a shRNA in a breast cancer cell line (ZR-75-1). CONCLUSIONS: Altogether, these results revealed a significant association between the mRNA editing in genes related to cancer-relevant pathways and clinical outcomes, suggesting an important role of ADAR1 expression and function in breast cancer.

Adenosine: an endogenous mediator in the pathogenesis of psoriasis

Abstract: It is known that inflammatory and immune responses protect us from the invasion of micro-organisms and eliminate "wastes" from the injured sites, but they may also be responsible for significant tissue damage. Adenosine, as a purine nucleoside, which is produced in inflamed or injured sites, fulfills its role in limiting tissue damage. Although, it may have a pleiotropic effect, which signals it with a proinflammatory state in certain situations, it can be considered a potent anti-inflammatory mediator. The effects of adenosine, which acts through its receptors on T cell, on mast cell and macrophages, on endothelial cells, on neutrophils and dendritic cells, as they indicate TNF-alpha and cytokines, show that this mediator has a central role in the pathogenesis of psoriasis. The way it acts in psoriasis will be reviewed in this study.

Modulatory effect of iron chelators on adenosine deaminase activity and gene expression in Trichomonas vaginalis

Trichomonas vaginalis is a flagellate protozoan that parasitises the urogenital human tract and causes trichomoniasis. During the infection, the acquisition of nutrients, such as iron and purine and pyrimidine nucleosides, is essential for the survival of the parasite. The enzymes for purinergic signalling, including adenosine deaminase (ADA), which degrades adenosine to inosine, have been characterised in T. vaginalis. In the evaluation of the ADA profile in different T. vaginalisisolates treated with different iron sources or with limited iron availability, a decrease in activity and an increase in ADA gene expression after iron limitation by 2,2-bipyridyl and ferrozine chelators were observed. This supported the hypothesis that iron can modulate the activity of the enzymes involved in purinergic signalling. Under bovine serum limitation conditions, no significant differences were observed. The results obtained in this study allow for the assessment of important aspects of ADA and contribute to a better understanding of the purinergic system in T. vaginalis and the role of iron in establishing infection and parasite survival.

Adenosin deaminasa en la tripanosomiasis experimental: futuras implicaciones / Adenosine deaminase in experimental trypanosomiasis: future implications

La adenosin deaminasa representa un punto de control en la regulación de los niveles extracelulares de adenosina, desempeñando así un papel fundamental en la modulación de las respuestas purinérgicas a ciertos eventos patofisiológicos. Diversos estudios señalan que los niveles séricos y plasmáticos de la enzima se elevan en algunas enfermedades causadas por microorganismos, lo cual podría representar un mecanismo compensatorio como consecuencia de la elevación de las concentraciones de adenosina y la liberación de mediadores inflamatorios. Recientes investigaciones indican que la actividad de la adenosin deaminasa disminuye e influye en los parámetros hematológicos de animales infectados con Trypanosoma evansi, de manera que tales alteraciones podrían tener implicaciones en la patogénesis de la enfermedad. Adicionalmente, la enzima ha sido detectada en este parásito; lo que permite inferir que podría estar asociada a las funciones vitales del mismo, de manera similar a lo que ocurre en los mamíferos. Este conocimiento puede ser útil al asociar la quimioterapia con inhibidores específicos de la enzima en futuros estudios.

The adenosine deaminase represents a control point in the regulation of extracellular adenosine levels, thus playing a critical role in the modulation of purinergic responses to certain pathophysiological events. Several studies have shown that serum and plasma enzyme levels are elevated in some diseases caused by microorganisms, which may represent a compensatory mechanism due to the elevated levels of adenosine and the release of inflammatory mediators. Recent research indicates that adenosine deaminase activity decreases and affects hematological parameters of infected animals with Trypanosoma evansi, so that such alterations could have implications in the pathogenesis of the disease. In addition, the enzyme has been detected in this parasite; allowing the inference that it could be associated with the vital functions of the same, similar to what occurs in mammals. This knowledge may be useful in the association of chemotherapy with specific inhibitors of the enzyme in future studies.

Utility of adenosine deaminase (ADA), PCR & thoracoscopy in differentiating tuberculous & non-tuberculous pleural effusion complicating chronic kidney disease.

Background & objectives: Pleural effusion is a common occurrence in patients with late-stage chronic kidney disease (CKD). In developing countries, many effusions remain undiagnosed after pleural fluid analysis (PFA) and patients are empirically treated with antitubercular therapy. The aim of this study was to evaluate the role of adenosine deaminase (ADA), nucleic acid amplification tests (NAAT) and medical thoracoscopy in distinguishing tubercular and non-tubercular aetiologies in exudative pleural effusions complicating CKD. Methods: Consecutive stage 4 and 5 CKD patients with pleural effusions underwent PFA including ADA and PCR [65 kDa gene; multiplex (IS6110, protein antigen b, MPB64)]. Patients with exudative pleural effusion undiagnosed after PFA underwent medical thoracoscopy. Results: All 107 patients underwent thoracocentesis with 45 and 62 patients diagnosed as transudative and exudative pleural effusions, respectively. Twenty six of the 62 patients underwent medical thoracoscopy. Tuberculous pleurisy was diagnosed in six while uraemic pleuritis was diagnosed in 20 subjects. The sensitivity and specificity of pleural fluid ADA, 65 kDa gene PCR, and multiplex PCR were 66.7 and 90 per cent, 100 and 50 per cent, and 100 and 100 per cent, respectively. Thoracoscopy was associated with five complications in three patients. Interpretation & conclusions: Uraemia remains the most common cause of pleural effusion in CKD even in high TB prevalence country. Multiplex PCR and thoracoscopy are useful investigations in the diagnostic work-up of pleural effusions complicating CKD while the sensitivity and/or specificity of ADA and 65 kDa gene PCR is poor.

Estigma e discriminação: experiências de mulheres HIV positivo nos bairros populares de Maputo, Moçambique / Stigma and discrimination: the experiences of HIV-positive women in poor neighborhoods of Maputo, Mozambique / Estigma y discriminación: las experiencias de las mujeres VIH positivas en los barrios populares de Maputo, Mozambique

A epidemia de HIV/AIDS é um sério problema de saúde pública em Moçambique, que convive com altas taxas de prevalência do HIV. O impacto da epidemia é agravado pelo forte estigma que atinge as pessoas soropositivas. O objetivo deste estudo foi investigar, com base em uma perspectiva socioantropológica, a experiência de mulheres HIV positivo nos bairros populares de Maputo e como lidam com o estigma e a discriminação. Foram realizadas entrevistas semiestruturadas com dez mulheres HIV positivo, residentes nos bairros populares de Maputo. Os resultados mostram como a desigualdade de gênero atua de forma importante na construção da vulnerabilidade das mulheres ao HIV, assim como em sua estigmatização e discriminação. No enfrentamento do estigma, as mulheres procuram preservar o sigilo do diagnóstico buscando apoio na reunião em grupos de pares HIV positivo. É fundamental que se implementem políticas públicas voltadas para o empoderamento das mulheres e redução do estigma associado ao HIV/AIDS.

The HIV/AIDS epidemic is a serious public health problem in Mozambique. The country has high prevalence rates, and the epidemic's impact is aggravated by the stigma affecting HIV-positive persons. This study takes a socio-anthropological perspective to analyze the experience of HIV-positive women in poor neighborhoods of Maputo and the ways they cope with stigma and discrimination. Semi-structured interviews were conducted with 10 HIV-positive women. The results show how gender inequalities increase women's vulnerability to HIV and contribute to their stigmatization and discrimination. In dealing with stigma, women try to keep their diagnosis confidential, seeking support in group meetings with others living with HIV. Public policies should focus on women's empowerment and the reduction of HIV/AIDS-related stigma.

El VIH/SIDA es un problema de salud pública grave en Mozambique, que convive con altas tasas de prevalencia del VIH. El impacto de la epidemia se ve agravada por el fuerte estigma que afecta a las personas con VIH. El objetivo de este estudio fue investigar, desde una perspectiva antropológica, la experiencia de las mujeres VIH positivas en los barrios populares de Maputo y cómo enfrentan el estigma y la discriminación. Se realizaron entrevistas semi-estructuradas con 10 mujeres VIH positivas que viven en barrios pobres de Maputo. Los resultados muestran cómo la desigualdad de género juega un papel importante en la construcción de la vulnerabilidad de las mujeres frente al VIH, así como en la estigmatización y discriminación. Para hacer frente el estigma, las mujeres buscan preservar la confidencialidad del diagnóstico y buscar apoyo en la reunión de grupos de pares con VIH. Es imprescindible implementar políticas públicas enfocadas al empoderamiento de las mujeres y a la reducción del estigma asociado con el VIH/SIDA.

Carvedilol atenua o estresse oxidativo na cardiopatia chagásica crônica / Carvedilol attenuates oxidative stress in chronic chagasic cardiomyopathy / Carvedilol atenúa el estrés oxidativo en la cardiopatía chagásica crónica

FUNDAMENTO: Há cada vez mais evidências sugerindo que doença de Chagas envolve dano oxidativo e contribui para a progressão da doença cardíaca. OBJETIVO: Avaliar o efeito do carvedilol sobre marcadores de estresse oxidativo na doença de Chagas crônica. MÉTODOS: A população de estudo incluiu 42 pacientes com cardiopatia chagásica e os biomarcadores de estresse oxidativo foram medidos antes e após um período de seis meses de tratamento com carvedilol (37,5 mg/dia). Os pacientes foram considerados de acordo com a classificação de Los Andes, e a atividade da superóxido dismutase, catalase, glutationa peroxidase, S-transferase e redutase, mieloperoxidase e adenosina deaminase; e os níveis de glutationa reduzida, de espécies reativas do ácido tiobarbitúrico, proteína carbonil, vitamina E e óxido nítrico foram medidos no sangue. RESULTADOS: Após o tratamento com carvedilol, todos os grupos apresentaram reduções significativas nos níveis de proteína carbonil e glutationa reduzida, enquanto os níveis de óxido nítrico e atividade da adenosina aumentaram significativamente somente no grupo IA. Além disso, a maioria das enzimas antioxidantes apresentou diminuição de suas atividades, nos grupos IA e IB. CONCLUSÃO: Os dados sugerem que o tratamento com carvedilol foi eficaz na atenuação do dano oxidativo, um efeito que pode ser particularmente importante em doença de Chagas crônica com cardiopatia.

BACKGROUND: There is increasing evidence suggesting that Chagas disease involves oxidative damage and contributes to heart disease progression. OBJECTIVE: To evaluate the effect of carvedilol on oxidative stress markers in chronic Chagas disease. METHODS: The study population included 42 patients with Chagas cardiomyopathy and oxidative stress biomarkers were measured before and after a period of six months of treatment with carvedilol (37.5 mg/day). Patients were considered according to the Los Andes classification and the activity of superoxide dismutase, catalase, glutathione peroxidase, S-transferase and reductase, myeloperoxidase and adenosine deaminase; levels of reduced glutathione, thiobarbituric acid reactive species, carbonyl protein, vitamin E and nitric oxide were measured in blood. RESULTS: After treatment with carvedilol, all groups showed significant reductions in levels of carbonyl protein and reduced glutathione, whereas the levels of nitric oxide and adenosine activity increased significantly only in group IA. Moreover, most of the antioxidant enzymes showed decrease in activity in groups IA and IB. CONCLUSION: The data suggest that treatment with carvedilol was effective in attenuating oxidative damage, an effect that may be particularly important in patients with chronic Chagas' disease cardiomyopathy.

FUNDAMENTO: Hay cada vez más evidencias sugiriendo que la enfermedad de Chagas envuelve daño oxidativo y contribuye a la progresión de la enfermedad cardíaca. OBJETIVO: Evaluar el efecto del carvedilol sobre marcadores de estrés oxidativo en la enfermedad de Chagas crónica. MÉTODOS: La población de estudio incluyó 42 pacientes con cardiopatía chagásica y los biomarcadores de estrés oxidativo fueron medidos antes y después de un período de seis meses de tratamiento con carvedilol (37,5 mg/día). Los pacientes fueron considerados de acuerdo con la clasificación de Los Andes, y la actividad de la superóxido dismutasa, catalasa, glutatión peroxidasa, S-transferasa y reductasa, mieloperoxidasa y adenosina deaminasa; y los niveles de glutatión reducida, de especies reactivas del ácido tiobarbitúrico, proteína carbonil, vitamina E y óxido nítrico fueron medidos en la sangre. RESULTADOS: Después del tratamiento con carvedilol, todos los grupos presentaron reducciones significativas en los niveles de proteína carbonil y glutatión reducida, mientras que los niveles de óxido nítrico y actividad de la adenosina aumentaron significativamente solamente en el grupo IA. Además de eso, la mayoría de las enzimas antioxidantes presentó disminución de sus actividades, en los grupos IA e IB. CONCLUSIONES: Los datos sugieren que el tratamiento con carvedilol fue eficaz en la atenuación del daño oxidativo, un efecto que puede ser particularmente importante en enfermedad de Chagas crónica con cardiopatía.

Myelomatous Pleural Effusion: A Case Series in a Single Institution and Literature Review / 대한진단검사의학회지

BACKGROUND: Myelomatous pleural effusion (MPE) is rare in myeloma patients. We present a consecutive series of patients with MPE in a single institution. METHODS: We retrospectively reviewed the medical records of 19 patients diagnosed with MPE between 1989 and 2008 at the Asan Medical Center. Diagnoses were confirmed by cytologic identification of malignant plasma cells in the pleural fluid. RESULTS: Our patients showed dominance of IgA (36.8%) and IgD (31.6%) subtypes. Of 734 myeloma patients, the incidence of MPE was remarkably high for the IgD myeloma subtype (16.7%), compared to the other subtypes (1.4% for IgG and 4.6% for IgA). At the time of diagnosis of MPE, elevated serum beta2-microglobulin, anemia, elevated serum lactate dehydrogenase, and elevated creatinine levels were found in 100%, 89.5%, 83.3%, and 57.9% of the patients, respectively. Approximately one-third (31.3%) of the patients had adenosine deaminase (ADA) activities in their pleural fluid exceeding the upper limit of the reported cutoff values for tuberculous pleural effusion (55.8 U/L). Chromosome 13 abnormality was seen in 77.8% of the tested patients. The median survival period from the development of MPE was 2.8 months. CONCLUSIONS: Patients with MPE have aggressive clinical and laboratory characteristics. The preponderance of IgD myeloma in MPE patients is a noteworthy finding because IgD myeloma is a rare subtype. Elevated ADA activity in the pleural fluid is also noteworthy, and may be helpful for detecting MPE. Physicians treating myeloma patients should monitor the development of MPE and consider the possibility of a worse clinical course.

Serum biochemical markers in rheumatoid arthritis.

Rheumatoid arthritis (RA) characterized by local and systemic effects of inflammation has a wide range of biochemical markers implicated directly or indirectly to its pathogenesis. In the present study, homocysteine, cortisol, adenosine deaminase (ADA), ferritin, malondialdehyde (MDA) and -tocopherol in serum of RA patients and healthy individuals were estimated to assess if they contribute to the disease process. The markers of disease activity such as erythrocyte sedimentation rate (ESR) and rheumatoid factor (RF) were also measured. The study group included a total of 45 subjects, including 30 RA patients and the rest being healthy individuals. RA group showed a significant increase in the levels of homocysteine, ADA and MDA, and a significant decrease in α-tocopherol compared to the healthy individuals. However, cortisol and ferritin levels did not show any significant change. Also, there was no significant correlation between the studied serum markers and markers of disease activity. Our results indicate that these biochemical markers contribute independently to the pathogenesis of RA.

Los niveles séricos de adenosin deaminasa y ácido úrico se correlacionan en pacientes gestantes con trastornos hipertensivos / Serum levels correlations of the enzyme adenosine deaminase (ADA) and uric acid in pregnant with hypertensive disorders

Objetivo: Evaluar los niveles séricos de la enzima adenosin-deaminasa (ADA) en pacientes gestantes normales y en pacientes con trastornos hipertensivos del embarazo, para determinar su relación con la gravedad del trastorno hipertensivo y con los niveles séricos de marcadores bioquímicos. Método: Se evaluaron pacientes con preeclampsia leve, preeclampsia grave, hipertensión gestacional y embarazadas sanas (n=10 por cada grupo). Se determinaron los niveles de ADA, ácido úrico, creatinina, amonio y enzimas hepáticas. Resultados: Se detectó una elevación en los niveles séricos de ADA en pacientes con preeclampsia y con hipertensión gestacional, en comparación con aquellas que cursaron con un embarazo normal. Los niveles ADA se correlacionaron positivamente con los niveles de ácido úrico y creatinina, más no con la severidad clínica. A su vez los niveles de ácido úrico se asociaron con la creatinina sérica y con la severidad clínica de los trastornos hipertensivos. Se encontró un incremento en los niveles de amonio en los pacientes con preeclampsia, el cual no se correlacionó con los otros marcadores bioquímicos, mientras que los niveles de TGO, TGP y LDH se encontraron significativamente elevados en la preeclampsia grave. Conclusión: Este estudio permite relacionar la actividad de ADA con los trastornos hipertensivos del embarazo, los niveles elevados de amonio con la preeclampsia y los niveles de ácido úrico, TGO, TGP y LDH con la severidad de los trastornos hipertensivos.

Objective: To evaluate serum levels of the enzyme adenosine deaminase (ADA) in normal pregnant and patients with hypertensive disorders induced by pregnancy, in order to determine their relationship with the severity of the hypertensive disorder and with serum biochemical markers. Method: We evaluated patients with mild preeclampsia, severe preeclampsia, gestational hypertension and healthy pregnancy (n=10 per group). We determined the serum levels of ADA, uric acid, creatinine, ammonia and liver enzymes. Results: In patients with preeclampsia and gestational hypertension we detected a rise in serum ADA as compared with those who had undergone a normal pregnancy. ADA levels were positively correlated with uric acid and creatinine serum levels, but not with clinical severity. Uric acid levels were associated with serum creatinine and the clinical severity of hypertensive disorders. We also found an increase in ammonia levels in patients with preeclampsia, which did not correlate with other biochemical markers, while the levels of SGOT, SGPT, and LDH were significantly elevated in severe preeclampsia. Conclusion: This study establishes a link between the activity of ADA with hypertensive disorders of pregnancy, high levels of ammonium with preeclampsia and uric acid, SGOT, SGPT and LDH levels with the severity of hypertensive disorders.

Auxiliary tools in tuberculosis: The hemolysis in pleural fluids underestimate the values of adenosine deaminase activity determined by the method of Giusti / La hemólisis en los fluidos pleurales subestima los valores de actividad de adenosina desaminasa determinados por el método de Giusti

The increase of adenosine deaminase (ADA) activity in pleural fluids (PF) is considered a useful tool in the diagnosis of pleural tuberculosis. It is known that numerous photometric methods are interfered by the hemolysis, as a result, hemolyzed specimens -or with blood- received in the laboratory are frequently rejected. In order to establish if the values of ADA were affected by the hemolysis or blood, ADA was determined in individual and pooled PF samples with the aggregate of erythrocyte lysate (H) or hemolyzed whole blood (HWB) from 312 mg/l to 12 g/l (final concentrations of hemoglobin in the samples), and plasma in appropriate dilutions. Negative interferences were caused by the H and HWB, starting already of 500 mg/l with relative errors until 50% in some cases, depending on the ADA activity. Increments of hemoglobin increased the negative interference. The aggregate of plasma increased slightly the ADA activity although it was insufficient for neutralize the negative effect of hemolysis. The clinical significance of the negative interference is in relation to the amount of hemoglobin present in the sample and the ADA activity. Near the cutoff (40 U/l) this interference can lead to discard erroneously the diagnosis of pleural tuberculosis.

El aumento de la adenosina desaminasa (ADA) en los líquidos pleurales (LP) es considerado una herramienta útil para el diagnóstico de la tuberculosis pleural. Se sabe que numerosos métodos fotométricos son interferidos por la hemólisis, y que muestras hemolisadas o con sangre son frecuentemente rechazadas por el laboratorio. Procuramos establecer si la hemólisis o la presencia de sangre interfieren en la determinación de ADA. Se determinó la ADA en muestras individuales y en mezclas de muestras de LP luego del agregado de lisado eritrocitario o sangre hemolisada, desde 312 mg/l hasta 12 g/l (concentraciones finales de hemoglobina en las muestras), y de plasma en diluciones adecuadas. El lisado eritrocitario y la sangre hemolisada interfirieron negativamente a partir de los 500 mg/l de hemoglobina, con errores relativos de hasta un 50% en algunos casos. El incremento de hemoglobina aumentó la interferencia negativa. El agregado de plasma aumentó levemente los valores de ADA, aunque fue insuficiente para neutralizar el efecto negativo de la hemólisis. La significación clínica de la interferencia negativa está relacionada con la cantidad de hemoglobina presente en la muestra y su actividad de ADA. Cerca del punto de corte (40 U/l), la interferencia negativa de la hemólisis puede conducir a descartar erróneamente el diagnóstico de tuberculosis pleural.

Serum nitrite level and adenosine deaminase activity is altered in visceral leishmaniasis.

In this study we sought to determine if there is alteration in nitric oxide (NO) production and adenosine deaminase (ADA) activity among patients with visceral leishmaniasis (VL) and the effect of four weeks of chemotherapy on these levels. Fifty-three VL patients diagnosed clinically and by direct demonstration of the LD bodies in the bone marrow smear were studied. They were treated with Sodium Stibogluconate and sampled at the baseline and four weeks. Forty-three healthy individuals coming from the same endemic area were taken as control. Total nitrite (NO2- and NO3-) as an index of NO production and ADA activity was measured spectrophotometrically. Serum nitrite level decreased significantly in patients as compared to the healthy individuals but significantly increased following 4 weeks of chemotherapy. Conversely, Increased ADA activity was observed in the beginning of treatment and decreased significantly with successive 4 weeks of chemotherapy. It seems a negative correlation between NO level and ADA activity. This result indicates parasite induced evasion of NO and activation of T lymphocytes during immunopathogenesis of VL. Therefore, assessment of NO metabolites may be useful marker in the evaluation of the effector mechanism of macrophages and clinical manifestation of patients.

Characterization of adenosine deaminase (ADA) in Hemolymph of Biomphalaria glabrata

A adenosina é uma molécula sinalizadora de muitos eventos celulares. A adenosina desaminase (ADA) é enzima chave para o controle dos níveis intra e extra celulares de adenosina. A atividade da ADA foi detectada em hemolinfa de B. glabrata e suas condições ótimas de ensaio foram determinadas experimentalmente. A variação do pH de 6,2 até 7,8 não causou mudança significativa na atividade. O Km aparente foi de 734 µmoles L-1, usando adenosina como substrato. A maior atividade foi encontrada usando 37ºC como temperatura de incubação. As condições de ensaio padrão foram então estabelecidas como sendo 15 minutos de tempo de incubação, 0,4 µL de hemolinfa por ensaio, pH 6.8 e 37ºC de temperatura de incubação. A enzima apresentou atividades de 834 ± 67 µmols.min-1.L-1 (25ºC) e 2029 ± 74 µmols.min-1.L-1 (37ºC), em torno de 40 e 100 vezes maiores que os níveis encontrados em soro de humanos sadios. Em temperaturas superiores, essa atividade cai 20% a 43ºC e 70% a 50ºC, em 15 minutos. A ADA perde 26 a 78% de sua atividade quando a hemolinfa é pré-incubada a 50ºC de 2 a 15 minutos, respectivamente. Considerando os altos níveis de ADA encontrados pode-se inferir que, em animais sadios e alimentados, a adenosina é mantida em baixas concentrações na hemolinfa. Tendo a atividade da enzima permanecido constante frente à larga faixa de pH testada, sugere-se que a ADA pode atuar com eficiência mesmo em situações adversas que determinem variações no pH da hemolinfa.

Diagnostic yield of adenosine deaminase in bronchoalveolar lavage.

Adenosine deaminase (ADA) activity rises in various body fluids in patients with tuberculosis. A prospective study was conducted to determine the diagnostic value of ADA activity in bronchoalveolar lavage. Between March 2001 and February 2003, 148 patients were enrolled in our study, mean age 55.6 years (SD 14.6), and a male to female ratio of 2.4:1. The mean duration of symptoms was 66.2 days. All patients were either sputum-smear negative for AFB or failed to produce sputum. The final diagnosis resulted in three patient groups: 43 with pulmonary tuberculosis, 70 malignancy, and 35 miscellaneous causes. The mean ADA activity in the bronchoalveolar lavage for the pulmonary tuberculosis, malignancy, and miscellaneous causes groups was 8.98 (95% CI, 3.79-14.17), 7.63 (95% CI, 4.12-11.14), and 11.61 U/l (95% CI, 3.59-19.62), respectively. No difference was detected in the ADA level in the pulmonary tuberculosis vs other groups (p=0.56, one-way ANOVA). A high level of ADA activity was found in non-tuberculous conditions such as bronchogenic carcinoma, pulmonary hemosiderosis, chronic pneumonia with empyema thoracis and chronic myeloid leukemia. We concluded that ADA activity in the bronchoalveolar lavage was not clearly diagnostic of smear-negative pulmonary tuberculosis. Early diagnosis required histopathology of biopsied transbronchial specimens obtained by fiberoptic bronchoscopy.

The Diagnostic Value of the Adenosine Deaminase Activity in the Pleural Fluid of Renal Transplant Patients with Tuberculous Pleural Effusion

The assessment of the adenosine deaminase activity (ADA) in the pleural effusion is used for the diagnosis of tuberculous pleural effusion (TPE). To examine whether the procedure can be applied to immunocompromised patients, we analyzed the ADA activity in the pleural fluid of renal transplant recipients. We studied 23 renal transplant patients with TPE (21 men and 2 women; the mean age, 33 years). They were treated at the Yonsei University Hospital between January 1985 and December 2001. Patients with granuloma in the pleural biopsy specimen or positive for Mycobacterium tuberculosis in the pleural fluid culture were recruited. The ADA activity in the pleural effusion of 23 renal transplant patients with TPE was compared with 23 immunocompetent patients with TPE. The mean ADA activity was 69.5 +/- 4.6 in renal transplant patients and 65.0 +/- 4.9 U/L in immunocompetent patients. Applying the 40 U/L cut-off point, the positivity of ADA was 91.3% in renal transplant patients, and 86.9% in immunocompetent patients. We thus concluded that the measurement of ADA in the pleural fluid is a useful means in the diagnosis of TPE in renal transplant patients.

The Serum Vitamin C Levels in Behcet's Disease

Behcet's disease (BD) is a chronic inflammatory disorder of unknown aetiology, and recognised as a multi-system vasculitis. It has been postulated that an imbalance of the oxidant and antioxidant systems related to the disease are important in its pathogenesis. Previous publications have reported increased levels of enzymatic antioxidant defence systems in patients with BD. The non-enzymatic antioxidant systems, including vitamin C and uric acid, were looked for in the present study. For this aim, the serum malondialdehyde (MDA), an end product of lipid peroxidation, and vitamin C and uric acid, as endogenous antioxidants, were determined in 20 patients with BD (11 in active and 9 in inactive periods) and 20 healthy subjects. The MDA level was significantly higher in both the active and inactive period patients compared with the control group (p 0.05). There was also no significant difference in uric acid levels between the groups (p > 0.05). In the patients group, a negative correlation was found between the levels of serum MDA and vitamin C (r=-0.517; p < 0.05). Our results indicate that decreased vitamin C and increased MDA levels reflect the increased levels of oxidative stress in BD patients, and this situation may be important in relation with its pathogenesis.

Expression of adenosine deaminase and 5'-nucleotidase in artificially induced deciduoma in rat and hamster.

Enzymes adenosine deaminase (ADA) and 5-nucleotidase (5-'NT) are known to play active role in tissue/cell proliferation and differentiation. To validate this the two enzymes were studied in artificially induced deciduoma of rat and hamster. The deciduoma was induced by traumatizing one of the uterine horns of progesterone primed animals. Non traumatized horn served as control. The animals were later maintained on progesterone, given alone (Gr.I) or conjointly with estrogen (Gr.II). The weight of each uterine horn was recorded to determine the formation of deciduoma. There was no marked difference between the weights of traumatized and control horn on day 2 post-traumatization (PT), but a progressive rise was noticed after this day in both species. The ADA activity however differed, day and species wise. While in the rats of Gr.I it was low in the traumatized horn on all the days, in the hamsters it was remarkably high from day 2 to 6 PT. In the rats of Gr.II also the activity though was low in the traumatized horn, but on day 2 and 4 only; on day 6 and 7 PT it increased markedly. In hamster, on the contrary, again the enzyme activity was remarkably high on all the three days. The 5'-NT activity, however, did not show any marked difference between the two horns under Gr.I and II in both species. It was rather high in the control horn of each group. The results suggest: (I) the progesterone alone though produces a significant rise in the uterine weight of traumatized horn in both species, the ADA activity increases only in hamster, (2) under the conjoint treatment also the enzyme activity remains high in hamster; and (3) the activity of enzyme 5'-NT does not alter during the deciduoma formation in both the species.

Diagnóstico da tuberculose pleural pela ADA, isolada ou combinada a outras variáveis, inclusive em HIV-positivos / Diagnosis of pleural tuberculosis by ADA, isolated or combined with other variables, incluing in HIV positive patients

A tuberculose pleural (TBP) é a de maior incidência entre as formas extrapulmonares e, provavelmente, o diagnóstico mais freqüente dos derrames pleurais observados no Brasil, justificando os estudos para se estabelecerem métodos diagnósticos rápidos e acessíveis. Neste trabalho foi dimensionado o volume da atividade da adenosina deaminase (ADA), isolada ou combinada a outras variáveis de fácil acesso, baixo custo e simples obtenção no diagnóstico da TBP, inclusive em infectados pelo HIV. Trata-se de estudo retrospectivo e seqüencial, avaliando os resultados de punção pleural diagnóstico realizada em dois tempos (toracocentese e biópsia pleural). Pleuris não-tuberculoso (PNT) foi diagnosticado quando presentes três parâmetros entre história clínica, radiologia, bioquímica e citologia do líqüido pleural, anatomopatologia e outros. As três variáveis selecionadas para avaliação combinada com a ADA foram: concentração de proteínas (CPT), taxa de linfócitos (TLT) no líquido pleural e idade dos doentes (IDD). Os resultados foram analisados estatisticamente pelos testes do qui-quadrado (X2) e t de Student. Os níveis de corte foram definidos pela maximização da sensibilidade e especificidade por meio da curva ROC, avaliados os valores diagnósticos e a contribuição independente no diagnóstico etiológico da TBP pela regressão logística múltipla das quatro variáveis. Foram estudados 417 pacientes maiores de 14 anos portadores de derrame pleural (excluídos os empiemas) matriculados no Instituto Clemente Ferreira (ICF) em São Paulo no período de 1990 a 1994. Destes pacientes 268 estavam com TBP, dos quais 47 HIV-negativos e 21 positivos e 149 com PNT, sendo 33 HIV-negativos e 8 positivos. Os valores médios das variáveis analisadas apresentaram diferenças estatisticamente significantes entre os pacientes com TBP e PNT. Os níveis de corte estabelecidos pela curva ROC foram de > 30 U/L para a ADA, de > 4,5 g/dL para a CPT, de > 90 por cento para a TLF e < 45 anos para a IDD. A sensibilidade, a especificidade o valor preditivo positivo, o valor preditivo negativo e a acurácia encontrados foram, respectivamente: para a ADA = 94, 95, 97, 89 e 94 por cento; para a CPT = 80, 64, 80, 64 e 74 por cento para a TLF = 73, 67, 80, 58 e 71 por cento e para a IDD = 77, 69, 77, 69 e 75 por cento. No modelo de regressão logística múltipla a ADA > 30 U/L se associou ao diagnóstico etiológico da TBP com uma razão de chance de 184 vezes...

Utility of polymerase chain reaction using two probes for rapid diagnosis of tubercular pleuritis in comparison to conventional methods.

We have used polymerase chain reaction (PCR) with IS6110 and a new set of primers from an insertion element like repetitive sequence, (TRC4) to detect Mycobacterium tuberculosis in pleural effusion samples from 50 patients having pleuritis. The results of PCR were compared with the results of conventional methods like smear, culture and adenosine deaminase activity. Thirty six specimens were positive and 14 were negative by PCR. Among the 36 samples, 33 were from patients with clinical evidence of tuberculosis including response to anti-tuberculosis therapy. Only six samples were positive by the gold standard which is culture, and three were positive by smear. The measurement of adenosine deaminase activity classified 19 samples as positives. The overall sensitivity and specificity of PCR was 100 and 85 per cent respectively. PCR using IS6110 and TRC4 primers is a sensitive test as compared to conventional tests for detection of M. tuberculosis from pleural fluid samples of patients with tubercular pleuritis.

Evaluación de la actividad de la adenosin deaminasa en suero de pacientes con eritema indurado de Bazin / Adenosin deaminase activity in sera of patients with erithema induratum of Bazin

La Adenosín Deaminasa (ADA) es una enzima del catabolismo de las purinas. Puede encontrarse actividad ADA en toda célula, pero alcanza niveles especialmente altos en monocitos y linfocitos T, que son importantes en la inmunidad celular. Nuestro objetivo fue medir la actividad sérica de ADA y la respuesta de hipersensibilidad a tuberculina (PPD) en pacientes con eritema indurado de Bazin (EIB). Se seleccionaron en forma prospectiva 15 pacientes con EIB a las que se les midió actividad ADA (según método de Giusti modificado) y reacción de hipersensibilidad cutánea con 2U de tuberculina (PPD) durante la fase activa de la enfermedad. Las mediciones también se realizaron en 18 mujeres control. El promedio de actividad ADA sérica en la serie en estudio fue de 29,5 U/L y en la serie control fue de 15,6 U/L. El promedio de PPD en la serie en estudio fue de 20,7 mm y en la serie control fue de 7,2 mm. Concluimos que las pacientes con EIB tuvieron niveles de ADA séricos significativamente mayores que las del grupo control, (p<0,0002). También tuvieron valores de PPD significativamente más altos que el control (p<0,00001). No se observó una asociación estadísticamente significativa entre un mayor nivel de ADA y una mayor respuesta cutánea a la tuberculina