Evaluation of the commercial kit SIRE nitratase for detecting resistant mycobacterium tuberculosis in Brazil

Abstract INTRODUCTION: This study aimed to evaluate a new commercial kit, Kit SIRE Nitratase-PlastLabor, for testing the drug susceptibility of clinical Mycobacterium tuberculosis isolates. METHODS: The accuracy of the Kit SIRE Nitratase was evaluated by examining the susceptibility (streptomycin, isoniazid, rifampicin, and ethambutol) of 40 M. tuberculosis isolates, using the proportion method with Lowenstein-Jensen medium or the BACTEC MGIT 960 system. RESULTS: The detection accuracy for streptomycin, isoniazid, rifampicin, and ethambutol was 95%, 97.5%, 100%, and 80%, respectively. CONCLUSIONS: The exceptional accuracy demonstrated by Kit SIRE Nitratase for isoniazid and rifampicin makes the kit an attractive option for screening M. tuberculosis strain resistance.

Transient Elastography vs Aspartate Aminotransferase to Platelet Ratio Index in Hepatitis C: A Meta-Analysis

ABSTRACT Background and rationale. Many different non-invasive methods have been studied with the purpose of staging liver fibrosis. The objective of this study was verifying if transient elastography is superior to aspartate aminotransferase to platelet ratio index for staging fibrosis in patients with chronic hepatitis C. Material and methods. A systematic review with meta-analysis of studies which evaluated both non-invasive tests and used biopsy as the reference standard was performed. A random-effects model was used, anticipating heterogeneity among studies. Diagnostic odds ratio was the main effect measure, and summary receiver operating characteristic curves were created. A sensitivity analysis was planned, in which the meta-analysis would be repeated excluding each study at a time. Results. Eight studies were included in the meta-analysis. Regarding the prediction of significant fibrosis, transient elastography and aspartate aminotransferase to platelet ratio index had diagnostic odds ratios of 11.70 (95% confidence interval = 7.13-19.21) and 8.56 (95% confidence interval = 4.90-14.94) respectively. Concerning the prediction of cirrhosis, transient elastography and aspartate aminotransferase to platelet ratio index had diagnostic odds ratios of 66.49 (95% confidence interval = 23.71- 186.48) and 7.47 (95% confidence interval = 4.88-11.43) respectively. Conclusion. In conclusion, there was no evidence of significant superiority of transient elastography over aspartate aminotransferase to platelet ratio index regarding the prediction of significant fibrosis, but the former proved to be better than the latter concerning prediction of cirrhosis.

Clinical and laboratory description of a series of cases of acute viral myositis / Descrição clínico-laboratorial de uma série de casos de Miosite Aguda Viral

ABSTRACT OBJECTIVE: Describe the clinical and laboratory profile, follow-up, and outcome of a series of cases of acute viral myositis. METHOD: A retrospective analysis of suspected cases under observation in the emergency department was performed, including outpatient follow-up with the recording of respiratory infection and musculoskeletal symptoms, measurement of muscle enzymes, creatine phosphokinase (CPK), lactate dehydrogenase (LDH), transaminases (AST and ALT), blood count, C-reactive protein, and erythrocyte sedimentation rate in the acute phase and during follow-up until normalization. RESULTS: Between 2000 and 2009, 42 suspected cases were identified and 35 (27 boys) were included. The median age was 7 years and the diagnosis was reported in 89% in the first emergency visit. The observed respiratory symptoms were cough (31%), rhinorrhea (23%), and fever (63%), with a mean duration of 4.3 days. Musculoskeletal symptoms were localized pain in the calves (80%), limited ambulation (57%), gait abnormality (40%), and muscle weakness in the lower limbs (71%), with a mean duration of 3.6 days. There was significant increase in CPK enzymes (5507 ± 9180 U/L), LDH (827 ± 598 U/L), and AST (199 ± 245 U/L), with a tendency to leukopenia (4590 ± 1420) leukocytes/mm3. The complete recovery of laboratory parameters was observed in 30 days (median), and laboratory and clinical recurrence was documented in one case after 10 months. CONCLUSION: Typical symptoms with increased muscle enzymes after diagnosis of influenza and self-limited course of the disease were the clues to the diagnosis. The increase in muscle enzymes indicate transient myotropic activity related to seasonal influenza, which should be considered, regardless of the viral identification, possibly associated with influenza virus or other respiratory viruses.

RESUMO OBJETIVO: Descrever o perfil clínico-laboratorial, o acompanhamento e o desfecho de uma série de casos de miosite aguda viral. MÉTODO: Foi conduzida uma análise retrospectiva de casos suspeitos, em observação em unidade de emergência, e seguimento ambulatorial com o registro de sintomas de infecção respiratória, sintomas músculo-esqueléticos, determinação de enzimas musculares, creatina-fosfoquinase (CPK), desidrogenase lática (DHL), transaminases (AST e ALT), hemograma, proteína C reativa e velocidade de hemossedimentação, na fase aguda e no acompanhamento, até a normalização. RESULTADOS: Entre 2000 e 2009, 42 casos suspeitos foram identificados e 35 (27 meninos) foram incluídos. A mediana de idade foi de sete anos e o diagnóstico relatado em 89%, na primeira visita de emergência. Os sintomas respiratórios observados foram: tosse (31%), coriza (23%) e febre (63%), com duração média de 4,3 dias. Os sintomas músculo-esqueléticos foram: dor localizada nas panturrilhas (80%), deambulação limitada (57%), marcha anormal (40%) e fraqueza muscular nos membros inferiores (71%), com duração média de 3,6 dias. Observou-se elevação importante das enzimas CPK (5.507 ± 9.180) U/l, DHL (827 ± 598) U/l e AST (199 ± 245) U/l e tendência a leucopenia (4.590 ± 1.420) leucócitos/mm3. A recuperação completa dos parâmetros laboratoriais foi observada em 30 dias (mediana) e a recaída clínica e laboratorial em um caso após 10 meses. CONCLUSÃO: Os sintomas típicos com enzimas musculares elevadas após diagnóstico de influenza e o curso autolimitado foram os indícios para o diagnóstico. A elevação de enzimas musculares indica a atividade miotrópica transitória relacionada à influenza sazonal que deve ser considerada, a despeito da identificação viral, possivelmente associada com o vírus influenza ou outros vírus respiratórios.

Prevalência de carbapenemases em enterobactérias resistentes a carbapenêmicos em quatro hospitais terciários de Porto Alegre / Prevalence of carbapenemases in carbapenem-resistant Enterobacteriaceae in four tertiary care hospitals in Porto Alegre

INTRODUÇÃO: A disseminação de Klebsiella pneumoniae carbapenemase (KPC) no Brasil e a recente detecção de bactérias produtoras de New Delhi metalo-β-lactamase (NDM-1) em hospital terciário do sul do Brasil indicam a necessidade da avaliação da presença destas enzimas em enterobactérias resistentes a carbapenêmicos (ERC).OBJETIVO: Avaliar prevalência de carbapenemases nas ERC em quatro hospitais terciários de Porto Alegre, por meio de PCR multiplex em tempo real. MÉTODOS: Estudo descritivo, período de abril a dezembro de 2013. Isolados bacterianos de pacientes internados foram identificados pelo sistema automatizado VITEK 2, com realização do teste de suscetibilidade aos antimicrobianos. Amostras com isolados de ERC foram encaminhadas ao laboratório de referência para análise por PCR em tempo real para identificação de carbapenemases. RESULTADOS: Total de 701 isolados. As ERC predominantes foram K. pneumoniae (47% das amostras positivas) e Enterobacter cloacae (18%). As carbapenemases mais frequentes foram KPC (48%), OXA-48-like (3%) e NDM (2%). Em 47% das amostras não foi identificado o mecanismo de resistência. Isolados originados de culturas de vigilância foram associados com maior positividade para carbapenemases do que isolados de amostras clínicas (p<0,0001). Isolados de ERC pertencentes ao grupo Proteae (Proteus spp., Morganella spp., Providencia spp.) foram associados a menor positividade para carbapenemase do que isolados de outras ERC (p<0,0001). CONCLUSÃO: KPC foi a carbapenemase mais frequentemente detectada. A circulação de uma enzima OXA-48-like foi demonstrada, um achado novo e preocupante. O achado da carbapenemase NDM também é preocupante devido ao seu potencial de disseminação. Esses dados e outros estudos poderão contribuir para um entendimento maior da epidemiologia das ERC.

BACKGROUND: The spread of Klebsiella pneumoniae carbapenemase (KPC) in Brazil and the recent detection of bacteria producing New Delhi metallo-β-lactamase (NDM-1) in a tertiary care hospital in Porto Alegre indicate the need to evaluate the presence of these enzymes in carbapenem-resistant Enterobacteriaceae (CRE). AIM: To evaluate the prevalence of carbapenemases in CRE in four tertiary care hospitals in Porto Alegre using multiplex real-time PCR.METHODS: Descriptive study from April to December 2013. Bacterial isolates from hospitalized patients were identified by VITEK 2 automated system, with antimicrobial susceptibility testing. Samples with CRE isolates were sent to the reference laboratory for analysis using real-time PCR for identification of carbapenemases. RESULTS: Total of 701 isolates. The predominant CRE were K. pneumoniae (46% of positive samples) and Enterobacter cloacae (18%). The most frequent carbapenemases were KPC (48%), OXA-48-like (3%), and NDM-1 (2%). In 47% of the samples no carbapenemase was identified. Isolates originated from surveillance cultures were associated with higher positivity for carbapenemases than isolates from clinical samples (p<0.0001). CRE isolates belonging to the Proteae group (Proteus spp., Morganella spp., Providencia spp.) were associated with less positivity for carbapanemase than isolates of other CRE (p<0.0001). CONCLUSION: KPC was the most frequently detected carbapenemase. The movement of an OXA-48-like enzyme was demonstrated, a novel and worrisome finding. The finding of carbapenemase NDM is also worrisome due to its dissemination potential. These data and further studies may contribute to a better understanding of the epidemiology of CRE.

Enzymes in clinical medicine: An overview.

Enzymes are biocatalysts and because of their remarkable properties, they are extensively used in medical diagnosis. Researches in the last two decades have concentrated more on enzymes such as creatine kinase–MB, alanine transaminase, aspartate transaminase, acid phosphatase, alkaline phosphatase etc. for clinical applications. Enzymes are the preferred markers in various disease states such as myocardial infarction, jaundice, pancreatitis, cancer, neurodegenerative disorders, etc. They provide insight into the disease process by diagnosis, prognosis and assessment of response therapy. Even though the literature on the use of enzymes in various disease conditions has accumulated, a comprehensive analysis is lacking and hence this review.

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

Diagnosis of Malaria by Detection of Parasite Lactate Dehydrogenase with Advantage Mal Card Test.

Background : Malaria is a global problem. Rapid diagnosis is essential for effective treatment and reducing mortality and morbidity of malaria. Diagnosis of malariaby peripheral smear is labor-intensive and requires considerable expertise for its interpretation. A rapid test , Advantage MAL card test is based on detection of parasite lactate dehydrogenase (pLDH) and has the ability to differentiate the four major Plasmodium species in 20 minutes. Objectives: 1) To evaluate utility of parasite lactate dehydrogenase for diagnosis of malaria with Advantage Mal card test.2) To compare the results of Advantage Mal card test with peripheral smear findings. Materials and Methods: In this retrospective study, total 5242 patients with malaria like symptoms attending OPD and admitted in wards at Acharya Vinoba Bhave Rural Hospital (AVBRH) from January 2008 to August 2011 were studied. Result: The age of patients ranged from < 1 year- >80 years. The commonest age group affected was 21-30 years. Male to female ratio was 1.04: 1. Prevalence rate of malaria was 101/1000 population in AVBRH. Malarial parasites were detected in PS in10.11% patients (P.falciparum 27.73% , P.vivax 71.32% , mixed infection 0.94%) and in 10.07% patients with Advantage Mal test (P. falciparum 28.03%, P.vivax 71.02%, mixed infection 0.95%). 3 cases of P.vivax and 1case of P.falciparum detected by PS were not detected by Advantage Mal test. 2 cases of P.falciparum detected by Advantage Mal and not by PS. Compared to PS, the Advantage Mal had sensitivity 99.24%, specificity 100%, positive predictive value 100%, negative predictive value was 89.92%. Conclusion: Diagnosis of malaria by detection of pLDH with Advantage Mal card test is simple ,rapid, reliable and cheap method. Results are comparable to blood films. It can detect P.flciparum infection when parasites are sequestered.

Planejamento racional de novos tuberculostáticos com base na estrutura das enzimas enoil redutase e timidina monofosfato quinase do Mycobacterium tuberculosis / Rational design of new antituberculosis agents based on the structures of Mycobacterium tuberculosis enzymes enoyl reductase and thymidylate kinase

Nos últimos quinze anos, a tuberculose (TB) ressurgiu tanto em países em desenvolvimento quanto naqueles desenvolvidos. Em 1993, a Organização Mundial da Saúde (OMS) declarou a TB como emergência global de saúde pública. Atualmente, um terço da população mundial está infectada pelo Mycobacterium tuberculosis e mais de 10% destes indivíduos desenvolverão a doença ativa. Em 2006, estimaram-se 9 milhões de novos casos de TB em todo o mundo. No Brasil, aproximadamente 95.000 novos casos são registrados anualmente, com incidência de 50 casos em cada 100.000 habitantes. Tendo em vista o quadro alarmante da TB no mundo e, em especial no Brasil, e considerando os índices elevados de resistência do microrganismo aos fármacos convencionalmente utilizados na terapêutica, há necessidade urgente do desenvolvimento de novos tuberculostáticos. Além disso, a busca por novos alvos de ação se faz necessária, já que os antimicobacterianos utilizados na terapia anti-TB têm como alvo apenas pequeno número de enzimas relacionadas a funções essenciais do microrganismo. A biossíntese bacteriana de ácidos graxos tem despertado atenção especial como alvo atraente no desenvolvimento de novos agentes antibacterianos. Diferenças bioquímicas e funcionais fazem com que as enzimas envolvidas em tal processo sejam alvos potencialmente atraentes para o desenvolvimento de novos agentes antibacterianos/antimicobacterianos. As enoil-acp redutases são enzimas determinantes na etapa de alongamento de ácidos graxos, produtos intermediários na biossíntese dos principais constituintes da parede celular micobacteriana, os ácidos micólicos...

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.

Optimization of b-galactosidase production using Kluyveromyces lactis NRRL Y-8279 by response surface methodology

This paper investigates the production and optimization of b-galactosidase enzyme using synthetic medium by Kluyveromyces lactis NRRL Y-8279 in shake flask cultures. Among the different cell disintegration methods used, the highest specific activity was obtained when the cells were permeabilized using isoamyl alcohol. Response surface methodology was used to investigate the effects of four fermentation parameters (agitation speed, pH, initial substrate concentration and incubation time) on b-galactosidase enzyme production. Results of the statistical analysis showed that the fit of the model was good in all cases. Maximum specific enzyme activity of 4218.4 U g-1 was obtained at the optimum levels of process variables (pH 7.35, agitation speed 179.2 rpm, initial sugar concentration 24.9 g l-1 and incubation time 50.9 hrs). The response surface methodology was found to be useful in optimizing and determining the interactions among process variables in b-galactosidase enzyme production.

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.

Actividad de la glutatión reductasa en el embarazo diabético / Glutathione reductase activity in diabetic pregnancy

ANTECEDENTES: El predominio de la diabetes gestacional está aumentando en nuestra población y sus efectos en el metabolismo celular y estatus oxidativo están siendo estudiados. OBJETIVO: Determinar si existe una relación entre la actividad de la glutatión reductasa eritrocitaria, evaluada a través del test CAGRE y la diabetes gestacional. MÉTODO: Estudio transversal de casos y controles, incluyó a 30 embarazadas con diagnóstico de diabetes gestacional y 30 sin patologías asociadas, pertenecientes a la Maternidad del Hospital Barros Luco-Trudeau, Santiago de Chile. La actividad de la glutatión reductasa de eritrocitos fue determinada espectrofotométricamente a través del test CAGRE, la que fue relacionada con variables maternas como: edad, hematocrito, presencia de antecedentes de enfermedades familiares, estado nutricional materno e índice de masa corporal. Para comparar las variables entre ambos grupos, se realizaron medidas de disparidad, posición y análisis de la correlación. RESULTADOS: El grupo de madres diabéticas presentó edad materna, índice de masa de corporal y antecedentes de diabetes gestacional mayores que el grupo control, aunque sin diferencia estadísticamente significativa. El predominio de anemia y la respuesta al suplemento del cofactor FAD en la actividad de la glutatión reductasa eritrocitaria fue similar entre los grupos. La mayor incidencia de diabetes familiar en el grupo control fue estadísticamente significativa. La distribución de los valores de CAGRE, utilizado también como un indicador de los niveles de riboflavina, mostró en el grupo de casos riesgo medio a alto de malnutrición, mientras que en el grupo control la tendencia fue normal o de riesgo bajo. CONCLUSIÓN: Las embarazadas diabéticas gestacionales, presentaron mal nutrición y un estrés oxidativo mayor que el grupo control, evidenciado por el test de CAGRE.

BACKGROUND: The prevalence of the gestational diabetes is increasing in our population and its effects in the cellular metabolism and oxidative status had been studied. OBJECTIVE: Determine if exists a relationship between the erythrocyte glutathione reductase activity evaluated by EGRAC test and the gestational diabetes. METHODS: This traversal study of cases and controls, included at 30 pregnant with diagnostic of gestational diabetes and 30 without associate pathologies, belonging to the Maternity of the Hospital Barros Luco-Trudeau, Santiago-Chile. The activity of the glutathione reductase was determined by spectrophotometric assay through the EGRAC test, and their values were related with maternal variables as: age, hematocrite, presence of antecedents of family illnesses, maternal nutritional status and the body mass index. To compare the variables between both groups, they were carried out measures of disparity, position and analysis of the correlation. RESULTS: We determine that the group of diabetic mothers was older, with higher body mass index and a bigger frequency of antecedents of gestational diabetes that the control group, although without significant difference. The prevalence of anemia and the answer to the supplement with FAD in the activity of the glutathione reductase was similar among the groups. On the other hand, the incidence of familiar diabetes in the group control was bigger. The distribution of the values of EGRAC, also used as an indicator of the riboflavin levels, showed in the group of cases half risk to high of malnutrition, while in the group control the tendency was normal or low risk. CONCLUSION: We can conclude that the gestational diabetics pregnant presented malnutrition and higher oxidative stress that the control group, evidenced by means of the EGRAC test.

Estimation of postmortem intervals by using terminal deoxynucleotidyl transferase / 法医学杂志

OBJECTIVE@#To investigate the relationship between DNA degradeation and postmortem interval.@*METHODS@#dUTP was transferred to 3'terminal of DNA by using terminal deoxynucleotidyl transferase (TDT), then the reminders of dUTP after experimental reaction, as indicator of quantity of DNA degradation, were detected.@*RESULTS@#The reminders of dUTP were decreasing along with the postmortem interval.@*CONCLUSION@#Postmortem DNA degradation may be used in postmortem interval judgment.

The effect of blood on rapid urease test for Helicobacter pylori detection: an in vitro study.

The sensitivity of rapid urease test (RUT) for detecting Helicobacter pylori (H. pylori) is limited in patients presented with bleeding peptic ulcers. Blood contents are potentially responsible for the lack of sensitivity of RUT in patients with upper gastrointestinal bleeding. This study was designed to determine the efficacy of RUT in detecting H. pylori when exposed to the patients' own blood at varying durations. Four gastric antral biopsy specimens were collected from dyspeptic patients who underwent gastroscopic examination. The first specimen was immediately placed into the urea broth as the control group. The second, third and fourth specimens were immersed in the patients' own blood for one, two and four hours respectively before testing for the RUT. The results of these RUT were compared to those of the control group. Gastric antral biopsy specimens tested with the RUT of twenty patients were evaluated in this study. The 1-hour RUT and 2-hour RUT was not significantly different in sensitivity (92.31% vs 100%: p>0.05, 84.62% vs 100%: p>0.05 respectively) and accuracy (95.0% vs 100%: p>0.05, 90.0% vs 100%: p>0.05 respectively) compared to the unexposed blood RUT for H. pylori diagnosis. However, the study demonstrated that the 4-hour RUT had significantly lower sensitivity (53.85% vs 100%: p<0.05) and accuracy (70.00% vs 100%: p<0.05) than that of the unexposed blood RUT for H. pylori diagnosis. It is concluded that exposure of gastric biopsy specimens to their own blood for four hours significantly decreased the sensitivity of the biopsy rapid urease test for H. pylori detection.

A simple, rapid fluorometric assay for the determination of glucose 6-phosphate dehydrogenase activity in dried blood spot specimens.

Glucose 6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathy of human beings and is the most common cause of jaundice and acute hemolytic anemia in South East Asia. The deficiency causes acute hemolytic anemia following ingestion of 6-amino quinoline antimalarials, phenacetin, and other substances. The rapid identification of infants or patients with this deficiency would help to prevent their exposure to these substances and subsequent risk to health. The assay is relatively simple. A 3mm punch from a dried blood spot sample is placed in a well of a black fluorescent microtiter plate containing calibrators and controls in duplicate. 100 microl of reagent is added and the sample is allowed to react for 30 minutes at ambient temperature after which 200 microl of stop reagent is added. The plate may be read immediately or up to one hour in a fluorescent reader (ex 355 nm: em 460 nm). Glutathione. ascorbate and bilirubin do not affect the assay. hemoglobin does quench the fluorescence by about 1.1 fluorescence units/g/dHb. This would not cause any false negatives and deficients would not be missed. G6PD activity in whole blood normal samples was examined at -20, 6 and 37 degrees C over 14 days. The samples lost about 20% activity after 48 hours and 31% by the end of 14 days. The samples stored at -20 degrees C and 6 degrees C remained relatively stable over this period. In a preliminary study eight diagnosed G6PD deficient samples had a mean value of 2.0 U/gHb (range 0.8 to 4.4) and fell within 3 SD units of the mean. Forty one normal samples had a mean of 6.6 micromol/min/gHb. Only one sample with a low hemoglobin level fell outside of 3 SD units of the mean. The Wallac assay was compared to the Sigma G6PD assay and although the values appeared lower at normal levels, the deficient samples compared well.

Provisional identification of haemophilus influenzae from sputum cultures within one hour by rapid enzyme test

In this study, Identification of possible H. Influenzae Isolates from cultures of sputum samples by a 5 min test for production of indole and Beta-galactosidase followed by 1 h porphyrin test was compared with tests for dependence on[X] and [V] factors. A commercially produced form of the rapid tests [Haemstrip Lab. M. Bury, Lancs] was used. The study was conducted on 300 patients with respiratory symptoms from Banha and Zagazig University Hospitals where 75 potential Haemophilus colonies from cultures of sputum samples on heated blood agar incubated in humified atmosphere with CO[2] were identified. When the results of the two tests were in disagreement, both tests were repeated. Colonies that were Beta-galactosidase positive after 5 min were considered to be non H. lnfluenzae and those that were Beta-galactosidase negative but indole positive were considered to be H. Influenzae. At this stage the test had a sensitivity of 85.3% and specificity of 85.7%. After 1 h, only colonies that were Beta-galactosidase and porphyrin negative were considered to be H. Influenzae, the sensitivity was then 100% and specificity 100%. Accordingly, Haemstrip results may permit provisional identification of H. influenzae, a potential pathogen of the lower respiratory tract, after overnight culutre of primary isolation plate

Pattern of certain enzyme activities in serum and urine in different types and stages of bilharzial patients

The study comprised 116 bilharzial adult male patients with age range 18-65 years. None of the selected individuals was complaining from other diseases and proved to have no treatment for bilharziasis prior to their inclusion in this study. Data for age, weight and degree of liver and /or spleen enlargement were registered. Besides the bilharzial cases, 15 healthy adult normal individuals [group C] with the same age range as control, were included in the study. The bilharzial cases were: a] 15 patients with hematobiasis [group H], b] 74 subjects of mansoniasis, further subdivided into 4 subgroups [23 cases in the early stage of the disease where the liver was not enlarged and the spleen was not palpable [m1], 21 subjects in the 2nd stage of the disease [m2], the main clinical feature was the occurrence of hepatomegaly, the spleen was not enlarged, 15 cases in the 3rd stage of the disease [m3], the hepatosplenomegalic stage of bilharziasis without ascites, 15 subjects in the 4th stage of the disease [m4], hepatosplenomegaly with ascites], c] 27 cases with mixed schistosomiasis were subdivided into 4 subgroups: 14 cases of mixed infection in early stage of mansoniasis [Mix1], 5 cases with hepatosplenomegalic stage of the disease [Mix2], 4 bilharzial subjects presented the hepatosplenomegaly without ascites [Mix3], 4 patients with hepatosplenomegaly and ascites [Mix4]. The results of this study were given in detail

Evaluaciòn de un método modificado para la determinación de fibra dietética en alimentos / Assessment of a modified method for determination of dietary fiber in food

Se evaluaron 2 modificaciones del método enzimático de Hellendoorn para la determinación de fibra dietética en alimentos, el cual simula las condiciones en que el alimento es ingerido por el hombre. Estas modificaciones permiten gelatinizar el almidòn de los alimentos y logran una mejor acción de las enzimas gastrointestinales empleadas. Se comprobó efectividad al realizar la prueba del yodo para revelar la presencia de almidón. Los resultados de fibra dietética se informan como residuo indigestible