The effect of probiotic use on ABO antibody titers.

The use of probiotics brings numerous benefits to the immune system, including an increase in antibody production. The development of ABO antibodies may occur naturally due to the bacteria of the intestinal microbiota. However, high titers of ABO antibodies can lead to hemolytic disease of the fetus and newborn and can cause immune transfusion reactions. In this context, this study aimed to evaluate the effect of probiotic consumption on ABO antibody titers in humans. ABO blood group, ABO antibody titer, and fecal pH and Bifidobacteria concentration were determined for 126 healthy individuals before and after daily consumption of yogurt containing Lactobacillus acidophilus and Bifidobacterium lactis over a 1-month period. No changes in fecal pH were observed after probiotic consumption, regardless of ABO blood group. There was, however, an increase in the fecal concentration of Bifidobacteria in individuals with blood group A but not for those with group B or O. A decrease in the titer of anti-B was observed, despite the increase in the concentration of Bifidobacteria in feces being unrelated to fecal pH, in blood group A individuals. Our study, therefore, sought to understand the relationship between probiotics and the antibody titer of the ABO blood system. Despite our findings, further human studies are needed with other probiotic strains and molecular analyses of the intestinal microbiota.

How to Really Understand and Improve the System of Internal Quality Control and External Quality Assessment in the Accreditation Process of the Medical Laboratory?

Internal quality control (IQC) regarding process to monitor analytical stability has a long tradition in laboratory medicine. The satisfactory results with different quality specifications of the IQC ensure the acceptability of the examination results. Although the statistical IQC is satisfactory some problems exist, resulting in unreliable patients' results due several reasons (non-commutable control materials, lot to lot difference of reagents, false interpreting test results regarding autovalidation or autoverification, different analytical and clinical specifications or goals etc.). Therefore, the results and findings of IQC have to be connected with external quality assessment (EQA) in order to provide the system of measurement of uncertainty (MU) with correct interpretation of laboratory result and detection relevant and significant shifts and drifts in medical laboratory.

Acurácia diagnóstica dos métodos sorológicos de detecção da COVID-19 / Diagnostic accuracy of serological methods for detecting COVID-19

A rápida caracterização do genoma do Coronavírus-2 (SARS-CoV-2) mobilizou a produção em larga escala de métodos diagnósticos. Agências reguladoras aprovaram condicionalmente o uso emergencial de vários deles. Na emergência de saúde, inúmeros exames foram utilizados sem o devido conhecimento da qualidade. O objetivo desta revisão narrativa foi destacar a acurácia diagnóstica dos métodos sorológicos de diagnóstico da COVID-19 em termos de sensibilidade e especificidade clínicas. A sensibilidade diagnóstica variou até 50% entre estudos, enquanto a especificidade apresentou menor variação; no entanto, uma mesma marca chegou a apresentar variação de 32%. Considerando-se o critério de especificidade >95% definido pelo FDA, apenas uma marca de ensaio para detecção de IgG e outra para IgM atingiram a meta. Para a detecção de IgA e de IgA+IgM, as únicas marcas apresentadas não atingiram a meta. Entre as cinco marcas para detecção de IgM+IgG, apenas uma não atingiu a especificidade clínica. Para Ig total, as duas marcas apresentaram especificidade aceitável. Considerando-se o critério de sensibilidade clínica >90%, apenas 6 dos 25 estudos com diferentes marcas de ensaios para detecção de IgG apresentam o desempenho especificado; destas, nenhuma é brasileira. Nenhuma das marcas de ensaios para detecção de IgM e IgM+IgG apresentaram o desempenho especificado. A única marca apresentada para detecção de IgA+IgM atingiu a meta especificada pelo FDA. Os ensaios imunocromatográficos apresentaram menor sensibilidade. Os resultados demonstraram o pobre valor diagnóstico dos imunoensaios, mas com potencial para estudos soroepidemiológicos. Mais estudos de validação analítica e acurácia diagnóstica de ensaios são essenciais, principalmente dos reagentes nacionais.

The rapid characterization of the coronavirus-2 (SARS-CoV-2) genome has mobilized the large-scale production of diagnostic methods. Regulatory agencies have conditionally approved the emergency use of several of them. In the health emergency, numerous tests were used without proper knowledge of quality. The aim of this narrative review was to highlight the diagnostic accuracy of serological methods for the diagnosis of COVID-19 in terms of clinical sensitivity and specificity. The diagnostic sensitivity varied up to 50% between studies, while the specificity showed less variation, however, the same brand even showed a variation of 32%. Considering the >95% specificity criteria defined by the FDA, only one test brand for detection of IgG and one for IgM reached the goal. For the detection of IgA and IgA+IgM, the only brands presented did not reach the goal. Among the 5 marks for detecting IgM+IgG, only one did not reach clinical specificity. For total Ig, both brands had acceptable specificity. Considering the criterion of clinical sensitivity >90%, only 6 of the 25 studies with different brands of assays for detection of IgG present the specified performance, none of which are Brazilian. None of the brands of assays for detecting IgM and IgM+IgG showed the specified performance. The only brand presented for detection of IgA+IgM reached the target specified by the FDA. Immunochromatographic assays showed lower sensitivity. The results demonstrated the poor diagnostic value of immunoassays, but with potential for seroepidemiological studies. More studies on analytical validation and diagnostic accuracy of assays are essential, especially for national reagents.

Biossegurança laboratorial na pandemia do SARS-CoV-2 / Laboratory biosafety in the SARS-CoV-2 pandemic

Devido à emergência do SARS-CoV-2, os laboratórios necessitaram se adequar na mes-ma velocidade em que a pandemia se instalou para atender com segurança à crescentedemanda pelos testes diagnósticos. Com o alto potencial de disseminação do vírus, o contato com pacientes e o manuseio laboratorial das amostras tornou-se um desafio sem precedentes para os laboratórios. A necessidade de práticas de biossegurança nunca foi globalmente tão enfatizada como nas circunstâncias atuais da pandemia. O objetivo desta revisão narrativa foi destacar medidas para prevenção da contaminação pelo SARS-CoV-2 nos laboratórios clínicos, utilizando como referência a literatura publicada em livros, artigos científicos, orientações técnicas de autoridades sanitárias e científicas, e na análise crítica e pessoal da autora. Alguns temas abordados foram: compreensão dos riscos, medidas de biossegurança, níveis de biossegurança, barreiras de contenção, uso correto dos equipamentos de proteção individual (EPI), desinfecção das áreas de laboratório, descarte seguro de resíduos, e biossegurança nas fases pré-analítica e analítica. As orientações são baseadas em evidências limitadas e frequentemente fracas, oriundas de opiniões, estudos observacionais ou extrapolações de epidemias anteriores causadas por outros coronavírus. As boas práticas de biossegurança destacadas foram estabelecidas muito antes do surgimento da COVID-19. No entanto, a pandemia trouxe à tona, aos profissionais de laboratório e à população em geral, boas práticas que estavam esquecidas, como a higienização das mãos, a etiqueta respiratória e a forma correta de paramentação e desparamentação dos EPI. Na pandemia, os laboratórios com poucos recursos necessitaram adaptar soluções seguras e econômicas para garantir a segurança laboral.

Due to the emergence of SARS-CoV-2 the laboratories had to adapt, as quickly as the pandemic was installed, to safely meet the growing demand for the diagnostic tests. The high potential for virus spread, contact with patients and laboratory handling of samples has become an unprecedented challenge for laboratories. The need for biosafety practices has never been more globally emphasized as in the current circumstances of the pandemic. The purpose of this narrative review was to highlight strategies to prevent contamination by SARS-CoV-2 in clinical laboratories, using as reference the literature published in books, scientific articles, technical guides from health and scientific authorities, and the critical and personal analysis of the author. Some topics that were covered: understanding risks, biosafety strategies, biosafety levels, containment barriers, correct use of personal protective equipment (PPE), disinfection of laboratory areas, safe disposal of waste, and biosafety in the pre-analytical and analytical phases.The orientations are based on limited and often weak evidence arising from opinions, observational studies or extrapolations from the previous epidemics coronaviruses. The highlighted good practices on biosafety were established long before the emergence of COVID-19. However, the pandemic brought up to the laboratory professionals and population in general, good practices that had been forgotten, such as hands hygiene, respiratory etiquette and the correct way of donning and doffing PPE. In the pandemic, laboratories with limited resources had to adapt safe and economical solutions to ensure safety in the clinical laboratory.

Diagnóstico laboratorial do SARS-CoV-2 por transcrição reversa seguida de reação em cadeia da polimerase em tempo real (RT-PCR) / Laboratory diagnosis of SARS-CoV-2 by real time reverse transcriptase-polymerase chain reaction (RT-PCR)

O diagnóstico da COVID-19 está alicerçado na clínica do paciente, nos exames de imagem e no diagnóstico laboratorial. O exame de detecção do ácido nucleico viral por transcrição reversa (RT) seguido da reação em cadeia da polimerase em tempo real (PCR) foi rapidamente o primeiro método de diagnóstico laboratorial estabelecido e permanece como o padrão ouro. Esta narrativa descritiva é resultado de uma busca referenciada onde o ponto focal foi descrever o diagnóstico laboratorial do SARS-CoV-2 por RT-PCR. O diagnóstico laboratorial do SARS-CoV-2 por RT-PCR envolve as etapas de extração do RNA, transcrição reversa para obtenção do DNA complementar e a reação em cadeia da polimerase. A detecção da amplificação do material genético é realizada pela medida de fluorescência emitida. Entre as várias amostras biológicas que podem ser utilizadas, aquela que tem apresentado mais praticidade e precisão é a de swab da nasofaringe. A coleta da amostra deve ser, idealmente, realizada até sete dias a partir do início dos sintomas. Quando o SARS-CoV-2 é detectado na RT-PCR, o diagnóstico de COVID-19 é confirmado. No entanto, um único resultado de SARS-CoV-2 não detectado em paciente sintomático não exclui o diagnóstico. O exame não tem apresentado reações cruzadas com outros patógenos respiratórios. Contudo, o exame é caro e demorado, e pode resultar em falso negativo devido ao momento inadequado da coleta da amostra, coleta e manuseio impróprio de amostras e material genético viral insuficiente no sítio de coleta. Lacunas diagnósticas ainda permanecem na triagem de assintomáticos e na detecção de vírus vivos na convalescença.

The diagnosis of COVID-19 is based on the patient's clinic, imaging tests and laboratory diagnosis. The detection of viral nucleic acid by reverse transcription (RT) followed by real-time polymerase chain reaction (PCR) was quickly the first established laboratory diagnosis method and remains the gold standard. This descriptive narrative is a result of a referenced search where the focal point was to describe the laboratory diagnosis of SARS-CoV-2 by RT-PCR. The laboratory diagnosis of SARS-CoV-2 by RT-PCR involves the RNA extraction, reverse transcription to obtain complementary DNA, and the polymerase chain reaction steps. The detection of genetic material amplification is carried out by measuring the emitted fluorescence. Among the various biological samples that can be used, the one that has shown the most practicality and precision is the nasopharyngeal swab. Sample collection should be performed, ideally, within 7 days from the symptoms onset. When SARS-CoV-2 is detected by RTPCR, the diagnosis of COVID-19 is confirmed. However, a single result of undetected SARS-CoV-2 in a symptomatic patient does not exclude the diagnosis. The test has not shown cross-reactions with common respiratory pathogens. However, the test is costly and timeconsuming, a false-negative result may arise due to inadequate sample collection time, improper samples collection and handling, and insufficient viral genetic material at the collection site. Diagnostic gaps remain on asymptomatic patients screening, and in the detection of live viruses in convalescence.

A relação entre o sistema sanguíneo ABO e a COVID-19: uma revisão sistemática / The relationship between the ABO blood system and COVID-19: a systematic review

A atribuição do sistema sanguíneo ABO às infecções não é recente e não é exclusiva de infecções virais. A relação entre a COVID-19 e o grupo sanguíneo ABO de pacientes infectados tem sido investigada. O objetivo desta revisão foi avaliar se a associação do sistema sanguíneo ABO com SARS-CoV-2 envolve a isomeria ABO tecidual e subgrupos sanguíneos. Realizou-se uma revisão sistemática com busca de artigos e publicações até 28 de julho de 2020 na base PubMed. Foram obtidos 311 manuscritos dos quais 15 atenderam os critérios de inclusão e foram incluídos no estudo. Quarenta por cento dos estudos discutiram a possibilidade dos antígenos teciduais ABO influenciar na transmissão ou gravidade da COVID-19. Nenhum manuscrito mencionou que a isomeria antigênica ABO tecidual poderia predispor indivíduos às infecções por SARS-CoV-2 ou agravamento da COVID-19. Um manuscrito discutiu a possibilidade de o impedimento estérico afetar a saturação do receptor de diferentes isótipos de anticorpos anti-A1IgG em pacientes que desenvolveram COVID-19. Se rejeitássemos cartas e/ou comentários, análises profundas sobre o aspecto do sistema sanguíneo ABO e infecções por SARS-CoV-2 teriam sido excluídos e prejudicariam a discussão científica e as conclusões da revisão.

The assignment of the ABO blood system to infection is not new and is not exclusive to viral infections. The relationship between COVID-19 and the ABO blood group of infected patients has been investigated. The purpose of this review was to assess whether the association between ABO blood system and SARS-CoV-2 involves tissue ABO isomerism and blood subgroups. A systematic review was carried out with search until July 28, 2020 in PubMed database. Three hundred and eleven manuscripts were obtained, of which 15 met the inclusion criteria and were included in the study. Forty percent of the studies discussed the possibility of ABO tissue antigens influence the transmission or severity of COVID-19. No manuscript mentioned that ABO tissue antigenic isomerism could predispose individuals to SARSCoV-2 infections or severe COVID-19. A manuscript discussed the possibility of steric impediment affect receptorsaturation of the different antibodies A1IgG isotypes in COVID-19 patients. If the letters, correspondences or comments were rejected, deep analyzes of ABO blood system and SARS-CoV-2 infections relationship would have been excluded, and would undermine the scientific discussion and conclusions of the review.

The top-down approach to measurement uncertainty: which formula should we use in laboratory medicine?

INTRODUCTION: By quantifying the measurement uncertainty (MU), both the laboratory and the physician can have an objective estimate of the results' quality. There is significant flexibility on how to determine the MU in laboratory medicine and different approaches have been proposed by Nordtest, Eurolab and Cofrac to obtain the data and apply them in formulas. The purpose of this study is to compare three different top-down approaches for the estimation of the MU and to suggest which of these approaches could be the most suitable choice for routine use in clinical laboratories. MATERIALS AND METHODS: Imprecision and bias of the methods were considered as components of the MU. The bias was obtained from certified reference calibrators (CRC), proficiency tests (PT), and inter-laboratory internal quality control scheme (IQCS) programs. The bias uncertainty, the combined and the expanded uncertainty were estimated using the Nordtest, Eurolab and Cofrac approaches. RESULTS: Using different approaches, the expanded uncertainty estimates ranged from 18.9-40.4%, 18.2-22.8%, 9.3-20.9%, and 7.1-18.6% for cancer antigen (CA) 19-9, testosterone, alkaline phosphatase (ALP), and creatinine, respectively. Permissible values for MU and total error ranged from 16.0-46.1%, 13.1-21.6%, 10.7-26.2%, and 7.5-17.3%, respectively. CONCLUSION: The bias was highest using PT, followed by CRC and IQCS data, which were similar. The Cofrac approach showed the highest uncertainties, followed by Eurolab and Nordtest. However, the Eurolab approach requires additional measurements to obtain uncertainty data. In summary, the Nordtest approach using IQCS data was therefore found to be the most practical formula.

Evaluation of the clinicai analysis service provided to an emergency department / Avaliação do serviço de análises clínicas prestado a uma unidade de emergência

ABSTRACT Introduction: Time for releasing test results and critical value communication by the clinical laboratory are considered important determinants of patients' length of stay in the Emergency Department (ED). As well as physician satisfaction, they are used as quality indicators of the Clinical Analysis Service (CAS). Objective: The aim of this study was to evaluate the time to return a test result, the communication of critical values and the level of satisfaction of the ED physicians of a university hospital with the CAS. Material and methods: A physician satisfaction survey was conducted using a questionnaire. The test turnaround time for emergency requests was analyzed by observation of the pre-analytical phase and monitoring the analytical and post-analytical phases through the laboratory information system. In order to evaluate the communication of critical values by the CAS, a document analysis of the process was performed. Results: Physicians' overall satisfaction with the CAS was considered average. Greater satisfaction was observed with reliability of the results and the staff courtesy and lesser with the delivery time of results. The test turnaround time exceeded clinicians' expectations and the average described in the literature. No experience with critical-value communication was perceived. Conclusion: The results show several opportunities for progress in the relationship between ED and CAS. The improvement of laboratory quality indicators also depends on the involvement of ED physicians and nurses and embraces monitoring, planning, education and investment in system computerization.

RESUMEN Introducción: El tiempo de entrega de pruebas y la comunicación de valores críticos por el laboratorio clínico son considerados factores determinantes del tiempo de permanencia del paciente en la Sección de Urgencias (SU). Por lo tanto, así como la satisfacción de los médicos, estos datos también son utilizados como indicadores de calidad del Servicio de Análisis Clínicos (SAC). Objetivo: Evaluar el tiempo de entrega de pruebas, la comunicación de valores críticos y el grado de satisfacción de los médicos de la SU de un hospital universitario con el SAC. Material y método: Se realizó una encuesta de satisfacción de los médicos, utilizando un cuestionario. El tiempo de reporte de resultados en la SU ha sido analizado por medio de observación presencial de la fase preanalítica y monitoreo en el sistema computorizado de las fases analítica y postanalítica del laboratorio. Para evaluar la comunicación de valores críticos por el SAC, un análisis documental del proceso ha sido realizado. Resultados: La satisfacción general con el SAC fue considerada regular por los médicos participantes; hubo mayor satisfacción con la confianza en los resultados y con la atención del personal, y menor con el tiempo de entrega de resultados. Los tiempos de reporte han sido más largos que las expectativas de los médicosy la media descripta en la literatura. No se ha observado una cultura de comunicación de valores críticos. Conclusión: Los resultados han ensenado varias oportunidades deprogreso en la relación entre SUy SAC. La mejoría de los indicadores de calidad de laboratorio evaluados depende también del envolvimiento de los médicos y enfermeros del SU e incluye monitoreo, planeamiento, educación e investimento en informatización del sistema.

RESUMO Introdução: O tempo de liberação de exames e a comunicação de valores críticos pelo laboratório clínico são considerados importantes determinantes do tempo de permanência do paciente no Serviço de Emergência (SE). Portanto, assim como a satisfação dos médicos, esses dados também são utilizados como indicadores de qualidade do Serviço de Análises Clínicas (SACL). Objetivo: Avaliar o tempo de liberação de exames, a comunicação de valores críticos e o nível de satisfação dos médicos do SE de um hospital universitário em relação ao SACL. Material e métodos: Foi realizada uma pesquisa de satisfação dos médicos, utilizando um questionário. O tempo de liberação dos resultados de exames de emergência foi analisado por meio de observação presencial da fase pré-analítica e monitoramento no sistema informatizado das fases analítica e pós-analítica do laboratório. Para avaliar a comunicação de valores críticos pelo SACL, foi realizada análise documental do processo. Resultados: A satisfação geral com o SACL foi considerada regular pelos médicos participantes, com maior satisfação quanto à confiança nos resultados e à cortesia dos servidores e menor quanto ao tempo de entrega de resultados. Os tempos de liberação de resultados dos exames ultrapassaram as expectativas dos médicos e a média descrita na literatura. Não foi observada cultura de comunicação de valores críticos. Conclusão: Os resultados demonstraram várias oportunidades de progresso na relação entre o SE e o SACL. A melhoria dos indicadores da qualidade laboratorial avaliados depende também do envolvimento dos médicos e dos enfermeiros do SE e inclui monitoramento, planejamento, educação e investimento em informatização do sistema.

Analytical validation of an in-house method for adenosine deaminase determination.

BACKGROUND: The adenosine deaminase (ADA) enzyme is a marker of inflammatory processes whose activity can be measured through a colorimetric method developed as an in-house assay. This validation can reduce costs and expand the alternatives for laboratory diagnosis. METHODS: The ADA analysis was achieved through a modified form of Giusti and Galanti's (1984) method. The following parameters were characterized: calibration curve, linearity, analytical sensitivity, limit of detection, limit of quantification, method working range, precision (within-assay and between-assay), bias, total analytical error, and sample stability. The results were statistically evaluated and compared with quality specifications based on biological variations and the performance of commercial tests. RESULTS: The analytical sensitivity and limit of detection (0.013 and 3.0 U/L, respectively) were lower than those of commercial tests. The method's working range was 3.2-100.0 U/L. According to the quality specification, the method showed optimum performance with a bias <3.5%. However, repeatability (2.2% and 1.7% for normal- and high-activity samples, respectively) and reproducibility achieved worse results when compared to commercial tests. The method demonstrated an inappropriate between-assay precision for low enzymatic activity (10.4%) and the minimum and desirable performance for medium (8.8%) and high (5.0%) activities, respectively. It also presented at least a minimum performance (<25%) for the total analytical error of the three analyzed samples. The pleural fluid samples were found to be stable at -20°C for six days. CONCLUSION: The findings show that the in-house method displays an acceptable performance and is capable of generating results comparable to existing commercial tests.

Dez anos da RDC 302/2005: avaliação da implantação em laboratórios de análises clínicas do estado de Santa Catarina / Ten years of RDC 302/2005: evaluation of implantation in laboratories of clinical analysis of Santa Catarina state

Objetivo: Avaliar dez anos após a publicação se os laboratórios de análises clínicas do estado de Santa Catarina atendem aos requisitos da RDC 302/2005. A população de laboratórios foi obtida no CNES. Métodos: Como ferramenta de pesquisa foi elaborado um questionário baseado na RDC 302/2005 e enviado aos laboratórios. Resultados: Dos 523 laboratórios clínicos aptos a participar do estudo, o contato foi exitoso com apenas 198 e, desses, somente 20 participaram. Os participantes relataram que a RDC 302/2005 é essencial para o funcionamento dos laboratórios. O tempo para iniciar a implantação da RDC levou, em média, um ano, e o tempo para a conclusão delongou mais um ano. Doze laboratórios relataram cumprir todos os requisitos da resolução. No entanto, apenas dois laboratórios cumprem integralmente os 38 requisitos da RDC pesquisados. Os requisitos de rotulagem de reagentes, registro de temperatura de equipamentos, gerenciamento de resíduos, comunicação de valores críticos, arquivamento de laudos e dados brutos e notificação de resultados que indiquem doença notificável foram cumpridos por todos os laboratórios. O registro da temperatura de transporte das amostras foi o requisito menos cumprido pelos laboratórios. As principais dificuldades relatadas para implantação dos requisitos foram nas áreas de gestão e garantia de qualidade, aplicação dos conhecimentos de biossegurança e resistência dos colaboradores ao processo de mudança. Conclusão: A formação dos profissionais voltada para conceitos e ferramentas de gestão, de sistema de qualidade e padronização de processos se faz necessária para os laboratórios e pode ser o início da solução para as dificuldades apresentadas pelos laboratórios pesquisados.

Assessment of the pre-analytical phase of a clinical analyses laboratory / Avaliação da fase pré-analítica de um laboratório de análises clínicas

ABSTRACT Introduction: The pre-analytical phase encompasses the procedures prior to the performance of the laboratory tests, which are the physician's requisition, patient and tests registration, collection and transport of samples. This phase involves different professionals and is responsible for the majority of the laboratory errors. Objective: To assess the pre-analytical phase of a public laboratory through quality indicators (QI). Method: Nonconformities (NC) in the physician's requisitions and in the patients and tests registrations were evaluated during the quality checking process. A questionnaire was applied to evaluate the service and the satisfaction of the laboratory's client. The QI were calculated considering the number of NC in the processes in relation to the total opportunities, compared with specifications described in the literature and evaluated by the Sigma metric. Results: The pre-analytical phase was evaluated by 34 QI. From these, 18 presented Sigma score lower than 3.0, six between 3.0 and 4.0, six above 4.0 and four presented Sigma score of 6.0. The completiton of requisitions presented worse performance than the process of tests registration. Regarding the three stages of the pre-analytical phase evaluated, the scheduling presented the worst performance, followed by the service at the reception and the sample collection. Conclusion: The evaluation of the QI allowed organizing the improvements that should be prioritized in the laboratory. The Sigma metric was useful for assessing the QIs considered important to the laboratory, for which there are no published specifications. The questionnaire for evaluating the laboratory's service and client's satisfaction was not a reliable tool for assessing the quality of the pre-analytical phase.

RESUMO Introdução: A fase pré-analítica abrange os procedimentos antes da execução dos testes laboratoriais, sendo eles a requisição, o cadastro de clientes e exames, a coleta e o transporte de amostras. Essa fase envolve diferentes profissionais e é responsável pela maioria dos erros de laboratório. Objetivo: Analisar a fase pré-analítica de um laboratório público por meio de indicadores da qualidade (IQ). Método: As não conformidades (NC) nas requisições e no cadastro dos clientes e dos exames foram avaliadas durante o processo de checagem. Foi aplicado um questionário para avaliar o atendimento e a satisfação dos clientes do laboratório. Os IQ foram calculados considerando o número de NC nos processos em relação ao total de oportunidades, comparando-os com as especificações encontradas na literatura e avaliando-os pela métrica Sigma. Resultados: A fase pré-analítica foi avaliada por 34 IQ. Desses, 18 apresentaram Sigma menor que 3,0; seis, entre 3,0 e 4,0; seis, acima de 4,0; e quatro, Sigma 6,0. O preenchimento das requisições apresentou pior desempenho do que o processo de cadastro dos exames. Em relação às três etapas da fase pré-analítica avaliadas, a que apresentou pior desempenho foi o agendamento, seguido do atendimento na recepção e da coleta. Conclusão: A avaliação dos IQ permitiu ordenar as melhorias que devem ser priorizadas no laboratório. A métrica Sigma foi útil para avaliar IQ, considerados importantes para o laboratório, para os quais não há especificações publicadas. O questionário de avaliação do atendimento e da satisfação do cliente não se mostrou uma ferramenta confiável para avaliar a qualidade da fase pré-analítica.

Chemopreventive effects of a Tamarindus indica fruit extract against colon carcinogenesis depends on the dietary cholesterol levels in hamsters.

Tamarind has significant antioxidant potential. We showed that tamarind protects hypercholesterolemic hamsters from atherosclerosis. Hypercholesterolemia might increase the risk of colon cancer. We investigated whether tamarind extract modulates the risk of colon cancer in hypercholesterolemic hamsters. Hamsters (n = 64) were given tamarind and a hypercholesterolemic diet for 8 weeks. The groups were the control, tamarind treatment, hypercholesterolemic, and hypercholesterolemic treated with tamarind groups. Half of each group was exposed to the carcinogen dimethylhydrazine (DMH) at the 8th week. All hamsters were euthanatized at the 10th week. In carcinogen-exposed hypercholesterolemic hamsters, tamarind did not alter the cholesterol or triglyceride serum levels, but it reduced biomarkers of liver damage (alanine transaminase [ALT], and aspartate aminotransferase [AST]). Tamarind decreased DNA damage in hepatocytes, as demonstrated by analysis with an anti-γH2A.X antibody. In liver and serum samples, we found that this fruit extract reduced lipid peroxidation (thiobarbituric acid reactive substances [TBARS]) and increased endogenous antioxidant mechanisms (glutathione peroxidase [GPx] and superoxide dismutase [SOD]). However, tamarind did not alter either lipid peroxidation or antioxidant defenses in the colon, which contrasts with DMH exposure. Moreover, tamarind significantly increased the stool content of cholesterol. Although tamarind reduced the risk of colon cancer in hypercholesterolemic hamsters that were carcinogenically exposed to DMH by 63.8% (Metallothionein), it was still ∼51% higher than for animals fed a regular diet. Staining colon samples with an anti-γH2A.X antibody confirmed these findings. We suggest that tamarind has chemoprotective activity against the development of colon carcinogenesis, although a hypercholesterolemic diet might impair this protection.

Effects of probiotic intake on intestinal bifidobacteria of celiac patients / Efeitos do consumo de probióticos sobre as bifidobactérias intestinais de pacientes celíacos

ABSTRACT BACKGROUND Healthy individuals exhibit a significantly higher concentration of faecal bifidobacteria in comparison to celiac patients. Even though there are potential benefits in probiotic usage, they have been little explored as an adjunctive therapy in celiac disease. OBJECTIVE This study aimed at the comparison of faecal bifidobacteria concentration and pH among celiac patients and healthy subjects before and after the daily intake of 100 g of yogurt containing probiotic for a thirty-day period. METHODS Feces from 17 healthy subjects and 14 celiac patients were analyzed, in which stool culture was performed for the isolation and quantification of faecal bifidobacteria. Furthermore, Gram’s method was employed for the microscopic analysis of the colonies, while the identification of the Bifidobacterium genus was made through determination of the fructose-6-phosphate phosphoketolase enzyme. Faecal pH was measured using a calibrated pHmeter. RESULTS Faecal bifidobacteria concentration before probiotic consumption was significantly higher in healthy individuals (2.3x108±6.3x107 CFU/g) when compared to celiac patients (1.0x107±1.7x107 CFU/g). Faecal pH values did not show a significant difference. After the daily consumption of probiotic-containing yogurt both groups showed a significant increase in the concentration of faecal bifidobacteria, but healthy subjects presented significantly higher bifidobacteria concentrations (14.7x108±0.2x108 CFU/g) than the celiac group (0.76x108±0.1x108 CFU/g). The obtained pH values from both groups were not significantly different, being 7.28±0.518 for the celiac patients and 7.07±0.570 for healthy individuals after the probiotic intake. CONCLUSION The probiotic supplementation significantly increased the number of bifidobacteria in the feces of celiac patients, although it was not sufficient to reach the concentration found in healthy individuals prior to its consumption.

RESUMO CONTEXTO Indivíduos saudáveis apresentam uma concentração de bifidobactérias fecais significativamente maior em comparação a pacientes celíacos. Apesar de haver benefícios potenciais no uso de probióticos na doença celíaca, estes têm sido pouco explorados como uma terapia adjuvante. OBJETIVO Este estudo objetivou a comparação do pH e concentração fecal de bifidobactérias entre pacientes celíacos e indivíduos saudáveis antes e após o consumo diário de 100 g de iogurte contendo probiótico por um período de 30 dias. MÉTODOS Foram analisadas fezes de 17 pessoas saudáveis e 14 pacientes celíacos, tendo sido realizada a coprocultura para o isolamento e quantificação de bifidobactérias fecais. Além disso, o método de Gram foi empregado na análise microscópica das colônias, enquanto a identificação do gênero Bifidobacterium foi feita através da determinação da enzima frutose-6-fosfato fosfocetolase. O pH fecal foi medido usando um pHmetro calibrado. RESULTADOS A concentração de bifidobactérias fecais antes do consumo do iogurte probiótico foi significativamente maior em indivíduos saudáveis (2.3x108±6.3x107 UFC/g) quando comparada aos celíacos (1.0x107±1.7x107 CFU/g). Por outro lado, o pH fecal de ambos os grupos não apresentou diferença significativa. Após o consumo diário de iogurte contendo probiótico, ambos os grupos tiveram um aumento significativo na concentração de bifidobactérias fecais, entretanto indivíduos saudáveis apresentaram concentrações de bifidobactérias significativamente maiores (14.7x108±0.2x108 UFC/g) do que o grupo celíaco (0.76x108±0.1x108 UFC/g). Os valores de pH obtidos de ambos os grupos não foram significativamente diferentes, sendo de 7.28±0.518 para os pacientes celíacos e de 7.07±0.570 para os indivíduos saudáveis após o consumo do probiótico. CONCLUSÃO A suplementação com probiótico aumentou significativamente o número de bifidobactérias nas fezes dos pacientes celíacos apesar de não ter sido suficiente para alcançar a concentração encontrada em indivíduos saudáveis antes do consumo de probióticos.

EFFECTS OF PROBIOTIC INTAKE ON INTESTINAL BIFIDOBACTERIA OF CELIAC PATIENTS.

BACKGROUND: Healthy individuals exhibit a significantly higher concentration of faecal bifidobacteria in comparison to celiac patients. Even though there are potential benefits in probiotic usage, they have been little explored as an adjunctive therapy in celiac disease. OBJECTIVE: This study aimed at the comparison of faecal bifidobacteria concentration and pH among celiac patients and healthy subjects before and after the daily intake of 100 g of yogurt containing probiotic for a thirty-day period. METHODS: Feces from 17 healthy subjects and 14 celiac patients were analyzed, in which stool culture was performed for the isolation and quantification of faecal bifidobacteria. Furthermore, Gram's method was employed for the microscopic analysis of the colonies, while the identification of the Bifidobacterium genus was made through determination of the fructose-6-phosphate phosphoketolase enzyme. Faecal pH was measured using a calibrated pHmeter. RESULTS: Faecal bifidobacteria concentration before probiotic consumption was significantly higher in healthy individuals (2.3x108±6.3x107 CFU/g) when compared to celiac patients (1.0x107±1.7x107 CFU/g). Faecal pH values did not show a significant difference. After the daily consumption of probiotic-containing yogurt both groups showed a significant increase in the concentration of faecal bifidobacteria, but healthy subjects presented significantly higher bifidobacteria concentrations (14.7x108±0.2x108 CFU/g) than the celiac group (0.76x108±0.1x108 CFU/g). The obtained pH values from both groups were not significantly different, being 7.28±0.518 for the celiac patients and 7.07±0.570 for healthy individuals after the probiotic intake. CONCLUSION: The probiotic supplementation significantly increased the number of bifidobacteria in the feces of celiac patients, although it was not sufficient to reach the concentration found in healthy individuals prior to its consumption.

Avaliação da Gestão da Qualidade de uma Agência Transfusional / Quality Management Assessment of a Transfusion Agency

Introdução: O controle da qualidade, quando implantado, deve assegurar o uso dos hemocomponentes livre de possíveis reações transfusionais. Inúmeras regulamentações na área de hemoterapia vêm direcionando as metodologias que devem ser utilizadas pelas agências transfusionais de forma a minimizar os riscos ao paciente/receptor. Objetivo: Este estudo teve como objetivo realizar a avaliação da gestão da qualidade de uma agência transfusional localizada no município de Balneário Camboriú-SC. Materiais e Métodos: Como metodologia central foi utilizado o Ciclo PDCA, do inglês Plan, Do, Check, Act, com o objetivo de planejar e organizar as ações do gerenciamento da qualidade, observar as melhorias e quando necessário auxiliar na atuação sobre as inconformidades encontradas. Resultados: As informações da rotina hemoterápica da instituição foram obtidas por meio de 15 visitas técnicas. Através dos Roteiros de Inspeção Técnica em Serviços de Hemoterapia foram detectadas conformidades em 80% dos requisitos, destacando-se as questões de biossegurança, área física e reagentes, enquanto as não-conformidades foram observadas principalmente no item Gestão da Qualidade. Neste sentido, foram utilizadas as Ferramentas da Qualidade para correções das não-conformidades. Após a execução de 11 ações de melhoria, o percentual de conformidades no item Gestão da Qualidade foi de 100%. Conclusão: Inconformidades foram encontradas na agência transfusional avaliada, entretanto, não foram observados riscos nos serviços de saúde fornecidos à população através dos itens avaliados. (AU)

Introduction: Quality control, when implemented, should ensure the use of blood components without the presence of transfusion reactions. Numerous regulations in hemotherapy area have driven the approaches used by transfusion agencies to minimize the risks for the patient/ receptor. Objective: To carry out the evaluation of the quality management of a transfusion agency located in the city of Camboriú, SC, Brazil. Material and Methods: The PDCA Cycle (Plan, Do, Check, Act) was used to plan and organize the quality management actions, to observe the improvements and, where necessary, to assist in the resolution of detected issues. Results: Information from the hemotherapic routine of the institution was obtained through 15 technical visits. Technical Inspection Checklists in the Hemotherapy Service indicated a compliance of 80% with the norms. related to biosafety , physical area and reagents, while most of the non-conformities were observed in the Quality Management sector. Thus, Quality Tools were used for correction of nonconformities. After the execution of 11 improvement actions, the compliance percentage of the Quality Management item was found to be 100%. Conclusion: Despite the nonconformities found in the transfusion agency under study, no significant risks in the health care provided to the population were observed. (AU)

Lower bifidobacteria counts in adult patients with celiac disease on a gluten-free diet.

CONTEXT: The ingestion of gluten is responsible for the symptoms of Celiac disease, but other environmental factors can also influence. Strains of the Bifidobacterium genus have been shown to afford protection against the inflammatory response and mucosal damage caused by gliadin peptides in vitro. OBJECTIVES: This study was designed to compare the concentration of fecal bifidobacteria and pH of patients with celiac disease on gluten-free diet and control subjects in order to identify if the imbalance on fecal microbiota still remain during the treatment of celiac disease and identify the necessity of dietary supplementation with pre- or probiotics. METHODS: It was analyzed the feces of 42 healthy subjects and 14 celiac patients. The bifidobacteria count in feces was done in selective medium BIM-25. Microscopic analysis of the colonies was performed by Gram stain. The identification of the genus Bifidobacterium was performed by determination of fructose-6-phosphate phosphoketolase. Fecal pH was measured using a pH meter. RESULTS: The concentration of bifidobacteria per gram of feces was significantly higher in healthy subjects (controls) (1.5 ± 0.63 x108 CFU/g) when compared to celiac patients (2.5 ± 1.5 x107 CFU/g). The fecal pH was not different between celiac patients (7.19 ± 0.521) and controls (7.18 ± 0.522). CONCLUSIONS: These results suggest that with lower levels of bifidobacteria, celiac patients have an imbalance in the intestinal microbiota, regardless of pH, even while on a gluten-free diet. This fact could favor the pathological process of the disorder.

Lower bifidobaceeria counts in adult patients with celiac disease on a gluten-free deet / Baixa contagem de bifidobactérias em pacientes adultos com doença celíaca, em dieta isenta de glúten

Context The ingestion of gluten is responsible for the symptoms of Celiac disease, but other environmental factors can also influence. Strains of the Bifidobacterium genus have been shown to afford protection against the inflammatory response and mucosal damage caused by gliadin peptides in vitro. Objectives This study was designed to compare the concentration of fecal bifidobacteria and pH of patients with celiac disease on gluten-free diet and control subjects in order to identify if the imbalance on fecal microbiota still remain during the treatment of celiac disease and identify the necessity of dietary supplementation with pre- or probiotics. Methods It was analyzed the feces of 42 healthy subjects and 14 celiac patients. The bifidobacteria count in feces was done in selective medium BIM-25. Microscopic analysis of the colonies was performed by Gram stain. The identification of the genus Bifidobacterium was performed by determination of fructose-6-phosphate phosphoketolase. Fecal pH was measured using a pH meter. Results The concentration of bifidobacteria per gram of feces was significantly higher in healthy subjects (controls) (1.5 ± 0.63 x108 CFU/g) when compared to celiac patients (2.5 ± 1.5 x107 CFU/g). The fecal pH was not different between celiac patients (7.19 ± 0.521) and controls (7.18 ± 0.522). Conclusions These results suggest that with lower levels of bifidobacteria, celiac patients have an imbalance in the intestinal microbiota, regardless of pH, even while on a gluten-free diet. This fact could favor the pathological process of the disorder. .

Contexto A ingestão do glúten é responsável pelos sintomas da doença celíaca, mas outros fatores ambientais também podem influenciar. Tem sido mostrado que as cepas do género Bifidobacterium proporcionam proteção contra a resposta inflamatória, lesão da mucosa causada por péptidos da gliadina in vitro. Objetivos Este estudo foi desenvolvido para comparar as concentrações de bifidobactérias e pH fecal de pacientes com doença celíaca em dieta isenta de glúten e indivíduos controles, a fim de identificar se o desequilíbrio na microbiota fecal ainda permanece durante o tratamento da doença celíaca e, identificar a necessidade de suplementação alimentar com pré ou probióticos. Métodos Foram analisadas as fezes de 42 indivíduos saudáveis e 14 pacientes com doença celíaca. A contagem de bifidobactérias nas fezes foi feita em meio seletivo BIM-25. A análise microscópica das colônias foi realizada por coloração de Gram. A identificação do género Bifidobacterium foi realizada por determinação de phosphoketolase frutose-6-fosfato. O pH fecal foi medido usando um medidor de pH. Resultados As concentrações de bifidobactérias por grama de fezes foi significativamente mais elevada nos indivíduos saudáveis (controles) (1,5 ± 0,63 x108 UFC/g), quando comparada com pacientes com doença celíaca (2,5 ± 1,5 x107 UFC/g). O pH fecal não foi diferente entre pacientes celíacos (7,19 ± 0,521) e controles (7,18 ± 0,522). Conclusões Estes resultados sugerem que, com concentrações inferiores de bifidobactérias, pacientes com doença celíaca tem um desequilíbrio na microbiota intestinal, independentemente do pH, mesmo durante uma dieta isenta de ...

The effect of dietary oils on the development of aberrant crypt foci, bifidobacteria and fecal pH.

The present study investigated the relationship between intestinal bifidobacteria and intake of commercial dietary oils of different fatty acids compositions on the development of aberrant crypt foci (ACF). Wistar rats were grouped according to diet and treatment with dimethylhydrazine (DMH): standard diet (CN), canola oil (CAN), olive oil (OLI), corn oil (COR), standard diet and DMH (CNDMH), canola oil and DMH (CANDMH), olive oil and DMH (OLIDMH) and corn oil and DMH (CORDMH). Diets and DMH did not alter the amount of bifidobacteria, fecal pH and serum total cholesterol level. DMH-treated groups had lower serum triglyceride levels compared to respective controls without DMH. Olive and corn oil diets resulted in higher hepatic cholesterol levels than standard diet under treatment with DMH. The numbers of ACF/field and cell proliferation were lower under treatment with CANDMH and OLIDMH, suggesting a protective effect of these oils on colorectal carcinogenesis.