A ISO 9000 e outras exigências da maior potência econômica do mundo

Exigências de certificação, normas obrigatórias, selo de conformidade da Comunidade Européia e suas novas regulamentações restritivas sobre sistemas de gerenciamento da qualidade e ambiental.

Detection of parvovirus B19 infection in formalin-fixed and paraffin-embedded placenta and fetal tissues

Parvovirus B19 infection was first discovered in 1975 and it is implicated in fetal death from hydrops fetalis the world over. Diagnosis is usually made through histological identification of the intranuclear inclusion in placenta and fetal organs. However, these cells may be scarce or uncharacteristic, making definitive diagnosis difficult. We analyzed histologically placentas and fetal organs from 34 cases of non-immune hydrops fetalis, stained with Hematoxylin and Eosin (HE) and submitted to immunohistochemistry and polymerase chain reaction (PCR). Of 34 tissue samples, two (5.9%) presented typical intranuclear inclusion in circulating normoblasts seen in Hematoxylin and Eosin stained sections, confirmed by immunohistochemistry and PCR. However, PCR of fetal organs was negative in one case in which the placenta PCR was positive. We concluded that parvovirus B19 infection frequency is similar to the literature and that immunohistochemistry was the best detection method. It is highly specific and sensitive, preserves the morphology and reveals a larger number of positive cells than does HE with the advantage of showing cytoplasmic and nuclear positivity, making it more reliable. Although PCR is more specific and sensitive in fresh or ideally fixed material it is not so in formalin-fixed paraffin-embedded tissues, frequently the only one available in such cases.

Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples

Development and standardization of reliable methods for detection of Mycobacterium tuberculosis in clinical samples is an important goal in laboratories throughout the world. In this work, lung and spleen fragments from a patient who died with the diagnosis of miliary tuberculosis were used to evaluate the influence of the type of fixative as well as the fixation and paraffin inclusion protocols on PCR performance in paraffin embedded specimens. Tissue fragments were fixed for four h to 48 h, using either 10% non-buffered or 10% buffered formalin, and embedded in pure paraffin or paraffin mixed with bee wax. Specimens were submitted to PCR for amplification of the human beta-actin gene and separately for amplification of the insertion sequence IS6110, specific from the M. tuberculosis complex. Amplification of the beta-actin gene was positive in all samples. No amplicons were generated by PCR-IS6110 when lung tissue fragments were fixed using 10% non-buffered formalin and were embedded in paraffin containing bee wax. In conclusion, combined inhibitory factors interfere in the detection of M. tuberculosis in stored material. It is important to control these inhibitory factors in order to implement molecular diagnosis in pathology laboratories.

HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES

This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.

Detection of Hepatitis B Virus Antigens in Paraffin-embedded Liver Specimens from the Amazon Region, Brazil

Hepatic viscerotomy of paraffin-preserved old specimens, collected in the period from 1934 to 1967, were analyzed by immunohistochemical assays to detect hepatitis B, hepatitis D, dengue and yellow fever virus antigens. The material belongs to the Yellow Fever Collection, Department of Pathology, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil and the cases were diagnosed at that time according to clinical aspects and histopathological findings reporting viral hepatitis, yellow fever, focal necrosis and hepatic atrophy. From the 79 specimens, 69 were collected at the Labrea Region and the other 10 in different other localities in the Amazon Region. The five micra thick histological slices were analyzed for the presence of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) by immunoperoxidase technique. An immunofluorescence assay was applied to the detection of hepatitis D, yellow fever and dengue virus antigens. Nine (11.4%) histological samples were HBsAg reactive and 5 (6.3%) were HBcAg reactive. The oldest reactive sample was from 1934. Viral antigens related to the other pathologies were not detected in this study. Our results confirm that the methodology described may be used to elucidate the aetiology of hepatitis diseases even after a long time of conservation of the specimens.

Detection of Leishmania braziliensis in human paraffin-embedded tissues from Tucumán, Argentina by polymerase chain reaction

American cutaneous leishmaniasis (ACL) is an endemic disease in Northern Argentina. We applied the polymerase chain reaction (PCR) followed by a hybridization labelled probe to 21 paraffin embedded human skin biopsies, already analyzed histologically, from leishmaniasis endemic areas in the province of Tucumán, Argentina. We used primers previously designed to detect a Leishmania-specific 120-base-pair fragment of kinetoplast DNA minicircle, other two primer pairs that amplify kDNA minicircles belonging to the L. braziliensis and L. mexicana complexes respectively, and specific oligonucleotide primers to detect L. (V.) braziliensis which amplify the sequence of the ribosomal protein L-14 of this species. The PCR-hybridization showed a sensitivity of 90.5% when compared to the histopathology test which was 61.9%. Five of the total samples analyzed were positive for the L. braziliensis complex whilst none was positive for the L. mexicana complex. The specific primers for L. (V.) braziliensis detected the parasite in four samples. These results are consistent with those reported for close endemic areas and demonstrate that the causative agent of human leishmaniasis in the analyzed cases was L. (V.) braziliensis. PCR should be used as a diagnostic tool for tegumentary leishmaniasis, especially in the mucosal form, and as a valuable technique for the identification of the Leishmania species that causes the disease in certain areas.

Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues

In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.

Las revistas científico-técnicas españolas de ciencias de la actividad física y el deporte: adecuación a las normas ISO y grado de normalización

Este artículo tiene por objeto conocer el grado de normalización y la adecuación de las revistas científico-técnicas españolas del campo de las Ciencias de la Actividad Física y el Deporte a diversos apartados clave de las normas ISO 8-1977 e ISO 215-1986. A partir de la observación directa de los fascículos, se han confeccionado dos índices para calcular el Grado General de Normalización (GGN) y el Grado Fundamental de Normalización (GFN). Los resultados indican que el ajuste de las publicaciones a la norma ISO 8-1977 es equivalente a la de otros campos españoles como el de Economía y el de Tecnología, mientras que es menor en la norma ISO 215-1986. La mayoría de las revistas presentan un GFN inferior a su respectivo GGN. Como principal conclusión se señala la existencia de un margen considerable de mejora en las revistas estudiadas, especialmente en los aspectos fundamentales de las normas.

O enfoque por processos da NBR ISO 9001 e sua aplicação nos serviços de informação

Este artigo tem como objetivo principal apresentar e discutir os fundamentos da gestão da qualidade e da nova versão da NBR ISO 9001, incluindo a aplicação do enfoque por processos nos serviços de informação, com o objetivo de apoiar a melhoria dos processos implantados nestes serviços e principalmente apoiar seu gerenciamento, visando à otimização de recursos e à elevação dos níveis de satisfação dos clientes, itens considerados fundamentais pelas organizações que optam por programas relacionados à gestão da qualidade.

Implantação da Norma ISO 9001: 2000 na Biblioteca Ministro Victor Nunes Leal do Supremo Tribunal Federal

Relato da experiência de implantação da Norma ISO 9001:2000 - sistemas de gestão da qualidade - requisitos na Biblioteca Ministro Victor Nunes Leal do Supremo Tribunal Federal, ocorrida em 2003. O processo selecionado foi o de atendimento a necessidades de informação de usuários internos e externos referentes às pesquisas de doutrina jurídica e de legislação. Destaca-se a importância da gestão da qualidade em unidades de informação. São descritas as etapas de implantação da Norma ISO 9001:2000, os requisitos de documentação de procedimentos, as alterações de rotinas e a padronização do trabalho, e os resultados obtidos são apresentados.

Comparison of USEPA 3050B and ISO 14869-1: 2001 digestion methods for sediment analysis by using FAAS and ICP-OES quantification techniques

A study of the partial USEPA 3050B and total ISO 14869-1:2001 digestion methods of sediments was performed. USEPA 3050B was recommended as the simpler method with less operational risk. However, the extraction ability of the method should be taken in account for the best environmental interpretation of the results. FAAS was used to quantify metal concentrations in sediment solutions. The alternative use of ICP-OES quantification should be conditioned by a previous detailed investigation and eventual correction of the matrix effect. For the first time, the EID method was employed for the detection and correction of the matrix effect in sediment ICP-OES analysis. Finally, some considerations were made about the level of metal contamination in the area under study.

Avaliação da resistência à corrosão de implantes em aços inoxidáveis ISO 5832-9 e F138 removidos de pacientes

This work has compared the surfaces of two different steel samples used as orthopedical implants, classified as ASTM F138 and ISO5832-9, through optical emission spectroscopy, by means of SEM and EDS. The samples (implants) were also submitted to potentiodynamic cyclic polarization in Ringer lactate and NaCl 0.9 M L-1 solutions; ISO5832-9 sample did not show any kind of localized corrosion, but in the case of F138 steel was observed a pit localized corrosion in both solutions. In Ringer lactate solution it was observed a loss of about 63% for nickel and 26% for iron for F138 stell, compared to the initial composition.

Quantificação dos isômeros ácido L-ascórbico e ácido D-iso-ascórbico em geleias de frutas por cromatografia líquida de alta eficiência

Ascorbic acid has important nutritional characteristics such as high antioxidant potential, preventing diverse damage and diseases in the tissues and the process of aging. Different isomeric forms of the ascorbic acid can be found in nature and each one have different potential antioxidant and different activity pro-vitamin C. This work examined a method to detect and quantify the isomers L-ascorbic acid (LAA) and D-iso-ascorbic acid (DIAA) in jelly fruit. The method showed acceptable selectivity, linearity, repeatability and recovery. DIAA was not found in the analyzed samples, but LAA was found up to 605 mg in 100 g of sample.