Dissemination of bla(KPC-2) by the Spread of Klebsiella pneumoniae Clonal Complex 258 Clones (ST258, ST11, ST437) and Plasmids (IncFII, IncN, IncL/M) among Enterobacteriaceae Species in Brazil

This article reports the spread of bla(KPC-2) in the Sao Paulo and Rio de Janeiro states, facilitated by globally spread K. pneumoniae clonal complex 258 (CC258) clones (ST258, ST11, and ST437) and a diversity of plasmids (IncFII, IncN, and IncL/M, two untypeable plasmids carrying Tn4401a or Tn4401b) successfully disseminated among species of the Enterobacteriaceae (Enterobacter cloacae, Serratia marcescens, and Citrobacter freundii). It also constitutes the first description of sequence type 258 (ST258) in Brazil, which was associated with a nosocomial hospital outbreak in Ribeirao Preto city.

Mathematical modelling of supercritical CO2 extraction of volatile oils from aromatic plants

The modelling of the experimental data of the extraction of the volatile oil from six aromatic plants (coriander, fennel, savoury, winter savoury, cotton lavender and thyme) was performed using five mathematical models, based on differential mass balances. In all cases the extraction was internal diffusion controlled and the internal mass transfer coefficienty (k(s)) have been found to change with pressure, temperature and particle size. For fennel, savoury and cotton lavender, the external mass transfer and the equilibrium phase also influenced the second extraction period, since k(s) changed with the tested flow rates. In general, the axial dispersion coefficient could be neglected for the conditions studied, since Peclet numbers were high. On the other hand, the solute-matrix interaction had to be considered in order to ensure a satisfactory description of the experimental data.

Ensino da Matemática : as primeiras explorações da ordem das dezenas

O nosso sistema de numeração decimal é um sistema de natureza posicional: os números são representados por sequências de símbolos, sendo que o valor de cada símbolo depende da posição que ocupa nessa sequência. Por exemplo, quando escrevemos o numeral relativo ao número treze, “13”, estamos na realidade a utilizar uma numeração mista: “1” vale uma dezena e “3” vale três unidades. Treze, na sua escrita matemática atual, traduz a organização uma dezena mais três unidades; dez unidades de uma ordem numérica são alvo de uma composição para uma unidade da ordem numérica seguinte, o que traduz a essência de um sistema posicional de base 10. Por isso, o “10” desempenha um papel de extrema importância e a forma como as crianças desenvolvem as primeiras explorações do nosso sistema de numeração é determinante para as suas aprendizagens futuras. (...) Para estimular uma verdadeira compreensão da ordem das dezenas, as atividades típicas são: (a) Separa 10 e diz o número; (b) Pinta 10 e diz o número; (c) Utilização de dispositivos com algarismos móveis (presentes em todos os manuais do bem sucedido método de Singapura). Vejamos como podemos promover a compreensão da ordem das dezenas e ultrapassar com eficácia a “barreira” do 10. (...)

Transversalidade e continuidade em educação: um olhar sobre os instrumentos de registo e observação na educação pré-escolar e no 1º ciclo do ensino básico

Tomar consciência do papel do educador de infância e do professor do 1º CEB manifesta-se uma tarefa essencial à prática, permitindo uma orientação da intervenção educativa, no sentido de ir ao encontro do que é esperado por parte destes profissionais. O trabalho espelha as experiências nos contextos de 1º Ciclo do Ensino Básico e de Educação Pré-Escolar, sendo que se encontram evidenciadas as especificidades, transversalidade e continuidade existentes em ambos os contextos. A abordagem do estudo apresenta um caráter misto. Isto é, por um lado um caráter qualitativo, na medida em que procuramos perceber mecanismos, comportamentos e atitudes, por outro lado um caráter quantitativo, na medida em que a informação, recolhida através dos questionários e entrevistas, pode espelhar uma perspetiva quantitativa. O trabalho, comum a ambas as valências, com vista a um incremento na melhoria da intervenção educativa, visa compreender as necessidades, motivações e comportamentos dos sujeitos de estudo. Este estudo tem caraterísticas próximas de uma investigação-ação que permitiram uma avaliação constante da intervenção com o objetivo de a tornar mais eficaz. Os instrumentos e técnicas de investigação utilizados foram a análise documental, as listas de verificação ou controlo, as grelhas de observação, a observação participante, os registos de observação, o inquérito por questionário, a entrevista, os instrumentos de avaliação das aprendizagens, o portfólio de criança, o registo de projeto lúdico e arede curricular. O conhecimento dos grupos de crianças permite atitudes adequadas, assim como uma planificação mais eficaz, na medida em que são promovidas atividades significativas e diferenciadas que vão de encontro às necessidades e interesses de cada um. Estes reguladores da prática educativa são instrumentos que permitem a reflexão, prática sine-qua-non para uma intervenção de qualidade, pois esta ação vai permitir uma avaliação dos acontecimentos e suas consequências e, ainda, a remodelação de práticas e atitudes.

Deteção automática de incêndios. Detetores lineares de calor e de fumos

Existem situações particulares de deteção de incêndio nas quais os tradicionais detetores pontuais de fumo e calor podem não ser a solução mais adequada. São exemplo destas situações, a proteção de grandes áreas e/ou grandes distâncias. Nestas situações, poder-se-á equacionar a utilização de detetores lineares de calor e de fumos.

L’hospitalié est un moment d’interaction culturelle

Espacio y hospitalidad, KHÔRA II-3, 2004, Junho, org. Danielle Provansal, (19-26)

Zones lliures de transgènics a Europa : origen, desenvolupament i implantació a l'Estat Espanyol i a Catalunya

El nou moviment de les Zones Lliures de Transgènics (ZLT) sorgeix dels moviments d’oposició als OMGAs, paral·lement al debat que està tenint lloc en el sí de la UE, entorn la viabilitat de la coexistència entre l’agricultura transgènica i la convencional o ecològica. Aquest moviment que s’articula des de la dimensió local però amb un marcat simbolisme i rellevància en l’àmbit global, constitueix una important eina de desobediència civil dins el marc regulatori de la UE. Actualment, dins el context europeu, és a l’Estat Espanyol on es concentra la major superfície de blat de moro MG a escala comercial, cosa que dificulta l’establiment de ZLT. Aquest moviment, que reivindica la sobirania de les regions dins el gegant europeu, constitueix una alternativa real al model de producció, consum i dedesenvolupament actuals.

A MODEL OF CREATINE DEFICIENCY SYNDROMES IN 3D BRAIN CELL CULTURES BY KNOCKDOWN OF GAMT AND SLC6A8 GENES

La créatine joue un rôle essentiel dans le métabolisme cellulaire par sa conversion, par la creatine kinase, en phosphocreatine permettant la régénération de l'ATP. La synthèse de créatine, chez les mammifères, s'effectue par une réaction en deux étapes impliquant Γ arginine: glycine amidinotransférase (AGAT) et la guanidinoacétate méthyltransférase (GAMT). L'entrée de créatine dans les cellules s'effectue par son transporteur, SLC6A8. Les déficiences en créatine, dues au déficit en GAMT, AGAT ou SLC6A8, sont fréquentes et caractérisées par une absence ou une forte baisse de créatine dans le système nerveux central. Alors qu'il est connu que AGAT, GAMT et SLC6A8 sont exprimés par le cerveau, les conséquences des déficiences en créatine sur les cellules nerveuses sont peu comprises. Le but de ce travail était de développer de nouveaux modèles expérimentaux des déficiences en Cr dans des cultures 3D de cellules nerveuses de rat en agrégats au moyen de l'interférence à l'ARN appliquée aux gènes GAMT et SLC6A8. Des séquences interférentes (shRNAs) pour les gènes GAMT et SLC6A8 ont été transduites par des vecteurs viraux AAV (virus adéno-associés), dans les cellules nerveuses en agrégats. Nous avons ainsi démontré une baisse de l'expression de GAMT au niveau protéique (mesuré par western blot), et ARN messager (mesuré par qPCR) ainsi qu'une variation caractérisitique de créatine et guanidinoacétate (mesuré par spectrométrie de masse). Après avoir validé nos modèles, nous avons montré que les knockdown de GAMT ou SLC6A8 affectent le développement des astrocytes et des neurones ou des oligodendrocytes et des astrocytes, respectivement, ainsi qu'une augmentation de la mort cellulaire et des modifications dans le pattern d'activation des voies de signalisation impliquant caspase 3 et p38 MAPK, ayant un rôle dans le processus d'apoptose. - Creatine plays essential roles in energy metabolism by the interconversion, by creatine kinase, to its phosphorylated analogue, phosphocreatine, allowing the regeneration of ATP. Creatine is synthesized in mammals by a two step mechanism involving arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT). Creatine is taken up by cells by a specific transporter, SLC6A8. Creatine deficiency syndromes, due to defects in GAMT, AGAT and SLC6A8, are among the most frequent inborn errors of metabolism, and are characterized by an absence or a severe decrease of creatine in central nervous system, which is the main tissue affected. While it is known that AGAT, GAMT and SLC6A8 are expressed in CNS, many questions remain on the specific effects of AGAT, GAMT and SLC6A8 deficiencies on brain cells. Our aim was to develop new experimental models of creatine deficiencies by knockdown of GAMT and SLC6A8 genes by RNAi in 3D organotypic rat brain cell cultures in aggregates. Specific shRNAs for the GAMT and SLC6A8 genes were transduced in brain cell aggregates by adeno-associated viruses (AAV). The AAV-transduced shRNAs were able to efficiently knockdown the expression of our genes of interest, as shown by a strong decrease of protein by western blotting, a decrease of mRNA by qPCR or characteristic variations of creatine and guanidinoacetate by tandem mass spectrometry. After having validated our experimental models, we have also shown that GAMT and SLC6A8 knockdown affected the development of astrocytes and neurons or oligodendrocytes and astrocytes, respectively. We also observed an increase of cell death and variations in activation pattern of caspase 3 and p38 MAPK pathways, involved in apoptosis, in our experimental model.

Laboratory detection of respiratory viruses by automated techniques.

Advances in clinical virology for detecting respiratory viruses have been focused on nucleic acids amplification techniques, which have converted in the reference method for the diagnosis of acute respiratory infections of viral aetiology. Improvements of current commercial molecular assays to reduce hands-on-time rely on two strategies, a stepwise automation (semi-automation) and the complete automation of the whole procedure. Contributions to the former strategy have been the use of automated nucleic acids extractors, multiplex PCR, real-time PCR and/or DNA arrays for detection of amplicons. Commercial fully-automated molecular systems are now available for the detection of respiratory viruses. Some of them could convert in point-of-care methods substituting antigen tests for detection of respiratory syncytial virus and influenza A and B viruses. This article describes laboratory methods for detection of respiratory viruses. A cost-effective and rational diagnostic algorithm is proposed, considering technical aspects of the available assays, infrastructure possibilities of each laboratory and clinic-epidemiologic factors of the infection.

Human Papillomavirus (HPV) Type Distribution in Females with Abnormal Cervical Cytology. A Correlation with Histological Study

The aim of this study was to determine human papillomavirus (HPV) types distribution in cervical preneoplasic lesions in a Southern Spanish population and their relationship between HPV type and grade of histopathological abnormality. Finally, 232 cervical samples from 135 women with previous cytological abnormalities were included in this study. Colposcopy studies and biopsies were performed. Haematoxylin-eosin stained slides were observed and detection of HPV DNA in cervical swabs was carried out with use of a polymerase chain reaction and microarrays technology. The relationship between the presence of HPV infection and diagnostic variables was evaluated. HPV 16 was the most common type followed by HPV 58, 51, 33 and 31. However, the two HPV types targeted in the prophylactic vaccines such as HPV type 16 and 18 were detected in only 37 (21.2%) and 2 (1.1%) cases respectively. Thirty-three (18.9%) of samples were infected with multiple types, the majority of them with two types. In addition, during the follow-up of patients many changes in type distribution were observed. Several studies will be necessary in order to evaluate the HPV type distribution for therapeutically and prophylactic purposes such as vaccine treatment. Also, because of the differences obtained depending of use of various DNA technologies, the performance of some comparative studies of the different methods from detection of HPV would be advisable in a high population of patients and with the most homogeneous conditions possible.

Strategies for the successful implementation of viral laboratory automation.

It has been estimated that more than 70% of all medical activity is directly related to information providing analytical data. Substantial technological advances have taken place recently, which have allowed a previously unimagined number of analytical samples to be processed while offering high quality results. Concurrently, yet more new diagnostic determinations have been introduced - all of which has led to a significant increase in the prescription of analytical parameters. This increased workload has placed great pressure on the laboratory with respect to health costs. The present manager of the Clinical Laboratory (CL) has had to examine cost control as well as rationing - meaning that the CL's focus has not been strictly metrological, as if it were purely a system producing results, but instead has had to concentrate on its efficiency and efficacy. By applying re-engineering criteria, an emphasis has had to be placed on improved organisation and operating practice within the CL, focussing on the current criteria of the Integrated Management Areas where the technical and human resources are brought together. This re-engineering has been based on the concepts of consolidating and integrating the analytical platforms, while differentiating the production areas (CORE Laboratory) from the information areas. With these present concepts in mind, automation and virological treatment, along with serology in general, follow the same criteria as the rest of the operating methodology in the Clinical Laboratory.

Application of Molecular Diagnostic Techniques for Viral Testing.

Nucleic acid amplification techniques are commonly used currently to diagnose viral diseases and manage patients with this kind of illnesses. These techniques have had a rapid but unconventional route of development during the last 30 years, with the discovery and introduction of several assays in clinical diagnosis. The increase in the number of commercially available methods has facilitated the use of this technology in the majority of laboratories worldwide. This technology has reduced the use of some other techniques such as viral culture based methods and serological assays in the clinical virology laboratory. Moreover, nucleic acid amplification techniques are now the methods of reference and also the most useful assays for the diagnosis in several diseases. The introduction of these techniques and their automation provides new opportunities for the clinical laboratory to affect patient care. The main objectives in performing nucleic acid tests in this field are to provide timely results useful for high-quality patient care at a reasonable cost, because rapid results are associated with improvements in patients care. The use of amplification techniques such as polymerase chain reaction, real-time polymerase chain reaction or nucleic acid sequence-based amplification for virus detection, genotyping and quantification have some advantages like high sensitivity and reproducibility, as well as a broad dynamic range. This review is an up-to-date of the main nucleic acid techniques and their clinical applications, and special challenges and opportunities that these techniques currently provide for the clinical virology laboratory.

Introduction of an automated system for the diagnosis and quantification of hepatitis B and hepatitis C viruses.

Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections pose major public health problems because of their prevalence worldwide. Consequently, screening for these infections is an important part of routine laboratory activity. Serological and molecular markers are key elements in diagnosis, prognosis and treatment monitoring for HBV and HCV infections. Today, automated chemiluminescence immunoassay (CLIA) analyzers are widely used for virological diagnosis, particularly in high-volume clinical laboratories. Molecular biology techniques are routinely used to detect and quantify viral genomes as well as to analyze their sequence; in order to determine their genotype and detect resistance to antiviral drugs. Real-time PCR, which provides high sensitivity and a broad dynamic range, has gradually replaced other signal and target amplification technologies for the quantification and detection of nucleic acid. The next-generation DNA sequencing techniques are still restricted to research laboratories.The serological and molecular marker methods available for HBV and HCV are discussed in this article, along with their utility and limitations for use in Chronic Hepatitis B (CHB) diagnosis and monitoring.