Abortive Lytic Reactivation of KSHV in CBF1/CSL Deficient Human B Cell Lines

Since Kaposi's sarcoma associated herpesvirus (KSHV) establishes a persistent infection in human B cells, B cells are a critical compartment for viral pathogenesis. RTA, the replication and transcription activator of KSHV, can either directly bind to DNA or use cellular DNA binding factors including CBF1/CSL as DNA adaptors. In addition, the viral factors LANA1 and vIRF4 are known to bind to CBF1/CSL and modulate RTA activity. To analyze the contribution of CBF1/CSL to reactivation in human B cells, we have successfully infected DG75 and DG75 CBF1/CSL knock-out cell lines with recombinant KSHV.219 and selected for viral maintenance by selective medium. Both lines maintained the virus irrespective of their CBF1/CSL status. Viral reactivation could be initiated in both B cell lines but viral genome replication was attenuated in CBF1/CSL deficient lines, which also failed to produce detectable levels of infectious virus. Induction of immediate early, early and late viral genes was impaired in CBF1/CSL deficient cells at multiple stages of the reactivation process but could be restored to wild-type levels by reintroduction of CBF1/CSL. To identify additional viral RTA target genes, which are directly controlled by CBF1/CSL, we analyzed promoters of a selected subset of viral genes. We show that the induction of the late viral genes ORF29a and ORF65 by RTA is strongly enhanced by CBF1/CSL. Orthologs of ORF29a in other herpesviruses are part of the terminase complex required for viral packaging. ORF65 encodes the small capsid protein essential for capsid shell assembly. Our study demonstrates for the first time that in human B cells viral replication can be initiated in the absence of CBF1/CSL but the reactivation process is severely attenuated at all stages and does not lead to virion production. Thus, CBF1/CSL acts as a global hub which is used by the virus to coordinate the lytic cascade

The antimicrobial effects of the alginate oligomer OligoG CF-5/20 are independent of direct bacterial cell membrane disruption

Concerns about acquisition of antibiotic resistance have led to increasing demand for new antimicrobial therapies. OligoG CF-5/20 is an alginate oligosaccharide previously shown to have antimicrobial and antibiotic potentiating activity. We investigated the structural modification of the bacterial cell wall by OligoG CF-5/20 and its effect on membrane permeability. Binding of OligoG CF-5/20 to the bacterial cell surface was demonstrated in Gram-negative bacteria. Permeability assays revealed that OligoG CF-5/20 had virtually no membrane-perturbing effects. Lipopolysaccharide (LPS) surface charge and aggregation were unaltered in the presence of OligoG CF-5/20. Small angle neutron scattering and circular dichroism spectroscopy showed no substantial change to the structure of LPS in the presence of OligoG CF-5/20, however, isothermal titration calorimetry demonstrated a weak calcium-mediated interaction. Metabolomic analysis confirmed no change in cellular metabolic response to a range of osmolytes when treated with OligoG CF-5/20. This data shows that, although weak interactions occur between LPS and OligoG CF-5/20 in the presence of calcium, the antimicrobial effects of OligoG CF-5/20 are not related to the induction of structural alterations in the LPS or cell permeability. These results suggest a novel mechanism of action that may avoid the common route in acquisition of resistance via LPS structural modification

Permanent education in health: a review

OBJECTIVE : To undertake a meta-synthesis of the literature on the main concepts and practices related to permanent education in health. METHODS : A bibliographical search was conducted for original articles in the PubMed, Web of Science, LILACS, IBECS and SciELO databases, using the following search terms: “public health professional education”, “permanent education”, “continuing education”, “permanent education health”. Of the 590 articles identified, after applying inclusion and exclusion criteria, 48 were selected for further analysis, grouped according to the criteria of key elements, and then underwent meta-synthesis. RESULTS : The 48 original publications were classified according to four thematic units of key elements: 1) concepts, 2) strategies and difficulties, 3) public policies and 4) educational institutions. Three main conceptions of permanent education in health were found: problem-focused and team work, directly related to continuing education and education that takes place throughout life. The main strategies for executing permanent education in health are discussion, maintaining an open space for permanent education, and permanent education clusters. The most limiting factor is mainly related to directly or indirect management. Another highlight is the requirement for implementation and maintenance of public policies, and the availability of financial and human resources. The educational institutions need to combine education and service aiming to form critical-reflexive graduates. CONCLUSIONS : The coordination between health and education is based as much on the actions of health services as on management and educational institutions. Thus, it becomes a challenge to implement the teaching-learning processes that are supported by critical-reflexive actions. It is necessary to carry out proposals for permanent education in health involving the participation of health professionals, teachers and educational institutions.OBJETIVO : Realizar metasíntesis de la literatura sobre los principales conceptos y prácticas relacionados con la educación permanente en salud. MÉTODOS : Se realizó búsqueda bibliográfica de artículos originales en las bases de datos PubMed, Web of Science, Lilacs, IBECS y SciELO, utilizando los siguientes descriptores: “ public health professional education”, “permanent education”, “continuing education”, “permanent education health ”. De un total de 590 artículos identificados, posterior a los criterios de inclusión y exclusión, fueron seleccionados 48 para análisis, los cuales fueron sometidos al análisis individual, análisis comparativo, análisis con criterios de agrupamiento de elementos-clave y sometidos a metasíntesis. RESULTADOS : Los 48 artículos originales fueron clasificados como elementos-clave en cuatro unidades temáticas: 1) Concepciones; 2) Estrategias y dificultades; 3) Políticas públicas e 4) Instituciones formadoras. Se encontraron tres concepciones principales de educación permanente en salud: ubicación del problema y enfocarlo en el trabajo en equipo, directamente relacionado con la educación continua y educación que se da a lo largo de la vida. Las principales estrategias para efectivar la educación permanente fueron la ubicación del problema, mantenimiento de espacios para la educación permanente y polos de educación permanente. El mayor factor limitante estuvo relacionado con la gerencia directa o indirecta. Fueron mencionadas la necesidad de implementación y mantenimiento de políticas públicas, así como la disponibilidad de recursos financieros y humanos. Las instituciones formadoras tendrían la necesidad de articular educación y servicio para la formación de egresados críticos-reflexivos. CONCLUSIONES : La articulación educación y salud se encuentra pautada tanto en las acciones de los servicios de salud, cuanto en la gestión y de instituciones formadoras. Así, se torna un desafío implementar procesos de educación-aprendizaje que sean respaldados por acciones crítico-reflexivas. Es necesario realizar propuestas de educación permanente en salud con la participación de profesionales de los servicios, profesores y profesionales de las instituciones de educación.OBJETIVO : Realizar metassíntese da literatura sobre os principais conceitos e práticas relacionados à educação permanente em saúde. MÉTODOS : Foi realizada busca bibliográfica de artigos originais nas bases de dados PubMed, Web of Science, Lilacs, IBECS e SciELO, utilizando os seguintes descritores: “ public health professional education ”, “ permanent education”, “continuing education ”, “ permanent education health ”. De um total de 590 artigos identificados, após os critérios de inclusão e exclusão, foram selecionados 48 para análise, os quais foram submetidos à análise individual, análise comparativa, análise com critérios de agrupamentos de elementos-chave e submetidos à metassíntese. RESULTADOS : Os 48 artigos originais foram classificados como elementos-chave em quatro unidades temáticas: 1) Concepções; 2) Estratégias e dificuldades; 3) Políticas públicas; e 4) Instituições formadoras. Foram encontradas três concepções principais de educação permanente em saúde: problematizadora e focada no trabalho em equipe, diretamente relacionada à educação continuada e educação que se dá ao longo da vida. As principais estratégias para efetivação da educação permanente foram a problematização, manutenção de espaços para a educação permanente e polos de educação permanente. O maior fator limitante foi relacionado à gerência direta ou indireta. Foram indicadas a necessidade de implementação e manutenção de políticas públicas, além de disponibilidade de recursos financeiros e de recursos humanos. As instituições formadoras teriam necessidade de articular ensino e serviço para a formação de egressos críticos-reflexivos. CONCLUSÕES : A articulação educação e saúde encontra-se pautada tanto nas ações dos serviços de saúde, quanto de gestão e de instituições formadoras. Assim, torna-se um desafio implementar processos de ensino-aprendizagem que sejam respaldados por ações crítico-reflexivas. É necessário realizar propostas de educação permanente em saúde com a participação de profissionais dos serviços, professores e profissionais das instituições de ensino

Root canal filling with cements based on mineral aggregates: an in vitro analysis of bacterial microleakage

The aim of this study was to assess bacterial microleakage through 2 different cements used as root canal filling materials: mineral-trioxide-aggregate (White MTA, Angelus, Londrina, PR, Brazil); and an experimental material containing calcium aluminate plus additives (EndoBinder (R), University of Sao Carlos [UFSCar-Brazil], patent number PI0704502-6). Forty incisors were divided into 4 groups: (1) white-MTA, (2) calcium aluminate cement, and (3) positive and (4) negative control group. Falcon tubes were prepared for this experiment, divided into 2 separated chambers; the lower part was filled with RTF-transport media so that only the root apex was in contact with the transport media, whereas the coronal part was immersed in BHI containing Enterococcus faecalis to investigate bacterial microleakage. The evaluated materials used as root canal filling materials showed an absence of microbial growth. Both materials when used as the root canal filling were efficient in sealing root canals and preventing E. faecalis microleakage for the 30-day evaluation period using an in vitro model. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: 140-144)108114014

An application of visibile human database in radiotherapy: tutorial for image guided externalradiotherapy (TIGER) Radiother Oncol. PMID: 15028403Luogo e data di pubblicazione: ELSEVIER SCI IRELAND LTD, CLARE, IRELAND, 2004

Abstract: Background and purpose: Three-dimensional conformal radiotherapy and intensity modulated radiotherapy allow accurate dose delivery on target volumes. Due to the different background among specialists involved in target volume definition, the contouring emerges as one of the most questionable steps in treatment planning procedures. A software tool devoted to contouring training, named tutorial for image guided external radiotherapy ('TIGER'), based on the Visible Human Project images data-set, is described. Materials and methods: TIGER is addressed to facilitate the learning of axial anatomical images, to promote the training and reproducibility in contouring process, to allow the availability of a tool to enhance the 'drill and practice' approach in training programs. TIGER includes three different environments: Anatomic tutorial devoted to facilitate a self-learning approach to axial body sections; Contouring tutorial addressed to practice contouring process of anatomical structures and to undergo a test program prepared by tutors; Teacher's tools to offer to tutors the opportunity to insert new outlines in TIGER-database, according to local needs or conventions, and to use them in tutorial programs. TIGER-database is grouped in six main anatomical sections: head and neck, male thorax, female thorax, abdomen, male pelvis, and female pelvis. Overall 432 corresponding CT-VH images and 1189 contours of 134 different anatomical structures and lymphatic drainage areas are available. The access to the TIGER software is allowed by ESTRO web site (http://www.estro.be). Conclusions: TIGER provides an interactive human anatomy cross-sectional oriented source to facilitate the interpretation of CT scan images usually contoured in daily practice. It offers a drill tool to facilitate the learning of a reproducible contouring procedur