Nop17 is a key R2TP factor for the assembly and maturation of box C/D snoRNP complex.

BACKGROUND: Box C/D snoRNPs are responsible for rRNA methylation and processing, and are formed by snoRNAs and four conserved proteins, Nop1, Nop56, Nop58 and Snu13. The snoRNP assembly is a stepwise process, involving other protein complexes, among which the R(2)TP and Hsp90 chaperone. Nop17, also known as Pih1, has been shown to be a constituent of the R(2)TP (Rvb1, Rvb2, Tah1, Pih1) and to participate in box C/D snoRNP assembly by its interaction with Nop58. The molecular function of Nop17, however, has not yet been described. RESULTS: To shed light on the role played by Nop17 in the maturation of snoRNP, here we analyzed the interactions domains of Nop58 - Nop17 - Tah1 and the importance of ATP to the interaction between Nop17 and the ATPase Rvb1/2. CONCLUSIONS: Based on the results shown here, we propose a model for the assembly of box C/D snoRNP, according to which R(2)TP complex is important for reducing the affinity of Nop58 for snoRNA, and for the binding of the other snoRNP subunits.

Primordial germ cells (spermatogonial stem cells) of bullfrogs express sex hormone-binding globulin and steroid receptors during seasonal spermatogenesis.

In vertebrate species, testosterone seems to inhibit spermatogonial differentiation and proliferation. However, this androgen can also be converted, via aromatase, into estrogen which stimulates spermatogonial differentiation and mitotic activity. During seasonal spermatogenesis of adult bullfrogs Lithobates catesbeianus, primordial germ cells (PGCs) show enhanced testosterone cytoplasm immunoexpression in winter; however, in summer, weak or no testosterone immunolabelling was observed. The aim of this study was to confirm if PGCs express stem cell markers - alkaline phosphatase (AP) activity and GFRα1 (glial-cell-line-derived neurotrophic factor) - and verify whether testosterone is maintained in these cells by androgen receptors (ARs) and/or sex hormone-binding globulin (SHBG) in winter. Furthermore, regarding the possibility that testosterone is converted into estrogen by PGCs in summer, the immunoexpression of estrogen receptor (ER)ß was investigated. Bullfrog testes were collected in winter and in summer and were embedded in glycol methacrylate for morphological analyses or in paraffin for the histochemical detection of AP activity. GFRα1, AR, SHBG and ERß expression were detected by Western blot and immunohistochemical analyses. The expression of AP activity and GFRα1 in the PGCs suggest that these cells are spermatogonial stem cells. In winter, the cytoplasmic immunoexpression of ARs and SHBG in the PGCs indicates that testosterone is maintained by these proteins in these cells. The cytoplasmic immunoexpression of ERß, in summer, also points to an ER-mediated action of estrogen in PGCs. The results indicate a participation of testosterone and estrogen in the control of the primordial spermatogonia during the seasonal spermatogenesis of L. catesbeianus.

Immunoexpression of aromatase and estrogen receptors ß in stem spermatogonia of bullfrogs indicates a role of estrogen in the seasonal spermatogonial mitotic activity.

Bullfrog stem spermatogonia, also named primordial germ cells (PGCs), show strong testosterone immunolabeling in winter, but no or weak testosterone immunoexpression in summer. Thus, the role of testosterone in these cells needs to be clarified. In this study, we proposed to evaluate whether PGCs express aromatase and estrogen receptors, and verify a possible role of estrogen in PGCs seasonal proliferation. Testes of male adult bullfrogs, collected in winter (WG) and summer (SG), were fixed and embedded in historesin, for quantitative analysis, or paraffin for immunohistochemistry (IHC). The number of haematoxylin/eosin stained PGCs/lobular area was obtained. Proliferating cell nuclear antigen (PCNA), aromatase, estrogen receptor ß (ERß) and PCNA/ERß double immunolabeling were detected by IHC. The number of PCNA-positive PGCs and the histological score (HSCORE) of aromatase and ERß immunolabeled PGCs were obtained. Although the number of PGCs increased significantly in WG, a high number of PCNA-positive PGCs was observed in summer. Moreover, aromatase and ERß HSCORE was higher in SG than WG. The results indicate that PGCs express a seasonal proliferative activity; the low mitotic activity in winter is related to the maximal limit of germ cells which can be supported in the large lobules. In SG, the increased ERß and aromatase HSCORE suggests that testosterone is converted into estrogen from winter to summer. Moreover, the parallelism between the high PGCs mitotic activity and ERß immunoexpression suggest a participation of estrogen in the control of the PGCs seasonal proliferative activity which guarantee the formation of new germ cysts from summer to next autumn.

Identification of archaeal proteins that affect the exosome function in vitro.

BACKGROUND: The archaeal exosome is formed by a hexameric RNase PH ring and three RNA binding subunits and has been shown to bind and degrade RNA in vitro. Despite extensive studies on the eukaryotic exosome and on the proteins interacting with this complex, little information is yet available on the identification and function of archaeal exosome regulatory factors. RESULTS: Here, we show that the proteins PaSBDS and PaNip7, which bind preferentially to poly-A and AU-rich RNAs, respectively, affect the Pyrococcus abyssi exosome activity in vitro. PaSBDS inhibits slightly degradation of a poly-rA substrate, while PaNip7 strongly inhibits the degradation of poly-A and poly-AU by the exosome. The exosome inhibition by PaNip7 appears to depend at least partially on its interaction with RNA, since mutants of PaNip7 that no longer bind RNA, inhibit the exosome less strongly. We also show that FITC-labeled PaNip7 associates with the exosome in the absence of substrate RNA. CONCLUSIONS: Given the high structural homology between the archaeal and eukaryotic proteins, the effect of archaeal Nip7 and SBDS on the exosome provides a model for an evolutionarily conserved exosome control mechanism.

Heterobinuclear copper(II)­platinum(II) complexes with oxindolimine ligands: Interactions with DNA, and inhibition of kinase and alkaline phosphatase proteins.

Two mononuclear copper(II) compounds, [Cu(isad)(H2O)Cl]Cl 1 and [Cu(isah)(H2O)Cl]Cl 2, and its corresponding heterobinuclear species containing also platinum(II), [CuCl(isad)Pt(NH3)Cl2] 3 and [CuCl(isah)Pt(NH3)Cl2] 4 (where isad and isah are oxindolimine ligands, (E)-3-(2-(3-aminopropylamino)ethylimino)indolin-2-one, and (E)-3-(3-amino-2-hydroxypropylimino)indolin-2-one, respectively), have been previously synthesized and characterized by different spectroscopic techniques in our laboratory. Cytotoxicity assays performed with B16F10 murine cancer cells, and MES-SA human uterine sarcoma cells, showed IC50 values lower or in the same order of cisplatin. Herein, in order to better elucidate their probable modes of action, possible interaction and damage to DNA, as well as their effect on the activity of crucial proteins were verified. Both mononuclear complexes and the binuclear compound 4 displayed a significant cleavage activity toward plasmid DNA, while compound 3 tends to protect DNA from oxidative damage, avoiding degradation. Complementary experiments indicated a significant inhibition activity toward cyclin-dependent kinase (CDK1/cyclinB) activity in the phosphorylation of histone H1, and only moderate inhibition concerning alkaline phosphatase. Results also revealed that the reactivity is reliant on the ligand structure and on the nature of the metal present, in a synergistic effect. Simulation studies complemented and supported our results, indicating different bindings of the binuclear compounds to DNA. Therefore, the verified cytotoxicity of these complexes comprises multiple modes of action, including modification of DNA conformation, scission of DNA strands by reactive oxygen species, and inhibition of selected proteins that are crucial to the cellular cycle.

Sdo1p, the yeast orthologue of Shwachman-Bodian-Diamond syndrome protein, binds RNA and interacts with nuclear rRNA-processing factors.

The Shwachman-Bodian-Diamond syndrome protein (SBDS) is a member of a highly conserved protein family of not well understood function, with putative orthologues found in different organisms ranging from Archaea, yeast and plants to vertebrate animals. The yeast orthologue of SBDS, Sdo1p, has been previously identified in association with the 60S ribosomal subunit and is proposed to participate in ribosomal recycling. Here we show that Sdo1p interacts with nucleolar rRNA processing factors and ribosomal proteins, indicating that it might bind the pre-60S complex and remain associated with it during processing and transport to the cytoplasm. Corroborating the protein interaction data, Sdo1p localizes to the nucleus and cytoplasm and co-immunoprecipitates precursors of 60S and 40S subunits, as well as the mature rRNAs. Sdo1p binds RNA directly, suggesting that it may associate with the ribosomal subunits also through RNA interaction.

Double-strand DNA cleavage induced by oxindole-Schiff base copper(II) complexes with potential antitumor activity.

Some oxindole-Schiff base copper(II) complexes have already shown potential antitumor activity towards different cells, inducing apoptosis in a process modulated by the ligand, and having nuclei and mitochondria as main targets. Here, three novel copper(II) complexes with analogous ligands were isolated and characterized by spectroscopic techniques, having their reactivity compared to the so far most active complex in this class. Cytotoxicity experiments carried out toward human neuroblastoma SH-SY5Y cells confirmed its pro-apoptosis property. DNA cleavage studies were then performed in the presence of these complexes, in order to verify the influence of ligand structural features in its nuclease activity. All of them were able to cause double-strand DNA scissions, giving rise to nicked circular Form II and linear Form III species, in the presence of hydrogen peroxide. Additionally, DNA Form II was also detected in the absence of peroxide when the most active complex, [Cu(isaepy)2]2+ 1, was used. In an effort to better elucidate their interactions with DNA, solutions of the different complexes titrated with DNA had their absorption spectra monitored. An absorbance hyperchromism observed at 260 nm pointed to the intercalation of these complexes into the DNA structure. Further, investigations of 2-deoxy-d-ribose (DR) oxidation catalyzed by each of those complexes, using 2-thiobarbituric acid reactive species (TBARS) method, and detection of reactive oxygen species (ROS) formation by spin-trapping EPR, suggested that their mechanism of action in performing efficiently DNA cleavage occurs preferentially, but not only by oxidative pathways.

Characterization of Saccharomyces cerevisiae Nop17p, a novel Nop58p-interacting protein that is involved in Pre-rRNA processing.

In eukaryotes, pre-rRNA processing depends on cis-acting elements and on a large number of non-ribosomal trans-acting factors, including endonucleases and exonucleases, RNA helicases, rRNA modifying enzymes and components of snoRNPs. The exosome is a conserved eukaryotic protein complex containing multiple 3'-5' exonucleases, which has been implicated in pre-rRNA, snoRNA and snRNA processing, as well as in mRNA degradation. In order to identify new proteins involved in rRNA processing, we have screened a yeast two-hybrid cDNA library, to isolate proteins interacting with the exosome subunit Rrp43p. In this screen, a novel nucleolar protein, Nop17p, was identified which also interacts with the box C/D snoRNP protein Nop58p. The NOP17 gene is not essential for cell viability but its deletion causes a temperature-sensitive phenotype. Pre-rRNA processing analyses revealed that rRNA formation is affected in the Deltanop17 strain subjected to the non-permissive temperature, although it is not blocked completely. In addition, primer extension analyses of RNA isolated from Nop17p-depleted cells subjected to the non-permissive temperature indicates that the pre-rRNA is undergoing different modification or degradation processes in these cells as compared to the parental strain. Nop17p was recently described in the same complex as Nop58p and, interestingly, its depletion leads to mislocalization of Nop1p, Nop56p, Nop58p and Snu13p, which are the core proteins of the box C/D ribonucleoprotein (snoRNP), indicating that Nop17p function is required either for nucleolar retention or for the proper assembly of the box C/D snoRNP.

Nop53p, an essential nucleolar protein that interacts with Nop17p and Nip7p, is required for pre-rRNA processing in Saccharomyces cerevisiae.

In eukaryotes, pre-rRNA processing depends on a large number of nonribosomal trans-acting factors that form large and intriguingly organized complexes. A novel nucleolar protein, Nop53p, was isolated by using Nop17p as bait in the yeast two-hybrid system. Nop53p also interacts with a second nucleolar protein, Nip7p. A carbon source-conditional strain with the NOP53 coding sequence under the control of the GAL1 promoter did not grow in glucose-containing medium, showing the phenotype of an essential gene. Under nonpermissive conditions, the conditional mutant strain showed rRNA biosynthesis defects, leading to an accumulation of the 27S and 7S pre-rRNAs and depletion of the mature 25S and 5.8S mature rRNAs. Nop53p did not interact with any of the exosome subunits in the yeast two-hybrid system, but its depletion affects the exosome function. In pull-down assays, protein A-tagged Nop53p coprecipitated the 27S and 7S pre-rRNAs, and His-Nop53p also bound directly 5.8S rRNA in vitro, which is consistent with a role for Nop53p in pre-rRNA processing.

[Understanding the meaning of being hospitalized in daily lives of children and teenager with chronic diseases]. / Compreendendo o significado de estar hospitalizado no cotidiano de crianças e adolescentes com doenças crônicas.

The study tried to understand the meaning of hospitalization for children and teenagers with chronic diseases substantiated on Symbolic Interactionism and developed from the qualitative Assistential Converging Research whose participants were a child and three teenagers. Data collection took place through a nursing interaction process, participating observation, sensible listening and child drawing. During the analysis, the meaning of hospitalization was expressed as being ill, away from home, feeling pain, sadness, suffering, wishing to cry, nervousness, aggressiveness and losing freedom. It was also understood as treatment, cure, support, friends, reciprocity with nature and "pleasant experiences" through care, passion, love, happiness and games. The participants' ability to show their view of the situation in which they are inserted was highlighted in their drawings, imbued with meanings and life lessons.

Currículos de cursos de graduação em enfermagem: revisão integrativa de literatura / Curriculum para cursos de grado en enfermería: revisión integradora de literatura / Curriculum for undergraduate courses in nursing: integrative review of literature

Objective: to know what issues have been addressed and discussed in research on curriculum in undergraduate courses in nursing in Brazil, in the period 2000-2013. Method: Used the assumptions of integrative literature review based on the LILACS, SciELO, BDENF and ERIC databases. Data analysis was organized from the preanalysis and exploration of material, processing and interpretation of results. Results: 36 articles were selected, which allowed the definition of three categories: Assumptions of curriculum reform; Characteristics of the training of nurses and National Curriculum Guidelines for Undergraduate Nursing courses and curricula. Conclusion: There was adherence of the studies will be relevant legislation, and the emphasis on the need for vocational training based on the principles of the Unified Health System.

Objetivo: conhecer quais assuntos tem sido abordados e debatidos nas investigações sobre currículo nos cursos de graduação em enfermagem no Brasil, no período de 2000 a 2013. Método: Utilizaram-se os pressupostos da revisão integrativa de literatura nas bases de dados LILACS, SciELO, BDENF e ERIC. A análise dos dados foi organizada a partir da pré-análise, exploração do material e, tratamento e interpretação dos resultados. Resultados: Foram selecionados 36 artigos, que possibilitaram a definição de três categorias: Pressupostos das reformas curriculares; Características da formação profissional de enfermeiros e Diretrizes Curriculares Nacionais dos cursos de graduação em Enfermagem e os currículos. Conclusão: Verificou-se aderência dos estudos à legislação pertinente,e a ênfase na necessidade de formação profissional fundamentada nos princípios do Sistema Único de Saúde.

Objetivo: conocer qué temas se han abordado y discutido en la investigación sobre el currículo en los cursos de licenciatura en enfermería en Brasil, en el período 2000-2013. Método: Se utilizó los supuestos de revisión integradora de la literatura en las bases de datos LILACS, SciELO, BDENF y ERIC. El análisis de datos se organizó desde el preanálisis y exploración de materiales, procesamiento e interpretación delos resultados. Resultados: Se seleccionaron 36 artículos, lo que permitió la definición de tres categorías: los supuestos de la reforma curricular; Características de la formación de enfermeras y Directrices Curriculares Nacionales para Pregrado Enfermería cursos y curriculum. Conclusión: Se encontro adhesión de los estudios con la legislación, y el énfasis en la necesidad de formación profesional basado en los principios del Sistema Único de Salud.

WASTE MANAGEMENT OF HEALTHCARE SERVICES FROM THE PERSPECTIVE OF NURSING PROFESSIONALS / LA ADMINISTRACIÓN DE RESIDUOS DE SERVICIO DE SALUD BAJO LA ÓPTICA DE LOS PROFESIONALES DE ENFERMERÍA / O GERENCIAMENTO DE RESÍDUOS DE SERVIÇO DE SAÚDE SOB A ÓTICA DOS PROFISSIONAIS DE ENFERMAGEM

ABSTRACT Objective: to identify the knowledge of nursing professionals at pediatric inpatient units about waste management of health services. Method: this descriptive, exploratory research was developed at two inpatient units of a children's hospital in Southern Brazil. Data collection was carried out between July and August 2014 through questionnaires to 30 nursing team professionals. The data were analyzed in a quantitative and qualitative perspective. Results: the results show that few professionals at the investigated institution know the Health Service Waste Management Program. The majority performs care related to the waste, but does not participate in training on the subject. Conclusion: the training of nursing professionals is critical for the effectiveness of the waste management program at the health institution, favoring proper disposal, facilitating the collection and contributing to environmental preservation.

RESUMEN Objetivo: identificar el conocimiento de los profesionales de enfermería de unidades de internamiento pediátrica sobre la administración de residuos de servicios de salud. Método: investigación exploratoria, descriptiva, desarrollada en dos unidades de internamiento de un hospital infantil del Sur de Brasil. La recolección de datos fue realizada entre julio y agosto de 2014, a través de cuestionarios aplicados junto a 30 profesionales del equipo de enfermería. Los datos fueron analizados bajo una perspectiva cuanti-cualitativa. Resultados: los resultados apuntan que pocos profesionales de la institución investigada conocen el Programa de Administración de Residuos de Servicios de Salud. La mayoría realiza cuidados en relación a los residuos, más no participan de capacitaciones sobre el tema. Conclusión: la capacitación de los profesionales de enfermería es fundamental para la efectivación del programa de administración de los residuos de la institución de salud, favoreciendo el descarte adecuado, facilitando la recolección y contribuyendo para la preservación del medio ambiente.

RESUMO Objetivo: identificar o conhecimento dos profissionais de enfermagem de unidades de internação pediátrica sobre o gerenciamento de resíduos do serviço de saúde Método: pesquisa exploratória, descritiva, desenvolvida em duas unidades de internação de um hospital infantil do Sul do Brasil. A coleta de dados foi realizada entre julho e agosto de 2014, através de questionários aplicados junto a 30 profissionais da equipe de enfermagem. Os dados foram analisados em uma perspectiva quanti-qualitativa Resultados: os resultados apontam que poucos profissionais da instituição investigada conhecem o Programa de Gerenciamento de Resíduos de Serviço de Saúde. A maioria realiza cuidados em relação aos resíduos, mas não participam de capacitações sobre o tema Conclusão: a capacitação dos profissionais de enfermagem é fundamental para a efetivação do programa de gerenciamento dos resíduos da instituição de saúde, favorecendo o descarte adequado, facilitando a coleta e contribuindo para preservação do meio ambiente.

Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems.

Phase diagrams of poly(ethylene glycol)/polyacrylate/Na(2)SO(4) systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coli can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coli homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Na(2)SO(4)-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH.

Binding of oxindole-Schiff base copper(II) complexes to DNA and its modulation by the ligand.

Previous studies on copper(II) complexes with oxindole-Schiff base ligands have shown their potential antitumor activity towards different cells, inducing apoptosis through a preferential attack to DNA and/or mitochondria. Herein, we better characterize the interactions between some of these copper(II) complexes and DNA. Investigations on its binding ability to DNA were carried out by fluorescence measurements in competitive experiments with ethidium bromide, using plasmidial or calf-thymus DNA. These results indicated an efficient binding process similar to that observed with copper(II)-phenanthroline species, [Cu(o-phen)(2)](2+), with binding constants in the range 3 to 9×10(2) M(-1). DNA cleavage experiments in the presence and absence of distamycin, a recognized binder of DNA, indicated that this binding probably occurs at major or minor groove, leading to double-strand DNA cleavage, and being modulated by the imine ligand. Corroborating these data, discrete changes in EPR spectra of the studied complexes were observed in the presence of DNA, while more remarkable changes were observed in the presence of nucleotides (AMP, GMP, CMP or UMP). Additional evidence for preferential coordination of the copper centers to the bases guanine or cytosine was obtained from titrations of these complexes with each nucleotide, monitored by absorption spectral changes. Therefore, the obtained data point out to their action as groove binders to DNA bases, rather than as intercalators or covalent cross-linkers. Further investigations by SDS PAGE using (32)P-ATP or (32)P-oligonucleotides attested that no hydrolysis of phosphate linkage in DNA or RNA occurs, in the presence of such complexes, confirming their main oxidative mechanism of action.

Lúdico no cuidado à criança e ao adolescente com câncer: perspectivas da equipe de de enfermagem / Playful activities in health care for children and adolescents with cancer: the perspectives of the nursing staff

Objetivo: Descrever a perspectiva da equipe de enfermagem sobre a utilização do lúdico no cuidado à criança e ao adolescente com câncer hospitalizada. Metodologia: Pesquisa descritiva exploratória de abordagem qualitativa, realizada entre agosto de 2013 e julho de 2015, com 29 profissionais de enfermagem de uma Unidade de Internação Onco-hematológica de um Hospital Pediátrico do Sul do Brasil. Os dados foram coletados por meio de entrevistas semiestruturadas, analisados e categorizados segundo Minayo, constituindo-se duas categorias, sendo apresentada aqui a segunda categoria denominada "no cuidar eu aproveito para brincar: tornando o hospital mais agradável". Resultados: Os profissionais relataram a vivência do lúdico no cotidiano da enfermagem, enfocando seus benefícios, dificuldades e possibilidades no cuidado. Conclusão: Salienta-se que o lúdico é uma ferramenta indispensável para o cuidado, contudo necessita de maior preparação teórica e prática para efetivá-lo e vivenciá-lo como uma estratégia de cuidado.

Objetivo: Describir la perspectiva del equipo de enfermería sobre la utilización del juego lúdico en el proceso de cuidado de niños y adolescentes hospitalizados con cáncer. Metodología: Investigación descriptiva exploratoria de abordaje cualitativo, realizada entre Agosto de 2013 y Julio de 2015 con 29 profesionales de enfermería de una Unidad de Hospitalización Onco-hematológica de un Hospital Pediátrico del Sur de Brasil. Los datos fueron recolectados por medio de entrevistas semiestructuradas, analizados y categorizados según Minayo, estableciendo dos categorías, siendo presentada aquí la segunda: "en el proceso de cuidar, yo aprovecho para jugar: tornando el hospital más agradable". Resultados: Los profesionales relataron la experiencia del lúdico en el cotidiano de la enfermería, enfocando sus beneficios, dificultades y posibilidades en el cuidado. Conclusión: Se observa el juego lúdico como herramienta indispensable para el cuidado, pero que necesita más preparación teórica y práctica para se convertir en una estrategia más efectiva.

Objective: To describe the perspective of the nursing staff on the use of playful activities in health care for children and adolescents hospitalized with cancer. Methodology: An exploratory descriptive study with a qualitative approach was performed between August 2013 and July 2015, including 29 nursing professionals from an Onco-hematology Inpatient Unit of a Pediatric Hospital in Southern Brazil. The data were collected through semi-structured interviews, analyzed and categorized according to Minayo, when two categories were formed, although only one was presented here: "When providing care, I take advantage to play, allowing the hospital environment to be more agreeable". Results: Professionals reported the experience of playful activities in routine nursing practice, focusing on their benefits, difficulties and opportunities in health care. Conclusion: Playful activities are an indispensable tool for care, but further theoretical and practical preparation is required to accomplish and experience it as a care strategy.

Expression, purification and structural analysis of the Pyrococcus abyssi RNA binding protein PAB1135.

BACKGROUND: The gene coding for the uncharacterized protein PAB1135 in the archaeon Pyrococcus abyssi is in the same operon as the ribonuclease P (RNase P) subunit Rpp30. FINDINGS: Here we report the expression, purification and structural analysis of PAB1135. We analyzed the interaction of PAB1135 with RNA and show that it binds efficiently double-stranded RNAs in a non-sequence specific manner. We also performed molecular modeling of the PAB1135 structure using the crystal structure of the protein Af2318 from Archaeoglobus fulgidus (2OGK) as the template. CONCLUSIONS: Comparison of this model has lead to the identification of a region in PAB1135 that could be involved in recognizing double-stranded RNA.

A informática na formação e qualificação dos profissionais de saúde: uma revisão integrativa / Information tecnology in education and training of health professionals: an integrative review

Objetivo: buscou-se conhecer as tendências das produções científicas sobre tecnologias na área da informática, como recurso para a formação e qualificação dos profissionais da área da saúde. Método: trata-se de uma revisão integrativa, que incluiu artigos indexados na Biblioteca Virtual de Saúde, entre 2007 e 2011. Resultados: identificaram-se 46 artigos que sinalizam para pontencialidade do uso da informática como recurso pedagógico, apontando vantagens, fragilidades e desafios da sua utilização, especialmente na formação na área da saúde. Considerações Finais: os achados sinalizam a necessidade de envolvimento e capacitação dos docentes, mudanças na postura dos acadêmicos e investimentos institucionais, bem como acompanhamento do uso dos recursos que envolvem as Tecnologias de Informação e Comunicação.

Aim: to know the trends of scientific production on technologies in the field of information technology as a resource for the training and qualification of health professionals. Method: an integrative review that included scientific articles indexed in the Virtual health Library in 2007 at 2011. Results: 46 articles were identified that signal potentiality in the use of computers as a teaching resource, pointing out advantages and weaknesses, especially in training in healthcare. Considerations: the findings suggest the need for involvement and empowerment of teachers, changes in posture of academic and institutional investments, as well as monitoring the use of resources that involve the Information Technologies and Communication.

Objetivo: este estudio objetivó conocer las tendencias de las producciones científicas sobre tecnologías en el área de la informática como recurso para la formación y calificación de los profesionales en el área de la salud. Método: fue realizada una revisión integradora, con la inclusión de artículos indexados en la Biblioteca Virtual de Salud entre los años 2007 y 2011. Resultados: fueron identificados 46 artículos que muestran para potencialidad del uso de la informática como recurso didáctico, señalando sus ventajas, debilidades y desafíos de su utilización, especialmente en la formación en salud. Consideraciones finales: los resultados señalan la necesidad de la participación y la capacitación de los docentes, los cambios en la postura de los académicos e inversiones institucionales, así como el acompañamiento del uso de los recursos relacionados con las Tecnologías de la Información y la Comunicación.

Compreendendo o significado de estar hospitalizado no cotidiano de crianças e adolescentes com doenças crônicas / Understanding the meaning of being hospitalized in daily lives of children and teenager with chronic diseases / Comprendiendo el significado del estar hospitalizado en el cotidiano de niños y adolescentes con enfermedades crónicas

O estudo buscou compreender o significado da hospitalização para crianças e adolescentes com doenças crônicas, fundamentado no Interacionismo Simbólico, desenvolvido a partir da Pesquisa Convergente Assistencial de abordagem qualitativa, cujos participantes foram uma criança e três adolescentes. Os dados foram coletados através do processo de interação em enfermagem, observação participante, escuta sensível e desenho infantil. O significado da hospitalização expressou doença, distância de casa, dor, tristeza, sofrimento, choro, nervosismo, agressividade, perda da liberdade, bem como tratamento, cura, apoio, amigos, reciprocidade com a natureza e "experiências agradáveis" através do cuidado, paixão, amor, felicidade e brincadeiras. Destacou-se a capacidade dos participantes em transformar o olhar da situação na qual estão inseridos, expressos em seus desenhos, permeados de significados e lições de vida.

The study tried to understand the meaning of hospitalization for children and teenagers with chronic diseases substantiated on Symbolic Interactionism and developed from the qualitative Assistential Converging Research whose participants were a child and three teenagers. Data collection took place through a nursing interaction process, participating observation, sensible listening and child drawing. During the analysis, the meaning of hospitalization was expressed as being ill, away from home, feeling pain, sadness, suffering, wishing to cry, nervousness, aggressiveness and losing freedom. It was also understood as treatment, cure, support, friends, reciprocity with nature and "pleasant experiences" through care, passion, love, happiness and games. The participants' ability to show their view of the situation in which they are inserted was highlighted in their drawings, imbued with meanings and life lessons.

El estudio buscó comprender el significado de la hospitalización para niños y adolescentes con enfermedades crónicas, fundamentado en el Interaccionismo Simbólico y desarrollado a partir de una Investigación Convergente Asistencial de abordaje cualitativa cuyos participantes fueron un niño y tres adolescentes. La cosecha de datos fue realizada a través del proceso de interacción en enfermería, observación participante, escucha sensible y dibujo infantil. El significado de hospitalización ha expresado el estar enfermo, lejos de casa, sentir dolor, tristeza, sufrimiento, ganas de llorar, nervosismo, agresividad e pérdida de libertad, pasando a ser comprendido también como tratamiento, cura, apoyo, amigos, estar en reciprocidad con la naturaleza y "experiencias agradables" a través del cuidado, pasión, amor, felicidad y juegos. La capacidad de los participantes de transformar su mirar de la situación donde están inseridos fue expresada en sus dibujos, permeados de significados y lecciones de vida.

Structural insights into the interaction of the Nip7 PUA domain with polyuridine RNA.

The conserved protein Nip7 is involved in ribosome biogenesis, being required for proper 27S pre-rRNA processing and 60S ribosome subunit assembly in Saccharomyces cerevisiae. Yeast Nip7p interacts with nucleolar proteins and with the exosome subunit Rrp43p, but its molecular function remains to be determined. Solution of the Pyrococcus abyssi Nip7 (PaNip7) crystal structure revealed a monomeric protein composed by two alpha-beta domains. The N-terminal domain is formed by a five-stranded antiparallel beta-sheet surrounded by three alpha-helices and a 310 helix while the C-terminal, a mixed beta-sheet domain composed by strands beta8 to beta12, one alpha-helix, and a 310 helix, corresponds to the conserved PUA domain (after Pseudo-Uridine synthases and Archaeosine-specific transglycosylases). By combining structural analyses and RNA interaction assays, we assessed the ability of both yeast and archaeal Nip7 orthologues to interact with RNA. Structural alignment of the PaNip7 PUA domain with the RNA-interacting surface of the ArcTGT (archaeosine tRNA-guanine transglycosylase) PUA domain indicated that in the archaeal PUA domain positively charged residues (R151, R152, K155, and K158) are involved in RNA interaction. However, equivalent positions are occupied by mostly hydrophobic residues (A/G160, I161, F164, and A167) in eukaryotic Nip7 orthologues. Both proteins can bind specifically to polyuridine, and RNA interaction requires specific residues of the PUA domain as determined by site-directed mutagenesis. This work provides experimental verification that the PUA domain mediates Nip7 interaction with RNA and reveals that the preference for interaction with polyuridine sequences is conserved in Archaea and eukaryotic Nip7 proteins.

Tumor necrosis factor-á signaling cascades in apoptosis, necrosis, necroptosis and cell proliferation

Tumor necrosis factor-á (TNF-á) is a multifunctional cytokine involved in host defense, inflammation, apoptosis, autoimmunity, organogenesis and lymphoid microarchitecture. Many of these activities may be explained by the ability of this cytokine to induce distinct signal transduction pathways that recruit regulatory proteins involved in differentiation, cell death or cell proliferation. In this review, we discuss the contribution of caspases -3, -6, -7 and -8, and of cyclin-dependent kinases (CDKs), cyclin B and cyclin-dependent kinase inhibitors (CKI p21 and p27), as well as retinoblastoma tumor suppressor in the signaling cascades triggered by TNF-á to induce apoptosis, necrosis and cellular proliferation in the murine cell lines NIH3T3 and WEHI-164 and the human cervical carcinoma cell line HeLa-S3. Based on the findings of many literature reports and our own data, we discussed a model in which caspases are continuously activated throughout the cell cycle and kept at a critical threshold level by IAP (inhibitor of apoptosis) antagonists. Following the release of Smac/Diablo and HtrA2/OMI from mitochondria in response to diverse stimuli, this threshold is overcome and results in amplified caspase activation and cell death. An alternative, caspase-independent mechanism of cell death is induced in NIH3T3 fi broblasts by a combination of TNF and the pan-caspase inhibitor z-VADfmk. This cell death phenotype, known as necroptosis, displays some morphological features of apoptosis and necrosis. Although caspases are critical regulators of the TNF signaling pathway during cellular life and death, the mechanisms involved in the fine regulation of their dual effects remain to be fully elucidated.