Significant redox insensitivity of the functions of the SARS-CoV spike glycoprotein - Comparison with HIV envelope

The capacity of the surface glycoproteins of enveloped viruses to mediate virus/cell binding and membrane fusion requires a proper thiol/disulfide balance. Chemical manipulation of their redox state using reducing agents or free sulfhydryl reagents affects virus/cell interaction. Conversely, natural thiol/disulfide rearrangements often occur during the cell interaction to trigger fusogenicity, hence the virus entry. We examined the relationship between the redox state of the 20 cysteine residues of the SARS-CoV (severe acute respiratory syndrome coronavirus) Spike glycoprotein S1 subdomain and its functional properties. Mature S1 exhibited similar to 4 unpaired cysteines, and chemically reduced S1 displaying up to similar to 6 additional unpaired cysteines still bound ACE2 and enabled fusion. In addition, virus/cell membrane fusion occurred in the presence of sulfhydryl-blocking reagents and oxidoreductase inhibitors. Thus, in contrast to various viruses including HIV (human immunodeficiency virus) examined in parallel, the functions of the SARS-CoV Spike glycoprotein exhibit a significant and surprising independence of redox state, which may contribute to the wide host range of the virus. These data suggest clues for molecularly engineering vaccine immunogens.

SARS coronavirus unique domain: three-domain molecular architecture in solution and RNA binding

Nonstructural protein 3 of the severe acute respiratory syndrome (SARS) coronavirus includes a "SARS-unique domain" (SUD) consisting of three globular domains separated by short linker peptide segments. This work reports NMR structure determinations of the C-terminal domain (SUD-C) and a two-domain construct (SUD-MC) containing the middle domain (SUD-M) and the C-terminal domain, and NMR data on the conformational states of the N-terminal domain (SUD-N) and the SUD-NM two-domain construct. Both SUD-N and SUD-NM are monomeric and globular in solution; in SUD-NM, there is high mobility in the two-residue interdomain linking sequence, with no preferred relative orientation of the two domains. SUD-C adopts a frataxin like fold and has structural similarity to DNA-binding domains of DNA-modifying enzymes. The structures of both SUD-M (previously determined) and SUD-C (from the present study) are maintained in SUD-MC, where the two domains are flexibly linked. Gel-shift experiments showed that both SUD-C and SUD-MC bind to single-stranded RNA and recognize purine bases more strongly than pyrimidine bases, whereby SUD-MC binds to a more restricted set of purine-containing RNA sequences than SUD-M. NMR chemical shift perturbation experiments with observations of (15)N-labeled proteins further resulted in delineation of RNA binding sites (i.e., in SUD-M, a positively charged surface area with a pronounced cavity, and in SUD-C, several residues of an anti-parallel beta-sheet). Overall, the present data provide evidence for molecular mechanisms involving the concerted actions of SUD-M and SUD-C, which result in specific RNA binding that might be unique to the SUD and, thus, to the SARS coronavirus.

Septic Shock, Necrotizing Pneumonitis, and Meningoencephalitis Caused by Mycoplasma pneumoniae in a Child: A Case Report

Mycoplasma pneumoniae is an important causative agent of respiratory infection in childhood. Although the infection caused by M. pneumoniae is classically described as benign, severe and life-threatening pulmonary and extrapulmonary complications can occur. This study describes the first case of septic shock related to M. pneumoniae in a child with necrotizing pneumonitis, severe encephalitis, and multiple organs involvement, with a favorable outcome after lobectomy and systemic corticosteroids

Angiotensin II modulates CD40 expression in vascular smooth muscle cells

The signalling pathway CD40/CD40L (CD40 ligand) plays an important role in atherosclerotic plaque formation and rupture. AngII (angiotensin II), which induces oxidative stress and inflammation, is also implicated in the progression of atherosclerosis. In the present study, we tested the hypothesis that AngII increases CD40/CD40L activity in vascular cells and that ROS (reactive oxygen species) are part of the signalling cascade that controls CD40/CD40L expression. Human CASMCs (coronary artery smooth muscle cells) in culture exposed to IL (interleukin)-1 beta or TNF-alpha (tumour necrosis factor-a) had increased superoxide generation and enhanced CD40 expression, detected by EPR (electron paramagnetic resonance) and immunoblotting respectively. Both phenomena were abolished by previous incubation with membrane-permeant antioxidants or cell transfection with P22(phox) antisense. AngII (50-200 nmol/l) induced an early and sustained increase in CD40 mRNA and protein expression in CASMCs, which was blocked by treatment with antioxidants. Increased CD40 expression led to enhanced activity of the pathway, as AngII-treated cells stimulated with recombinant CD40L released higher amounts of IL-8 and had increased COX-2 (cyclo-oxygenase-2) expression. We conclude that AngII stimulation of vascular cells leads to a ROS-dependent increase in CD40/CD40L signalling pathway activity. This phenomenon may be an important mechanism modulating the arterial injury observed in atherosclerosis-related vasculopathy.

Perfuração espontânea de colédoco em adolescente

Spontaneous perforation of the common bile duct is rare. It happens predominantly in children and it is related to obstructive disease of the biliary tract. We present a case of an 18 year-old male patient, with ulcerative rectocolitis associated with malignant tumor of the head of pancreas. The patient developed an acute abdomen syndrome and laparotomy, a spontaneous perforation of common bile duct was evidenced. The authors make a revision of the clinical aspects of that pathology

Viral infections in workers in hospital and research laboratory settings: a comparative review of infection modes and respective biosafety aspects

Objectives: To compare modes and sources of infection and clinical and biosafety aspects of accidental viral infections in hospital workers and research laboratory staff reported in scientific articles. Methods: PubMed, Google Scholar, ISI Web of Knowledge, Scirus, and Scielo were searched (to December 2008) for reports of accidental viral infections, written in English, Portuguese, Spanish, or German; the authors' personal file of scientific articles and references from the articles retrieved in the initial search were also used. Systematic review was carried out with inclusion criteria of presence of accidental viral infection's cases information, and exclusion criteria of absence of information about the viral etiology, and at least probable mode of infection.Results: One hundred and forty-one scientific articles were obtained, 66 of which were included in the analysis. For arboviruses, 84% of the laboratory infections had aerosol as the source; for alphaviruses alone, aerosol exposure accounted for 94% of accidental infections. of laboratory arboviral infections, 15.7% were acquired percutaneously, whereas 41.6% of hospital infections were percutaneous. For airborne viruses, 81% of the infections occurred in laboratories, with hantavirus the leading causative agent. Aerosol inhalation was implicated in 96% of lymphocytic choriomeningitis virus infections, 99% of hantavirus infections, and 50% of coxsackievirus infections, but infective droplet inhalation was the leading mode of infection for severe acute respiratory syndrome coronavirus and the mucocutaneous mode of infection was involved in the case of infection with influenza B. For blood-borne viruses, 92% of infections occurred in hospitals and 93% of these had percutaneous mode of infection, while among laboratory infections 77% were due to infective aerosol inhalation. Among blood-borne virus infections there were six cases of particular note: three cases of acute hepatitis following hepatitis C virus infection with a short period of incubation, one laboratory case of human immunodeficiency virus infection through aerosol inhalation, one case of hepatitis following hepatitis G virus infection, and one case of fulminant hepatitis with hepatitis B virus infection following exposure of the worker's conjunctiva to hepatitis B virus e antigen-negative patient saliva. of the 12 infections with viruses with preferential mucocutaneous transmission, seven occurred percutaneously, aerosol was implicated as a possible source of infection in two cases, and one atypical infection with Macacine herpesvirus 1 with fatal encephalitis as the outcome occurred through a louse bite. One outbreak of norovirus infection among hospital staff had as its probable mode of infection the ingestion of inocula spread in the environment by fomites.Conclusions: The currently accepted and practiced risk analysis of accidental viral infections based on the conventional dynamics of infection of the etiological agents is insufficient to cope with accidental viral infections in laboratories and to a lesser extent in hospitals, where unconventional modes of infection are less frequently present but still have relevant clinical and potential epidemiological consequences. Unconventional modes of infection, atypical clinical development, or extremely severe cases are frequently present together with high viral loads and high virulence of the agents manipulated in laboratories. In hospitals by contrast, the only possible association of atypical cases is with the individual resistance of the worker. Current standard precaution practices are insufficient to prevent most of the unconventional infections in hospitals analyzed in this study; it is recommended that special attention be given to flaviviruses in these settings. (C) 2011 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Advances in sickle cell disease treatment: From drug discovery until the patient monitoring

Sickle Cell Disease (SCD) is one of the most prevalent hematological diseases in the world. Despite the immense progress in molecular knowledge about SCD in last years few therapeutical sources are currently available. Nowadays the treatment is performed mainly with drugs such as hydroxyurea or other fetal hemoglobin inducers and chelating agents. This review summarizes current knowledge about the treatment and the advancements in drug design in order to discover more effective and safe drugs. Patient monitoring methods in SCD are also discussed. © 2011 Bentham Science Publishers Ltd.

Evaluación preliminar del impacto en México de la influenza AH1N1: documento elaborado por el equipo conjunto CEPAL

Incluye Bibliografía

Estratificação de risco e desfechos clínicos em pacientes com síndrome coronariana aguda em classe funcional de Killip I ou II

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Obstrução experimental de jejuno em eqüinos: efeito da hidrocortisona nos parâmetros clínicos e laboratoriais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência do metapneumovírus humano nas infecções respiratórias agudas em estados da região nordeste do Brasil

As doenças do trato respiratório são responsáveis por uma significativa taxa de absenteísmo laboral bem como por elevados índices de morbidade e morte, entre as quais as infecções respiratórias aguda (IRA) representam as maiores queixas nos serviços de atendimento médico-ambulatorial em todo o mundo. Os vírus são considerados os principais agentes etiológicos das IRA, atuando seja como patógeno principal ou predispondo às infecções bacterianas secundárias, entre eles encontra-se o Metapneumovirus Humano (HMPV). Este vírus foi identificado em 2001 apresentando-se como um importante agente causador de IRA adquirida na comunidade. É um vírus cosmopolita que causa doenças respiratórias semelhantes ao Vírus Respiratório Sincicial. No Brasil, são relativamente escassos os relatos da ocorrência do HMPV na população. O objetivo deste estudo é investigar a ocorrência de Metapneumovírus Humano (HMPV) em pacientes com diagnóstico clínico de infecção respiratória aguda (IRA) na Região Nordeste do Brasil. Entre o período de Junho de 2009 a Setembro de 2010, pacientes oriundos de atendimentos em unidades de atenção básica ou hospitalar de cinco estados da Região Nordeste, foram submetidos a coleta de espécimes para detecção a partir de técnicas de biologia molecular. Análises estatísticas foram utilizadas para escolha do tamanho amostral (545) e tratamento dos resultados obtidos. O estudo mostrou uma positividade de 4.7% para HMPV, sem a existência de uma faixa etária específica para a ocorrência da infecção. Ocorreu uma prevalência do sexo feminino entre os casos positivos, entretanto, sem significado estatístico. O pico de positividade para o vírus (n=16) mostrou existir no terceiro trimestre do ano em todos os Estados investigados. Neste estudo, foi possível descrever a ocorrência de HMPV na Região Nordeste, afetando pacientes portadores de infecção respiratória aguda, tanto acompanhados ambulatorialmente como hospitalizados, que preencheram critério clínico para Síndrome Respiratória Aguda Grave.

Caracterização dos genes codificadores da hemaglutinina e PB2 do vírus Influenza A (H1N1) pandêmico isolado na mesorregião metropolitana de Belém

A recente pandemia de gripe de 2009/2010 causada pelo vírus A (H1N1) pandêmico mostrou um perfil de gravidade diferente da gripe sazonal, pois um percentual considerável de casos graves e fatais ocorreu em indivíduos adultos jovens, sem comorbidade. A virulência dos vírus Influenza A (H1N1) pandêmico resulta de interações protéicas complexas e depende essencialmente de alguns genes virais. O objetivo deste estudo foi caracterizar os genes codificadores da hemaglutinina (H1) e polimerase básica 2 (PB2) do vírus Influenza A (H1N1) pandêmico mediante a obtenção de cepas provenientes de pacientes com gripe procedente da mesorregião metropolitana de Belém-PA. O tamanho amostral foi constituído de 87 amostras aleatórias de ambos os sexos de 0 a 96 anos, com síndrome respiratória aguda grave (SRAG) sem nenhuma comorbidade relatada, no período de maio de 2009 a agosto de 2010. As amostras foram isoladas em cultura de célula MDCK e analisadas por técnicas de biologia molecular que compreenderam três etapas principais: a) extração do RNA viral (RNAv) a partir do sobrenadante celular; b) amplificação do RNAv pela técnica de Reação em Cadeia mediada pela Polimerase precedida de Transcrição Reversa (RT-PCR); c) sequenciamento completo dos genes codificadores da H1 e PB2. Das 87 cepas amplificadas pelo RT-PCR, em 82 tornou-se possível a obtenção e análise de sequências para o gene HA, enquanto que de 81 amostras virais obteve-se sequências para o gene PB2. A análise comparativa das sequências obtidas com a sequência da cepa vacinal (A/California/07/2009(H1N1)) revelou substituições aminoacídicas na HA (P83S; D97N; S203T; D222G; Q293H e I321V) e na PB2 (K340N; K526R e M631L), no entanto sem associação a hospitalização. Ao nível de substituição na HA, a D97N isolada ou associada com a S203T, foi detectada com mais frequência na primeira onda. Já ao nível da PB2 a substituição K526R foi mais encontrada em cepas que circularam na primeira onda, enquanto que, a M631L foi mais evidenciada na segunda. A substituição D222G na HA só foi encontrada em casos de óbitos. Por fim, observou-se uma tendência de alterações nos sítios antigênicos da HA. Sendo assim, a contínua vigilância genética e antigênica do vírus Influenza A (H1N1) pdm em circulação, bem como o compartilhamento de informações é de extrema importância para a melhor recomendação possível para os vírus que entram na composição vacinal evitando assim maior risco de epidemias severas no futuro.

Avaliação de neurotoxicidade de formas enantioméricas de praguicidas organifosforados: estudos in vitro, in vivo e de neuroproteção

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Determinantes genéticos, bioquímicos e clínicos na resposta ao uso de hidroxiureia na doença falciforme

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Farmacocinética e farmacodinâmica de derivado ftalimídico planejado para o tratamento da anemia falciforme

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)