RESUMO
Purpose Cerebral venous sinus thrombosis (CVST) is an unusual and potentially life-threatening condition with variable and nonspecific clinical symptoms and high morbimortality rates. Standard therapy consists of systemic anticoagulation; although there is no clear evidence about the best choice for treatment, intravenous heparin is used as the first-line treatment modality. Intravenous sinus thrombolysis can be an effective and relatively safe treatment for acutely deteriorating patients who have not responded to conventional therapy. This case report presents the possibility of endovascular treatment in multiple steps with mechanical thrombolysis with balloon, local pharmacological thrombolysis and stenting, in a patient with a severe form of CVST. Case summary A 67-year-old woman presented severe headache, agitation and confusion with diagnosis of venous sinus dural thrombosis in both lateral sinus and torcula. After 24 h there was neurological worsening evolving with seizures and numbness even after starting heparin, without sinus recanalization; CT scan showed left temporal intracerebral hemorrhage. We decided to take an endovascular approach in multiple steps. The first step was mechanical static thrombolysis with balloon; the second step was dynamic mechanical thrombolysis with a balloon partially deflated and "pulled"; the third step was local thrombolysis with Actilyse™; finally, the fourth step was angioplasty and reconstruction of the sinuses using multiple carotid stents and complete angiographic recanalization of both sinuses and torcula. After 24 h of endovascular treatment there was full clinical recovery and no tomographic complications. Conclusion This result shows that mechanical clot disruption, intrasinus thrombolysis and reconstruction of wall sinuses with stenting can be an endovascular option in the severe form of CVST with intracerebral hemorrhage and rapid worsening of neurological symptoms. Although this type of treatment can re-channel the occluded sinuses, further comparative and randomized studies are needed to clarify its efficacy versus other therapeutic modalities.
Assuntos
Trombólise Mecânica , Trombose dos Seios Intracranianos/diagnóstico por imagem , Trombose dos Seios Intracranianos/terapia , Stents , Terapia Trombolítica , Idoso , Angiografia Digital , Angiografia Cerebral , Diagnóstico Diferencial , Feminino , Escala de Coma de Glasgow , Humanos , Tomografia Computadorizada por Raios XRESUMO
INTRODUCTION: There has been a great improvement in transplantation medicine in Brazil in the last 2 decades. However, there remain several barriers regarding notification of brain and cardiac death as well as completion of the donation process. METHODS: This retrospective study was performed between January 2008 and December 2010. We reviewed all deaths in a University Hospital, observing the causes of non-notification to the State Transplantation Authority and non-donations. RESULTS: There were 41 notifications of brain death resulting in donation in only 19.5% of those cases. Cardiac death was diagnosed in 21 patients, resulting in 52.4% donations. The main cause for non-donation were family refusal (37.2%), infectious diseases (30.2%), and clinical contraindications (32.6%). Most of the missed possible donors occurred during the night (54.8%) and in the emergency room (80.9%). CONCLUSION: There is an urgent need for better education of the Brazilian population about organ donation and brain death definitions. Other identified problems include lack of uniformity in brain death determinations among hospitals, rigid contraindications to donation in the State of Parana, physician unawareness or disbelief about brain death diagnostic criteria, and lack of structure of our Hospital.
Assuntos
Morte Encefálica/diagnóstico , Seleção do Doador , Família/psicologia , Consentimento do Representante Legal , Obtenção de Tecidos e Órgãos , Altruísmo , Atitude do Pessoal de Saúde , Conscientização , Brasil , Causas de Morte , Doenças Transmissíveis/mortalidade , Serviço Hospitalar de Emergência , Doações , Conhecimentos, Atitudes e Prática em Saúde , Hospitais Universitários , Humanos , Motivação , Papel do Médico , Estudos Retrospectivos , Fatores de TempoRESUMO
The weight-of-evidence - WOE approach was used to assess the environmental impact of sediments contaminated by metals in the São Francisco river and one of its tributaries, Consciência creek, both affected by anthropic activities, in the region of Três Marias (Minas Gerais/Brazil). The assessment provided support to a risk management decision. The WOE was based on bulk metal analysis, AVS-SEM assays, elutriate tests, ecotoxicity assays, benthic community assessment and a comparison for the reference area. Brazilian legislation and other available literature were used as criteria to evaluate the lines of evidence. All samples, except for the reference area, presented some contamination. However, geochemical testing for bioavailability studies showed that toxicity is unlikely as suggested by the chemical results. Ecotoxicity and benthic structure studies provided further information to support decision making. Metal acid volatile sulfide formation mechanisms were identified, which can eventually attenuate metal toxicity observed. The removal of active sources of contamination (for example, from tailings dumps) associated with Monitoring Natural Recovery could be sufficient to eventually lessen the risk of the biota in São Francisco river sediments.
Esse artigo teve como objetivo avaliar o impacto da contaminação por metais dos sedimentos de corrente do Rio São Francisco e de um de seus tributários, Córrego Consciência, na região de Três Marias (Minas Gerais/Brasil), ambos impactados por atividades antrópicas, a partir da metodologia de Balanço das Evidências - BDE, e, dessa forma, prover embasamento para uma decisão de gerenciamento de risco. Para o BDE se utilizou resultados de: concentração de metal nos sedimentos, ensaios de SVA-MES, ensaios de elutriação, ensaios de ecotoxicidade, estudo da comunidade bentônica. Legislação nacional e outras literaturas disponíveis foram utilizadas como critério de avaliação das linhas de evidências. Todas as amostras, exceto pela área de referência, apresentou alguma contaminação. Entretanto, ensaios geoquímicos para estudo da biodisponibilidade demonstraram que a toxicidade não é provável como sugeriram os resultados químicos. Estudos de ecotoxicidade e de estrutura da comunidade bentônica forneceram informações complementares importantes para embasar a tomada de decisão quanto ao gerenciamento ambiental dos sedimentos. Mecanismos de formação de metais volatilizáveis por acidificação foram identificados, podendo atenuar a toxicidade eventualmente verificada. A remoção de fontes ativas de contaminação associada ao Monitoramento da Recuperação Natural pode, portanto, ser suficiente para diminuir o risco à biota, eventualmente identificado nos sedimentos do Rio São Francisco.
Assuntos
Poluição da Água/análise , Fauna Bentônica/análise , Sedimentos , Metais/toxicidadeRESUMO
A new tool called System for Automated Bacterial Integrated Annotation--SABIA (SABIA being a very well-known bird in Brazil) was developed for the assembly and annotation of bacterial genomes. This system performs automatic tasks of assembly analysis, ORFs identification/analysis, and extragenic region analyses. Genome assembly and contig automatic annotation data are also available in the same working environment. The system integrates several public domains and newly developed software programs capable of dealing with several types of databases, and it is portable to other operational systems. These programs interact with most of the well-known biological database/softwares, such as Glimmer, Genemark, the BLAST family programs, InterPro, COG, Kegg, PSORT, GO, tRNAScan and RBSFinder, and can also be used to identify metabolic pathways
Assuntos
Biologia Computacional/métodos , Chromobacterium/genética , Bases de Dados Genéticas , Genoma Bacteriano , Software , Brasil , Biologia Computacional/instrumentaçãoRESUMO
Characterization of enterotoxigenic Escherichia coli (ETEC) has been based almost exclusively on the detection of phenotypic traits such as serotypes and virulence-associated factors: heat-labile (LT) and heat-stable (ST) toxins and colonization factors (CFs). In the present work we show that the analysis of band patterns generated by randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE) of digested chromosomal DNA can be used to detect genetic diversity among ETEC strains expressing identical phenotypic traits. The study included 29 ETEC isolates from Latin America and Spain expressing the phenotype O153:H45 CFA/I ST plus 1 rough derivative, 2 nonmotile derivatives, and 1 O78:H12 CFA/I ST isolate, and a representative of a genetically distinct ETEC group. The results showed that the O153:H45 CFA/I ST ETEC isolates belong to a single clonal cluster whose isolates share on average, 84% of the RAPD bands and 77% of the PFGE restriction fragments, while the O78:H12 isolate shared only 44 and 4% of the RAPD bands and PFGE fragments, respectively, with the isolates of the O153:H45 group. More relevantly, RAPD and PFGE fingerprints disclosed the presence of different clonal lineages among the isolates of the O153:H45 cluster. Some of the genetic variants were isolated from defined geographic areas, while places like São Paulo City in Brazil and the middle-eastern part of Argentina were populated by several genetic variants of related, but not identical, ETEC strains. These results show that molecular biology-based typing methods can disclose strain diversity, which is usually missed in studies restricted to phenotypic typing of ETEC.
Assuntos
Técnicas de Tipagem Bacteriana , Infecções por Escherichia coli/microbiologia , Escherichia coli/classificação , Escherichia coli/genética , Proteínas de Fímbrias , Variação Genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado , Enterotoxinas/metabolismo , Escherichia coli/patogenicidade , Proteínas de Escherichia coli , Humanos , Fenótipo , Sorotipagem , VirulênciaRESUMO
The CFA/I fimbria promotes the attachment of enterotoxigenic Escherichia coli (ETEC) to the surface of human enterocytes. The generation of a protective immune response requires the induction of antibodies able to block the CFA/I-mediated binding of ETEC to receptors located on the small intestine epithelium or on the surface of human red blood cells, in hemagglutination tests. An eukaryotic expression plasmid, pBLCFA, encoding the CFA/I gene under the control of the human cytomegalovirus major immediate-early promoter was constructed as a prototype DNA vaccine against ETEC. pBLCFA-tranfected BHK-21 cells secreted a peptide cross-reacting with a monoclonal antibody raised against CFA/I subunits. BALB/c mice immunized intramuscularly with one or two doses of purified pBLCFA developed CFA/I-specific serum antibodies for at least 52 weeks, composed predominantly of the IgG1 subclass. pBLCFA-induced antibodies bind mainly to epitopes exposed on the surface of intact CFA/I fimbriae and do not react with immune recessive epitopes found in other ETEC fimbra sharing amino acid homologies with CFA/I. Furthermore, pBLCFA-induced antibodies were able to block the adhesive properties of the CFA/I fimbriae, as evaluated by the ability to inhibit the hemagglutination promoted by CFA/I-expressing ETEC cells. These results suggest that secretion of CFA/I encoded by pBLCFA preserves important conformational epitopes required for the generation of protective antibodies against the adhesive properties of the CFA/I fimbriae and open new perspectives for the development of DNA vaccines against enteric bacterial pathogens.
Assuntos
Proteínas de Bactérias/imunologia , Vacinas Bacterianas/farmacologia , Escherichia coli/imunologia , Proteínas de Fímbrias , Fímbrias Bacterianas/imunologia , Vacinas de DNA/farmacologia , Animais , Anticorpos Antibacterianos/sangue , Especificidade de Anticorpos , Antígenos de Bactérias/genética , Aderência Bacteriana/imunologia , Proteínas de Bactérias/genética , Vacinas Bacterianas/genética , Sequência de Bases , Linhagem Celular , Cricetinae , Primers do DNA/genética , Epitopos , Escherichia coli/genética , Fímbrias Bacterianas/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Transfecção , Vacinas de DNA/genéticaRESUMO
Systemic and mucosal antibody responses against both the major subunit of colonization factor antigen I (CFA/I) of enterotoxigenic Escherichia coli (ETEC) and the somatic lipopolysaccharide expressed by recombinant bivalent Salmonella vaccine strains were significantly enhanced by coadministration of a detoxified derivative with preserved adjuvant effects of the ETEC heat-labile toxin, LT((R192G)). The results further support the adjuvant effects of LT((R192G)) and represent a simple alternative to improve responses against passenger antigens expressed by orally delivered Salmonella vaccine strains.
Assuntos
Adjuvantes Imunológicos/farmacologia , Anticorpos Antibacterianos/biossíntese , Toxinas Bacterianas/farmacologia , Enterotoxinas/farmacologia , Proteínas de Escherichia coli , Escherichia coli/imunologia , Proteínas de Fímbrias , Imunidade nas Mucosas , Animais , Antígenos de Bactérias/administração & dosagem , Antígenos de Bactérias/genética , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/genética , Enterotoxinas/genética , Escherichia coli/genética , Lipopolissacarídeos/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Salmonella typhimurium/genética , Salmonella typhimurium/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genéticaRESUMO
Cooperativity in the interactions among proteins subunits and DNA is crucial for DNA recognition. LexA repressor was originally thought to bind DNA as a monomer, with cooperativity leading to tighter binding of the second monomer. The main support for this model was a high value of the dissociation constant for the LexA dimer (micromolar range). Here we show that the protein is a dimer at nanomolar concentrations under different conditions. The reversible dissociation of LexA dimer was investigated by the effects of hydrostatic pressure or urea, using fluorescence emission and polarization to monitor the dissociation process. The dissociation constant lies in the picomolar range (lower than 20 pM). LexA monomers associate with an unusual large volume change (340 ml/mol), indicating the burial of a large surface area upon dimerization. Whereas nonspecific DNA has no stabilizing effect, specific DNA induces tightening of the dimer and a 750-fold decrease in the K(d). In contrast to the previous model, a tight dimer rather than a monomer is the functional repressor. Accordingly, the LexA dimer only loses its ability to recognize a specific DNA sequence by RecA-induced autoproteolysis. Our work provides insights into the linkage between protein-protein interactions, DNA recognition, and DNA repair.
Assuntos
Proteínas de Bactérias/química , Serina Endopeptidases/química , Proteínas de Bactérias/metabolismo , Sequência de Bases , Cromatografia em Gel , DNA/metabolismo , Dimerização , Eletroforese em Gel de Poliacrilamida , Serina Endopeptidases/metabolismo , Soluções , Espectrometria de Fluorescência , TermodinâmicaRESUMO
MOTIVATION: The importance of the various kinds of repetitive nucleotide sequences for the workings of bacterial DNA has been widely recognized. This work is concerned with the distribution of a particular group of repetitive sequences, the short-sequenced interrupted extragenic palindromes, on the genetic maps of Escherichia coli K-12, Haemophilus influenzae Rd and Neisseria meningitidis Z2491 and MC58. A tool has been developed based upon a statistical hypothesis test taking into account the markovian structure of random sequences in order to determine the non-random character of extragenic palindromes. RESULTS: Totals of 7631, 12904, 4722 and 5477 non-random short interrupted palindromes have been found on the E.coli, H.influenzae, and N.meningitidis serogroup A and serogroup B genomes, respectively. Their distribution patterns on the respective genomes vary according to the bacterial species considered. Based on their position on the genome, palindromes could be distinguished as those which integrate longer, repetitive sequences; those which stand in isolation, and still others are associated to specific genome sites. AVAILABILITY: The complete list of the observed palindromes is available at the site http://www/lncc.br/~atrv. CONTACT: atrv@lncc.br
Assuntos
DNA Bacteriano/genética , Escherichia coli/genética , Haemophilus influenzae/genética , Neisseria meningitidis/genética , Biologia Computacional , Técnicas Genéticas , Genoma Bacteriano , Análise de Regressão , Sequências Repetitivas de Ácido NucleicoRESUMO
The colonization factor antigen I (CFA/I) is one of the most epidemiologically relevant enterotoxigenic Escherichia coli (ETEC) fimbrial adhesins, which mediates the binding to human small intestine epithelium. A recombinant eukaryotic expression plasmid, pRECFA, encoding the CFA/I protein fused to the glycoprotein D of herpes simplex type 1 virus, was used to generate an antibody response in a murine model following intramuscular inoculation of purified DNA. Eukaryotic cells (BHK-21) transfected with pRECFA expressed the CFA/I protein in vitro, as revealed by Western blot and immunofluorescence microscopy. Administration of a single pRECFA 100-microg dose induced a long-term CFA/I-specific antibody response in BALB/c mice composed mainly of IgG and, to a lesser extent, IgA isotypes. The major CFA/I-specific IgG subclass was IgG2a, suggesting a Th-1-type immune response. A second dose with the same amount of purified DNA, given 2 weeks later, caused a booster effect on the immunoglobulin levels, but did not qualitatively alter the isotypes and subclasses of the induced antibody response. Immunization with different amounts of purified DNA and/or number of doses showed that maximal transient CFA/I-specific antibody levels could be obtained after two 100-microg doses of pRECFA given 2 weeks apart, but long-term antibody levels were similar.