Severity of the cardiac impairment determines whether digitalis prolongs or reduces survival of rats with heart failure due to myocardial infarction

Background: the aim of the present study was to evaluate if the influence of digitalis on survival depends on the severity of cardiac dysfunction in heart failure (HF).Methods and results: Doppler echocardiogram (DE) parameters were analyzed in 84 Wistar control (C) and 80 Wistar rats treated with 0.1 mg/100 g/day of digitoxin (D) five days after coronary occlusion. the de variables correlated with the survival of the animals were: myocardial infarction size, left chamber dimensions, fractional area change and E/A ratio. the animals were observed for up to 280 days. Mortality was worsened in rats in the D group with a myocardial infarction (MI) = 37% and worse de predictors.Conclusion: for the first time our study has shown experimentally that the action of digitalis glycosides can positively or negatively influence survival during treatment of HF. It prolongs survival of those in advanced state and compromises survival when there is less severity of the disease. in fact, the greater benefits occur when digitoxin was used in heightened ventricular dilatations and worse ventricular performance. (c) 2011 Elsevier Ireland Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Div Cardiol, São Paulo, BrazilUniversidade Federal de São Paulo, Div Cardiol, São Paulo, BrazilFAPESP: 09/54225-8CNPq: 300737/2009-2Web of Scienc

Modelo experimental para o estudo de fibras de colágeno no músculo esquelético

PURPOSE: To examine histological and histomorphometric techniques for measuring collagen in skeletal muscle. METHODS: The following staining methods were used in the study: hematoxylin and eosin, Masson's trichrome, reticulin, and picrosirius red, and immunostaining for collagen types I, II, III, IV, and V. Histomorphometric measurements were performed using Corel PhotoPaint and UTHSCSA Image Tool 3.0 software. RESULTS: Both the Masson's trichrome and picrosirius red staining provided the best visualization for the measurement of collagen content. CONCLUSION: This methodology is important for the identification and quantification of the different types of collagen in muscles and can be used in the investigation of the qualitative and quantitative influence of collagen on physical activities, aging, and diseases.OBJETIVO: Examinar os procedimentos empregados na quantificação do colágeno por métodos histológicos e histomorfométricos. MÉTODOS: Foram utilizadas colorações histológicas por HE, Tricrômico de Masson, Reticulina, Picrossírius e reação imuno-histoquímica para colágeno I, II, III, IV e V. A quantificação histomorfométrica foi realizada utilizando-se os programas Corel PhotoPaint e Image Tool versão 3.0. RESULTADOS: Os métodos histológicos de Masson e Picrossírius apresentaram uma maior facilidade na quantificação do colágeno. CONCLUSÃO: Este modelo é importante para que possa ser identificado e quantificado os diferentes tipos de colágenos nos músculos e relacionar com a atividade física, envelhecimento e doenças.UNIFESPUNIFESPSciEL

Signal transduction-related responses to phytohormones and environmental challenges in sugarcane

BACKGROUND: Sugarcane is an increasingly economically and environmentally important C4 grass, used for the production of sugar and bioethanol, a low-carbon emission fuel. Sugarcane originated from crosses of Saccharum species and is noted for its unique capacity to accumulate high amounts of sucrose in its stems. Environmental stresses limit enormously sugarcane productivity worldwide. To investigate transcriptome changes in response to environmental inputs that alter yield we used cDNA microarrays to profile expression of 1,545 genes in plants submitted to drought, phosphate starvation, herbivory and N(2)-fixing endophytic bacteria. We also investigated the response to phytohormones (abscisic acid and methyl jasmonate). The arrayed elements correspond mostly to genes involved in signal transduction, hormone biosynthesis, transcription factors, novel genes and genes corresponding to unknown proteins. RESULTS: Adopting an outliers searching method 179 genes with strikingly different expression levels were identified as differentially expressed in at least one of the treatments analysed. Self Organizing Maps were used to cluster the expression profiles of 695 genes that showed a highly correlated expression pattern among replicates. The expression data for 22 genes was evaluated for 36 experimental data points by quantitative RT-PCR indicating a validation rate of 80.5% using three biological experimental replicates. The SUCAST Database was created that provides public access to the data described in this work, linked to tissue expression profiling and the SUCAST gene category and sequence analysis. The SUCAST database also includes a categorization of the sugarcane kinome based on a phylogenetic grouping that included 182 undefined kinases. CONCLUSION: An extensive study on the sugarcane transcriptome was performed. Sugarcane genes responsive to phytohormones and to challenges sugarcane commonly deals with in the field were identified. Additionally, the protein kinases were annotated based on a phylogenetic approach. The experimental design and statistical analysis applied proved robust to unravel genes associated with a diverse array of conditions attributing novel functions to previously unknown or undefined genes. The data consolidated in the SUCAST database resource can guide further studies and be useful for the development of improved sugarcane varieties

Influence of calcium hydroxide addition to AH Plus sealer on its biocompatibility

Objective. the objective of this study was to determine whether the addition of 5% calcium hydroxide to AH Plus sealer improves its biocompatibility in subcutaneous connective tissue of rats.Study design. Thirty female rats distributed into 3 groups of 10 animals each received subcutaneous dorsal implants of silicone tubes filled with AH Plus (Group 1), AH Plus containing 5% (wt/wt) calcium hydroxide (Group 2), or no material (Group 3: control). the animals were killed after 14 days and the subcutaneous tissue containing the tubes was removed and processed for histological analysis. Biocompatibility was assessed by evaluating the inflammatory response to the implants qualitatively and quantitatively. Data were analyzed statistically by the Wilcoxon and Kruskal-Wallis tests (alpha = 0.05).Results. the area adjacent to the implant was characterized by nonspecific chronic inflammation and was qualitatively similar in the 3 groups. Animals implanted with AH Plus/calcium hydroxide showed significantly less intense inflammatory response when compared with the animals implanted with AH Plus alone.Conclusion. the addition of calcium hydroxide to AH Plus root canal sealer improved its histopathological behavior within 14 days of analysis, producing less severe inflammatory response and less cytotoxicity when implanted in the subcutaneous tissue of rats. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109:e50-e54)Universidade Federal de São Paulo, UNIFESP, Discipline Plast Surg, São Paulo, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Dent Dent Mat & Endodont, Bauru Dent Sch, São Paulo, BrazilUniversidade Federal de São Paulo, UNIFESP, Discipline Plast Surg, São Paulo, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Dent Dent Mat & Endodont, Bauru Dent Sch, São Paulo, BrazilWeb of Scienc

DNA extraction and molecular analysis of non-tumoral liver, spleen, and brain from autopsy samples: the effect of formalin fixation and paraffin embedding

The use of molecular biology in combination with morphological analysis is increasing because of the treatments by target therapies. However, to improve the methods for obtaining DNA for molecular analyses from formalin-fixed, paraffin-embedded (FFPE) tissue is a challenge. the aim of this study was to evaluate the DNA extracted from FFPE tissue blocks (non-tumoral liver, spleen, and brain), obtained from autopsy, 8-24h post mortem, using three methods of DNA extraction. PCR of the beta-actin (136pb) and human amelogenin (AMEL 212-218 bp/106-112 bp) genes, as well as short tandem repeat (STR) (100-400 bp fragments), reported in forensic scientific analysis, was performed to evaluate the effectiveness of the methods of DNA extraction. We used 28 archived (1 and 5 years) and 12 recent autopsy cases. the commercial kit showed reproducible and consistent results in the PCR amplification of the beta-actin and AMEL genes and in analysis by SIR used in forensic analysis. This is the first report using non-tumoral samples from FFPE autopsy tissues, comparing the three most common methods of DNA extraction and using the SIR previously described in forensics. Our study has clarified the challenges for pathologists in applying the molecular biology approach in combination with methods suited for morphology, which must be improved. the data provided here should be used in other molecular studies in FFPE samples. (C) 2012 Elsevier GmbH. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo, Escola Paulista Med, Dept Pathol, EPM UNIFESP, BR-04023062 São Paulo, BrazilInst Criminalist São Paulo, Lab Biol & Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Pathol, EPM UNIFESP, BR-04023062 São Paulo, BrazilFAPESP: 2008/11233-8Web of Scienc

Deletion of AA9 Lytic Polysaccharide Monooxygenases Impacts A. nidulans Secretome and Growth on Lignocellulose

Lytic polysaccharide monooxygenases (LPMOs) are oxidative enzymes found in viruses, archaea, and bacteria as well as eukaryotes, such as fungi, algae and insects, actively contributing to the degradation of different polysaccharides. In Aspergillus nidulans, LPMOs from family AA9 (AnLPMO9s), along with an AA3 cellobiose dehydrogenase (AnCDH1), are cosecreted upon growth on crystalline cellulose and lignocellulosic substrates, indicating their role in the degradation of plant cell wall components. Functional analysis revealed that three target LPMO9s (AnLPMO9C, AnLPMO9F and AnLPMO9G) correspond to cellulose-active enzymes with distinct regioselectivity and activity on cellulose with different proportions of crystalline and amorphous regions. AnLPMO9s deletion and overexpression studies corroborate functional data. The abundantly secreted AnLPMO9F is a major component of the extracellular cellulolytic system, while AnLPMO9G was less abundant and constantly secreted, and acts preferentially on crystalline regions of cellulose, uniquely displaying activity on highly crystalline algae cellulose. Single or double deletion of AnLPMO9s resulted in about 25% reduction in fungal growth on sugarcane straw but not on Avicel, demonstrating the contribution of LPMO9s for the saprophytic fungal lifestyle relies on the degradation of complex lignocellulosic substrates. Although the deletion of AnCDH1 slightly reduced the cellulolytic activity, it did not affect fungal growth indicating the existence of alternative electron donors to LPMOs. Additionally, double or triple knockouts of these enzymes had no accumulative deleterious effect on the cellulolytic activity nor on fungal growth, regardless of the deleted gene. Overexpression of AnLPMO9s in a cellulose-induced secretome background confirmed the importance and applicability of AnLPMO9G to improve lignocellulose saccharification. IMPORTANCE Fungal lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that boost plant biomass degradation in combination with glycoside hydrolases. Secretion of LPMO9s arsenal by Aspergillus nidulans is influenced by the substrate and time of induction. These findings along with the biochemical characterization of novel fungal LPMO9s have implications on our understanding of their concerted action, allowing rational engineering of fungal strains for biotechnological applications such as plant biomass degradation. Additionally, the role of oxidative players in fungal growth on plant biomass was evaluated by deletion and overexpression experiments using a model fungal system

Allo- and autoantibodies in human cardiac allograft recipients

UNIFESP, Escola Paulista Med, BR-04025002 São Paulo, BrazilUNESP, Fac Odontol, Sao Jose Dos Campos, BrazilUNIFESP, Escola Paulista Med, BR-04025002 São Paulo, BrazilWeb of Scienc

Recomendações para a proteção da privacidade do paciente

Resumo A preocupação sobre aspectos bioéticos da privacidade do indivíduo e da privacidade dos dados de seus atendimentos é crescente no meio médico. Processos propedêuticos e terapêuticos atuais requerem envolvimento multidisciplinar de uma série de indivíduos, especialmente em se tratando de internações hospitalares. A transmissão e o armazenamento das informações clínicas e laboratoriais dos pacientes envolvem diferentes mídias, com problemas inerentes de acessibilidade e proteção da informação. Os autores sugerem situações hipotéticas que exemplificam problemas comumente abordados na atuação de comitê de bioética clínica, contextualizando-os no Brasil e no mundo, e sugerindo passos para minimizar potenciais problemas de quebra de privacidade e confidencialidade