Hydroxamic acid derivatives: a promising scaffold for rational compound optimization in Chagas disease.

This work describes the antitrypanocidal activity of two hydroxamic acid derivatives containing o-ethoxy (HAD1) and p-ethoxy (HAD2) as substituent in the aromatic ring linked to the isoxazoline ring. HAD1 and HAD2 induced a significant reduction in the number of intracellular parasites and consequently showed activity on the multiplication of the parasite. Treatment of cardiomyocytes and macrophages with the compounds revealed no significant loss in cell viability. Ultrastructural alterations after treatment of cardiomyocytes or macrophages infected by Trypanosoma cruzi with the IC50 value of HAD1 revealed alterations to amastigotes, showing initial damage seen as swelling of the kinetoplast. This gave a good indication of the ability of the drug to permeate through the host cell membrane as well as its selectivity to the parasite target. Both compounds HAD1 and 2 were able to reduce the cysteine peptidases and decrease the activity of metallopeptidases

4-aminopyridyl-based lead compounds targeting CYP51 prevent spontaneous parasite relapse in a chronic model and improve cardiac pathology in an acute model of Trypanosoma cruzi infection.

BackgroundChagas disease, caused by the protozoan Trypanosoma cruzi, is the leading cause of heart failure in Latin America. The clinical treatment of Chagas disease is limited to two 60 year-old drugs, nifurtimox and benznidazole, that have variable efficacy against different strains of the parasite and may lead to severe side effects. CYP51 is an enzyme in the sterol biosynthesis pathway that has been exploited for the development of therapeutics for fungal and parasitic infections. In a target-based drug discovery program guided by x-ray crystallography, we identified the 4-aminopyridyl-based series of CYP51 inhibitors as being efficacious versus T.cruzi in vitro; two of the most potent leads, 9 and 12, have now been evaluated for toxicity and efficacy in mice.Methodology/principal findingsBoth acute and chronic animal models infected with wild type or transgenic T. cruzi strains were evaluated. There was no evidence of toxicity in the 28-day dosing study of uninfected animals, as judged by the monitoring of multiple serum and histological parameters. In two acute models of Chagas disease, 9 and 12 drastically reduced parasitemia, increased survival of mice, and prevented liver and heart injury. None of the compounds produced long term sterile cure. In the less severe acute model using the transgenic CL-Brenner strain of T.cruzi, parasitemia relapsed upon drug withdrawal. In the chronic model, parasitemia fell to a background level and, as evidenced by the bioluminescence detection of T. cruzi expressing the red-shifted luciferase marker, mice remained negative for 4 weeks after drug withdrawal. Two immunosuppression cycles with cyclophosphamide were required to re-activate the parasites. Although no sterile cure was achieved, the suppression of parasitemia in acutely infected mice resulted in drastically reduced inflammation in the heart.Conclusions/significanceThe positive outcomes achieved in the absence of sterile cure suggest that the target product profile in anti-Chagasic drug discovery should be revised in favor of safe re-administration of the medication during the lifespan of a Chagas disease patient. A medication that reduces parasite burden may halt or slow progression of cardiomyopathy and therefore improve both life expectancy and quality of life

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

Background: Leishmania (V.) braziliensis is a causative agent of cutaneous leishmaniasis in Brazil. During the parasite life cycle, the promastigotes adhere to the gut of sandflies, to avoid being eliminated with the dejection. The Lulo cell line, derived from Lutzomyia longipalpis (Diptera: Psychodidae), is a suitable in vitro study model to understand the features of parasite adhesion. Here, we analyze the role of glycosaminoglycans (GAGs) from Lulo cells and proteins from the parasites in this event. Methods: Flagellar (F f) and membrane (Mf) fractions from promastigotes were obtained by differential centrifugation and the purity of fractions confirmed by western blot assays, using specific antibodies for cellular compartments. Heparin-binding proteins (HBP) were isolated from both fractions using a HiTrap-Heparin column. In addition, binding of promastigotes to Lulo cells or to a heparin-coated surface was assessed by inhibition assays or surface plasmon resonance (SPR) analysis. Results: The success of promastigotes subcellular fractionation led to the obtainment of Ff and Mf proteins, both of which presented two main protein bands (65.0 and 55.0kDa) with affinity to heparin. The contribution of HBPs in the adherence of promastigotes to Lulo cells was assessed through competition assays, using HS or the purified HBPs fractions. All tested samples presented a measurable inhibition rate when compared to control adhesion rate (17 ± 2.0% of culture cells with adhered parasites): 30% (for HS 20?g/ml) and 16% (for HS 10μg/ml); HBP Mf (35.2% for 10μg/ml and 25.4% for 20μg/ml) and HBP Ff (10.0% for 10μg/ml and 31.4% for 20μg/ml). Additionally, to verify the presence of sulfated GAGs in Lulo cells surface and intracellular compartment, metabolic labeling with radioactive sulfate was performed, indicating the presence of an HS and chondroitin sulfate in both cell sections. The SPR analysis performed further confirmed the presence of GAGs ligands on L. (V.) braziliensis promastigote surfaces. Conclusions: The data presented here point to evidences that HBPs present on the surface of L. (V.) braziliensis promastigotes participate in adhesion of these parasites to Lulo cells through HS participation. © 2012 de Castro Crtes et al.; licensee BioMed Central Ltd

Quando chega o fim? uma revisão narrativa sobre terminalidade do período escolar para alunos deficientes mentais

The process of inclusion of disabled students is possibly one of the biggest dilemmas faced by professionals in the field of education today. It is necessary to create different educational or assistive devices, due to the termination of the school term, precisely so that future opportunities may be real, and not designed as an exclusionary practice. This article aims to present and understand the concept of specific terminality, allowing reflection on the education of disabled persons. This is a qualitative study, which used the technique of narrative review and methodological contributions. The results indicate the need of effective school termination, which enables social inclusion of disabled people.O processo de inclusão de alunos deficientes possivelmente é um dos maiores impasses enfrentados pelos profissionais da área da educação, na atualidade. Faz-se necessária a criação de diferentes dispositivos educativos ou assistenciais, em face da terminalidade do período escolar, justamente para que as oportunidades futuras possam ser reais, e não concebidas como prática excludente. Este artigo objetivou apresentar e compreender o conceito de terminalidade específica, possibilitando reflexão sobre o processo educativo de pessoas deficientes. Trata-se de um estudo qualitativo, o qual utilizou a técnica de revisão narrativa como aporte metodológico. Os resultados apontam a necessidade da efetivação da terminalidade escolar, que possibilita a inclusão social de pessoas deficientes.El proceso de inclusión de alumnos deficientes posiblemente es uno de los mayores impases enfrentados por los profesionales del área de la educación, en la actualidad. Se hace necesaria la creación de diferentes dispositivos educativos o asistenciales, en faz del fin del período escolar, justamente para que las oportunidades futuras puedan ser reales, y no concebidas como práctica excluyente. Este artículo objetivó presentar y comprender el concepto de fin específico, posibilitando ponderación sobre el proceso educativo de personas deficientes. Se trata de un estudio cualitativo, lo cual utilizó la técnica de revisión narrativa como aporte metodológico. Los resultados apuntan la necesidad de la efectuación del fin escolar, que posibilita la inclusión social de personas deficientes

Peróxido de hidrogênio no controle de patógenos e do escurecimento enzimático de vagem minimamente processada

The objective of this study was to evaluate the efficacy of sanitization with hydrogen peroxide in reducing the enzymatic browning and pathogens of fresh cut snap bean. Fresh cut snap beans were inoculated by immersion in an Escherichia coli and Salmonella enteritidis contaminant solution Then, the vegetable was sanitized with hydrogen peroxide (0; 5; 10 and 20 mL L-1) for 10 minutes. The pathogens were monitored immediately after sanitization and after 4 days of storage at 10ºC. In other similar experiment, but without inoculation of pathogens, the snap bean were evaluated periodically. The surface color, pH, activity of phenylalanine ammonia-lyase, peroxidase and polyphenoloxidase enzymes, chlorophyll and carotenoids contents were evaluated. The sanitization with hydrogen peroxide reduced the count of E. coli and S. enteritidis, being similar for different hydrogen peroxide concentrations. The browning of the section surface of snap bean was controlled by hydrogen peroxide in the concentration of 20 mL L-1. The activity of the enzymes as well as the pigments content (chlorophyll and carotenoids) were not affected by treatments, except the polyphenoloxidase enzyme. The pH increased during storage and was greatest in snap bean sanitized with 20 mL L-1. The sanitization with hydrogen peroxide is effective in reducing population of E.coli and S. enteritidis and minimizing browning in fresh cut snap bean, but caused spots on the vegetable skin, precluding its use.O objetivo deste trabalho foi avaliar a eficácia da sanificação com peróxido de hidrogênio em minimizar o escurecimento enzimático e reduzir patógenos em vagem minimamente processada. Vagens minimamente processadas foram inoculadas por imersão em solução contaminante de Escherichia coli e Salmonella enteritidis e sanificadas com peróxido de hidrogênio (0, 5, 10 e 20 mL L-1) por 10 minutos. A população de patógenos foi monitorada imediatamente após a sanificação e no quarto dia de armazenamento a 10ºC. Em outro experimento similar, porém sem a inoculação dos patógenos, as vagens foram avaliadas periodicamente quanto à: coloração da superfície da casca e corte; pH; atividade das enzimas fenilalanina-amônia-liase, peroxidase e polifenoloxidase; teor de clorofila e carotenoides. A sanificação reduziu a contagem de E. coli e S. enteritidis, sendo essa redução similar nas diferentes concentrações de peróxido. O escurecimento da superfície de corte das vagens foi minimizado pelo peróxido de hidrogênio na concentração de 20 mL L-1. As atividades das enzimas, assim como o teor de clorofila e carotenoides não foram afetados pelos tratamentos, exceto a polifenoloxidase. O pH aumentou durante o armazenamento, sendo maior nas vagens sanitizadas com 20 mL L-1 de peróxido. Conclui-se que a sanificação com peróxido de hidrogênio é eficaz em reduzir a população de E. coli e S. enteritidis bem como minimizar o escurecimento na superfície de corte da vagem, porém ocasiona manchas na casca das vagens minimamente processadas, inviabilizando sua utilização

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

Background: Leishmania (V.) braziliensis is a causative agent of cutaneous leishmaniasis in Brazil. During the parasite life cycle, the promastigotes adhere to the gut of sandflies, to avoid being eliminated with the dejection. the Lulo cell line, derived from Lutzomyia longipalpis (Diptera: Psychodidae), is a suitable in vitro study model to understand the features of parasite adhesion. Here, we analyze the role of glycosaminoglycans (GAGs) from Lulo cells and proteins from the parasites in this event.Methods: Flagellar (F-f) and membrane (M-f) fractions from promastigotes were obtained by differential centrifugation and the purity of fractions confirmed by western blot assays, using specific antibodies for cellular compartments. Heparin-binding proteins (HBP) were isolated from both fractions using a HiTrap-Heparin column. in addition, binding of promastigotes to Lulo cells or to a heparin-coated surface was assessed by inhibition assays or surface plasmon resonance (SPR) analysis.Results: the success of promastigotes subcellular fractionation led to the obtainment of F-f and M-f proteins, both of which presented two main protein bands (65.0 and 55.0kDa) with affinity to heparin. the contribution of HBPs in the adherence of promastigotes to Lulo cells was assessed through competition assays, using HS or the purified HBPs fractions. All tested samples presented a measurable inhibition rate when compared to control adhesion rate (17 +/- 2.0% of culture cells with adhered parasites): 30% (for HS 20 mu g/ml) and 16% (for HS 10 mu g/ml); HBP M-f (35.2% for 10 mu g/ml and 25.4% for 20 mu g/ml) and HBP F-f (10.0% for 10 mu g/ml and 31.4% for 20 mu g/ml). Additionally, to verify the presence of sulfated GAGs in Lulo cells surface and intracellular compartment, metabolic labeling with radioactive sulfate was performed, indicating the presence of an HS and chondroitin sulfate in both cell sections. the SPR analysis performed further confirmed the presence of GAGs ligands on L. (V.) braziliensis promastigote surfaces.Conclusions: the data presented here point to evidences that HBPs present on the surface of L. (V.) braziliensis promastigotes participate in adhesion of these parasites to Lulo cells through HS participation.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Lab Biol Mol & Doencas Endem, BR-21040360 Rio de Janeiro, BrazilLab Ultraestrutura Celular, BR-21040360 Rio de Janeiro, BrazilFiocruz MS, IOC, Lab Bioquim & Fisiol Insetos, BR-21040360 Rio de Janeiro, BrazilFiocruz MS, IPEC, Lab Vigilancia Leishmanioses, BR-21040360 Rio de Janeiro, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Bioquim, São Paulo, BrazilUniv Rosario, Escuela Med, Bogota, DC, ColombiaUniversidade Federal de São Paulo, UNIFESP, Dept Bioquim, São Paulo, BrazilCNPq: 300731/2010-8CNPq: 509737/2010-2CAPES: EDITAL - 11/2009FAPERJ: E-26/103.060/2008FAPERJ: E-26/110.257/2010Web of Scienc

Physiotherapy Care of Patients with Coronavirus Disease 2019 (COVID-19) - A Brazilian Experience

Some patients with coronavirus disease (COVID-19) present with severe acute respiratory syndrome, which causes multiple organ dysfunction, besides dysfunction of the respiratory system, that requires invasive procedures. On the basis of the opinions of front-line experts and a review of the relevant literature on several topics, we proposed clinical practice recommendations on the following aspects for physiotherapists facing challenges in treating patients and containing virus spread: 1. personal protective equipment, 2. conventional chest physiotherapy, 3. exercise and early mobilization, 4. oxygen therapy, 5. nebulizer treatment, 6. noninvasive ventilation and high-flow nasal oxygen, 7. endotracheal intubation, 8. protective mechanical ventilation, 9. management of mechanical ventilation in severe and refractory cases of hypoxemia, 10. prone positioning, 11. cuff pressure, 12. tube and nasotracheal suction, 13. humidifier use for ventilated patients, 14. methods of weaning ventilated patients and extubation, and 15. equipment and hand hygiene. These recommendations can serve as clinical practice guidelines for physiotherapists. This article details the development of guidelines on these aspects for physiotherapy of patients with COVID-19

práticas artísticas no ensino básico e secundário

Educação Artística: integrar a inovação. A educação artística apresenta-se como um território a re-cartografar, numa atualização tão rápida quanto aquela que ocorre no campo artístico. As propostas publicadas neste número 11 da Revista Matéria-Prima trazem essa diversidade de abordagens, com novidades conceptuais que estabelecem as devidas relações entre educação e cidadania, participação, sustentabilidade, cultura visual, e também com alguma atenção sobre os debates pós-coloniais e as questões de género. Os 16 artigos reunidos neste 11º número da Revista Matéria-Prima trazem a realidade operativa quer na formação de professores e quer na formulação dos discursos pedagógicos, suas justificações e suas propostas alternativas.As propostas apresentadas devolvem o debate ao terreno, e alargam-no. Provocam as periferias, convocam abordagens diferenciadas sobre o tema da arte e da educação. Em todas elas a proposta de crescimento através da arte, que hoje implica cada vez mais cidadania, crítica, criatividade, interligação, comprometimento, participação.info:eu-repo/semantics/publishedVersio

Leishmania major: Parasite Interactions Suggesting Sexuality

In five experiments, Leishmania (Leishmania) major (MRHO/SU/59/P-strain) grew poorly when seeded in FYTS medium supplemented with 15% fetal calf serum, but presented several peculiar pairs of promastigotes diametrically opposed and attached at their posterior ends (5.8-13.5%). As seen in Giemsa-stained smears, a ring-like line and/or an enlargement, generally occurred at the parasite junction. A close proximity of nuclei, which sometimes were difficult to distinguish from each other, was also observed at this junction. Several of these pairs appeared to be composed of fused cells in which the nuclei could be apparently fused, as shown by fluorescence microscopy to detect ß-tubulin and DNA, and by scanning electron microscopy. Under other culture conditions these pairs were absent or occurred at very low rates (0.2-2.2%). Such pairs differ markedly from longitudinally dividing cells and resemble those described in two other Leishmania species, as well as in Herpetomonas megaseliae and Phytomonas davidi, suggesting steps of a putative sexual proces