Activity of pyridyl-pyrazolone derivatives against Trypanosoma cruzi.

New affordable drugs are needed for the treatment of infection with the protozoan parasite Trypanosoma cruzi responsible for the Chagas disease (CD). Only two old drugs are currently available, nifurtimox and benznidazole (Bz) but they exhibit unwanted side effects and display a weak activity in the late chronic phase of the disease. In this context, we evaluated the activity of a series of aryl-pyrazolone derivatives against T cruzi, using both bloodstream trypomastigote and intracellular amastigote forms of the parasite. The test compounds originate from a series of anticancer agents targeting the immune checkpoint ligand PD-L1 and bear an analogy with known anti-trypanosomal pyrazolones. A first group of 6 phenyl-pyrazolones was tested, revealing the activity of a single pyridyl-pyrazolone derivative. Then a second group of 8 compounds with a common pyridyl-pyrazolone core was evaluated. The in vitro testing process led to the identification of two non-cytotoxic and highly potent molecules against the intracellular form of T. cruzi, with an activity comparable to Bz. Moreover, one compound revealed an activity largely superior to that of Bz against bloodstream trypomastigotes, while being non-cytotoxic (selectivity index >1000). Unfortunately, the compound showed little activity in vivo, most likely due to its very limited plasma stability. However, the study opens novel perspectives for the design of new anti-trypanosomal products and the mechanism of action of the compounds is discussed.

N6-methyltubercidin gives sterile cure in a cutaneous Leishmania amazonensis mouse model.

Leishmania is a trypanosomatid parasite that causes skin lesions in its cutaneous form. Current therapies rely on old and expensive drugs, against which the parasites have acquired considerable resistance. Trypanosomatids are unable to synthesize purines relying on salvaging from the host, and nucleoside analogues have emerged as attractive antiparasitic drug candidates. 4-Methyl-7-ß-D-ribofuranosyl-7H-pyrrolo[2,3-d]pyrimidine (CL5564), an analogue of tubercidin in which the amine has been replaced by a methyl group, demonstrates activity against Trypanosoma cruzi and Leishmania infantum. Herein, we investigated its in vitro and in vivo activity against L. amazonensis. CL5564 was 6.5-fold (P = 0.0002) more potent than milteforan™ (ML) against intracellular forms in peritoneal mouse macrophages, and highly selective, while combination with ML gave an additive effect. These results stimulated us to study the activity of CL5564 in mouse model of cutaneous Leishmania infection. BALB/c female and male mice infected by L. amazonensis treated with CL5564 (10 mg kg−1, intralesional route for five days) presented a >93% reduction of paw lesion size likely ML given orally at 40 mg kg−1, while the combination (10 + 40 mg kg−1 of CL5564 and ML, respectively) caused >96% reduction. The qPCR confirmed the suppression of parasite load, but only the combination approach reached 66% of parasitological cure. These results support additional studies with nucleoside derivatives.

Arylimidamides Have Potential for Chemoprophylaxis against Blood-Transmitted Chagas Disease.

Chagas disease (CD) affects over 6 million people worldwide and can be transmitted iatrogenically. Crystal violet (CV) was previously used for pathogen reduction but has harmful side-effects. In the present study, three arylimidamides (AIAs) and CV were used to sterilize mice blood samples experimentally contaminated with bloodstream trypomastigotes (BT) of Trypanosoma cruzi, at non hemolytic doses. All AIAs were not toxic to mouse blood cells until the highest tested concentration (96 µM). The previous treatment of BT with the AIAs impaired the infection establishment of cardiac cell cultures. In vivo assays showed that pre-incubation of mouse blood samples with the AIAs and CV (96 µM) significantly suppressed the parasitemia peak, but only the AIA DB1831 gave ≥90% animal survival, while vehicle treated samples reached 0%. Our findings support further studies regarding the potential use of AIAs for blood bank purposes.

Overexpression of TcNTPDase-1 Gene Increases Infectivity in Mice Infected with Trypanosoma cruzi.

Ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) are enzymes located on the surface of the T. cruzi plasma membrane, which hydrolyze a wide range of tri-/-diphosphate nucleosides. In this work, we used previously developed genetically modified strains of Trypanosoma cruzi (T. cruzi), hemi-knockout (KO +/−) and overexpressing (OE) the TcNTPDase-1 gene to evaluate the parasite infectivity profile in a mouse model of acute infection (n = 6 mice per group). Our results showed significantly higher parasitemia and mortality, and lower weight in animals infected with parasites OE TcNTPDase-1, as compared to the infection with the wild type (WT) parasites. On the other hand, animals infected with (KO +/−) parasites showed no mortality during the 30-day trial and mouse weight was more similar to the non-infected (NI) animals. In addition, they had low parasitemia (45.7 times lower) when compared with parasites overexpressing TcNTPDase-1 from the hemi-knockout (OE KO +/−) group. The hearts of animals infected with the OE KO +/− and OE parasites showed significantly larger regions of cardiac inflammation than those infected with the WT parasites (p < 0.001). Only animals infected with KO +/− did not show individual electrocardiographic changes during the period of experimentation. Together, our results expand the knowledge on the role of NTPDases in T. cruzi infectivity, reenforcing the potential of this enzyme as a chemotherapy target to treat Chagas disease (CD).

Phenotypic Evaluation of Nucleoside Analogues against Trypanosoma cruzi Infection: In Vitro and In Vivo Approaches.

Chagas disease, caused by Trypanosoma cruzi (T. cruzi), is a serious public health problem. Current treatment is restricted to two drugs, benznidazole and nifurtimox, displaying serious efficacy and safety drawbacks. Nucleoside analogues represent a promising alternative as protozoans do not biosynthesize purines and rely on purine salvage from the hosts. Protozoan transporters often present different substrate specificities from mammalian transporters, justifying the exploration of nucleoside analogues as therapeutic agents. Previous reports identified nucleosides with potent trypanocidal activity; therefore, two 7-derivatized tubercidins (FH11706, FH10714) and a 3'-deoxytubercidin (FH8513) were assayed against T. cruzi. They were highly potent and selective, and the uptake of the tubercidin analogues appeared to be mediated by the nucleoside transporter TcrNT2. At 10 µM, the analogues reduced parasitemia >90% in 2D and 3D cardiac cultures. The washout assays showed that FH10714 sterilized the infected cultures. Given orally, the compounds did not induce noticeable mouse toxicity (50 mg/kg), suppressed the parasitemia of T. cruzi-infected Swiss mice (25 mg/kg, 5 days) and presented DNA amplification below the limit of detection. These findings justify further studies with longer treatment regimens, as well as evaluations in combination with nitro drugs, aiming to identify more effective and safer therapies for Chagas disease.

The Impact of the CTHRSSVVC Peptide Upon Experimental Models of Trypanosoma cruzi Infection.

Chagas disease (CD), caused by the hemoflagellate protozoan Trypanosoma cruzi, affects more than six million people worldwide and presents an unsatisfactory therapy, based on two nitroderivatives, introduced in clinical medicine for decades. The synthetic peptide, with CTHRSSVVC sequence (PepA), mimics the CD163 and TNF-α tripeptide "RSS" motif and binds to atheromatous plaques in carotid biopsies of human patients, spleen tissues, and a low-density lipoprotein receptor knockout (LDLr-/-) mouse model of atherosclerosis. CD163 receptor is present on monocytes, macrophages, and neutrophils, acting as a regulator of acute-phase processes and modulating aspects of the inflammatory response and the establishment of infections. Due to the potential theranostic role of PepA, our aim was to investigate its effect upon T. cruzi infection in vitro and in vivo. PepA and two other peptides with shuffled sequences were assayed upon different binomials of host cell/parasite, including professional [as peritoneal mouse macrophages (PMM)] and non-professional phagocytes [primary cultures of cardiac cells (CM)], under different protocols. Also, their impact was further addressed in vivo using a mouse model of acute experimental Chagas disease. Our in-vitro findings demonstrate that PepA and PepB (the peptide with random sequence retaining the "RS" sequence) reduced the intracellular parasitism of the PMM but were inactive during the infection of cardiac cells. Another set of in-vitro and in-vivo studies showed that they do not display a trypanocidal effect on bloodstream trypomastigotes nor exhibit in-vivo efficacy when administered after the parasite inoculation. Our data report the in-vitro activity of PepA and PepB upon the infection of PMM by T. cruzi, possibly triggering the microbicidal arsenal of the host professional phagocytes, capable of controlling parasitic invasion and proliferation.

7-Aryl-7-deazapurine 3'-deoxyribonucleoside derivative as a novel lead for Chagas' disease therapy: in vitro and in vivo pharmacology.

BACKGROUND: The protozoan Trypanosoma cruzi is auxotrophic for purines and causes Chagas' disease (CD), a neglected illness affecting >6 million people. Combining the 3-deoxyribofuranose part of cordycepin with the modified purine ring of a nucleoside 'hit' led to the discovery of 4-amino-5-(4-chlorophenyl)-N7-(3'-deoxy-ß-d-ribofuranosyl)-pyrrolo[2,3-d]pyrimidine (Cpd1), revealing promising anti-T. cruzi activity. OBJECTIVES: To further evaluate Cpd1 in vitro and in vivo to fully assess its therapeutic potential against CD, covering cell culture sterilization through washout assays, drug combination with benznidazole and long-term administration in T. cruzi-infected mice. RESULTS: Although less susceptible to Cpd1 than amastigotes, trypomastigotes present an impaired capacity to successfully establish intracellular infection of cardiac cultures. Combination of benznidazole with Cpd1 indicated no interaction (additive effect) (FIC index = 0.72) while administration to mice at one-tenth of the optimal dose (2.5 mg/kg and 10 mg/kg for Cpd1 and benznidazole, respectively) suppressed parasitaemia but failed to avoid mortality. Long-term treatment (60 days) gave a rapid drop of the parasitaemia (>98% decline) and 100% mice survival but only 16% cure. In vitro washout experiments demonstrated that although parasite release into the supernatant of infected cardiac cultures was reduced by >94%, parasite recrudescence did occur after treatment. CONCLUSIONS: Parasite recrudescence did occur after treatment corroborating the hypothesis of therapeutic failure due to subpopulations of dormant forms and/or genetic factors in persister parasites involved in natural drug resistance.

Analysis of an NGS retinopathy panel detects chromosome 1 uniparental isodisomy in a patient with RPE65-related leber congenital amaurosis.

Purpose: This study aims to demonstrate the possibility of detecting segmental uniparental isodisomy (iUPD) using a next-generation sequencing gene panel by reporting a Leber congenital amaurosis (LCA) case caused by a homozygous pathogenic variant in RPE65 (c.1022 T > C:p.Leu341Ser) inherited exclusively from the proband's mother.Methods: Samples from the trio (proband, mother, and father) were sequenced with a next-generation sequencing (NGS) retinopathy gene panel (224 genes) and the VCF file containing all variants was used in order to determine single nucleotide variant (SNV) counts from each sample across all chromosomes.Results: Trio analysis showed that of 81 Chr1 inherited variants 41 were exclusively maternal, including 21 homozygous. The other 40 variants were common to both parents. On remaining autosomal chromosomes (Chr2-22) 645 inherited variants were found, 147 of them were exclusively maternal and 132 exclusively paternal. Based on these NGS data, it was possible to note that the proband's chromosomes 1 are more similar to his mother's chromosome 1 than his father's, suggesting the pathogenic homozygous variant found in this patient was inherited exclusively from the mother due to uniparental maternal isodisomy.Conclusions: This study presents a secondary analysis pipeline to identify responsible variants for a phenotype and the correct inheritance pattern, which is a critical step to the proper and accurate genetic counseling of all family members. In addition, this approach could be used to determine iUPD in different Mendelian disorders if the sequencing panel identifies variants spread throughout the genome.

Benznidazole modulates release of inflammatory mediators by cardiac spheroids infected with Trypanosoma cruzi.

Chagas disease (CD) caused by Trypanosoma cruzi remains a serious public health problem in Latin America. The available treatment is limited to two old drugs, benznidazole (Bz) and nifurtimox, which exhibit limited efficacy and trigger side effects, justifying the search for new therapies. Also, more accurate and sensitive experimental protocols for drug discovery programs are necessary to shrink the translational gaps found among pre-clinical and clinical trials. Presently, cardiac spheroids were used to evaluate host cell cytotoxicity and anti-T.cruzi activity of benznidazole, exploring its effect on the release of inflammatory mediators. Bz presented low toxic profile on 3D matrices (LC50 > 200 µM) and high potency in vitro (EC50 = 0.99 µM) evidenced by qPCR analysis of T.cruzi-infected cardiac spheroids. Flow cytometry appraisal of inflammatory mediators released at the cellular supernatant showed increases in IL - 6 and TNF contents (≈190 and ≈ 25-fold) in parasitized spheroids as compared to uninfected cultures. Bz at 10 µM suppressed parasite load (92%) concomitantly decreasing in IL-6 (36%) and TNF (68%). Our findings corroborate the successful use of 3D cardiac matrices for in vitro identification of novel anti-parasitic agents and potential impact in host cell physiology.

Blog no ensino da ressuscitação cardiopulmonar: uma ferramenta para a formação do enfermeiro / Blog en la ensenanza de la reanimación cardioplumonar: una herramienta para la formación en enfermería / Blog in the teaching of cardiopulmonary resuscitation : a tool for nursing training

Objetivo: Desenvolver e validar um blog para o ensino e a aprendizagem da ressuscitação cardiopulmonar do adulto voltado à formação do enfermeiro. Método: Pesquisa aplicada, de produção tecnológica, que abordou o desenvolvimento de um blog sobre a ressuscitação cardiopulmonar do adulto no ambiente intra e extra-hospitalar para enfermagem. Na elaboração, cumpriram-se as fases de análise; design; desenvolvimento, implementação e avaliação do blog. A validação desse ambiente virtual de aprendizagem envolveu 11 profissionais enfermeiros da área de urgência e emergência e três especialistas de informática, totalizando 14 participantes. Resultados: Elaborou-se uma ferramenta virtual pedagógica de ensino e aprendizagem, denominada "Blog da Ressuscitação Cardiopulmonar". A avaliação dos três especialistas em informática abordou os domínios; tempo de resposta, qualidadede, interface e ferramentas e recursos, abrangendo 33 critérios, considerados excelentes pela maioria. Os experts em enfermagem avaliaram 32 critérios entre aspectos educacionais, a interface do ambiente virtual e os recursos didáticos, apontados, predominantemente, como excelentes. Conclusão: Neste estudo, elaborou-se e validou-se um blog para o ensino da ressuscitação cardiopulmonar de adultos, representando um arcabouço de evidências científicas atualizadas, fidedignas, interativas e tecnológicas para a enfermagem, que poderá ser replicado em outros ambientes de aprendizagem.(AU)

Objective: to develop and validate a blog for teaching and learning adult cardiopulmonary resuscitation to nurses. Method: applied research of technological production, which addressed the development of a blog about adult cardiopulmonary resuscitation in the intra and extra-hospital environment focused on nursing. For the elaboration, the analysis phases were accomplished; design; development, implementation and evaluation of the blog. The validation of this virtual learning environment involved 11 professional nurses in the urgency and emergency area and three computer specialists, totaling 14 participants. Results: A virtual teaching and learning tool was developed, named "Blog of Cardiopulmonary Resuscitation". The evaluation of the three computer experts covered the following domains: response time, interface quality and tools and resources, totaling 33 criteria considered excellent by most. The nursing experts evaluated 32 criteria between educational aspects, the interface of the virtual environment and didactic resources, which were predominantly indicated as excellent. Conclusion: This study designed and validated a blog for teaching adult cardiopulmonary resuscitation, representing an updated, reliable, interactive and technological scientific evidence for nursing, which can be replicated in other learning environments.(AU)

Objetivo: desarrollar y validar un blog para enseñar y aprender la reanimación cardiopulmonar de adultos para capacitar a los enfermeros. Método: investigación aplicada, de producción tecnológica, que abordó el desarrollo de un blog sobre reanimación cardiopulmonar para adultos en entorno intra y extrahospitalario, en enfermería. Para la elaboración, se realizaron las fases de análisis, diseño, desarrollo, implementación y evaluación del blog. La validación de este entorno virtual de aprendizaje involucró a 11 enfermeros profesionales del área de urgencias y emergencias y tres especialistas en informática, con un total de 14 participantes. Resultados: se desarrolló una herramienta virtual de enseñanza y aprendizaje, llamada "Blog de reanimación cardiopulmonar". La evaluación de los tres expertos en informática cubrió los dominios; tiempo de respuesta, calidad de interfaz y herramientas y recursos, cubriendo 33 criterios, considerados excelentes por la mayoría. Los expertos en enfermería evaluaron 32 criterios entre los aspectos educativos, la interfaz del entorno virtual y los recursos didácticos, que se indicaron predominantemente como excelentes. Conclusión: este estudio diseñó y validó un blog para la enseñanza de la reanimación cardiopulmonar para adultos, que representa un marco de evidencia científica actualizada, confiable, interactiva y tecnológica para enfermería, que puede replicarse en otros entornos de aprendizaje.(AU)

Efficacy of Novel Pyrazolone Phosphodiesterase Inhibitors in Experimental Mouse Models of Trypanosoma cruzi.

Pyrazolones are heterocyclic compounds with interesting biological properties. Some derivatives inhibit phosphodiesterases (PDEs) and thereby increase the cellular concentration of cyclic AMP (cAMP), which plays a vital role in the control of metabolism in eukaryotic cells, including the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease (CD), a major neglected tropical disease. In vitro phenotypic screening identified a 4-bromophenyl-dihydropyrazole dimer as an anti-T. cruzi hit and 17 novel pyrazolone analogues with variations on the phenyl ring were investigated in a panel of phenotypic laboratory models. Potent activity against the intracellular forms (Tulahuen and Y strains) was obtained with 50% effective concentration (EC50) values within the 0.17 to 3.3 µM range. Although most were not active against bloodstream trypomastigotes, an altered morphology and loss of infectivity were observed. Pretreatment of the mammalian host cells with pyrazolones did not interfere with infection and proliferation, showing that the drug activity was not the result of changes to host cell metabolism. The pyrazolone NPD-227 increased the intracellular cAMP levels and was able to sterilize T. cruzi-infected cell cultures. Thus, due to its high potency and selectivity in vitro, and its additive interaction with benznidazole (Bz), NPD-227 was next assessed in the acute mouse model. Oral dosing for 5 days of NPD-227 at 10 mg/kg + Bz at 10 mg/kg not only reduced parasitemia (>87%) but also protected against mortality (>83% survival), hence demonstrating superiority to the monotherapy schemes. These data support these pyrazolone molecules as potential novel therapeutic alternatives for Chagas disease.

Production and validation of a video lesson on cardiopulmonary resuscitation. / Construção e validação de videoaula sobre ressuscitação cardiopulmonar.

OBJECTIVE: To elaborate and validate a teaching virtual contemporary object, video-lesson, about resuscitation cardiopulmonary with adult in life support care using automatic external defibrillator in the hospital environment. METHOD: This is an applied research of techonlogical productions in witch the video-lesson elaboration was according to the methodological trajectory proposed by Fleming, Reynolds and Wallace. The research was accomplished in the Minas Gerais University State and in the Ribeirão Preto Nursing School (Brazil). Sixteen expertises nurses in the area of urgency and emergency participated of this research. The AC1 Gwet's statistic was used to the interobsevers agreement. RESULTS: The validation of script and storyboard to the video-lesson development was reached the interobsevers agreement, classified as "moderate agreenment" according to Landis and Kock, with AC1=0.59 and p<0.0001. CONCLUSIONS: The video-lesson elaborated and validated in this research represent an adequate contemporary important strategy to aplication in the teaching-learning process.

Symptoms and Vocal Risk Factors in Individuals with High and Low Anxiety.

AIMS: To assess the frequency of vocal symptoms and risk factors in individuals with high and low anxiety and to investigate the relationships among vocal symptoms, risk factors, and anxiety levels. PATIENTS AND METHODS: A total of 201 patients of both sexes with a mean age of 42.88 years (SD = 15.78) served as participants. The patients were divided into two groups according to the level of trait anxiety: a low-anxiety group (n = 108) and a high-anxiety group (HAG, n = 93). Data were collected using three self-assessment instruments: the Vocal Screening Protocol, the Voice Symptom Scale (VoiSS), and the State-Trait Anxiety Inventory (STAI). RESULTS: The HAG presented higher scores in all VoiSS domains. There were positive correlations between the total and emotional scores on the VoiSS and the STAI Trait subscale. There were significant associations between 13 vocal risk factors and anxiety levels. CONCLUSION: Individuals with high anxiety present a higher frequency of vocal symptoms related to either vocal limitation, physical aspects, or the emotional impact of dysphonia. The higher the trait anxiety, the greater the total and emotional scores on the VoiSS. Anxiety levels are associated with certain vocal risk factors.

Blog no ensino da ressuscitação cardiopulmonar: uma ferramenta para a formação do enfermeiro / Cardiopulmonary resuscitation teaching blog: A tool for nurse education

ntrodução: As mídias sociais são constituídas de ferramentas primárias como os blogs que podem ser utilizadas para fins educacionais por permitirem deferentes modalidades de interação, com maior flexibilidade e reflexão, para formação do enfermeiro. Objetivo: Desenvolver e avaliar um blog para o ensino e aprendizagem da ressuscitação cardiopulmonar (RCP) no adulto para formação do enfermeiro Metodologia:Trata-se de uma pesquisa aplicada, de produção tecnológica, que envolve o desenvolvimento de um blog sobre a RCP. O planejamento e o desenvolvimento do conteúdo tiveram como base o design instrucional contextualizado compreendendo cinco fases: análise, design, desenvolvimento, implementação e avaliação. Resultados: Criação do ambiente virtual de aprendizagem denominado Blog da RCP, sob o endereço: www.blogdarcp.com.br. Este ambiente multimídia encontra-se hospedado na locaweb e apresenta textos, hipertextos, recursos de vídeos e imagens que permitem a interação do conteúdo e pode ser acessado por meio de plataformas como smatphones, computadores e tablets. A avaliação foi realizada por três especialistas de informática e onze experts em enfermagem. Os especialistas de informática avaliaram aspectos relacionados ao tempo de resposta, qualidade de interface, ferramentas e recursos. Os experts enfermeiros avaliaram os aspectos educacionais, interface do ambiente e recursos didáticos. Conclusão: O Blog da RCP foi avaliado positivamente pelos especialistas da informática quanto tempo resposta e qualidade de interface; para os experts em enfermagem a avaliação também se demonstrou positiva nos aspectos educacionais e interface de ambiente. O item fórum e chats foi avaliado como insatisfatório, pois a opção discussão que permite comentários não foi selecionado pela web desing na fase construção. O blog é dinâmico e pode ser utilizado como mais um recurso educacional no ensino de enfermagem; é inovador, desperta curiosidades voltadas para o campo científico e abre espaço para informação, reflexão e colaboração entre os usuários e o blogger, propagando boas práticas com base em evidências

Introduction: Social media are made up of primary tools such as blogs and can be used for educational purposes because they allow different forms of interaction, with greater flexibility and reflection, to form opinion. Purpose: Develop and evaluate a blog for teaching and learning adult cardiopulmonary resuscitation (CPR) for nurse education. Methodology: This is an applied research based on technology production which involves the development of a blog about CPR. Content planning and development was based on contextualized instructional design comprising five phases: analysis, design, development, implementation, and evaluation. Results: Creation of the virtual learning environment called RCP Blog, at the address: www.blogdarcp.com.br. This multimedia environment is hosted on locaweb and features text, hypertext, video and image resources that allow the content interaction and can be accessed through platforms such as smartphones, computers, and tablets. The evaluation was performed by three computer specialists and eleven nursing experts. Computer specialists evaluated the aspects related to response time, interface quality, tools, and resources. The nurse experts evaluated the educational aspects, environment interface, and didactic resources. Conclusions: The RCP Blog was positively evaluated by computer experts regarding response time and interface quality; for the nursing experts, the evaluation was also positive in the educational and environmental interface aspects. The item 'Forum and Chats' was evaluated as unsatisfactory because the discussion option that allows comments was not selected by the web design in the construction phase. The blog is dynamic and can be used as another educational resource in nursing education. It is innovative, arouses curiosities focused on the scientific field and opens space for information, reflection, and collaboration among users and the blogger, disseminating good practices based on evidence

Construção e validação de videoaula sobre ressuscitação cardiopulmonar / Elaboración y validación de video-lección acerca de reanimación cardiopulmonar / Production and validation of a video lesson on cardiopulmonary resuscitation

Resumo OBJETIVO Construir e validar um objeto contemporâneo virtual de ensino, videoaula, sobre ressuscitação cardiopulmonar no adulto em suporte básico de vida com o uso do desfibrilador externo automático no ambiente hospitalar. MÉTODOS Trata-se de uma pesquisa aplicada, de produção tecnológica, com produção de vídeoaula de acordo com a trajetória metodológica proposta por Fleming, Reynolds e Wallace. Desenvolvido na Universidade do Estado de Minas Gerais e na Escola de Enfermagem de Ribeirão Preto no período de janeiro de 2017 a março de 2018. Participaram 16 enfermeiros expertises na área de urgência e emergência. Para concordância inter-avaliadores foi utilizado a estatística AC1 de Gwet. RESULTADOS A validação do roteiro/script e storyboard foi alcançada a concordância inter-avaliadores, de acordo com Landis e Kock, classificada em "concordância moderada", com AC1=0,59 e p<0,0001. CONCLUSÕES A videoaula construída e validada neste estudo, representa importante estratégia contemporânea adequada para aplicação no processo de ensino-aprendizagem.

Resumen OBJETIVO Construir y validar un objeto contemporáneo virtual de enseñanza, vídeo-lección, acerca de la reanimación cardiopulmonar con adultos en cuidados para prolongar la vida, utilizando el desfibrilador externo automático en ambiente hospitalario. MÉTODO Se trata de una investigación aplicada, de producción tecnológica en que la elaboración del vídeo-lección se llevó a cabo según la trayectoria metodológica propuesta por Fleming, Reynolds y Wallace. El estudio se desarrolló en la Universidad del Estado de Minas Gerais y en la Escuela de Enfermería de Ribeirão Preto (Brasil). Participaron de esta investigación 16 enfermeros especialistas en el área de urgencia y emergencia. Para la concordancia 'interevaluadores' se usó la estadística AC1 de Gwet. RESULTADOS Se alcanzó la concordancia 'interevaluadores' para el desarrollo del vídeo-lección a través de la validación del script y storyboard que, según Landis y Kock, es clasificada como "concordancia moderada" con AC1=0,59 y p<0,0001. CONCLUSIONES El vídeo-lección, elaborado y validado en este estudio, representa una importante estrategia contemporánea apropiada para la aplicación en el proceso de enseñanza-aprendizaje.

Abstract OBJECTIVE To elaborate and validate a teaching virtual contemporary object, video-lesson, about resuscitation cardiopulmonary with adult in life support care using automatic external defibrillator in the hospital environment. METHOD This is an applied research of techonlogical productions in witch the video-lesson elaboration was according to the methodological trajectory proposed by Fleming, Reynolds and Wallace. The research was accomplished in the Minas Gerais University State and in the Ribeirão Preto Nursing School (Brazil). Sixteen expertises nurses in the area of urgency and emergency participated of this research. The AC1 Gwet's statistic was used to the interobsevers agreement. RESULTS The validation of script and storyboard to the video-lesson development was reached the interobsevers agreement, classified as "moderate agreenment" according to Landis and Kock, with AC1=0.59 and p<0.0001. CONCLUSIONS The video-lesson elaborated and validated in this research represent an adequate contemporary important strategy to aplication in the teaching-learning process.

Construção e validação de questionário para avaliação de conhecimento sobre ressuscitação cardiopulmonar / Construcción y validación de cuestionario para avaliación de conocimiento sobre reanimación cardiopulmonar / Construction and validation of a questionnaire for cardiopulmonary resuscitation knowledge assessment

RESUMO OBJETIVO: construir e validar um questionário sobre Ressuscitação Cardiopulmonar no adulto em Suporte Básico de Vida, com o uso do Desfibrilador Externo Automático, no ambiente hospitalar. METODOLOGIA: pesquisa aplicada, realizada na Escola de Enfermagem de Ribeirão Preto da Universidade de São Paulo, no período de janeiro de 2017 a março de 2018. Composta por 16 experts em Urgência e Emergência, para selecioná-los foi adotado os critérios de Fehring. Foram aplicadas as regras do manual do Conselho Nacional de Examinadores Médicos e diretrizes da American Heart Association. Utilizou-se estatística descritiva e análise de concordância inter-avaliadores por AC1 de Gwet. Validado em relação à organização, objetividade e clareza. RESULTADOS: questionário validado com 20 questões de Múltipla Escolha com concordância inter-avaliadores "quase perfeita". Conclusão: o questionário mostrou-se válido para utilização como instrumento de avaliação sobre o assunto abordado.

RESUMEN OBJETIVO: construir y validar un cuestionario sobre Reanimación de adulto, en Soporte Básico de Vida, con el uso de Desfibrilador Externo Automático, en ambiente hospitalario. METODOLOGÍA: investigación aplicada, realizada en la Escuela de Enfermería de Ribeirao Preto de la Universidad de Sao Paulo, en el período de enero de 2017 a marzo de 2018. Estuvo compuesta por 16 especialistas en Urgencia y Emergencia; para seleccionarlos fueron adoptados los criterios de Fehring. Se aplicaron las reglas del manual del Consejo Nacional de Examinadores Médicos y las directrices de la American Heart Association. Se utilizó la estadística descriptiva y el análisis de concordancia entre evaluadores AC1 de Gwet. El cuestionario fue validado en relación a la organización, objetividad y clareza. RESULTADOS: el cuestionario fue validado con 20 preguntas de Múltiple Elección con concordancia entre evaluadores "casi perfecta". Conclusión: el cuestionario se mostró válido para utilización como instrumento de evaluación sobre el asunto abordado.

ABSTRACT OBJECTIVE: To construct and validate a questionnaire related to adult cardiopulmonary resuscitation in Basic Life Support, using the Automatic External Defibrillator, in the hospital environment. METHODOLOGY: applied study, conducted at the University of São Paulo at Ribeirão Preto College of Nursing, from January 2017 to March 2018. Participants were 16 Urgency and Emergency experts, with Fehring's criteria used to select them. The rules of the National Council of Medical Examiners manual and guidelines of the American Heart Association were applied. Descriptive statistics and inter-rater agreement analysis through Gwet's AC1 were used. The questionnaire was validated in relation to organization, objectivity and clarity. RESULTS: a validated questionnaire with 20 multiple choice questions with "almost perfect" inter-rater agreement was produced. Conclusion: the questionnaire was shown to be valid for use as an assessment instrument on the subject addressed.

Efficacy of sertraline against Trypanosoma cruzi: an in vitro and in silico study.

BACKGROUND: Drug repurposing has been an interesting and cost-effective approach, especially for neglected diseases, such as Chagas disease. METHODS: In this work, we studied the activity of the antidepressant drug sertraline against Trypanosoma cruzi trypomastigotes and intracellular amastigotes of the Y and Tulahuen strains, and investigated its action mode using cell biology and in silico approaches. RESULTS: Sertraline demonstrated in vitro efficacy against intracellular amastigotes of both T. cruzi strains inside different host cells, including cardiomyocytes, with IC50 values between 1 to 10 µM, and activity against bloodstream trypomastigotes, with IC50 of 14 µM. Considering the mammalian cytotoxicity, the drug resulted in a selectivity index of 17.8. Sertraline induced a change in the mitochondrial integrity of T. cruzi, resulting in a decrease in ATP levels, but not affecting reactive oxygen levels or plasma membrane permeability. In silico approaches using chemogenomic target fishing, homology modeling and molecular docking suggested the enzyme isocitrate dehydrogenase 2 of T. cruzi (TcIDH2) as a potential target for sertraline. CONCLUSIONS: The present study demonstrated that sertraline had a lethal effect on different forms and strains of T. cruzi, by affecting the bioenergetic metabolism of the parasite. These findings provide a starting point for future experimental assays and may contribute to the development of new compounds.

Efficacy of sertraline against Trypanosoma cruzi: an in vitro and in silico study

Background: Drug repurposing has been an interesting and cost-effective approach, especially for neglected diseases, such as Chagas disease. Methods: In this work, we studied the activity of the antidepressant drug sertraline against Trypanosoma cruzi trypomastigotes and intracellular amastigotes of the Y and Tulahuen strains, and investigated its action mode using cell biology and in silico approaches. Results: Sertraline demonstrated in vitro efficacy against intracellular amastigotes of both T. cruzi strains inside different host cells, including cardiomyocytes, with IC50 values between 1 to 10 M, and activity against bloodstream trypomastigotes, with IC50 of 14 M. Considering the mammalian cytotoxicity, the drug resulted in a selectivity index of 17.8. Sertraline induced a change in the mitochondrial integrity of T. cruzi, resulting in a decrease in ATP levels, but not affecting reactive oxygen levels or plasma membrane permeability. In silico approaches using chemogenomic target fishing, homology modeling and molecular docking suggested the enzyme isocitrate dehydrogenase 2 of T. cruzi (TcIDH2) as a potential target for sertraline. Conclusions: The present study demonstrated that sertraline had a lethal effect on different forms and strains of T. cruzi, by affecting the bioenergetic metabolism of the parasite. These findings provide a starting point for future experimental assays and may contribute to the development of new compounds.(AU)

New 1,2,3-triazole-based analogues of benznidazole for use against Trypanosoma cruzi infection: In vitro and in vivo evaluations.

Chagas disease has spread throughout the world mainly because of the migration of infected individuals. In Brazil, only benznidazole (Bnz) is used; however, it is toxic and not active in the chronic phase, and cases of resistance are described. This work aimed at the synthesis and the trypanocidal evaluation in vitro and in vivo of six new Bnz analogues (3-8). They were designed by exploring the bioisosteric substitution between the amide group contained in Bnz and the 1,2,3-triazole ring. All the compounds were synthesized in good yields. With the exception of compound 7, the in vitro biological evaluation shows that all Bnz analogues were active against the amastigote form, whereas only compounds 3, 4, 5, and 8 were active against trypomastigote. Compounds 4 and 5 showed the most promising activities in vitro against the form of trypomastigote, being more active than Bnz. In vivo evaluation of compounds, 3-8 showed lower potency and higher toxicity than Bnz. Although the 1,2,3-triazole ring has been described in the literature as an amide bioisostere, its substitution here has reduced the activity of the compounds and made them more toxic. Thus, further molecular optimization could provide novel therapeutic agents for Chagas' disease.

Identification of Pyrazolo[3,4-e][1,4]thiazepin based CYP51 inhibitors as potential Chagas disease therapeutic alternative: In vitro and in vivo evaluation, binding mode prediction and SAR exploration.

American trypanosomiasis or Chagas disease (CD) is a vector borne pathology caused by the parasite Trypanosoma cruzi (T. cruzi), which remains a serious global health problem. The current available treatment for CD is limited to two nitroderivatives with limited efficacy and several side effects. The rational design of ergosterol synthetic route inhibitors (e.g. CYP51 inhibitors) represents a promising strategy for fungi and trypanosomatids, exhibiting excellent anti-T.cruzi activity in pre-clinical assays. In the present work, we evaluate through different approaches (molecular docking, structure activity relationships, CYP51 inhibitory assay, and phenotypic screenings in vitro and in vivo) the potency and selectivity of a novel CYP51 inhibitor (compound 1) and its analogues against T.cruzi infection. Regarding anti-parasitic effect, compound 1 was active in vitro with EC50 3.86 and 4.00 µM upon intracellular (Tulahuen strain) and bloodstream forms (Y strain), respectively. In vivo assays showed that compound 1 reduced in 43% the parasitemia peak but, unfortunately failed to promote animal survival. In order to promote an enhancement at the potency and pharmacological properties, 17 new analogues were purchased and screened in vitro. Our findings demonstrated that five compounds were active against intracellular forms, highlighting compounds 1e and 1f, with EC50 2.20 and 2.70 µM, respectively, and selectivity indices (SI) = 50 and 36, respectively. Against bloodstream trypomastigotes, compound 1f reached an EC50 value of 20.62 µM, in a similar range to Benznidazole, but with low SI (3). Although improved the solubility of compound 1, the analogue 1f did not enhance the potency in vitro neither promote better in vivo efficacy against mouse model of acute T.cruzi infection arguing for the synthesis of novel pyrazolo[3,4-e][1,4]thiazepin derivatives aiming to contribute for alternative therapies for CD.