Identificación del virus de hepatitis delta genotipo 3 en comunidades andinas y amazónicas del Perú

Introducción. El virus de la hepatitis delta (VHD) es el causante de la forma más severa de la hepatitis viral humana, se asocia con un riesgo alto de fibrosis al hígado y carcinoma hepatocelular (HCC). Existen 8 genotipos del VHD con diferente distribución geográfica. Objetivos. Identificar los genotipos del VHD circulante en Huanta y tres pueblos indígenas de la Amazonía peruana. Métodos. Estudio observacional y transversal, realizado en 582 muestras reactivas para anti-HBc del VHB. Por el método nRT-PCR se procesaron todos los anti VHD positivos, el genotipo fue determinado mediante secuenciamiento directo tipo Sanger y análisis filogenético del fragmento R0. Se utilizaron 111 secuencias de referencia del GenBank. Las 42 secuencias del estudio fueron editadas y ensambladas con programas bioinformáticos. El análisis filogenético y evolutivo se realizó con los programas: Beast V2.5.2, Jmodeltest v2.1.10, Tracer v1.7.1, Tree Annotator y Figtree v1.4.4. Se utilizaron los modelos Bayesianos Yule y Birth Death skyline serial, el MCMC en 30 y 80 millones respectivamente, con el relaxed uncorrelated Exponential molecular clock. Se calcularon las medidas de resumen y de tendencia central utilizando el programa en STATA 14.0. Resultados. La media de la edad fue de 38 años, el 52,8% fueron mujeres. 101 muestras fueron positivas para anticuerpos anti-VHD. El ARN del VHD fue detectado en el 49,5% de las muestras reactivas a ELISA anti-VHD. El análisis filogenético determinó la presencia del genotipo 3. Conclusiones. Se evidencia la presencia del genotipo 3 del VHD en comunidades andinas y amazónicas del Perú.

Introduction. The Hepatitis Delta Virus (HDV) is the cause of the most severe form of human viral hepatitis and is associated with a high risk of liver fibrosis and hepatocellular carcinoma (HCC). There are 8 HDV genotypes with different geographic distribution. Objectives. To identify the genotypes of VHD circulating in Huanta and three indigenous peoples of the Peruvian Amazon. Methods. Observational and cross-sectional study, from 582 reactive samples for anti-HBc-HBV. Anti-HDV positive samples were processed with the nRT-PCR method, genotype was determined by direct Sanger-type sequencing and phylogenetic analysis of the R0 fragment. 111 reference sequences from GenBank were used. The 42 sequences of the study were edited y assembled with the bioinformatics programs. Phylogenetic and evolutionary analysis was performed with the following software: Beast v2.5.2, Jmodeltest v2.1.10, Tracer v1.7.1, Tree Annotator and Figtree v1.4.4. The Bayesian Yule and Birth Death skyline serial models were used, the MCMC at 30 and 80 million respectively, with the relaxed uncorrelated Exponential molecular clock. Summary and central tendency measures were calculated using the program in STATA 14.0. Results. The mean age was 38 years, 52.8% were women. 101 samples were positive for anti-HDV antibodies. HDV RNA was detected in 49.5% of the anti-HDV ELISA reactive samples. Phylogenetic analysis determined the presence of genotype 3. Conclusions. The presence of HDV genotype 3 in Andean and Amazonian communities of Peru is evidenced.

Interprofessional clinical event debriefing-does it make a difference? Attitudes of emergency department care providers to INFO clinical event debriefings.

BACKGROUND: Debriefing is increasingly used in clinical environments. Surveys indicate staff support for debriefing clinical events, but little is known about the specific effects of debriefing on healthcare workers in the workplace. INFO (Immediate, Not for personal assessment, Fast facilitated feedback, and Opportunity to support and ask questions) is a charge nurse facilitated clinical event debriefing program implemented in 2016 and currently used in five Emergency Departments (ED) in Calgary, Alberta, Canada. There have been more than 840 documented INFO debriefings. METHODS: Thirty interprofessional ED healthcare workers were recruited through posters and email to take part in voluntary one-on-one interviews using a standardized question script that asked participants about their experience with INFO debriefing assessments. Specifically, participants were asked to provide demographic information, give feedback about their involvement in INFO clinical debriefings, impact of debriefings on their clinical practice, stress levels and wellbeing. Interviews were transcribed and analysed using NVivo software. RESULTS: Forty-five healthcare workers replied to the initial recruitment methods with fifteen not responding to follow-up communication. Overall, staff satisfaction with INFO debriefing was highly rated. A qualitative thematic analysis to saturation approach was used to analyse the data. Five main themes were identified: 1.Effect of debriefing on clinical practice and patient care. 2. Psychological safety and teamwork. 3. Emotional acknowledgment after critical events. 4. Managing work stress in the ED. 5. Barriers to debriefing. CONCLUSIONS: In this study, debriefing in the ED helped interprofessional healthcare workers manage stress, provide improved patient care and teamwork while acknowledging emotions. This study specifically involved INFO, however, there are similarities that make our findings applicable to other clinical event debriefing programs. We believe this study provides further evidence supporting debriefing in clinical care areas.

RéSUMé: CONTEXTE: Le débriefing est de plus en plus utilisé dans les environnements cliniques. Les enquêtes indiquent que le personnel est favorable au débriefing des événements cliniques, mais on sait peu de choses sur les effets spécifiques du débriefing sur les travailleurs de la santé sur le lieu de travail. INFO (Immediate, Not for personal assessment, Fast facilitated feedback, and Opportunity to support and ask questions) est un programme de débriefing d'événements cliniques animé par l'infirmière en chef, mis en œuvre en 2016 et actuellement utilisé dans cinq services d'urgence (SU) à Calgary, Alberta, Canada. Il y a eu plus de 840 débriefings INFO documentés. MéTHODES: Trente travailleurs interprofessionnels des services d'urgence ont été recrutés par le biais d'affiches et de courriels pour participer à des entretiens individuels volontaires à l'aide d'un script de questions standardisé qui demandait aux participants de parler de leur expérience des évaluations de débriefing INFO. Plus précisément, les participants ont été invités à fournir des informations démographiques, à donner leur avis sur leur participation aux débriefings cliniques INFO, sur l'impact des débriefings sur leur pratique clinique, sur leur niveau de stress et sur leur bien-être. Les entretiens ont été transcrits et analysés à l'aide du logiciel NVivo. RéSULTATS: Quarante-cinq travailleurs de la santé ont répondu aux méthodes de recrutement initiales, quinze n'ont pas répondu à la communication de suivi. Dans l'ensemble, la satisfaction du personnel à l'égard du compte rendu d'INFO a été très bonne. Une analyse thématique qualitative jusqu'à saturation a été utilisée pour analyser les données. Cinq thèmes principaux ont été identifiés : 1. l'effet du débriefing sur la pratique clinique et les soins aux patients. 2. La sécurité psychologique et le travail en équipe. 3. Reconnaissance émotionnelle après des événements critiques. 4. Gestion du stress au travail dans les services d'urgence. 5. Obstacles au débriefing. CONCLUSIONS: Dans cette étude, le débriefing aux urgences a aidé les travailleurs de la santé interprofessionnels à gérer le stress, à améliorer les soins aux patients et le travail d'équipe tout en reconnaissant les émotions. Cette étude a porté spécifiquement sur INFO, mais il existe des similitudes qui rendent nos résultats applicables à d'autres programmes de débriefing d'événements cliniques. Nous pensons que cette étude apporte des preuves supplémentaires en faveur du débriefing dans les domaines des soins cliniques.

[DESIGN AND EVALUATION OF A MULTIEPITOPIC PROTEIN AS A CANDIDATE FOR A CARRION DISEASE VACCINE]. / DISEÑO Y EVALUACIÓN DE UNA PROTEÍNA MULTIEPITÓPICA COMO CANDIDATA PARA VACUNA CONTRA LA ENFERMEDAD DE CARRIÓN.

OBJECTIVES.: To design and assess a multiepitopic protein as a candidate for a vaccine against Carrion disease. MATERIALS AND METHODS.: Using bioinformatics tools, epitopes of external membrane proteins were selected and a multiepitopic protein was designed. The multiepitopic protein gene was subcloned into the expression plasmid pET28b and transformed into E. coli BL21 pLys. The multiepitopic protein was expressed using isopropyl-ß-D-1-thiogalactopyranoside and purified using resin. This purified protein was used to immunize BALB/c mice obtaining polyclonal antibodies. In vitro invasion assays were conducted using a strain of Bartonella bacilliformis (B. bacilliformis) in human red blood cells. RESULTS.: The multiepitopic protein M1 presents preserved epitopes between isolates of B. bacilliformis with are non-toxic, and not homologous to human and surface proteins. Immunized mice presented IgG antibody levels capable of reducing in vitro the rate of invasion of B. bacilliformis into human red blood cells. CONCLUSIONS.: Multiepitopic protein M1 may serve as a candidate for a Carrion disease vaccine; however, more studies are needed to characterize the use of this antigen as a vaccine.

OBJETIVOS.: Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. MATERIALES Y MÉTODOS.: Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-ß-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. RESULTADOS.: La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. CONCLUSIONES.: La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.

Diseño y evaluación de una proteína multiepitópica como candidata para vacuna contra la enfermedad de carrión / Design and evaluation of a multiepitopic protein as a candidate for a carrion disease vaccine

RESUMEN Objetivos. Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. Materiales y métodos. Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-β-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. Resultados. La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. Conclusiones. La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.

ABSTRACT Objectives. To design and assess a multiepitopic protein as a candidate for a vaccine against Carrion disease. Materials and Methods. Using bioinformatics tools, epitopes of external membrane proteins were selected and a multiepitopic protein was designed. The multiepitopic protein gene was subcloned into the expression plasmid pET28b and transformed into E. coli BL21 pLys. The multiepitopic protein was expressed using isopropyl-β-D-1-thiogalactopyranoside and purified using resin. This purified protein was used to immunize BALB/c mice obtaining polyclonal antibodies. In vitro invasion assays were conducted using a strain of Bartonella bacilliformis (B. bacilliformis) in human red blood cells. Results. The multiepitopic protein M1 presents preserved epitopes between isolates of B. bacilliformis with are non-toxic, and not homologous to human and surface proteins. Immunized mice presented IgG antibody levels capable of reducing in vitro the rate of invasion of B. bacilliformis into human red blood cells. Conclusions. Multiepitopic protein M1 may serve as a candidate for a Carrion disease vaccine; however, more studies are needed to characterize the use of this antigen as a vaccine.