Methods of Cryptococcal Polysaccharide Analysis Using ELISA.

One of the standard assays for the fungal pathogen Cryptococcus neoformans is the glucuronoxylomannan (GXM) ELISA. This assay utilizes monoclonal antibodies targeted against the critical virulence factor, the polysaccharide (PS) capsule. GXM ELISA is one of the most used assays in the field used for diagnosis of cryptococcal infection, quantification of PS content, and determination of binding specificity for antibodies. Here we present three variations of the GXM ELISA used by our group-indirect, capture, and competition ELISAs. We have also provided some history, perspective, and notes on these methods, which we hope will help the reader choose, and implement, the best assay for their research.While it has long been referred to as the GXM ELISA, we also suggest a name update to better reflect our updated understanding of the polysaccharide antigens targeted by this assay. The Cryptococcal PS ELISA is a more accurate description of this set of methodologies and the antigens they measure. Finally, we discuss the limitations of this assay and put forth future plans for expanding the antigens assayed by ELISA.

Semisynthetic Glycoconjugate Vaccine Candidates against Cryptococcus neoformans.

Cryptococcus neoformans is a fungus classified by the World Health Organization as a critically important pathogen, which poses a significant threat to immunocompromised individuals. In this study, we present the chemical synthesis and evaluation of two semisynthetic vaccine candidates targeting the capsular polysaccharide glucuronoxylomannan (GXM) of C. neoformans. These semisynthetic glycoconjugate vaccines contain an identical synthetic decasaccharide (M2 motif) antigen. This antigen is present in serotype A strains, which constitute 95% of the clinical cryptococcosis cases. This synthetic oligosaccharide was conjugated to two proteins (CRM197 and Anthrax 63 kDa PA) and tested for immunogenicity in mice. The conjugates elicited a specific antibody response that bound to the M2 motif but also exhibited additional cross-reactivity toward M1 and M4 GXM motifs. Both glycoconjugates produced antibodies that bound to GXM in ELISA assays and to live fungal cells. Mice immunized with the CRM197 glycoconjugate produced weakly opsonic antibodies and displayed trends toward increased median survival relative to mice given a mock PBS injection (18 vs 15 days, p = 0.06). These findings indicate promise, achieving a successful vaccine demands further optimization of the glycoconjugate. This antigen could serve as a component in a multivalent GXM motif vaccine.

Semi-synthetic glycoconjugate vaccine candidate against Cryptococcus neoformans.

Cryptococcus neoformans is a fungus classified by the World Health Organization as a critically important pathogen, posing a significant threat to immunocompromised individuals. In this study, we present the chemical synthesis and evaluation of two semi-synthetic vaccine candidates targeting the capsular polysaccharide glucuronoxylomannan (GXM) of C. neoformans. These semi-synthetic glycoconjugate vaccines contain the identical synthetic decasaccharide (M2 motif) antigen. This motif is present in serotype A strains, which constitute 95% of clinical cryptococcosis cases. This synthetic oligosaccharide was conjugated to two proteins (CRM197 and Anthrax 63 kDa PA) and tested for immunogenicity in mice. The conjugates elicited a specific antibody response that bound to the M2 motif but also exhibited additional cross-reactivity towards M1 and M4 GXM motifs. Both glycoconjugates produced antibodies that bound to GXM in ELISA assays and to live fungal cells. Mice immunized with the CRM197 glycoconjugate produced opsonic antibodies and displayed trends toward increased median survival relative to mice given a mock PBS injection (18 vs 15 days, p = 0.06). While these findings indicate promise, achieving a successful vaccine demands further optimization of the glycoconjugate. It could serve as a component in a multi-valent GXM motif vaccine, enhancing both strength and breadth of immune responses.

Corrigendum: Home-based exercise program in the indeterminate form of Chagas disease (PEDI-CHAGAS study): a study protocol for a randomized clinical trial.

[This corrects the article DOI: 10.3389/fmed.2022.1087188.].

Bionized Nanoferrite Particles Alter the Course of Experimental Cryptococcus neoformans Pneumonia.

Cryptococcosis is a devastating fungal disease associated with high morbidity and mortality even when treated with antifungal drugs. Bionized nanoferrite (BNF) nanoparticles are powerful immunomodulators, but their efficacy for infectious diseases has not been investigated. Administration of BNF nanoparticles to mice with experimental cryptococcal pneumonia altered the outcome of infection in a dose response manner as measured by CFU and survival. The protective effects were higher at lower doses, with reductions in IL-2, IL-4, and TNF-α, consistent with immune modulation whereby reductions in inflammation translate into reduced host damage, clearance of infection, and longer survival.

Home-based exercise program in the indeterminate form of Chagas disease (PEDI-CHAGAS study): A study protocol for a randomized clinical trial.

Background: Chagas disease (CD) is a neglected endemic disease with worldwide impact due to migration. Approximately 50-70% of individuals in the chronic phase of CD present the indeterminate form, characterized by parasitological and/or serological evidence of Trypanosoma cruzi infection, but without clinical signs and symptoms. Subclinical abnormalities have been reported in indeterminate form of CD, including pro-inflammatory states and alterations in cardiac function, biomarkers and autonomic modulation. Moreover, individuals with CD are usually impacted on their personal and professional life, making social insertion difficult and impacting their mental health and quality of life (QoL). Physical exercise has been acknowledged as an important strategy to prevent and control numerous chronic-degenerative diseases, but unexplored in individuals with the indeterminate form of CD. The PEDI-CHAGAS study (which stands for "Home-Based Exercise Program in the Indeterminate Form of Chagas Disease" in Portuguese) aims to evaluate the effects of a home-based exercise program on physical and mental health outcomes in individuals with indeterminate form of CD. Methods and design: The PEDI-CHAGAS is a two-arm (exercise and control) phase 3 superiority randomized clinical trial including patients with indeterminate form of CD. The exclusion criteria are <18 years old, evidence of non-Chagasic cardiomyopathy, musculoskeletal or cognitive limitations that preclude the realization of exercise protocol, clinical contraindication for regular exercise, and regular physical exercise (≥1 × per week). Participants will be assessed at baseline, and after three and 6 months of follow-up. The primary outcome will be QoL. Secondary outcomes will include blood pressure, physical fitness components, nutritional status, fatigability, autonomic modulation, cardiac morphology and function, low back pain, depression and anxiety, stress, sleep quality, medication use and adherence, and biochemical, inflammatory and cardiac biomarkers. Participants in the intervention group will undergo a home-based exercise program whilst those in the control group will receive only general information regarding the benefits of physical activity. Both groups will receive the same general nutritional counseling consisting of general orientations about healthy diets. Conclusion: The findings from the present study may support public health intervention strategies to improve physical and mental health parameters to be implemented more effectively in this population. Clinical trial registration: [https://ensaiosclinicos.gov.br/rg/RBR-10yxgcr9/], identifier [U1111-1263-0153].

Push-out bond strength and failure mode of single adjustable and customized glass fiber posts.

PURPOSE: To evaluate the push-out bond strength (PBS) and failure mode of single adjustable (SAP) and customized (CP) posts cemented to root canal dentin using conventional (RelyX Ultimate) or self-adhesive (RelyX U200) dual-cure resin cements. METHODS: Herein, 40 bovine mandibular incisors were divided into four groups (n = 10): SAP cemented with RelyX Ultimate (SAP-UT), SAP cemented with RelyX U200 (SAP-U2), CP cemented with RelyX Ultimate (CP-UT), and CP cemented with RelyX U200 (CP-U2). PBS and failure modes were analyzed. Three-way repeated measures ANOVA test followed by Tukey's test and Fisher-Freeman-Halton exact test were used for data analysis (α = 5%). RESULTS: The PBS values for SAP (p < .05) were higher than those for CP and were not influenced by the root third and resin cement (p > .05). When conventional resin cement was used, the SAP showed significant differences compared to CP (p < .05). When cemented with RelyX Ultimate, a higher prevalence of mixed and adhesive failures for SAP and CP, respectively, was observed (p < .05). For the self-adhesive resin cement, the failures were mostly adhesive (p < .05). CONCLUSION: SAP showed better performance than CP. The root third and resin cements did not influence the PBS. The most prevalent failures were adhesive and mixed.

Amoeba Predation of Cryptococcus neoformans Results in Pleiotropic Changes to Traits Associated with Virulence.

Amoeboid predators, such as amoebae, are proposed to select for survival traits in soil microbes such as Cryptococcus neoformans; these traits can also function in animal virulence by defeating phagocytic immune cells, such as macrophages. Consistent with this notion, incubation of various fungal species with amoebae enhanced their virulence, but the mechanisms involved are unknown. In this study, we exposed three strains of C. neoformans (1 clinical and 2 environmental) to predation by Acanthamoeba castellanii for prolonged times and then analyzed surviving colonies phenotypically and genetically. Surviving colonies comprised cells that expressed either pseudohyphal or yeast phenotypes, which demonstrated variable expression of traits associated with virulence, such as capsule size, urease production, and melanization. Phenotypic changes were associated with aneuploidy and DNA sequence mutations in some amoeba-passaged isolates, but not in others. Mutations in the gene encoding the oligopeptide transporter (CNAG_03013; OPT1) were observed among amoeba-passaged isolates from each of the three strains. Isolates derived from environmental strains gained the capacity for enhanced macrophage toxicity after amoeba selection and carried mutations on the CNAG_00570 gene encoding Pkr1 (AMP-dependent protein kinase regulator) but manifested reduced virulence in mice because they elicited more effective fungal-clearing immune responses. Our results indicate that C. neoformans survival under constant amoeba predation involves the generation of strains expressing pleiotropic phenotypic and genetic changes. Given the myriad potential predators in soils, the diversity observed among amoeba-selected strains suggests a bet-hedging strategy whereby variant diversity increases the likelihood that some will survive predation.IMPORTANCECryptococcus neoformans is a ubiquitous environmental fungus that is also a leading cause of fatal fungal infection in humans, especially among immunocompromised patients. A major question in the field is how an environmental yeast such as C. neoformans becomes a human pathogen when it has no need for an animal host in its life cycle. Previous studies showed that C. neoformans increases its pathogenicity after interacting with its environmental predator amoebae. Amoebae, like macrophages, are phagocytic cells that are considered an environmental training ground for pathogens to resist macrophages, but the mechanism by which C. neoformans changes its virulence through interactions with protozoa is unknown. Our study indicates that fungal survival in the face of amoeba predation is associated with the emergence of pleiotropic phenotypic and genomic changes that increase the chance of fungal survival, with this diversity suggesting a bet-hedging strategy to ensure that some forms survive.

Influence of relevant cystic fibrosis bacteria on Scedosporium apiospermum and Scedosporium boydii growth and viability.

Cystic fibrosis (CF) causes a variety of symptoms in different organs, but the majority of the morbidity and mortality of CF is related with pulmonary conditions. Primary infections are usually bacterial, and when treated with antibiotics, yeast infections appear or become more evident. Studies show that different microorganisms can co-inhabit the same environment and the interactions could be synergistic or antagonistic. Using techniques including viable and non-viable cell-to-cell interactions, mixed culture in liquid, and solid media sharing or not the supernatant, this study has evaluated interactions between the fungal species Scedosporium apiospermum and Scedosporium boydii with the bacterial species Staphylococcus aureus, Pseudomonas aeruginosa, and Burkholderia cepacia. Cell-to-cell interactions in liquid medium showed that P. aeruginosa and B. cepacia were able to reduce fungal viability but only in the presence of alive bacteria. Interactions without cell contact using a semi-permeable membrane showed that all bacteria were able to inhibit both fungal growths/viabilities. Cell-free supernatants from bacterial growth reduced fungal viability in planktonic fungal cells as well as in some conditions for preformed fungal biomass. According to the chemical analysis of the bacterial supernatants, the predominant component is protein. In this work, we verified that bacterial cells and their metabolites, present in the supernatants, can play anti-S. apiospermum and anti-S. boydii roles on fungal growth and viability.

Dealing with the impact of the COVID-19 pandemic on a rapid response team operation in Brazil: Quality in practice.

QUALITY PROBLEM OR ISSUE: Up to 13 July 2020, >12 million laboratory-confirmed cases of coronavirus disease of 2019 (COVID-19) infection have been reported worldwide, 1 864 681 in Brazil. We aimed to assess an intervention to deal with the impact of the COVID-19 pandemic on the operations of a rapid response team (RRT). INITIAL ASSESSMENT: An observational study with medical record review was carried out at a large tertiary care hospital in Fortaleza, a 400-bed quaternary hospital, 96 of which are intensive care unit beds. All adult patients admitted to hospital wards, treated by the RRTs during the study period, were included, and a total of 15 461 RRT calls were analyzed. CHOICE OF SOLUTION: Adequacy of workforce sizing. IMPLEMENTATION: The hospital adjusted the size of its RRTs during the period, going from two to four simultaneous on-duty medical professionals. EVALUATION: After the beginning of the pandemic, the number of treated cases in general went from an average of 30.6 daily calls to 79.2, whereas the extremely critical cases went from 3.5 to 22 on average. In percentages, the extremely critical care cases went from 10.47 to 20%, with P < 0.001. Patient mortality remained unchanged. The number of critically ill cases and the number of treated patients increased 2-fold in relation to the prepandemic period, but the effectiveness of the RRT in relation to mortality was not affected. LESSONS LEARNED: The observation of these data is important for hospital managers to adjust the size of their RRTs according to the new scenario, aiming to maintain the intervention effectiveness.

Vulnerability factors associated with HIV/AIDS hospitalizations: a case-control study.

OBJECTIVES: to identify the association between HIV/AIDS hospitalizations and factors that integrate individual, social, and programmatic vulnerabilities. METHODS: a case-control study conducted in 2014 in a municipality in the state of São Paulo. "Cases" included people living with HIV (PLHIV) hospitalized and "control" those who were outpatients. Interviews were conducted using a tool with sociodemographic variables, clinical characteristics and other vulnerabilities. Data were analyzed by conditional logistic regression. RESULTS: fifty-six cases and 112 control participated. Risk factors for HIV hospitalization were: unemployed and retired individuals; homeless people; non-antiretroviral users; individuals who did not regularly attend returns. Access to social workers was a protective factor for hospitalization. CONCLUSIONS: this research contributed to measure the social, individual and programmatic vulnerabilities that interfere with HIV worsening and, consequently, unfavorable outcome such as hospitalization.

A comparative analysis between atraumatic and conventional anesthetic techniques in surgical removal of upper third molars.

Pain control is essential in dental practice, and can be accomplished through various techniques. This study seeks to compare atraumatic and conventional anesthetic techniques, applied during surgeries to remove upper third molars. The endpoints evaluated were pain indices, patient satisfaction and anesthetic efficacy. A random parallel split-mouth clinical trial was conducted with 14 patients. Group A received atraumatic anesthesia without a needle (Comfort-in®) and group B received conventional anesthesia by blocking the posterior superior alveolar nerve (PSAN) and Greater Palatine Nerve (GPN). A Visual Analogue Scale (VAS) was used to assess pain. A significantly (p<0.001) lower perception of pain was observed among individuals who received the atraumatic technique. In 71% of cases, it was necessary to supplement the anesthesia during the procedure. Even considering the need for additional anesthesia, the Comfort-in® technique was more accepted by patients with regard to pain perception than the conventional manual technique.

O controle da dor é fundamental na prática odontológica, podendo ser feito a partir de várias técnicas. Este estudo visa comparar as técnicas anestésicas atraumática e convencional, aplicadas durante cirurgias para remover terceiros molares superiores. Os pontos avaliados foram índice de dor, satisfação do paciente e eficácia anestésica. Um ensaio clínico randomizado, paralelo, boca dividida, foi conduzido com 14 participantes. O grupo A recebeu anestesia atraumática sem agulha (Comfort-in®) e o grupo B anestesia convencional por meio de bloqueio do nervo alveolar superior posterior (NASP) e nervo palatino maior (NPM). Uma Escala Visual Analógica (EVA) foi utilizada para avaliar a dor. Uma significativa (p<0,001) menor percepção de dor entre os indivíduos que receberam a técnica atraumática foi observada. Em 71% dos casos, fez-se necessária a complementação da anestesia durante a realização do procedimento. Mesmo considerando a necessidade de anestesia adicional, a técnica empregando o sistema Comfort- in® teve maior aceitação dos pacientes quando comparado à técnica manual convencional, no que se refere a percepção de dor, ainda que considerando a necessidade de complementação da anestesia.

Coordenação do cuidado às pessoas que vivem com HIV no sistema prisional / Coordinación del cuidado a las personas que viven con el VIH en el sistema penitenciario / Coordination of care for people living with HIV in the prison system

Resumo Objetivo: Analisar a coordenação do cuidado às pessoas que vivem com HIV, segundo unidade prisional. Métodos: Estudo transversal, realizado em seis unidades prisionais do Estado de São Paulo. Entrevistaram-se 85 detentos vivendo com HIV e seis diretores técnicos. Indicadores de coordenação foram criados a partir de perguntas com escala de 1 a 5, classificando-os em satisfatórios (>3,5 a 5,0), regulares (>2,5 a 3,5) e insatisfatórios (1,0 a 2,5). Utilizou-se ANOVA e Kruskal Wallis. Resultados: A coordenação foi classificada como insatisfatória (média 2,49). Indicadores insatisfatórios: "Questionar efeitos colaterais da terapia antirretroviral (TARV)"; "Questionar dificuldades na tomada da TARV"; "Observar tomada da TARV"; "Solicitar fracos/embalagens da TARV para monitorar a ingesta medicamentosa"; "Pedir explicações quanto ao uso da TARV"; "Questionar condições de acondicionamento da TARV na cela"; "Informar e discutir resultados T-CD4+ e carga viral"; "Informar agendamento da consulta no serviço de referência em HIV" e "Levar para atendimento em outras especialidades médicas". Obtiveram classificação regular: "Levar para atendimento médico de urgência quando necessário" e "Não perder consulta no serviço de referência em HIV". "Questionar sobre a regularidade no uso da TARV" foi o único indicador pior avaliado na comparação entre as unidades prisionais estudadas (p<0,05). Conclusão: O desempenho das unidades prisionais não difere em relação à grande parte dos indicadores de coordenação estudados, indicando que todas precisam melhorar o desempenho no que diz respeito ao desenvolvimento de ações de monitoramento do uso da TARV, informar e discutir resultados dos exames com os detentos e levar para atendimento fora da unidade prisional.

Resumen Objetivo: Analizar la coordinación del cuidado a las personas que viven con el VIH, según unidad penitenciaria. Métodos: Estudio transversal realizado en seis unidades penitenciarias del estado de São Paulo. Se realizó entrevista a 85 presos que viven con el VIH y seis directores técnicos. Fueron creados indicadores de coordinación a partir de preguntas con escala de 1 a 5 y se clasificaron en satisfactorios (>3,5 a 5,0), regulares (>2,5 a 3,5) e insatisfactorios (1,0 a 2,5). Se utilizó ANOVA y Kruskal Wallis. Resultados: La coordinación fue clasificada como insatisfactoria (promedio 2,49). Indicadores insatisfactorios: "Preguntar sobre efectos secundarios del tratamiento antirretroviral (TARV)", "Preguntar sobre dificultades en la toma del TARV", "Observar toma del TARV", "Solicitar frascos/envases del TARV para monitorear la ingesta de medicamentos", "Pedir explicaciones sobre el uso del TARV", "Preguntar sobre condiciones de almacenaje del TARV en la celda", "Informar y discutir resultados T CD4+ y carga viral", "Informar consultas agendadas en el servicio de referencia en VIH" y "Llevar para recibir atención en otras especialidades médicas". Obtuvieron clasificación regular los indicadores: "Llevar para recibir atención médica de urgencia cuando es necesario" y "No perder el turno en el servicio de referencia en VIH". "Preguntar sobre la regularidad de uso del TARV" fue el único indicador peor evaluado en la comparación entre las unidades penitenciarias estudiadas (p<0,05). Conclusión: El desempeño de las unidades penitenciarias no difiere con relación a la mayoría de los indicadores de coordinación estudiados, lo que indica que todas necesitan mejorar el desempeño respecto al desarrollo de acciones de monitoreo del uso del TARV, informar y discutir resultados de los análisis con los presos y llevarlos para recibir atención fuera de la unidad penitenciaria.

Abstract Objective: To analyze the care coordination for people living with HIV according to the prison unit. Methods: Cross-sectional study conducted in six prison units in the state of São Paulo. Eighty-five inmates living with HIV and six technical directors were interviewed. Coordination indicators were created from questions with a 1-5 scale and classified as satisfactory (>3.5 to 5.0), regular (>2.5 to 3.5) and unsatisfactory (1.0 to 2.5). ANOVA and Kruskal Wallis were used. Results: The coordination was classified as unsatisfactory (mean 2.49). Unsatisfactory indicators: "Questioning side effects of antiretroviral therapy (ART)"; "Questioning the difficulties in ART intake"; "Observing ART intake"; "Requesting ART bottles/packages to monitor medication intake"; "Asking for explanations regarding the use of ART"; "Questioning the storage conditions of ART in the prison cell"; "Informing and discussing T-CD4 + and viral loading results"; "Informing the scheduling of consultation at the HIV reference service" and "Take to care for other medical specialties". The following obtained regular classification: "Take to emergency medical care when needed" and "Not missing an appointment at the HIV reference service". "Questioning the regularity of the use of ART" was the single worst indicator evaluated in the comparison between the prison units studied (p<0.05). Conclusion: The performance of prison units does not differ in relation to most coordination indicators studied, which shows the need for improving the performance with regard to the development of actions to monitor the use of ART, inform and discuss test results with inmates and take them to care outside the prison unit.

Vulnerability factors associated with HIV/AIDS hospitalizations: a case-control study / Factores de vulnerabilidad asociados a la hospitalización del VIH/SIDA: estudio de control de casos / Fatores de vulnerabilidade associados às internações por HIV/aids: estudo caso controle

ABSTRACT Objectives: to identify the association between HIV/AIDS hospitalizations and factors that integrate individual, social, and programmatic vulnerabilities. Methods: a case-control study conducted in 2014 in a municipality in the state of São Paulo. "Cases" included people living with HIV (PLHIV) hospitalized and "control" those who were outpatients. Interviews were conducted using a tool with sociodemographic variables, clinical characteristics and other vulnerabilities. Data were analyzed by conditional logistic regression. Results: fifty-six cases and 112 control participated. Risk factors for HIV hospitalization were: unemployed and retired individuals; homeless people; non-antiretroviral users; individuals who did not regularly attend returns. Access to social workers was a protective factor for hospitalization. Conclusions: this research contributed to measure the social, individual and programmatic vulnerabilities that interfere with HIV worsening and, consequently, unfavorable outcome such as hospitalization.

RESUMEN Objetivos: identificar la asociación entre las hospitalizaciones por VIH/SIDA y los factores que integran las vulnerabilidades individuales, sociales y del programa. Métodos: estudio de casos y controles realizado en 2014 en un municipio del estado de São Paulo. Los "casos" incluyeron a personas que viven con VIH (PVVIH) hospitalizadas y "controla" a los pacientes ambulatorios. Las entrevistas se realizaron utilizando un instrumento con variables sociodemográficas, características clínicas y otras vulnerabilidades. Los datos fueron analizados por regresión logística condicional. Resultados: participaron 56 casos y 112 controles. Los factores de riesgo de hospitalización por VIH fueron: personas desempleadas y jubiladas; personas sin hogar; usuarios no antirretrovirales; individuos que no asistieron regularmente a las declaraciones. El acceso a los trabajadores sociales fue un factor protector para la hospitalización. Conclusiones: esta investigación contribuyó a medir las vulnerabilidades sociales, individuales y programáticas que interfieren con la agudización del VIH y, en consecuencia, el resultado desfavorable, como la hospitalización.

RESUMO Objetivos: identificar a associação entre as internações por HIV/aids e os fatores que integram as vulnerabilidades individuais, sociais e programáticas. Métodos: estudo caso-controle realizado em 2014 em um município do estado de São Paulo. "Casos" compreenderam pessoas que viviam com HIV (PVHIV) internadas e "controles" aquelas que faziam acompanhamento ambulatorial. Foram realizadas entrevistas utilizando um instrumento com variáveis sociodemográficas, características clínicas e outras vulnerabilidades. Os dados foram analisados por meio de regressão logística condicional. Resultados: participaram 56 casos e 112 controles. Constituíram fatores de risco para internação hospitalar por HIV: indivíduos desempregados e aposentados/do lar; pessoas em situação de rua; não usuários de antirretroviral; indivíduos que não compareciam regularmente aos retornos. Acesso à assistente social constituiu-se um fator de proteção para internação. Conclus ões: esta investigação contribuiu para mensurar as vulnerabilidades sociais, individuais e programáticas que interferem na agudização do HIV e, consequentemente, no desfecho desfavorável, como a internação hospitalar.

Sphingolipid biosynthetic pathway is crucial for growth, biofilm formation and membrane integrity of Scedosporium boydii.

Aim: Glycosphingolipids are conserved lipids displaying a variety of functions in fungal cells, such as determination of cell polarity and virulence. They have been considered as potent targets for new antifungal drugs. The present work aimed to test two inhibitors, myriocin and DL-threo-1-Phenyl-2-palmitoylamino-3-morpholino-1-propanol, in Scedosporium boydii, a pathogenic fungus which causes a wide range of disease. Materials & methods: Mass spectrometry, microscopy and cell biology approaches showed that treatment with both inhibitors led to defects in fungal growth and membrane integrity, and caused an increased susceptibility to the current antifungal agents. Conclusion: These data demonstrate the antifungal potential of drugs inhibiting sphingolipid biosynthesis, as well as the usefulness of sphingolipids as promising targets for the development of new therapeutic options.

Pessoas vivendo com HIV/Aids no cárcere: regularidade no uso da terapia antirretroviral / Personas que viven con el VIH/sida en la cárcel: regularidad del uso del tratamiento antirretroviral / People living with HIV/AIDS in prison: regular use of antiretroviral therapy

Resumo Objetivo Analisar os desafios relacionados à regularidade no uso da terapia antirretroviral pelas pessoas vivendo com HIV privadas de liberdade. Métodos Trata-se de um estudo exploratório, com abordagem quantitativa, realizado em seis unidades prisionais de dois municípios paulistas. A coleta de dados ocorreu no período de agosto a novembro de 2015, mediante entrevistas, com apoio de instrumento específico. Os dados foram analisados a partir de técnicas descritivas, análise univariada (testes de associação Qui-quadrado e Exato de Fisher) e teste não paramétrico U de Mann-Whitney. Resultados Participaram da pesquisa 67 indivíduos em uso da terapia antirretroviral. Identificou-se que, nos últimos sete dias, 80,6% dos indivíduos não deixaram de tomar os medicamentos antirretrovirais e 91% negaram tomar quantidade errada de comprimidos, de acordo com a prescrição médica. Quanto às ações de monitoramento da terapia antirretroviral, o questionamento sobre o uso contínuo dos medicamentos pela equipe de saúde das unidades prisionais foi considerado regular. O uso irregular da terapia antirretroviral apresentou associação estatisticamente significante com uso de drogas lícitas antes do encarceramento (p=0,006) e interrupção do acompanhamento médico (p=0,014). Conclusão Tais achados mostram a necessidade de estratégias de intervenção que favoreçam o acompanhamento e monitoramento do uso da terapia antirretroviral no contexto prisional.

Resumen Objetivo analizar los desafíos relacionados con la regularidad del uso del tratamiento antirretroviral por personas privadas de la libertad que viven con el VIH. Métodos se trata de un estudio exploratorio, con enfoque cuantitativo, realizado en seis unidades penitenciarias de dos municipios del estado de São Paulo. La recolección de datos se llevó a cabo en el período de agosto a noviembre de 2015 mediante entrevistas, con apoyo de instrumento específico. Los datos fueron analizados a partir de técnicas descriptivas, análisis univariado (prueba χ2 de Pearson y prueba exacta de Fisher) y prueba no paramétrica U de Mann-Whitney. Resultados participaron de la investigación 67 individuos en tratamiento antirretroviral. Se identificó que, en los últimos siete días, el 80,6% de los individuos no dejó de tomar los medicamentos antirretrovirales y un 91% negó haber tomado la cantidad equivocada de comprimidos, de acuerdo con la prescripción médica. Con relación a las acciones de monitoreo del tratamiento antirretroviral, el cuestionamiento sobre el uso continuo de los medicamentos por el equipo de salud de las unidades penitenciarias fue considerado regular. El tratamiento antirretroviral irregular presentó una relación estadísticamente significativa con el uso de drogas lícitas antes del encarcelamiento (p=0,006) y la interrupción del seguimiento médico (p=0,014). Conclusión tales descubrimientos demuestran la necesidad de estrategias de intervención que favorezcan el seguimiento y monitoreo del uso del tratamiento antirretroviral en el contexto penitenciario.

Abstract Objective To analyze the challenges related to regularity in the use of antiretroviral therapy by people living with HIV deprived of their liberty. Methods This is an exploratory study, with a quantitative approach, carried out in six prison units of two municipalities in São Paulo State. The data collection took place in the period from August to November 2015, through interviews, with the support of a specific tool. Data were analyzed using descriptive techniques, univariate analysis (Chi-Square association test and Fisher's Exact Test) and non-parametric Mann-Whitney U test. Results 67 individuals taking antiretroviral therapy participated in the study. It was identified that, in the last seven days, 80.6% of the subjects did not stop taking the antiretroviral drugs and 91% denied taking the wrong amount of tablets, according to the medical prescription. Regarding the antiretroviral therapy monitoring actions, questioning about continuous use of the drugs by the health team of the prison units was considered regular. Irregular use of antiretroviral therapy showed a statistically significant association with use of licit drugs prior to incarceration (p = 0.006) and interruption of medical follow-up (p = 0.014). Conclusion These findings show the need for intervention strategies that favor follow-up and monitoring of the use of antiretroviral therapy in the prison context.

Active Teaching Methodology Applied in the Training of Undergraduate Dental Students: Use of Video-Lesson / Metodologia Ativa de Ensino Aplicada na Formação de Graduandos de Odontologia: Utilização de Vídeo-Aula

The goal of the present study was to evaluate the effectiveness of the active video-class methodology in the teaching-learning process, ascertaining the understanding, clarity and absorption through a pre-structured questionnaire. The sample consisted of 35 students, who were enrolled in the fixed prosthesis discipline, between the age of 19 and 31 years old. A videotape with provisional crown content was applied to the sample and, after a pre-structured questionnaire with 5 questions with objective and subjective questions related to the class. In the results obtained two people did not miss any of the questions, corresponding to 5.7% of the total number of students. Meanwhile, 57.1% of the students erred only one question in that they are equivalent to 20 students of the sample, 34.3% did not answer right two questions of the questionnaire, in the total of twelve people and finally 2,9% missed 3 questions, represented by only 1 person. It was evidenced that most of the questions students achieved success above 80%. The use of videoconference as an active teaching methodology in the academic environment has shown satisfactory results when applied in the classroom as an aid material, since most of the students have had success in most of the questions with the use this tool. The active methodologies can be a satisfactory alternative in the teaching-learning process to assist the teacher in the classroom, and its use can yield good results in the training of health professionals. (AU)

O objetivo do presente trabalho foi avaliar a eficácia da metodologia ativa vídeo-aula aplicada aos alunos de odontologia de uma instituição de ensino superior no processo de ensino aprendizagem, averiguando o entendimento, clareza e absorção, através de um questionário préestruturado. A amostra constou de 35 alunos devidamente matriculados na disciplina de prótese fixa, com idades que variavam entre 19 a 31 anos. Foi administrado uma vídeo-aula com conteúdo de coroas provisória e logo após responderam um questionário pré-estruturado com 5 perguntas com questões objetivas e subjetivas relacionadas a aula. Nos resultados obtidos duas pessoas não erraram nenhuma das questões, correspondendo 5,7% do total de alunos. Já 57,1% dos alunos erraram apenas uma questão no que equivalem 20 alunos da amostra, 34,3% não acertaram duas questões do questionário, no total de 12 pessoas e por fim 2,9% erraram 3 questões, representado por apenas 1 pessoa. Ficou evidenciado que a maioria das questões os alunos obtiveram êxitos acima de 80%. O emprego da vídeo-aula como uma metodologia ativa de ensino, no ambiente acadêmico, mostrou resultados satisfatórios ao ser aplicada em sala de aula como material de auxílio, visto que a maioria dos alunos tiveram êxitos na maior parte das questões com a utilização desta ferramenta. As metodologias ativas podem ser uma alternativa satisfatória no processo de ensino-prendizagem para auxiliar o professor em sala de aula, e sua utilização pode render bons resultados na formação de profissionais da saúde. (AU)

Structural Differences Influence Biological Properties of Glucosylceramides from Clinical and Environmental Isolates of Scedosporium aurantiacum and Pseudallescheria minutispora.

Scedosporium/Lomentospora complex is composed of filamentous fungi, including some clinically relevant species, such as Pseudallescheria boydii, Scedosporium aurantiacum, and Scedosporium apiospermum. Glucosylceramide (GlcCer), a conserved neutral glycosphingolipid, has been described as an important cell surface molecule playing a role in fungal morphological transition and pathogenesis. The present work aimed at the evaluation of GlcCer structures in S. aurantiacum and Pseudallescheria minutispora, a clinical and an environmental isolate, respectively, in order to determine their participation in fungal growth and host-pathogen interactions. Structural analysis by positive ion-mode ESI-MS (electrospray ionization mass spectrometer) revealed the presence of different ceramide moieties in GlcCer in these species. Monoclonal antibodies against Aspergillus fumigatus GlcCer could recognize S. aurantiacum and P. minutispora conidia, suggesting a conserved epitope in fungal GlcCer. In addition, these antibodies reduced fungal viability, enhanced conidia phagocytosis by macrophages, and decreased fungal survival inside phagocytic cells. Purified GlcCer from both species led to macrophage activation, increasing cell viability as well as nitric oxide and superoxide production in different proportions between the two species. These results evidenced some important properties of GlcCer from species of the Scedosporium/Lomentospora complex, as well as the effects of monoclonal anti-GlcCer antibodies on fungal cells and host-pathogen interaction. The differences between the two species regarding the observed biological properties suggest that variation in GlcCer structures and strain origin could interfere in the role of GlcCer in host-pathogen interaction.

Usability and sensitivity of the risk of bias assessment tool for randomized controlled trials of pharmacist interventions.

Background The Cochrane collaboration risk of bias assessment (RoB) tool is used in several fields to evaluate the methodological quality of studies. Its strengths and challenges are discussed. Objective To assess the sensitivity of the RoB tool in studies of pharmacist interventions. Setting DEPICT database was used to pool randomized controlled trials (RCTs) of complex interventions. Method A Guide for RoB Judgment in Pharmacy Services was created to help in the interpretation and judgment of bias criteria. The evaluation of bias (low, unclear, high risk) was performed by RCT. Sensitivity analyses were performed to assess the influence of different interpretations of eight elements of judgment in the RoB tool. Paired analysis and estimations of the effect size (95% confidence interval) of the criteria modifications compared to the original analyses were calculated. Main outcome measure Changes in the interpretations of judgment in the RoB tool. Results Overall, 8.3, 45.4, and 46.3% of the studies were determined to have low, unclear, and high risk of bias, respectively. High risk of bias was caused by attrition and detection domains. The number of studies classified with high risk of bias significantly increased for five of the eight interpretations, while unclear risk of bias increased for three interpretations (with a negligible effect size in all of them). Lack of blinding, loss of participants, and the use of subjective and self-reported outcomes were the main elements resulting in high risk of bias. Conclusion The RoB tool is useful for evaluating RCTs of pharmacist interventions if adapted criteria for judgment are used. Ignoring these adjustments produces a floor-effect with studies classified with high risk of bias.

Health-related quality of life assessment among people living with HIV in Rio de Janeiro, Brazil: a cross-sectional study.

PURPOSE: To assess health-related quality of life (HRQoL) and its associated factors among people living with HIV/AIDS (PLWHA) in Rio de Janeiro, Brazil. METHODS: A cross-sectional study including PLWHA receiving usual HIV-care at Instituto Nacional de Infectologia Evandro Chagas (INI/Fiocruz) was conducted between 2014 and 2016 in Rio de Janeiro, Brazil. The EQ-5D-3L assessed HRQoL; PHQ-2 and ASSIST were used for screening depression and substance use, respectively. Clinical variables were obtained from the INI/Fiocruz cohort database, and structured questions evaluated intimate partner violence, sexual abstinence and relationship status. Data were analysed using multivariable Tobit regression model. RESULTS: A total of 1480 PLWHA were included: 64.7% were male at birth (38.4% men who have sex with men [MSM], 24.3% heterosexual men and 2% transgender women [TGW]); median age was 43.1 years, and 95.8% were receiving antiretroviral therapy. The median EQ-5D-3L utility score was 0.801. Results showed that the following factors: MSM and women; older age; lower educational level; no engagement in a relationship; depression screening positive; polysubstance use; and, detectable viral load were independently associated with worse HRQoL. CONCLUSIONS: PLWHA under care at INI/Fiocruz presented good HRQoL. Polysubstance use, depression and lower educational level were among the factors negatively associated with HRQoL. This was the first time that the EQ-5D-3L utility scores were calculated for a considerable number of PLWHA in Brazil, which is a fundamental piece of information for future cost-effectiveness analysis.