Lactic acid bacteria secreted proteins as potential Listeria monocytogenes quorum sensing inhibitors.

Listeria monocytogenes is an important human and animal pathogen able to cause an infection named listeriosis and is mainly transmitted through contaminated food. Among its virulence traits, the ability to form biofilms and to survive in harsh environments stand out and lead to the persistence of L. monocytogenes for long periods in food processing environments. Virulence and biofilm formation are phenotypes regulated by quorum sensing (QS) and, therefore, the control of L. monocytogenes through an anti-QS strategy is promising. This study aimed to identify, by in silico approaches, proteins secreted by lactic acid bacteria (LAB) potentially able to interfere with the agr QS system of L. monocytogenes. The genome mining of Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus NCFM revealed 151 predicted secreted proteins. Concomitantly, the three-dimensional (3D) structures of AgrB and AgrC proteins of L. monocytogenes were modeled and validated, and their active sites were predicted. Through protein-protein docking and molecular dynamic, Serine-type D-Ala-D-Ala carboxypeptidase and L,D-transpeptidase, potentially secreted by L. rhamnosus GG and L. acidophilus NCFM, respectively, were identified with high affinity to AgrB and AgrC proteins, respectively. By inhibiting the translocation of the cyclic autoinducer peptide (cyclic AIP) via AgrB, and its recognition in the active site of AgrC, these LAB proteins could disrupt L. monocytogenes communication by impairing the agr QS system. The application of the QS inhibitors predicted in this study can emerge as a promising strategy in controlling L. monocytogenes in food processing environment and as an adjunct to antibiotic therapy for the treatment of listeriosis.

Bacillus velezensis iturins inhibit the hemolytic activity of Staphylococcus aureus.

Bovine mastitis caused by S. aureus has a major economic impact on the dairy sector. With the crucial need for new therapies, anti-virulence strategies have gained attention as alternatives to antibiotics. Here we aimed to identify novel compounds that inhibit the production/activity of hemolysins, a virulence factor of S. aureus associated with mastitis severity. We screened Bacillus strains obtained from diverse sources for compounds showing anti-hemolytic activity. Our results demonstrate that lipopeptides produced by Bacillus spp. completely prevented the hemolytic activity of S. aureus at certain concentrations. Following purification, both iturins, fengycins, and surfactins were able to reduce hemolysis caused by S. aureus, with iturins showing the highest anti-hemolytic activity (up to 76% reduction). The lipopeptides showed an effect at the post-translational level. Molecular docking simulations demonstrated that these compounds can bind to hemolysin, possibly interfering with enzyme action. Lastly, molecular dynamics analysis indicated general stability of important residues for hemolysin activity as well as the presence of hydrogen bonds between iturins and these residues, with longevous interactions. Our data reveals, for the first time, an anti-hemolytic activity of lipopeptides and highlights the potential application of iturins as an anti-virulence therapy to control bovine mastitis caused by S. aureus.

Design of a Multi-Epitope Vaccine against Histoplasma capsulatum through Immunoinformatics Approaches.

Histoplasmosis is a widespread systemic disease caused by Histoplasma capsulatum, prevalent in the Americas. Despite its significant morbidity and mortality rates, no vaccines are currently available. Previously, five vaccine targets and specific epitopes for H. capsulatum were identified. Immunoinformatics has emerged as a novel approach for determining the main immunogenic components of antigens through in silico methods. Therefore, we predicted the main helper and cytotoxic T lymphocytes and B-cell epitopes for these targets to create a potential multi-epitope vaccine known as HistoVAC-TSFM. A total of 38 epitopes were found: 23 common to CTL and B-cell responses, 11 linked to HTL and B cells, and 4 previously validated epitopes associated with the B subunit of cholera toxin, a potent adjuvant. In silico evaluations confirmed the stability, non-toxicity, non-allergenicity, and non-homology of these vaccines with the host. Notably, the vaccine exhibited the potential to trigger both innate and adaptive immune responses, likely involving the TLR4 pathway, as supported by 3D modeling and molecular docking. The designed HistoVAC-TSFM appears promising against Histoplasma, with the ability to induce important cytokines, such as IFN-γ, TNF-α, IL17, and IL6. Future studies could be carried out to test the vaccine's efficacy in in vivo models.

Design of a multi-epitope vaccine (vme-VAC/MST-1) against cholera and vibriosis based on reverse vaccinology and immunoinformatics approaches.

Vibriosis and cholera are serious diseases distributed worldwide and caused by six marine bacteria of the Vibrio genus. Thousands of deaths occur each year due to these illnesses, necessitating the development of new preventive measures. Presently, the existing cholera vaccine demonstrates an effectiveness of approximately 60%. Here we describe a new multi-epitope vaccine, 'vme-VAC/MST-1' based on vaccine targets identified by reverse vaccinology and epitopes predicted by immunoinformatics, two currently effective tools for predicting new vaccines for bacterial pathogens. The vaccine was designed to combat vibriosis and cholera by incorporating epitopes predicted for CTL, HTL, and B cells. These epitopes were identified from six vaccine targets revealed through subtractive genomics, combined with reverse vaccinology, and were further filtered using immunoinformatics approaches based on their predicted immunogenicity. To construct the vaccine, 28 epitopes (24 CTL/B and 4 HTL/B) were linked to the sequence of the cholera toxin B subunit adjuvant. In silico analyses indicate that the resulting immunogen is stable, soluble, non-toxic, and non-allergenic. Furthermore, it exhibits no homology to the host and demonstrates a strong capacity to elicit innate, B-cell, and T-cell immune responses. Our analysis suggests that it is likely to elicit immune reactions mediated through the TLR5 pathway, as evidenced by the molecular docking of the vaccine with the receptor, which revealed high affinity and a favorable reaction. Thus, vme-VAC/MST-1 is predicted to be a safe and effective solution against pathogenic Vibrio spp. However, further experimental analyses are required to measure the vaccine's effects In vivo.Communicated by Ramaswamy H. Sarma.

[Continuity of care between care levels: experiences of users in a municipal health network]. / Continuidade da gestão clínica entre níveis assistenciais: experiências dos usuários de uma rede municipal de saúde.

This study analyzes the experiences of users on the continuity of clinical management between care levels. This is a cross-sectional quantitative study that uses data from a survey conducted with 407 users of a public health network in Recife, Pernambuco State, Brazil, in 2017 and 2018. The experiences on the continuity of clinical management were explored from two dimensions: coherence of care and accessibility between levels of care. Users presented more positive opinions about coherence of care than accessibility. Regarding coherence of care, most users reported that primary and specialized care physicians agree on diagnosis, treatment, and recommendations, and that the primary care physician refers them to a specialist when necessary. Only 43% of users reported collaboration between physicians to solve their health problems. Concerning accessibility, most users (77.2%) reported a long waiting time for the consultation with a specialist and less than half (48.9%) reported delay for primary care. The results of this study coincide with other investigations and highlight the need to promote strategies for achieving effective integration of care networks and thus provide users with greater continuity of health care.

Este estudo analisa as experiências dos usuários sobre a continuidade da gestão clínica entre níveis assistenciais. Trata-se de um estudo transversal, quantitativo, que utiliza dados de um inquérito realizado com 407 usuários de uma rede pública de saúde de Recife, Pernambuco, Brasil, nos anos de 2017 e 2018. As experiências sobre a continuidade da gestão clínica foram exploradas a partir de duas dimensões: coerência da atenção e acessibilidade entre níveis assistenciais. Os usuários apresentaram opiniões mais positivas sobre a coerência da atenção que sobre a acessibilidade. Quanto à coerência da atenção, a maioria dos usuários referiu que os médicos da atenção primária e da especializada concordam entre si quanto a diagnóstico, tratamento e recomendações, e que o médico da atenção primária encaminha ao especialista quando necessário. Apenas 43% dos usuários relataram existir colaboração entre os médicos para resolução dos seus problemas de saúde. Quanto à acessibilidade, a maioria dos usuários (77,2%) referiu um longo tempo de espera para a consulta com o especialista e menos da metade (48,9%) referiu demora para atendimento na atenção primária. Os resultados deste estudo coincidem com outras investigações e evidenciam a necessidade de fomentar estratégias para alcançar uma integração efetiva das redes assistenciais e assim conferir ao usuário uma maior continuidade dos cuidados em saúde.

Este estudio analiza las experiencias de los usuarios sobre la continuidad de la gestión clínica entre los niveles asistenciales. Se trata de un estudio transversal, de carácter cuantitativo, realizado con datos de una encuesta aplicada a 407 usuarios de una red pública de salud en Recife, Pernambuco, Brasil, en los años de 2017 y 2018. Las experiencias sobre la continuidad de la gestión clínica fueron exploradas desde dos dimensiones: la consistencia de la atención y la accesibilidad entre niveles asistenciales. Los usuarios tenían opiniones más positivas sobre la consistencia de la atención que sobre la accesibilidad. En cuanto a la consistencia de la atención, la mayoría de los usuarios reportó que los médicos de atención primaria y de la atención especializada concuerdan entre sí en el diagnóstico, tratamiento y recomendaciones, y que los médicos de atención primaria realizan la derivación al especialista cuando necesario. Solamente el 43% de los usuarios reportaron percibir una colaboración entre los médicos para la búsqueda de soluciones a sus problemas de salud. Respecto a la accesibilidad, la mayoría de los encuestados (77,2%) refirió un largo tiempo de espera para la consulta con el especialista y menos de la mitad (48,9%) mencionó que tardaba la atención primaria. Los resultados de este estudio coinciden con el de otras investigaciones y muestran la necesidad de promover estrategias para lograr una efectiva integración de las redes de atención y, así, brindarle al usuario una mayor continuidad de la atención en salud.

Insights into the Vibrio Genus: A One Health Perspective from Host Adaptability and Antibiotic Resistance to In Silico Identification of Drug Targets.

The genus Vibrio comprises an important group of ubiquitous bacteria of marine systems with a high infectious capacity for humans and fish, which can lead to death or cause economic losses in aquaculture. However, little is known about the evolutionary process that led to the adaptation and colonization of humans and also about the consequences of the uncontrollable use of antibiotics in aquaculture. Here, comparative genomics analysis and functional gene annotation showed that the species more related to humans presented a significantly higher amount of proteins associated with colonization processes, such as transcriptional factors, signal transduction mechanisms, and iron uptake. In comparison, those aquaculture-associated species possess a much higher amount of resistance-associated genes, as with those of the tetracycline class. Finally, through subtractive genomics, we propose seven new drug targets such as: UMP Kinase, required to catalyze the phosphorylation of UMP into UDP, essential for the survival of bacteria of this genus; and, new natural molecules, which have demonstrated high affinity for the active sites of these targets. These data also suggest that the species most adaptable to fish and humans have a distinct natural evolution and probably undergo changes due to anthropogenic action in aquaculture or indiscriminate/irregular use of antibiotics.

Continuidade da gestão clínica entre níveis assistenciais: experiências dos usuários de uma rede municipal de saúde / Continuity of care between care levels: experiences of users in a municipal health network / Continuidad de la atención entre niveles asistenciales: experiencias de usuarios de una red municipal de salud

Este estudo analisa as experiências dos usuários sobre a continuidade da gestão clínica entre níveis assistenciais. Trata-se de um estudo transversal, quantitativo, que utiliza dados de um inquérito realizado com 407 usuários de uma rede pública de saúde de Recife, Pernambuco, Brasil, nos anos de 2017 e 2018. As experiências sobre a continuidade da gestão clínica foram exploradas a partir de duas dimensões: coerência da atenção e acessibilidade entre níveis assistenciais. Os usuários apresentaram opiniões mais positivas sobre a coerência da atenção que sobre a acessibilidade. Quanto à coerência da atenção, a maioria dos usuários referiu que os médicos da atenção primária e da especializada concordam entre si quanto a diagnóstico, tratamento e recomendações, e que o médico da atenção primária encaminha ao especialista quando necessário. Apenas 43% dos usuários relataram existir colaboração entre os médicos para resolução dos seus problemas de saúde. Quanto à acessibilidade, a maioria dos usuários (77,2%) referiu um longo tempo de espera para a consulta com o especialista e menos da metade (48,9%) referiu demora para atendimento na atenção primária. Os resultados deste estudo coincidem com outras investigações e evidenciam a necessidade de fomentar estratégias para alcançar uma integração efetiva das redes assistenciais e assim conferir ao usuário uma maior continuidade dos cuidados em saúde.

This study analyzes the experiences of users on the continuity of clinical management between care levels. This is a cross-sectional quantitative study that uses data from a survey conducted with 407 users of a public health network in Recife, Pernambuco State, Brazil, in 2017 and 2018. The experiences on the continuity of clinical management were explored from two dimensions: coherence of care and accessibility between levels of care. Users presented more positive opinions about coherence of care than accessibility. Regarding coherence of care, most users reported that primary and specialized care physicians agree on diagnosis, treatment, and recommendations, and that the primary care physician refers them to a specialist when necessary. Only 43% of users reported collaboration between physicians to solve their health problems. Concerning accessibility, most users (77.2%) reported a long waiting time for the consultation with a specialist and less than half (48.9%) reported delay for primary care. The results of this study coincide with other investigations and highlight the need to promote strategies for achieving effective integration of care networks and thus provide users with greater continuity of health care.

Este estudio analiza las experiencias de los usuarios sobre la continuidad de la gestión clínica entre los niveles asistenciales. Se trata de un estudio transversal, de carácter cuantitativo, realizado con datos de una encuesta aplicada a 407 usuarios de una red pública de salud en Recife, Pernambuco, Brasil, en los años de 2017 y 2018. Las experiencias sobre la continuidad de la gestión clínica fueron exploradas desde dos dimensiones: la consistencia de la atención y la accesibilidad entre niveles asistenciales. Los usuarios tenían opiniones más positivas sobre la consistencia de la atención que sobre la accesibilidad. En cuanto a la consistencia de la atención, la mayoría de los usuarios reportó que los médicos de atención primaria y de la atención especializada concuerdan entre sí en el diagnóstico, tratamiento y recomendaciones, y que los médicos de atención primaria realizan la derivación al especialista cuando necesario. Solamente el 43% de los usuarios reportaron percibir una colaboración entre los médicos para la búsqueda de soluciones a sus problemas de salud. Respecto a la accesibilidad, la mayoría de los encuestados (77,2%) refirió un largo tiempo de espera para la consulta con el especialista y menos de la mitad (48,9%) mencionó que tardaba la atención primaria. Los resultados de este estudio coinciden con el de otras investigaciones y muestran la necesidad de promover estrategias para lograr una efectiva integración de las redes de atención y, así, brindarle al usuario una mayor continuidad de la atención en salud.

Avaliação das ações de vigilância sanitária: construção participativa de mecanismos para o monitoramento do desempenho da gestão / Assessment of health surveillance actions: participatory construction of mechanisms for monitoring management performance

Introdução: A avaliação focada na utilização constitui-se em referencial teórico importante a ser observado na proposição de um caminho entre tantos que o campo da avaliação oferece como suporte à qualificação da gestão em saúde. O monitoramento do desempenho é considerado uma estratégia impulsionadora à racionalização da gestão e das decisões. Objetivo: Apresentar o caminho percorrido pela Agência Nacional de Vigilância Sanitária e pelo Hospital Alemão Oswaldo Cruz no processo de construção participativa de mecanismos para o monitoramento do desempenho da gestão em instâncias subnacionais, como parte da implantação da ação ­ mecanismos estruturais e técnicos ­ integrante da primeira linha de execução do projeto Institucionalização de Práticas Avaliativas: a gestão estratégica da vigilância sanitária baseada em evidências. Método: Descrição do processo de construção participativa de mecanismos para o monitoramento do desempenho da gestão em instâncias subnacionais, considerando duas dimensões ­ o envolvimento dos principais interessados na perspectiva do uso do monitoramento e a estruturação do instrumental teórico e operacional e de estratégias de utilização. Resultados: Projeto implantado em quatro instituições de Vigilância Sanitária, permeado pelo caráter participativo, desde a elaboração da estratégia, à modelagem, à formulação dos dispositivos gerenciais e de instrumentos de análise e interpretação dos indicadores. Conclusões: O projeto viabilizou a instituição de um espaço de aprendizado institucional que valorizou não só a apreciação dos resultados, mas, também, o próprio processo de produção da informação, contribuindo, assim, com a instituição de mudanças e inovações na execução das ações.

Introduction: Evaluation focused on use is an important theoretical framework to be observed in proposing a pathway among those of the feld of evaluation to support the qualifcation of health management. Performance monitoring is considered a driving strategy for rationalizing management and decisions. Objective: To present the pathway carried out by the Agência Nacional de Vigilância Sanitária and the Hospital Alemão Oswaldo Cruz in the participatory construction process of mechanisms for monitoring management performance in subnational instances, as part of the implementation of the action ­ structural and technical mechanisms ­ that makes part of the frst line of execution of the Institutionalization of the project "Evaluative Practices: strategic management of the evidence-based health surveillance". Method: Description of the participatory construction process of mechanisms for monitoring management performance in subnational instances, considering two dimensions ­ the involvement of the main stakeholders in the perspective of the use of monitoring and the building of theoretical and operational tools and utilization strategies. Results: The project was implemented in four health surveillance institutions. It was permeated by the participatory feature since the strategy conception, to the modeling, to the formulation of the management devices and instruments of analysis and interpretation of the indicators. Conclusions: The project enabled the development of an institutional learning locus that valued not only the appreciation of results, but also the information production process itself. Therefore, it contributes to the institutionalization of changes and innovations in the execution of actions.