Anti-HIV Humoral Response Induced by Different Anti-Idiotype Antibody Formats: An In Silico and In Vivo Approach.

Despite advancements in vaccinology, there is currently no effective anti-HIV vaccine. One strategy under investigation is based on the identification of epitopes recognized by broadly neutralizing antibodies to include in vaccine preparation. Taking into account the benefits of anti-idiotype molecules and the diverse biological attributes of different antibody formats, our aim was to identify the most immunogenic antibody format. This format could serve as a foundational element for the development of an oligo-polyclonal anti-idiotype vaccine against HIV-1. For our investigation, we anchored our study on an established b12 anti-idiotype, referred to as P1, and proposed four distinct formats: two single chains and two minibodies, both in two different orientations. For a deeper characterization of these molecules, we used immunoinformatic tools and tested them on rabbits. Our studies have revealed that a particular minibody conformation, MbVHVL, emerges as the most promising candidate. It demonstrates a significant binding affinity with b12 and elicits a humoral anti-HIV-1 response in rabbits similar to the Fab format. This study marks the first instance where the minibody format has been shown to provoke a humoral response against a pathogen. Furthermore, this format presents biological advantages over the Fab format, including bivalency and being encoded by a monocistronic gene, making it better suited for the development of RNA-based vaccines.

The Question of HIV Vaccine: Why Is a Solution Not Yet Available?

Ever since its discovery, human immunodeficiency virus type 1 (HIV-1) infection has remained a significant public health concern. The number of HIV-1 seropositive individuals currently stands at 40.1 million, yet definitive treatment for the virus is still unavailable on the market. Vaccination has proven to be a potent tool in combating infectious diseases, as evidenced by its success against other pathogens. However, despite ongoing efforts and research, the unique viral characteristics have prevented the development of an effective anti-HIV-1 vaccine. In this review, we aim to provide an historical overview of the various approaches attempted to create an effective anti-HIV-1 vaccine. Our objective is to explore the reasons why specific methods have failed to induce a protective immune response and to analyze the different modalities of immunogen presentation. This trial is registered with NCT05414786, NCT05471076, NCT04224701, and NCT01937455.

The initial interplay between HIV and mucosal innate immunity.

Human Immunodeficiency Virus (HIV) is still one of the major global health issues, and despite significant efforts that have been put into studying the pathogenesis of HIV infection, several aspects need to be clarified, including how innate immunity acts in different anatomical compartments. Given the nature of HIV as a sexually transmitted disease, one of the aspects that demands particular attention is the mucosal innate immune response. Given this scenario, we focused our attention on the interplay between HIV and mucosal innate response: the different mucosae act as a physical barrier, whose integrity can be compromised by the infection, and the virus-cell interaction induces the innate immune response. In addition, we explored the role of the mucosal microbiota in facilitating or preventing HIV infection and highlighted how its changes could influence the development of several opportunistic infections. Although recent progress, a proper characterization of mucosal innate immune response and microbiota is still missing, and further studies are needed to understand how they can be helpful for the formulation of an effective vaccine.

Identification of a novel mutation involved in colistin resistance in Klebsiella pneumoniae through Next-Generation Sequencing (NGS) based approaches.

The spread of multidrug-resistant (MDR) K. pneumoniae carbapenemase-producing bacteria (KPC) is one of the most serious threats to global public health. Due to the limited antibiotic options, colis- tin often represents a therapeutic choice. In this study, we performed Whole-Genome Sequencing (WGS) by Illumina and Nanopore platforms on four colistin-resistant K. pneumoniae isolates (CoRKp) to explore the resistance profile and the mutations involved in colistin resistance. Mapping reads with reference sequence of the most com- mon genes involved in colistin resistance did not show the presence of mobile colistin resistance (mcr) genes in all CoRKp. Complete or partial deletions of mgrB gene were observed in three out of four CoRKp, while in one CoRKp the mutation V24G on phoQ was identified. Complementation assay with proper wild type genes restored colistin susceptibility, validating the role of the amino acid substitution V24G and, as already described in the literature, confirming the key role of mgrB alterations in colistin resistance. In conclusion, this study allowed the identification of the novel mutation on phoQ gene involved in colistin resistance phenotype.

Detection of low-level HCV variants in DAA treated patients: comparison amongst three different NGS data analysis protocols.

BACKGROUND: Notwithstanding the efforts of direct-acting antivirals (DAAs) for the treatment of chronically infected hepatitis C virus (HCV) patients, concerns exist regarding the emergence of resistance-associated substitutions (RAS) related to therapy failure. Sanger sequencing is still the reference technique used for the detection of RAS and it detects viral variants present up to 15%, meaning that minority variants are undetectable, using this technique. To date, many studies are focused on the analysis of the impact of HCV low variants using next-generation sequencing (NGS) techniques, but the importance of these minority variants is still debated, and importantly, a common data analysis method is still not defined. METHODS: Serum samples from four patients failing DAAs therapy were collected at baseline and failure, and amplification of NS3, NS5A and NS5B genes was performed on each sample. The genes amplified were sequenced using Sanger and NGS Illumina sequencing and the data generated were analyzed with different approaches. Three different NGS data analysis methods, two homemade in silico pipeline and one commercially available certified user-friendly software, were used to detect low-level variants. RESULTS: The NGS approach allowed to infer also very-low level virus variants. Moreover, data processing allowed to generate high accuracy data which results in reduction in the error rates for each single sequence polymorphism. The results improved the detection of low-level viral variants in the HCV quasispecies of the analyzed patients, and in one patient a low-level RAS related to treatment failure was identified. Importantly, the results obtained from only two out of the three data analysis strategies were in complete agreement in terms of both detection and frequency of RAS. CONCLUSIONS: These results highlight the need to find a robust NGS data analysis method to standardize NGS results for a better comprehension of the clinical role of low-level HCV variants. Based on the extreme importance of data analysis approaches for wet-data interpretation, a detailed description of the used pipelines and further standardization of the in silico analysis could allow increasing diagnostic laboratory networking to unleash true potentials of NGS.

Divergent Trends of Anti-JCPyV Serum Reactivity and Neutralizing Activity in Multiple Sclerosis (MS) Patients during Treatment with Natalizumab.

The association between natalizumab and progressive multifocal leukoencephalopathy (PML) is established, but a reliable clinical risk stratification flow-chart is lacking. New risk factors are needed, such as the possible role of the anti-JC polyomavirus (JCPyV) neutralizing antibody. In this pilot study, we analyzed this parameter during natalizumab treatment. Sequential sera of 38 multiple sclerosis patients during their first year of natalizumab treatment were collected, and grouped according to the number of infusions. For 11 patients, samples were also available after 24 infusions (T24), when progressive multifocal leukoencephalopathy (PML) risk is higher. The reactivity against VP1, the main JCPyV surface protein, and the anti-JCPyV neutralizing activity were evaluated. During the first year, a lack of correlation between anti-JCPyV antibody response and its neutralizing activity was observed: a significant decrease in anti-JCPyV antibody response was observed (p = 0.0039), not paralleled by a similar trend in the total anti-JCPyV neutralizing activity (p = 0.2239). This lack of correlation was even more evident at T24 when, notwithstanding a significant increase in the anti-JCPyV response (p = 0.0097), a further decrease of the neutralizing activity was observed (p = 0.0062). This is the first study evidencing, prospectively, the lack of correlation between the anti-JCPyV antibody response and its neutralizing activity during natalizumab treatment.

Medicina baseada em evidências: instrumento para educação médica permanente entre psiquiatras? / Evidence-based medicine: a tool for continuing medical education among psychiatrists?

INTRODUÇÃO: O médico atualizado pode oferecer o melhor cuidado ao paciente e evitar consequências negativas que a defasagem científica pode acarretar. OBJETIVOS: Identificar os meios de atualização utilizados pelos psiquiatras brasileiros; avaliar seus conhecimentos sobre Medicina Baseada em Evidências (MBE) e sua utilização na educação permanente. MÉTODO: Estudo transversal realizado no XXIV Congresso Brasileiro de Psiquiatria. Os participantes (n = 188) responderam um questionário anônimo autoaplicado, com 28 perguntas sobre características sociodemográficas, fontes e periodicidade de atualização, e conhecimentos sobre MBE. RESULTADOS: Para atualização de conhecimentos, 98,3 por cento utilizavam os congressos brasileiros; 97,9 por cento as revistas nacionais; 93,9 por cento os livros-texto; 89,9 por cento as revistas das indústrias farmacêuticas; 63,5 por cento os consensos brasileiros; 63,3 por cento a base de dados Medline; 56,7 por cento as revistas internacionais; e 35 por cento a Biblioteca Cochrane. Os fatores estatisticamente significativos associados com bom conhecimento sobre MBE foram estar graduados há menos de dez anos (p < 0,001), usar o Medline (p < 0,009) e a Biblioteca Cochrane (p < 0,03) como fonte de busca de literatura médica. CONCLUSÕES: Os psiquiatras fazem pouco uso da melhor fonte de evidência para educação médica permanente e continuada, havendo, assim, menor benefício aos pacientes na tomada de decisão clínica.

BACKGROUND: Up-to-date psychiatrists provide better patient care and avoid the potentially negative consequences of outdated scientific knowledge. OBJECTIVES: The aim was to identify how Brazilian psychiatrists update their skills, and to evaluate their knowledge of evidenced-based medicine (EBM) and its use in continuing medical education. METHOD: This was a cross-sectional study conducted at the 24th Brazilian Congress of Psychiatry. Participants (n = 188) answered an anonymous, selfapplied questionnaire with 28 questions on socio-demographic characteristics, sources and frequency of updating, and knowledge of EBM. RESULTS: To update their knowledge, 98.3 percent of the sample attended Brazilian congresses, 97.9 percent read Brazilian medical journals, 93.9 percent used textbooks, 89.9 percent consulted journals published by pharmaceutical companies, 63.5 percent accessed the Brazilian guidelines, 63.3 percent used the Medline database, 56.7 percent read international journals, and 35 percent accessed the Cochrane Library. Factors that showed statistically significant association with good knowledge of EBM were having graduated from medical school less than ten years before (p < 0.001) and use of Medline (p < 0.009) and Cochrane Library (p < 0.03) as sources for searching the medical literature. CONCLUSIONS: Brazilian psychiatrists make limited use of the best sources of evidence for continuing medical education, thus producing less benefit for patients in clinical decision-making.

Gravidez na adolescência e uso freqüente de álcool e drogas no contexto familiar / Teenage pregnancy and frequent use of alcohol and drugs in the home environment

OBJETIVO: Analisar fatores individuais e familiares associados à gravidez na adolescência, incluindo uso freqüente de álcool e drogas ilícitas por familiar. MÉTODOS: Estudo de caso-controle com amostra de 408 adolescentes (13-17 anos) escolares sexualmente ativas, de Marília, SP, 2003-2004. Os casos eram 100 primigestas atendidas em programas de pré-natal de unidades de saúde e os controles, 308 estudantes que nunca haviam engravidado, provenientes de oito escolas estaduais. Foram coletados com instrumentos padronizados dados sociodemográficos, educacionais, comportamento contraceptivo, problemas de saúde mental e características familiares. A análise estatística incluiu testes de qui-quadrado e modelos de regressão logística. RESULTADOS: A baixa escolaridade paterna (p=0,01), a falta de informação sobre sexualidade e fertilização (p=0,001) e o uso de drogas ilícitas por familiar residente no domicílio (p=0,006) foram fatores de risco independentemente dos demais. Renda familiar per capita e pedir ao parceiro que usasse preservativo foram fatores de confundimento. CONCLUSÕES: O uso freqüente de drogas ilícitas por familiar residente no domicílio é um fator fortemente associado à gravidez na adolescência, independentemente dos demais. A expectativa de cursar a faculdade constitui fator de proteção, principalmente na presença de baixa escolaridade materna.

[Teenage pregnancy and frequent use of alcohol and drugs in the home environment]. / Gravidez na adolescência e uso freqüente de álcool e drogas no contexto familiar.

OBJECTIVE: To assess individual and family factors associated to teenage pregnancy, including frequent use of alcohol and illicit drugs by family members. METHODS: Case-control study conducted with a sample of 408 sexually active female adolescents (aged 13-17 years) in school from the city of Marília (Southeastern Brazil) in 2003-2004. Cases consisted of 100 primigravid teenagers assisted in prenatal care programs in health units. Controls were 308 nulligravid students from state public schools. Standardized instruments identified demographic and educational factors, contraceptive behavior, mental health problems, and family characteristics. Statistical analysis included chi-square tests and logistic regression models. RESULTS: Low paternal education (p=0.01), lack of information on sexuality and fertilization (p=0.001) and the use of illicit drugs by a resident family member (p=0.006) were independent risk factors. Family income per capita and asking the partner to use a condom were confounders. CONCLUSIONS: The frequent use of illicit drugs by a resident family member is a factor strongly associated to teenage pregnancy, regardless of other risk factors. The expectation of going to college constitutes a protective factor, mainly in the presence of low maternal education.

Problemas de saúde mental entre jovens grávidas e não-grávidas / Mental health problems among pregnant and non-pregnant youth

OBJETIVO: Estimar a prevalência de problemas de saúde mental em adolescentes primigestas e comparar seu perfil de saúde mental com o daquelas sexualmente ativas que nunca engravidaram. MÉTODOS: Estudo de corte transversal, comparativo entre dois grupos de adolescentes de 13 a 17 anos, em Marília, Estado de São Paulo, 2003-2004. A amostra incluiu 207 primigestas atendidas em programas de pré-natal de serviços públicos municipais urbanos e 308 estudantes de escolas estaduais, sexualmente ativas, que nunca engravidaram. Foram aplicados um instrumento de rastreamento para problemas de saúde mental em adolescentes (versão brasileira do Youth Self-Report) e um questionário sobre fatores de risco para gravidez na adolescência. A análise estatística incluiu testes de qui-quadrado, exato de Fisher, U de Mann Whitney e modelos de regressão logística. RESULTADOS: Adolescentes grávidas e não-grávidas não diferiram quanto à prevalência do total de problemas de saúde mental (24,6 por cento vs. 27,3 por cento; p=0,50). Comparado às adolescentes não-grávidas, o grupo das primigestas apresentou maior prevalência de sintomas de ansiedade/depressão (24,2 por cento vs. 15,3 por cento; p=0,01) e sintomas de retraimento/depressão (13,0 por cento vs, 4,5 por cento; p<0,001), além de maior número de fumantes (21,3 por cento vs. 11,0 por cento; p=0,002). Estas diferenças foram confirmadas em modelos de regressão logística, controlados para escolaridade da mãe. CONCLUSÕES: Foram mais freqüentes os sintomas de ansiedade e depressão e uso de tabaco em adolescentes primigestas em comparação com as adolescentes não-grávidas. Esses problemas requerem especial atenção dos serviços de pré-natal a fim de evitar possíveis prejuízos para a saúde das mães e de seus filhos.

OBJECTIVE: To estimate the prevalence of mental health problems in primiparous adolescents and to compare their mental health profile with the profile of sexually active adolescents who have never gotten pregnant. METHODS: Cross-sectional study that compared two groups of adolescents aged 13 to 17 years from a city in Southern Brazil, 2003-2004. The sample included 207 primiparous who attended prenatal care programs in urban public services, and 308 sexually active students from state public schools who had never gotten pregnant. A screening instrument to identify mental health problems in adolescents (Brazilian version of the Youth Self-Report) was applied to the participants together with a questionnaire on risk factors for pregnancy in adolescence. Statistical analysis included Chi-square tests, Fisher's exact test, Mann Whitney's U and logistic regression models. RESULTS: Pregnant and non-pregnant adolescents did not differ in the prevalence of total mental health problems (24.6 percent vs. 27.3 percent; p=0.50). Compared to the group of never pregnant adolescents, the group of primiparous presented higher prevalence of anxious/depressive symptoms (24.2 percent vs. 15.3 percent; p=0.01) and withdrawn/depressed symptoms (13.0 percent vs., 4.5 percent; p<0.001) and there was a greater number of tobacco users (21.3 percent vs. 11.0 percent; p=0.002). These differences were confirmed by logistic regression models controlled for maternal education. CONCLUSIONS: Anxiety and depression symptoms and tobacco use were more frequent in pregnant adolescents in comparison to non-pregnant. These problems require special attention from prenatal care services in order to avoid potential damages for the health of mothers and their children.

[Mental health problems among pregnant and non-pregnant youth]. / Problemas de saúde mental entre jovens grávidas e não-grávidas.

OBJECTIVE: To estimate the prevalence of mental health problems in primiparous adolescents and to compare their mental health profile with the profile of sexually active adolescents who have never gotten pregnant. METHODS: Cross-sectional study that compared two groups of adolescents aged 13 to 17 years from a city in Southern Brazil, 2003-2004. The sample included 207 primiparous who attended prenatal care programs in urban public services, and 308 sexually active students from state public schools who had never gotten pregnant. A screening instrument to identify mental health problems in adolescents (Brazilian version of the Youth Self-Report) was applied to the participants together with a questionnaire on risk factors for pregnancy in adolescence. Statistical analysis included Chi-square tests, Fisher's exact test, Mann Whitney's U and logistic regression models. RESULTS: Pregnant and non-pregnant adolescents did not differ in the prevalence of total mental health problems (24.6% vs. 27.3%; p=0.50). Compared to the group of never pregnant adolescents, the group of primiparous presented higher prevalence of anxious/depressive symptoms (24.2% vs. 15.3%; p=0.01) and withdrawn/depressed symptoms (13.0% vs., 4.5%; p<0.001) and there was a greater number of tobacco users (21.3% vs. 11.0%; p=0.002). These differences were confirmed by logistic regression models controlled for maternal education. CONCLUSIONS: Anxiety and depression symptoms and tobacco use were more frequent in pregnant adolescents in comparison to non-pregnant. These problems require special attention from prenatal care services in order to avoid potential damages for the health of mothers and their children.

Aprendendo com a prática: experiência de estudantes da Famema / Learning with practice: an experience of Famema students

A Faculdade de Medicina de Marília, desde 1997, vem implementando mudanças curriculares nos cursos de Enfermagem e Medicina. Tais mudanças pressupõem articulação entre teoria e prática, processo de ensino baseado na aprendizagem significativa, centrado nos estudantes, em situações de aprendizagem que emergem da prática profissional, abordados de forma interdisciplinare multiprofissional. Para isto, os estudantes de Enfermagem e Medicina, de forma integrada, desenvolvem atividades em contato com usuários e trabalhadores da saúde, desde o primeiro ano, inseridos em equipes nas Unidades de Saúde da Família, num ciclo de dois anos,denominado Unidade de Prática Profissional. O presente relato apresenta a experiência de três estudantes que tiveram a oportunidade de avaliar o estado de saúde e acompanhar seis famílias. Esta atividade possibilitou identificar diferentes necessidades de saúde e levou os estudantes a buscar informações em diversas fontes a fim de propor estratégias de ação que contribuam para a melhora da qualidade de vida das pessoas. Foi possível intervenção por meio de orientações, encaminhamentos e realização de procedimentos. Os estudantes concluem que esta prática possibilita maior compreensão dos valores dos usuários e a aproximação de uma realidade bastante distante do seu modo de vida, além de uma aprendizagem significativa.

O agente comunitário de saúde e os estressores no contexto do trabalho / The Community Health Worker and the Stress in his/her Job

Considerando a importância do papel do agente comunitário de saúde (ACS) nas equipes do Programa de Saúde da Família (PSF), o presente estudo pretende explorar os estressores no contexto do trabalho dos mesmos, segundo sua própria percepção. Para tanto, 99 ACS responderam ao “Inventario de Malestar Percibido em el Ambiente Laborial” (IMPAL). Os resultados mostraram que os fatores mais freqüentemente reconhecidos como causadores de impacto negativo foram: 1) relacionados ao meio ambiente físico: falta de espaço próprio na unidade e andar longas distâncias sob sol; 2) fatores próprios da tarefa: excesso de trabalho burocrático e alta exigência do serviço; 3) relacionados à organização do tempo de trabalho: trabalhar com prazos muito curtos; 4) quanto aos aspectos institucionais: ter que lutar contra a ineficiência do trabalho de outros setores; 5) quanto aos aspectos pessoais: necessidade de enfrentar problemas que excedem sua capacidade como ACS e saber que seus erros podem prejudicar outras pessoas. Tais resultados apontam para a necessidade de adoção de estratégias capazes de minimizar os efeitos destes fatores, a fim de reforçar a proposta de vigilância à saúde dos PSF.