Standardized Brazilian green propolis extract (EPP-AF®) in COVID-19 outcomes: a randomized double-blind placebo-controlled trial.

SARS-CoV-2 and its different variants caused a "wave and wave" pandemic pattern. During the first wave we demonstrated that standardized Brazilian green propolis extract (EPP-AF®) reduces length of hospital stay in adult patients with COVID-19. Afterwards, we decided to evaluate the impact of EPP-AF in hospitalized patients during the third wave of the pandemic. BeeCovid2 was a randomized, double-blind, placebo-controlled clinical trial in hospitalized COVID-19 adult patients. Patients were allocated to receive an oral dose of 900 mg/day of EPP-AF® or placebo for 10 days. The primary outcome was length of hospital stay. Secondary outcomes included safety, secondary infection rate, duration of oxygen therapy dependency, acute kidney injury and need for intensive care. Patients were followed up for 28 days after admission. We enrolled 188 patients; 98 were assigned to the propolis group and 90 to the placebo group. The post-intervention length of hospital stay was of 6.5 ± 6.0 days in the propolis group versus 7.7 ± 7.1 days in the control group (95% CI - 0.74 [- 1.94 to 0.42]; p = 0.22). Propolis did not have significant impact on the need for oxygen supplementation or frequency of AKI. There was a significant difference in the incidence of secondary infection between groups, with 6.1% in the propolis group versus 18.9% in the control group (95% CI - 0.28 [0.1-0.76], p = 0.01). The use of EPP-AF was considered safe and associated with a decrease in secondary infections. The drug was not associated with a significant reduction in length of hospital stay. ClinicalTrials.gov (NCT04800224).

Differential Binding of NLRP3 to non-oxidized and Ox-mtDNA mediates NLRP3 Inflammasome Activation.

The NLRP3 inflammasome is a key mediator of the innate immune response to sterile tissue injury and is involved in many chronic and acute diseases. Physically and chemically diverse agents activate the NLRP3 inflammasome. Here, we show that NLRP3 binds non-oxidized and Ox-mtDNA differentially, with a half maximum inhibitory concentration (IC50) for non-oxidized and Ox-mtDNA of 4 nM and 247.2 nM, respectively. The NLRP3 Neonatal-Onset Multisystem Inflammatory Disease (NOMIDFCAS) gain of function mutant could bind non-oxidized mtDNA but had higher affinity for Ox-mtDNA compared to WT with an IC50 of 8.1 nM. NLRP3 lacking the pyrin domain can bind both oxidized and non-oxidized mtDNA. Isolated pyrin domain prefers Ox-mtDNA. The NLRP3 pyrin domain shares a protein fold with DNA glycosylases and generate a model for DNA binding based on the structure and sequence alignment to Clostridium acetobutylicum and human OGG1, an inhibitor of Ox-mtDNA generation, 8-oxoguanine DNA glycosylases. We provide a new model for how NLRP3 interacts with Ox-mtDNA supported by DNA binding in the presence of a monoclonal antibody against the pyrin domain. These results give new insights into the mechanism of inflammasome assembly, and into the function of reactive oxygen species in establishing a robust immune response.

Small molecule inhibitor binds to NLRP3 and prevents inflammasome activation.

Despite recent advances in the mechanism of oxidized DNA activating NLRP3, the molecular mechanism and consequence of oxidized DNA associating with NLRP3 remains unknown. Cytosolic NLRP3 binds oxidized DNA which has been released from the mitochondria, which subsequently triggers inflammasome activation. Human glycosylase (hOGG1) repairs oxidized DNA damage which inhibits inflammasome activation. The fold of NLRP3 pyrin domain contains amino acids and a protein fold similar to hOGG1. Amino acids that enable hOGG1 to bind and cleave oxidized DNA are conserved in NLRP3. We found NLRP3 could bind and cleave oxidized guanine within mitochondrial DNA. The binding of oxidized DNA to NLRP3 was prevented by small molecule drugs which also inhibit hOGG1. These same drugs also inhibited inflammasome activation. Elucidating this mechanism will enable design of drug memetics that treat inflammasome pathologies, illustrated herein by NLRP3 pyrin domain inhibitors which suppressed interleukin-1ß (IL-1ß) production in macrophages. One-Sentence Summary: NLRP3 cleaves oxidized DNA and small molecule drug binding inhibits inflammasome activation.

Violência contra trabalhadores de enfermagem na pandemia de COVID-19: revisão de escopo / Violence against nursing workers in the COVID-19 pandemic: scoping review

Resumo Objetivo Mapear e sintetizar as evidências científicas disponíveis sobre violência contra trabalhadores de enfermagem durante a pandemia de COVID-19. Métodos Revisão de escopo elaborada de acordo com as recomendações do Instituto Joanna Briggs feita nas bases de dados National Library of Medicine, Cumulative Index to Nursing and Allied Health Literature, Web of Science, Excerpa Medica DataBASE, PsycINFO - APA PsycNET e Literatura Latino-Americana e do Caribe em Ciências da Saúde. Foram incluídos estudos que abordaram o tema violência relacionada ao trabalho contra trabalhadores de enfermagem durante a pandemia de COVID-19, publicados nos idiomas português, espanhol, ou inglês, a partir de 2020, disponíveis na íntegra. Excluíram-se relatórios parciais de pesquisa, editoriais, cartas resposta e resumos de anais. Foi realizada análise descritiva. Resultados De um total de 56 publicações, nove estudos responderam aos critérios de seleção, sendo sete com qualidade metodológica elevada e dois moderada. O fenômeno ocorreu em diferentes países do mundo, sendo as vítimas usuais trabalhadores de enfermagem que atuavam na linha de frente, com menor tempo de formação, perpetrada por diferentes atores. Prejuízos psicossociais aos trabalhadores foram encontrados, além de riscos aos pacientes. Conclusão Trabalhadores de enfermagem tiveram elevada incidência de violência durante a pandemia. As consequências desta violência foram danosas à saúde, levando ao desejo de abandonar a profissão ou afetando a saúde física ou mental das vítimas. Como medidas de prevenção e controle foram sugeridas implementação de políticas públicas e estratégias administrativas para redução da violência contra essa população em condições de pandemias.

Resumen Objetivo Mapear y sintetizar las evidencias científicas disponibles sobre violencia contra trabajadores de enfermería durante la pandemia de COVID-19. Métodos Revisión de alcance elaborada de acuerdo con las recomendaciones del Instituto Joanna Briggs realizada en las bases de datos National Library of Medicine, Cumulative Index to Nursing and Allied Health Literature, Web of Science, Excerpa Medica DataBASE, PsycINFO - APA PsycNET y Literatura Latinoamericana y del Caribe en Ciencias de la Salud. Se incluyeron estudios que abordaron el tema violencia relacionada con el trabajo contra trabajadores de enfermería durante la pandemia de COVID-19, publicados en idioma portugués, español o inglés, a partir de 2020, disponibles en su totalidad. Se excluyeron informes parciales de investigación, editoriales, cartas de respuesta y resúmenes de ponencias. Se realizó análisis descriptivo. Resultados De un total de 56 publicaciones, nueve estudios respondieron a los criterios de selección, de los cuales siete tenían calidad metodológica elevada y dos moderada. El fenómeno ocurrió en diferentes países del mundo, donde las víctimas habituales eran trabajadores de enfermería que actuaban en la línea de frente, con menor tiempo de graduación y perpetuada por diferentes actores. Se observaron daños psicosociales en los trabajadores, además de riesgos hacia los pacientes. Conclusión Los trabajadores de enfermería tuvieron una elevada incidencia de violencia durante la pandemia. Las consecuencias de esta violencia fueron perjudiciales para la salud, provocaron un deseo de dejar la profesión y afectaron la salud física o mental de las víctimas. Como medida de prevención y control, se sugirió la implementación de políticas públicas y estrategias administrativas para reducir la violencia contra esta población en condiciones de pandemias.

Abstract Objective To map and synthesize the available scientific evidence on violence against nursing workers during the COVID-19 pandemic. Methods Scoping review prepared in accordance with recommendations of the Joanna Briggs Institute, carried out in the following databases; National Library of Medicine, Cumulative Index to Nursing and Allied Health Literature, Web of Science, Excerpa Medica DataBASE, PsycINFO - APA PsycNET and Latin-American and Caribbean Health Sciences Literature. Studies addressing the theme of work-related violence against nursing workers during the COVID-19 pandemic, published in Portuguese, Spanish, or English, from 2020 and available in full were included. Partial research reports, editorials, response letters and abstracts of annals were excluded. Descriptive analysis was performed. Results Nine out of a total of 56 publications met the selection criteria, seven with high methodological quality and two with moderate quality. The phenomenon occurred in different countries around the world, the usual victims were nursing workers working on the front line, with less training time, and it was perpetrated by different actors. Psychosocial damage to workers and risks to patients were found. Conclusion Nursing workers suffered a high incidence of violence during the pandemic. The consequences of this violence were harmful to health, leading to the desire to leave the profession or affecting the physical or mental health of victims. The implementation of public policies and administrative strategies to reduce violence against this population in pandemic conditions were suggested as prevention and control measures.

Cryo-EM for Small Molecules.

In recent years, protein structure analysis using cryo-electron microscopy(cryo-EM) has expanded and improved. In this review, we discuss many recent improvements to the field, the problems those improvements hope to solve, and some of the still unanswered questions. Most notably, this review will discuss improvements in resolving small or fragmented protein structures, as well as methods to improve the signal-to-noise ratio of the data by increasing image contrast using carbon-based systems. We will also describe how, in the last 5 years, methodological improvements have allowed for better 3D image resolution by capturing a continuum of 3D images. We will provide examples of these methods in practice and discuss how these improved methods may be used in small-molecule drug discovery and development. © 2022 Wiley Periodicals LLC.

Coadministration of tianeptine alters behavioral parameters and levels of neurotrophins in a chronic model of Maple Syrup Urine disease.

Maple Syrup Urine Disease (MSUD) is caused by the deficiency in the activity of the branched-chain α-ketoacid dehydrogenase complex (BCKDC), resulting in the accumulation of the branched-chain amino acids (BCAA) leucine, isoleucine, and valine, and their respective branched-chain α-keto acids. Patients with MSUD are at high risk of developing chronic neuropsychiatric disorders; however, the pathophysiology of brain damage in these patients remains unclear. We hypothesize that MSUD can cause depressive symptoms in patients. To test our hypothesis, Wistar rats were submitted to the BCAA and tianeptine (antidepressant) administration for 21 days, starting seven days postnatal. Depression-like symptoms were assessed by testing for anhedonia and forced swimming after treatments. After the last test, the brain structures were dissected for the evaluation of neutrophins. We demonstrate that chronic BCAA administration induced depressive-like behavior, increased BDNF levels, and decreased NGF levels, suggesting a relationship between BCAA toxicity and brain damage, as observed in patients with MSUD. However, the administration of tianeptine was effective in preventing behavioral changes and restoring neurotrophins levels.

The use of standardized Brazilian green propolis extract (EPP-AF) as an adjunct treatment for hospitalized COVID-19 patients (BeeCovid2): a structured summary of a study protocol for a randomized controlled trial.

BACKGROUND: The 2019 coronavirus disease (COVID-19) pandemic continues to spread and affects large numbers of people with unprecedented impacts. Experimental evidence has already been obtained for use of the standardized extract of Brazilian green propolis (EPP-AF) against viral targets, and clinical rationality has been demonstrated for testing this extract as an adjunct to treatment in patients affected by COVID-19. The BeeCovid2 study aims to assess whether EPP-AF has an impact on the improvement of patients hospitalized with COVID-19 by reducing the length of hospital stay. METHODS: BeeCovid2 is a randomized, double-blinded, placebo-controlled clinical study being conducted in Brazil to provide further evidence on the effectiveness of standardized green propolis extract as an adjunctive treatment for adults hospitalized with COVID-19. Hospitalized patients over 18 years of age with a confirmed diagnosis of COVID-19 and up to 14 days of symptoms were included. Patients under mechanical ventilation at randomization, pregnant women, cancer patients, transplanted or using immunosuppression, HIV patients, patients who used propolis in the last 30 days, bacterial or fungal infection at randomization, impossibility of using medication orally or enterally, and advanced chronic diseases (e.g., advanced heart failure, severe liver disease, and end-stage chronic kidney disease). Enrolled patients are randomized at a 1:1 ratio to receive placebo or standardized propolis extract (900 mg/day) for 10 days. The study treatments are administered in a double-blinded manner, and patients are followed for 28 days. The primary outcome is the difference in length of hospital stay in days between groups. Secondary outcomes include the need for mechanical ventilation, the rate of secondary infection, rate of acute kidney injury, the need for renal replacement therapy, the requirement for vasoactive drugs, the use of an intra-aortic balloon pump (IABP), and the use of extracorporeal membrane oxygenation (ECMO). DISCUSSION: This trial is very useful and will provide more data on the effectiveness of using the standardized Brazilian green propolis extract as an adjunctive treatment in association with standard care in adults hospitalized with moderate to severe acute COVID-19. TRIAL REGISTRATION: ClinicalTrials.gov NCT04800224 . Registered on March 16, 2021.

Evaluation of urine SARS-COV-2 RT-PCR as a predictor of acute kidney injury and disease severity in patients with critical COVID-19.

Coronavirus disease 2019 (COVID-19) is an emerging infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which began as an outbreak in Wuhan, China and has spread rapidly across the globe. Although most infections are mild, patients with severe and critical COVID-19 infections face deterioration of respiratory function and may also have extrapulmonary manifestations, mostly affecting the kidney, digestive tract, heart, and nervous system. Here, we prospectively evaluated the presence of SARS-CoV-2 genetic material using reverse-transcription polymerase chain reaction in urine samples obtained from patients with COVID-19 receiving critical care. Among 51 included patients, we found higher serum creatinine levels, a longer hospital stay, and more frequent need for dialysis in urine-positive patients. These findings could suggest that, in predisposed patients, a direct viral cytopathic effect may contribute to a more severe disease phenotype.

Inter- and intraobserver agreement in computed tomography enterography in inflammatory bowel disease.

AIM: To evaluate intra- and interobserver agreement in imaging features in inflammatory bowel disease and comparison with fecal calprotectin (FC) levels. METHODS: Our institutional computed tomography enterography (CTE) database was retrospectively queried to identify patients who underwent CTE from January 2014 to June 2015. Patient inclusion criteria were confirmed inflammatory bowel disease (IBD) and FC collected < 4 mo after CTE without any change in clinical treatment or surgical treatment during this interval. The exclusion criterion was poor image quality. Two blinded abdominal radiologists, with 12 and 3 years of experience analyzed the CTE regarding localization (small bowel, colonic, both, or no disease detected); type of IBD (inflammatory, stenosing, fistulizing, > 1 pattern, or normal); and signs of active disease (present or absent). In 42 of 44 patients evaluated, routine CTE reports were made by one of the readers who re-evaluated the CTEs ≥ 6 mo later, to determine the intraobserver agreement. FC was considered a sign of disease activity when it was higher than 250 µg/g. RESULTS: Forty-four patients with IBD (38 with Crohn's disease and 6 with ulcerative colitis) were included. There was a moderate interobserver agreement regarding localization of IBD (κ = 0.540), type of disease (κ = 0.410) and the presence of active signs in CTE (κ = 0.419). There was almost perfect intraobserver agreement regarding localization, type and signs of active disease in IBD. The κ values were 0.902, 0.937 and 0.830, respectively. After a consensus between both radiologists regarding inflammatory activity in CTE, we found that 24 (85.7%) of 28 patients who were classified with active disease had elevated FC, and six (37.5%) of 16 patients without inflammatory activity in CTE had elevated FC (P = 0.003). The correlation between elevated FC and the presence of active disease in CTE was significant (κ = 0.495, P = 0.001). CONCLUSION: We found almost perfect intraobserver and moderate interobserver agreement in the signs of active disease in CTE with concurrence of high FC levels.

Viability of probiotic micro-organism Lactobacillus acidophilus in dairy chocolate dessert and its action against foodborne pathogens / Viabilidade de microrganismo probiótico Lactobacillus acidophilus em sobremesaLáctea de chocolate e ação sobre patógenos alimentares

ABSTRACT: The ability to produce antimicrobial factors is considered an important feature of probiotic microorganisms. Bacteriocins, hydrogen peroxide, acetic acid and lactic acid are examples of these substances. The present research aimed to develop probiotic dairy desserts (DD) with Lactobacillus acidophilus and evaluate the viability of this strain, as well as its action on food pathogens. Treatments with and without interactions between L. acidophilus and pathogenic Gram-negative bacteria (Salmonella sp. and Escherichia coli O157:H7) and Gram positive (Bacillus cereus and Staphylococcus aureus) were produced. The products were stored at a temperature of 8°C and analyzed at the times 24, 48, 72 hours, 7 days and 28 days (at 28 days, only T1 was analyzed because the other products were deteriorated). In an analysis of the potential for development of new products, the dairy dessert with L. acidophilus was considered a probiotic product. Assessment of the counts of pathogens in dairy desserts with or without L. acidophilus showed different behaviors of these products in response to pathogens, which could be justified by a possible action of bacteriocins or microbial competition, but there has been no overall reduction or reduction up to a safe level. It is concluded that the probiotic products developed reduced significant food pathogens, but not up to safe levels. Thus, we emphasize the importance of the use of quality tools in the development and monitoring of dairy desserts.

RESUMO: A capacidade de produzir fatores antimicrobianos é considerada uma importante característica dos microrganismos probióticos. Bacteriocinas, peróxido de hidrogênio, ácido acético e ácido lático, são exemplos destas substâncias. Com o presente trabalho, objetivou-se desenvolver sobremesas lácteas (SL) probióticas, acrescidas de Lactobacillus acidophilus e avaliar a viabilidade desta cepa, além da ação frente a patógenos alimentares. Foram produzidos tratamentos com e sem interações entre o L. acidophilus e bactérias patogênicas Gram negativas (Salmonella sp e Escherichia coli O157:H7) e Gram positivas (Bacillus cereus e Staphylococcus aureus). Os produtos foram armazenados em temperatura de 8ºC e analisados nos tempos 24, 48, 72 horas, 7 dias e 28 dias (apenas T1 por deterioração dos demais produtos neste tempo). A sobremesa láctea com L. acidophilus foi considerada produto probiótico, verificando o potencial de desenvolvimento de um novo produto. Analisando as contagens dos patógenos nas sobremesas lácteas com e sem adição de L. acidophilus, observaram-se diferentes comportamentos diante dos patógenos, o que poderia ser justificado por uma possível ação de bacteriocinas ou competição microbiana, porém não houve uma redução total ou até um nível considerado seguro. Conclui-se que os produtos probióticos desenvolvidos reduziram os patógenos alimentares de importância, porém não a níveis considerados seguros. Dessa forma, ressalta-se a importância de ferramentas de qualidade no desenvolvimento e monitoramento de sobremesas lácteas.

Formação de biofilmes de escherichia coli produtora de toxina Shiga sorotipos O153: H25, O113:H21 e O111: H8 em superfície de aço inoxidável e eficácia de sanitizante / Biofilm formation of shiga toxin-producing escherichia coli O153: H25, O113:H21 and O111: H8 on the stainless steel surface and the effectiveness of sanitizer

Neste estudo as cepas de Escherichia coli produtora de toxina Shiga (STEC) O153:H25, O113:H21 e O111:H8, isoladas de rebanhos do país, foram avaliadas quanto à capacidade de formar biofilmes em superfície de aço inoxidável utilizada na indústria de alimentos, bem como a eficácia de diferentes concentrações de hipoclorito de sódio na inativação desses biofilmes. A capacidade de formação de biofilme foi detectada em todas as cepas de E. coli produtoras de toxina Shiga. Na avaliação da eficáciado sanitizante hipoclorito de sódio, nas concentrações de 100 mg.L-1 e 200 mg.L-1, observou-se a redução a <1 log UFC/cm2 em todas as cepas e nos tempos avaliados. Estes dados reforçam aimportância do correto procedimento de higienização, uma vez que biofilmes podem tornar-se importantes fontes de contaminação no ambiente de produção de alimentos.

This study aimed at evaluating the biofilm formation of Shiga toxin-producing Escherichia coli O153:H25, O113:H21 and O111:H8 (STEC non-O157), isolated from Brazilian cattle, on the stainless steel surface, and also the efficacy of different s concentrations of sodium hypochlorite for inactivating these biofilms. The ability to form biofilm was demonstrated in all of Shiga toxin-producing E. coli strains. In assessing the effectiveness of sodium hypochlorite sanitizer, a reduction to <1 log CFU/cm2 was observed in all of the evaluated strains and times. These data strengthen the relevance of the correct cleaning procedure, considering that biofilms formations might be an important source of food contamination.