Lipid Fraction from Agaricus brasiliensis as a Potential Therapeutic Agent for Lethal Sepsis in Mice.

Sepsis is a potentially fatal clinical condition that results from an immune imbalance in the host during an infection. It presents systemic alterations due to excessive activation of pro-inflammatory mediators that contribute to inflammation, formation of reactive species, and tissue damage. Anti-inflammatory mediators are then extensively activated to regulate this process, leading to immune exhaustion and, consequently, immunosuppression of the host. Considering the biological activities of the nutraceutical Agaricus brasiliensis (A. brasiliensis), such as immunomodulatory, antioxidant, and antitumor activities, the present study investigated the therapeutic potential of the lipid fraction of A. brasiliensis (LF) in a model of lethal sepsis in mice (Mus musculus), induced by cecal ligation and perforation (CLP). The results showed that treatment of septic animals with LF or LF associated with ertapenem (LF-Erta) reduced systemic inflammation, promoting improvement in clinical parameters and increased survival. The data show a reduction in pro-inflammatory and oxidative stress markers, regulation of the anti-inflammatory response and oxidizing agents, and increased bacterial clearance in the peritoneal cavity and liver. Thus, it can be concluded that LF as a treatment, and in conjunction with antibiotic therapy, has shown promising effects as a hepatoprotective, antioxidant, antimicrobial, and immunomodulatory agent.

A contribuição do pensamento de Paulo Freire para a Vigilância Popular em Saúde / The contribution of Paulo Freire's thought to Popular Health Surveillance

Resumo Paulo Freire pensava a Educação Popular (EP) como um processo emancipatório, por meio da problematização e agir crítico como dimensões da existência humana. A partir de suas contribuições aos campos da Educação e da Saúde, este texto tem como objetivo realizar uma revisão narrativa de literatura sobre as articulações das ideias de Paulo Freire com as ações de Vigilância Popular em Saúde (VPS). O texto apresenta como a contribuição de Paulo Freire e da Educação Popular em Saúde inspirou a construção de uma Vigilância Popular em Saúde, que busca promover a transformação da realidade local frente às situações de violações de direitos e na defesa da vida. Dessa maneira, experiências dos territórios e das populações em situação de vulnerabilidade, na maioria das vezes, lançam mão de estratégias pedagógicas da Educação Popular para se constituírem como práticas de VPS. É no território que a Educação Popular em Saúde se torna fundamental para o estímulo à transformação da percepção dos indivíduos, problematizando sua realidade. A promoção de uma práxis acerca da "situação limite" no cotidiano dos atores sociais permite um diagnóstico da realidade, baseado na informação científica em diálogo com a cultura e a organização popular, como possibilidade de construção de "inéditos viáveis".

Abstract Paulo Freire considered Popular Education (PE) as an emancipatory process, through debate and critical action, as a dimension of human existence. This text aims to conduct a narrative literature review on the articulations of Paulo Freire's ideas with Popular Health Surveillance (PHS) actions based on his contributions to Education and Health. The text presents how the contributions of Paulo Freire and Popular Health Education inspired the construction of Popular Health Surveillance, which seeks to promote the transformation of local reality in the face of rights violations and advocate for life. Thus, experiences from vulnerable territories and populations often use Popular Education pedagogical strategies to establish PHS practices. Popular Health Education becomes crucial in the territory to stimulate the transformation of individual perception and discuss their reality. Promoting a praxis about the "critical situation" in the daily lives of social stakeholders allows diagnosing reality based on scientific information in dialogue with culture and popular organization as a possibility of building the "viable unprecedented".

In Vitro Digestibility and Models of Cumulative Gas Production of Forage-Free Diet.

Our objectives were to evaluate the use of cottonseed cake in replacing corn silage in a diet without forage and to identify the model with higher precision and accuracy of adjustment of parameters of ruminal degradation kinetics. A diet containing corn silage and another with cottonseed cake as a fiber source were formulated. Gompertz, Dual-pool Logistic, Brody, and Ørskov models were evaluated for goodness of fit to gas production. There were significant differences in dry matter (DM), organic matter (OM), and neutral detergent fiber (NDF) in the in vitro digestibility for diets and fiber sources. The estimated values of the Gompertz (6.77), Brody (6.72), and Ørskov (6.73) models were similar to the observed mean of gas production in the corn silage diet (6.73 mL/100 mg DM). Similarly, the estimated values of the Brody (5.87) and Ørskov (5.89) models were similar to the observed mean of gas production in the cottonseed cake diet (5.87 mL/100 mg DM). The roughage-free diet containing cottonseed cake as a fiber source stimulated higher gas production. Brody and Ørskov models presented higher precision and accuracy in the fitting of kinetics of degradation independent of the fiber source in the diet.

Oleanolic Acid Glycosides from Scabiosa caucasica and Scabiosa ochroleuca: Structural Analysis and Cytotoxicity.

In the field of research on medicinal plants from the Armenian flora, the phytochemical study of two Scabiosa L. species, S. caucasica M. Bieb. and S. ochroleuca L. (Caprifoliaceae), has led to the isolation of five previously undescribed oleanolic acid glycosides from an aqueous-ethanolic extract of the roots: 3-O-α-L-rhamnopyranosyl-(1→3)-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-xylopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester. Their full structural elucidation required extensive 1D and 2D NMR experiments, as well as mass spectrometry analysis. For the biological activity of the bidesmosidic saponins and the monodesmosidic saponin, their cytotoxicity on a mouse colon cancer cell line (MC-38) was evaluated.

SPECT Imaging of Lysyl Oxidase-like 2 in a Model of Idiopathic Pulmonary Fibrosis.

Noninvasive imaging of idiopathic pulmonary fibrosis (IPF) remains a challenge. The aim of this study was to develop an antibody-based radiotracer targeting Lysyl Oxidase-like 2 (LOXL2), an enzyme involved in the fibrogenesis process, for SPECT/CT imaging of pulmonary fibrosis. The bifunctional chelator DOTAGA-PEG4-NH2 was chemoenzymatically conjugated to the murine antibody AB0023 using microbial transglutaminase, resulting in a degree of labeling (number of chelators per antibody) of 2.3. Biolayer interferometry confirmed that the binding affinity of DOTAGA-AB0023 to LOXL2 was preserved with a dissociation constant of 2.45 ± 0.04 nM. DOTAGA-AB0023 was then labeled with 111In and in vivo experiments were carried out in a mice model of progressive pulmonary fibrosis induced by intratracheal administration of bleomycin. [111In]In-DOTAGA-AB0023 was injected in three groups of mice (control, fibrotic, and treated with nintedanib). SPECT/CT images were recorded over 4 days p.i. and an ex vivo biodistribution study was performed by gamma counting. A significant accumulation of the tracer in the lungs of the fibrotic mice was observed at D18 post-bleomycin. Interestingly, the tracer uptake was found selectively upregulated in fibrotic lesions observed on CT scans. Images of mice that received the antifibrotic drug nintedanib from D8 up to D18 showed a decrease in [111In]In-DOTAGA-AB0023 lung uptake associated with a decrease in pulmonary fibrosis measured by CT scan. In conclusion, we report the first radioimmunotracer targeting the protein LOXL2 for nuclear imaging of IPF. The tracer showed promising results in a preclinical model of bleomycin-induced pulmonary fibrosis, with high lung uptake in fibrotic areas, and accounted for the antifibrotic activity of nintedanib.

Saúde e saneamento: uma avaliação das políticas públicas de prevenção, controle e contingência das arboviroses no Brasil / Health and sanitation: an evaluation of public policies for the prevention, control, and contingency of arboviruses in Brazil

Resumo As arboviroses, sobretudo as transmitidas pelo mosquito Aedes aegypti, têm-se constituído em grave problema de saúde pública no Brasil. Com o intuito de analisar como o saneamento básico é abordado em instrumentos norteadores das políticas públicas de controle das arboviroses no país, foi realizada uma análise de conteúdo em oito documentos governamentais de referência. Como resultados, foi possível identificar que aspectos relacionados à comunicação e mobilização social, controle vetorial e gestão são os temas mais abordados nos documentos analisados. Já as componentes do saneamento básico, destacando-se o manejo e a drenagem das águas pluviais e o esgotamento sanitário não são abordados nos instrumentos. A intersetorialidade é mencionada, no entanto, não existem proposições específicas que apontem e assegurem sua implementação. As Diretrizes Nacionais para a Prevenção e Controle de Epidemias de Dengue, do Ministério da Saúde, constitui-se no documento mais completo sobre o assunto. Conclui-se que o saneamento básico não está suficientemente abordado nos instrumentos de enfrentamento às arboviroses o que pode contribuir para a baixa efetividade de intervenção pública e que, portanto, tal contradição precisa ser superada pelas políticas públicas no Brasil.

Abstract Arboviruses, especially those transmitted by the Aedes aegypti mosquito, have become a serious public health problem in Brazil. In order to analyze how sanitation is addressed in the basic guidelines of public policies to control arboviruses in the country, content analysis was performed on eight governmental reference documents. As a result, it was possible to identify that aspects related to communication and social mobilization, vector control, and management are the themes most addressed in the documents analyzed. On the other hand, the components of basic sanitation, especially rainwater management and drainage, and sewage control, are not addressed in the instruments. Intersectorality is mentioned, however, there are no specific proposals to implement the plan and ensure its implementation. The Ministry of Health's National Guidelines for the Prevention and Control of Dengue Epidemics is the most complete document on the subject. The conclusion drawn is that basic sanitation is not sufficiently addressed in the instruments for combatting arboviruses, which can contribute to the low effectiveness of public intervention and, consequently, this discrepancy needs to be focused on by public policies in Brazil.

Genotype-first approach to identify associations between CDH1 germline variants and cancer phenotypes: a multicentre study by the European Reference Network on Genetic Tumour Risk Syndromes.

BACKGROUND: Truncating pathogenic or likely pathogenic variants of CDH1 cause hereditary diffuse gastric cancer (HDGC), a tumour risk syndrome that predisposes carrier individuals to diffuse gastric and lobular breast cancer. Rare CDH1 missense variants are often classified as variants of unknown significance. We conducted a genotype-phenotype analysis in families carrying rare CDH1 variants, comparing cancer spectrum in carriers of pathogenic or likely pathogenic variants (PV/LPV; analysed jointly) or missense variants of unknown significance, assessing the frequency of families with lobular breast cancer among PV/LPV carrier families, and testing the performance of lobular breast cancer-expanded criteria for CDH1 testing. METHODS: This genotype-first study used retrospective diagnostic and clinical data from 854 carriers of 398 rare CDH1 variants and 1021 relatives, irrespective of HDGC clinical criteria, from 29 institutions in ten member-countries of the European Reference Network on Tumour Risk Syndromes (ERN GENTURIS). Data were collected from Oct 1, 2018, to Sept 20, 2022. Variants were classified by molecular type and clinical actionability with the American College of Medical Genetics and Association for Molecular Pathology CDH1 guidelines (version 2). Families were categorised by whether they fulfilled the 2015 and 2020 HDGC clinical criteria. Genotype-phenotype associations were analysed by Student's t test, Kruskal-Wallis, χ2, and multivariable logistic regression models. Performance of HDGC clinical criteria sets were assessed with an equivalence test and Youden index, and the areas under the receiver operating characteristic curves were compared by Z test. FINDINGS: From 1971 phenotypes (contributed by 854 probands and 1021 relatives aged 1-93 years), 460 had gastric and breast cancer histology available. CDH1 truncating PV/LPVs occurred in 176 (21%) of 854 families and missense variants of unknown significance in 169 (20%) families. Multivariable logistic regression comparing phenotypes occurring in families carrying PV/LPVs or missense variants of unknown significance showed that lobular breast cancer had the greatest positive association with the presence of PV/LPVs (odds ratio 12·39 [95% CI 2·66-57·74], p=0·0014), followed by diffuse gastric cancer (8·00 [2·18-29·39], p=0·0017) and gastric cancer (7·81 [2·03-29·96], p=0·0027). 136 (77%) of 176 families carrying PV/LPVs fulfilled the 2015 HDGC criteria. Of the remaining 40 (23%) families, who did not fulfil the 2015 criteria, 11 fulfilled the 2020 HDGC criteria, and 18 had lobular breast cancer only or lobular breast cancer and gastric cancer, but did not meet the 2020 criteria. No specific CDH1 variant was found to predispose individuals specifically to lobular breast cancer, although 12 (7%) of 176 PV/LPV carrier families had lobular breast cancer only. Addition of three new lobular breast cancer-centred criteria improved testing sensitivity while retaining high specificity. The probability of finding CDH1 PV/LPVs in patients fulfilling the lobular breast cancer-expanded criteria, compared with the 2020 criteria, increased significantly (AUC 0·92 vs 0·88; Z score 3·54; p=0·0004). INTERPRETATION: CDH1 PV/LPVs were positively associated with HDGC-related phenotypes (lobular breast cancer, diffuse gastric cancer, and gastric cancer), and no evidence for a positive association with these phenotypes was found for CDH1 missense variants of unknown significance. CDH1 PV/LPVs occurred often in families with lobular breast cancer who did not fulfil the 2020 HDGC criteria, supporting the expansion of lobular breast cancer-centred criteria. FUNDING: European Reference Network on Genetic Tumour Risk Syndromes, European Regional Development Fund, Fundação para a Ciência e a Tecnologia (Portugal), Cancer Research UK, and European Union's Horizon 2020 research and innovation programme.

Novel FERMT3 and PTPRQ Mutations Associated with Leukocyte Adhesion Deficiency-III and Sensorineural Hearing Loss.

Leukocyte adhesion deficiency-III (LAD-III) is a rare genetic disease caused by defective integrin activation in hematopoietic cells due to mutations in the FERMT3 gene. The PTPRQ gene encodes the protein tyrosine phosphatase receptor Q and is essential for the normal maturation and function of hair bundle in the cochlea. Homozygous PTPRQ mutations impair the stereocilia in hair cells which lead to nonsyndromic sensorineural hearing loss (SNHL) with vestibular dysfunction. Here, we report two novel pathogenic homozygous mutations found in two genes, FERMT3 and PTPRQ , in a Brazilian patient with LAD-III and SNHL, which may develop our understanding of the phenotype-genotype correlation and prognosis of patients with these rare diseases.

Timed Up and Go na avaliação da fragilidade de pessoas idosas agricultoras do Rio Grande do Sul: estudo transversal / Timed Up and Go in assessing the frailty of older farmers in Rio Grande do Sul: cross-sectional study

Resumo Objetivos Avaliar a sensibilidade do teste Timed Up and Go test (TUG) como preditor da síndrome da fragilidade do idoso (SFI) da população rural idosa do Rio Grande do Sul (RS) e identificar a prevalência de SFI nessa população. Método Estudo transversal, realizado com 604 agricultores com mais de 60 anos de idade (321 homens e 283 mulheres) identificados por meio de conglomerados estruturados a partir das regionais da Federação dos Trabalhadores da Agricultura do Rio Grande do Sul (FETAG-RS) e respectivos sindicatos. Além de variáveis demográficas (sexo, idade), foi avaliada a mobilidade funcional mediante a realização do TUG e a fragilidade referida. A curva Receiver-Operating Characteristic (ROC) foi construída para avaliar um ponto de corte do teste TUG para fragilidade. Resultados A SFI ou fragilidade foi identificada em 52,5% (n=317) da população pesquisada; 35,1% (n=212) pré-frageis e 12,4% (n=75) não-frágeis. E o tempo médio de realização do TUG em relação ao sexo foi de 11,6 segundos para mulheres e 10,8 segundos para homens (p=0,0001). A progressão da idade esteve relacionada com maior tempo de realização do teste (idosos jovens - 60-64 anos; idosos mais velhos -75-79 e longevos - 80+ - p=0,0001). A curva ROC indicou 10 segundos na execução do teste TUG como melhor ponto de corte para diagnóstico da SF em idosos rurais. Conclusão A frequência de fragilidade e pré-fragilidade nesta pesquisa, indicam uma condição de vulnerabilidade do trabalhador rural do RS no seu processo de envelhecimento. Demonstrando, a partir do teste TUG, características de mobilidade funcional e risco de fragilidade dos agricultores mais velhos, importantes para considerações futuras sobre as singularidades da saúde dessa população e intervenções profissionais necessárias.

Abstract Objectives To evaluate the sensitivity of the Timed Up and Go test (TUG) as a predictor of frailty syndrome in the elderly (IFS) in the elderly rural population of Rio Grande do Sul (RS) and to identify the prevalence of IFS in this population. Method Cross-sectional study, carried out with 604 farmers over 60 years of age (321 men and 283 women) identified through clusters structured from the regions of the Federation of Agricultural Workers of Rio Grande do Sul (FETAG-RS) and respective unions. In addition to demographic variables (gender, age), functional mobility was assessed by performing the TUG and reported frailty. The Receiver-Operating Characteristic (ROC) curve was constructed to assess a TUG test cutoff point for frailty. Results IFS or frailty was identified in 52.5% (n=317) of the surveyed population; 35.1% (n=212) pre-frail and 12.4% (n=75) non-frail. And the mean time to perform the TUG varied according to gender was 11.6 seconds for women and 10.8 seconds for men - (p=0.0001). The progression of age was related to longer time spent on the age test (young elderly - 60-64 years old; older elderly -75-79 and oldest old - 80+ - p=0.0001). The ROC curve indicated 10 seconds in the execution of the TUG test as the best cutoff point for diagnosing the SF frailty syndrome in rural elderly. Conclusion The frequency of frailty and pre-frailty in this research indicates a condition of vulnerability of rural workers in RS in their aging process. Demonstrating, from the TUG test, characteristics of functional mobility and risk of frailty of older farmers, important for future considerations on the singularities of the health of this population and necessary professional interventions.

Aplicativos móveis desenvolvidos para a gestão intra-hospitalar de pacientes com acidente vascular cerebral: uma revisão sistemática / Mobile applications developed for the in-hospital management of patients with stroke: a systematic review

O uso de tecnologias móveis e sem fio para apoiar a realização dos objetivos de saúde é uma tendência dentro da e-saúde. Existem lacunas de conhecimento adicionais no uso emergente de smartphones para a gestão e monitoramento de doenças crônicas como o acidente vascular cerebral, principalmente dentro do ambiente hospitalar. O objetivo deste estudo foi o de revisar os estudos que utilizaram aplicativos móveis para a gestão de pacientes hospitalizados com acidente vascular cerebral. Esta revisão sistemática seguiu as recomendações preconizadas pelo Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), sendo realizada nas bases de dados eletrônicas Medline/PubMed e Embase, além de busca manual às referências de estudos já publicados sobre o assunto. Foram encontrados 207 estudos e após análise dos critérios de inclusão e exclusão, 6 foram incluídos para análise completa. O uso de aplicativos móveis pode acelerar o tempo de atendimento de pacientes internados com acidente vascular cerebral, diminuindo a morte neuronal, melhorando a mortalidade hospitalar, diminuindo a hemorragia intracraniana e contribuindo para um retorno precoce para a casa dos pacientes.

The use of mobile and wireless technologies to support the achievement of health goals is a trend within e-health. There are additional knowledge gaps in the emerging use of smartphones for the management and monitoring of chronic diseases such as stroke, particularly within the hospital setting. The objective this study was review studies developed with the interaction of mobile applications for the management of hospitalized patients with stroke. This systematic review followed the recommendations recommended by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and was carried out in the Medline/PubMed and Embase electronic databases, in addition to a manual search of references to studies already published on the subject, and 207 studies were found and after analyzing the inclusion and exclusion criteria, 6 were included for complete analysis. The use of mobile applications can accelerate the care times of hospitalized patients with stroke, decreasing neuronal death, improvement in hospital mortality, less intracranial hemorrhage, and contributing to an early return home of patients.

IL-1RAP, a Key Therapeutic Target in Cancer.

Cancer is a major cause of death worldwide and especially in high- and upper-middle-income countries. Despite recent progress in cancer therapies, such as chimeric antigen receptor T (CAR-T) cells or antibody-drug conjugate (ADC), new targets expressed by the tumor cells need to be identified in order to selectively drive these innovative therapies to tumors. In this context, IL-1RAP recently showed great potential to become one of these new targets for cancer therapy. IL-1RAP is highly involved in the inflammation process through the interleukins 1, 33, and 36 (IL-1, IL-33, IL-36) signaling pathways. Inflammation is now recognized as a hallmark of carcinogenesis, suggesting that IL-1RAP could play a role in cancer development and progression. Furthermore, IL-1RAP was found overexpressed on tumor cells from several hematological and solid cancers, thus confirming its potential involvement in carcinogenesis. This review will first describe the structure and genetics of IL-1RAP as well as its role in tumor development. Finally, a focus will be made on the therapies based on IL-1RAP targeting, which are now under preclinical or clinical development.

Superparamagnetic Iron Oxide Nanoparticles for Immunotherapy of Cancers through Macrophages and Magnetic Hyperthermia.

Cancer immunotherapy has tremendous promise, but it has yet to be clinically applied in a wider variety of tumor situations. Many therapeutic combinations are envisaged to improve their effectiveness. In this way, strategies capable of inducing immunogenic cell death (e.g., doxorubicin, radiotherapy, hyperthermia) and the reprogramming of the immunosuppressive tumor microenvironment (TME) (e.g., M2-to-M1-like macrophages repolarization of tumor-associated macrophages (TAMs)) are particularly appealing to enhance the efficacy of approved immunotherapies (e.g., immune checkpoint inhibitors, ICIs). Due to their modular construction and versatility, iron oxide-based nanomedicines such as superparamagnetic iron oxide nanoparticles (SPIONs) can combine these different approaches in a single agent. SPIONs have already shown their safety and biocompatibility and possess both drug-delivery (e.g., chemotherapy, ICIs) and magnetic capabilities (e.g., magnetic hyperthermia (MHT), magnetic resonance imaging). In this review, we will discuss the multiple applications of SPIONs in cancer immunotherapy, focusing on their theranostic properties to target TAMs and to generate MHT. The first section of this review will briefly describe immune targets for NPs. The following sections will deal with the overall properties of SPIONs (including MHT). The last section is dedicated to the SPION-induced immune response through its effects on TAMs and MHT.

Characterization and in vitro evaluations of Baru pulp from almond production waste ensiled with different additives.

The silage process is an efficient way of storing nutrients for animal nutrition. Our hypothesis was that the Baru pulp can be preserved in the form of silage, regardless of the use of additives to aid the process. Silages of Baru pulp containing different additives were evaluated through in vitro analysis, fermentation parameters, and composition and thermal analysis including differential scanning calorimetry and thermogravimetry. The treatments consisted of (1) silage with Baru pulp (BP) in natura without additive; (2) BP in natura with acetic acid; (3) BP in natura with formic acids; and (4) BP in natura added with microbial inoculums in a randomized experimental design with three replications per treatment and analyzed in duplicate. The ensiled material was kept in anaerobic conditions for a period of 30 days. BP before and after the silage process presented averages of 67.31 and 66.24% for in vitro digestibility of DM (IVDMD). Microbial inoculant additive was the most effective in reducing pH, followed by acetic acid and formic acid. There were effects of additives on silages for all degradation parameters in ruminal liquid in vitro. It was observed that BP before ensiling had the highest A fraction (7.9 mL gas/100 mg DM), without differing from the silage treated with formic acid (7.1 mL gas/100 mg DM). Similar effects were observed on mass loss (TG) and heat flux (DSC) between the silages. Our findings suggest that Baru pulp with formic acid is more efficient in the conservation and preservation of fermentable carbohydrates as well as in silage production.

Panorama dos Planos Municipais de Saneamento Básico e Planos Diretores de Drenagem Urbana em municípios de pequeno porte de Minas Gerais / Panorama of elaboration of Municipal Basic Sanitation Plans and Urban Drainage Master Plans of Minas Gerais' small municipalities

RESUMO No Brasil, os impactos socioambientais da crise hídrica são intensificados uma vez que a drenagem e o manejo das águas pluviais são marcados por ausências e limitações de políticas públicas. O saneamento básico é de titularidade municipal e, para o exercício desse direito e dever, é fundamental que os municípios estruturem a política municipal, sendo o planejamento seu principal instrumento. Nesse contexto, buscou-se neste artigo identificar e analisar o panorama da elaboração de Planos Municipais de Saneamento Básico e de Planos Diretores de Drenagem Urbana nos municípios do estado de Minas Gerais com população inferior a 50 mil habitantes e comparar esses resultados com as informações divulgadas pelo Sistema Nacional de Informações sobre Saneamento no ano de 2019. Realizaram-se buscas na internet e contatos com os municípios, por telefone, e foi possível identificar que, entre os 752 municípios de Minas Gerais de pequeno porte, 499 possuem Planos Municipais de Saneamento Básico, enquanto nenhum possui Planos Diretores de Drenagem Urbana. Ao comparar esses resultados com as informações divulgadas pelo Sistema Nacional de Informações sobre Saneamento, observou-se que 79 municípios prestaram informações inconsistentes relacionadas à existência de Planos Municipais de Saneamento Básico e 48 afirmaram possuir Planos Diretores de Drenagem Urbana. Conclui-se que muitos municípios mineiros ainda não possuem Planos Municipais de Saneamento Básico, e que o processo de coleta dos dados do Sistema Nacional de Informações sobre Saneamento precisa ser aprimorado para que haja melhor qualidade dos dados divulgados. A ausência de instrumentos de planejamento que auxiliem a efetivação da política municipal é um importante problema a ser superado para o enfrentamento dos complexos desafios que envolvem a drenagem e o manejo das águas pluviais municipal.

ABSTRACT In Brazil, the impacts of the water crisis are intensified since the Drainage and Rainwater Management are marked by absences and limitations of public policies. Basic sanitation is a municipal property and for the exercise of this right and duty, it is essential that the municipalities structure the municipal policy, with planning as their main instrument. In this context, this article aimed to identify and analyze the panorama of the elaboration of Municipal Basic Sanitation Plans and Master Plans for Urban Drainage in the municipalities of the state of Minas Gerais with a population of less than 50,000 inhabitants and to compare these results with the information released by the National Sanitation Information System, in 2019. Internet searches and contacts with the municipalities were carried out by telephone, so that it was possible to identify that among the 752 small-sized ones in Minas Gerais, 499 of them have Municipal Basic Sanitation Plans, while none of them have Master Plans for Urban Drainage. When comparing these results with the information released by the National Sanitation Information System, it was observed that 79 municipalities provided inconsistent information related to the existence of Municipal Basic Sanitation Plans and 48 municipalities claimed to have Master Plans for Urban Drainage. It is concluded that many municipalities in Minas Gerais do not yet have Municipal Basic Sanitation Plans and that the process of collecting National Sanitation Information System data needs to be improved so that there is a better quality of the data released. The absence of planning instruments to assist the implementation of municipal policy is an important problem to be overcome in order to face the complex challenges involving municipal Stormwater Drainage and Management.

Sarcopenia, oxidative stress and inflammatory process in muscle of cirrhotic rats - Action of melatonin and physical exercise.

Sarcopenia is one of the most common features of cirrhosis, contributing to morbidity and mortality in this population. We aimed to evaluate the effect of melatonin (MLT) and exercise (EX) on the quadriceps muscle in rats with biliary cirrhosis induced by bile duct ligation (BDL). We used 48 males (mean weight = 300 g), divided into eight groups. A 20 mg/Kg MLT dose was administered via i.p. (1 x daily), and the EX, the animals were set to swim in couples for 10 min each day. Upon completion, blood, liver, and quadriceps samples were taken for analysis. In the liver enzymes analysis and comet assay results, a reduction was observed in the groups treated with MLT with/or EX comparing to the BDL group. In the evaluation of substances that react to thiobarbituric acid (TBARS), nitric oxide levels (NO), and tumor necrosis factor-alpha levels (TNF-α), there was a significant increase in the BDL group and a reduction in the treated groups. In the activity of the superoxide dismutase enzyme (SOD) and interleukin-10 levels (IL-10) concentrations, there was a significant increase in the treated groups of the BDL group. Histological analysis revealed muscle hypotrophy in the BDL group in comparison with the control group (CO) and increased muscle mass in the treated groups. There was an increase in weight gain and phase angle in the groups treated with MLT with/or EX comparing to the BDL group. We suggest that treatments may contribute to the reduction of muscle changes in cirrhotic patients.

Fragment binding to the Nsp3 macrodomain of SARS-CoV-2 identified through crystallographic screening and computational docking.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) macrodomain within the nonstructural protein 3 counteracts host-mediated antiviral adenosine diphosphate-ribosylation signaling. This enzyme is a promising antiviral target because catalytic mutations render viruses nonpathogenic. Here, we report a massive crystallographic screening and computational docking effort, identifying new chemical matter primarily targeting the active site of the macrodomain. Crystallographic screening of 2533 diverse fragments resulted in 214 unique macrodomain-binders. An additional 60 molecules were selected from docking more than 20 million fragments, of which 20 were crystallographically confirmed. X-ray data collection to ultra-high resolution and at physiological temperature enabled assessment of the conformational heterogeneity around the active site. Several fragment hits were confirmed by solution binding using three biophysical techniques (differential scanning fluorimetry, homogeneous time-resolved fluorescence, and isothermal titration calorimetry). The 234 fragment structures explore a wide range of chemotypes and provide starting points for development of potent SARS-CoV-2 macrodomain inhibitors.

Nanofitins targeting heat shock protein 110: An innovative immunotherapeutic modality in cancer.

The presence of an inactivating heat shock protein 110 (HSP110) mutation in colorectal cancers has been correlated with an excellent prognosis and with the ability of HSP110 to favor the formation of tolerogenic (M2-like) macrophages. These clinical and experimental results suggest a potentially powerful new strategy against colorectal cancer: the inhibition of HSP110. In this work, as an alternative to neutralizing antibodies, Nanofitins (scaffold ~7 kDa proteins) targeting HSP110 were isolated from the screening of a synthetic Nanofitin library, and their capacity to bind (immunoprecipitation, biolayer interferometry) and to inhibit HSP110 was analyzed in vitro and in vivo. Three Nanofitins were found to inhibit HSP110 chaperone activity. Interestingly, they share a high degree of homology in their variable domain and target the peptide-binding domain of HSP110. In vitro, they inhibited the ability of HSP110 to favor M2-like macrophages. The Nanofitin with the highest affinity, A-C2, was studied in the CT26 colorectal cancer mice model. Our PET/scan experiments demonstrate that A-C2 may be localized within the tumor area, in accordance with the reported HSP110 abundance in the tumor microenvironment. A-C2 treatment reduced tumor growth and was associated with an increase in immune cells infiltrating the tumor and particularly cytotoxic macrophages. These results were confirmed in a chicken chorioallantoic membrane tumor model. Finally, we showed the complementarity between A-C2 and an anti-PD-L1 strategy in the in vivo and in ovo tumor models. Overall, Nanofitins appear to be promising new immunotherapeutic lead compounds.

Crystallographic and electrophilic fragment screening of the SARS-CoV-2 main protease.

COVID-19, caused by SARS-CoV-2, lacks effective therapeutics. Additionally, no antiviral drugs or vaccines were developed against the closely related coronavirus, SARS-CoV-1 or MERS-CoV, despite previous zoonotic outbreaks. To identify starting points for such therapeutics, we performed a large-scale screen of electrophile and non-covalent fragments through a combined mass spectrometry and X-ray approach against the SARS-CoV-2 main protease, one of two cysteine viral proteases essential for viral replication. Our crystallographic screen identified 71 hits that span the entire active site, as well as 3 hits at the dimer interface. These structures reveal routes to rapidly develop more potent inhibitors through merging of covalent and non-covalent fragment hits; one series of low-reactivity, tractable covalent fragments were progressed to discover improved binders. These combined hits offer unprecedented structural and reactivity information for on-going structure-based drug design against SARS-CoV-2 main protease.

Gene expression profile suggesting immunological dysregulation in two Brazilian Bloom's syndrome cases.

BACKGROUND: Bloom syndrome (BS) is a rare autosomal recessive chromosome instability disorder. The main clinical manifestations are growth deficiency, telangiectasic facial erythema, immunodeficiency, and increased risk to develop neoplasias at early age. Cytogenetic test for sister chromatid exchanges (SCEs) is used as a diagnostic marker for BS. In addition, most patients also present mutations in the BLM gene, related to defects in the DNA repair mechanism. However, the molecular mechanism behind the pathogenicity of BS is still not completely understood. METHODS: We describe two patients confirmed with BS by SCE and molecular analysis. Also, we performed the gene expression profile by the RNA-seq methodology in mRNA transcripts for differential gene expression analysis using as a biological condition for comparison BS versus health controls. RESULTS: We detected 216 differentially expressed genes related to immunological pathways such as positive regulation and activation of B cells, immune effector process and absence of difference of DNA repair genes expression. In addition; we also observed differentially expressed genes associated with apoptosis control, such as BCL2L1, CASP7, CDKN1A, E2F2, ITPR, CD274, TNFAIP6, TNFRSF25, TNFRSF13C, and TNFRSF17. CONCLUSION: Our results suggest that the combination of altered expression of genes involved in signaling pathways of immune response and apoptosis control may contribute directly to the main characteristics observed in BS, such as recurrent infections, growth failure, and high risk of cancer. Transcriptome studies of other instability syndromes could allow a more accurate analysis of the relevant gene interactions associated with the destabilization of the genome. This is a first description of the profile of differential gene expression related to immunological aspects detected in patients with BS by RNA-seq.

Selecting the first chemical molecule inhibitor of HSP110 for colorectal cancer therapy.

Pro-survival stress-inducible chaperone HSP110 is the only HSP for which a mutation has been found in a cancer. Multicenter clinical studies demonstrated a direct association between HSP110 inactivating mutation presence and excellent prognosis in colorectal cancer patients. Here, we have combined crystallographic studies on human HSP110 and in silico modeling to identify HSP110 inhibitors that could be used in colorectal cancer therapy. Two molecules (foldamers 33 and 52), binding to the same cleft of HSP110 nucleotide-binding domain, were selected from a chemical library (by co-immunoprecipitation, AlphaScreening, Interference-Biolayer, Duo-link). These molecules block HSP110 chaperone anti-aggregation activity and HSP110 association to its client protein STAT3, thereby inhibiting STAT3 phosphorylation and colorectal cancer cell growth. These effects were strongly decreased in HSP110 knockdown cells. Foldamer's 33 ability to inhibit tumor growth was confirmed in two colorectal cancer animal models. Although tumor cell death (apoptosis) was noted after treatment of the animals with foldamer 33, no apparent toxicity was observed, notably in epithelial cells from intestinal crypts. Taken together, we identified the first HSP110 inhibitor, a possible drug-candidate for colorectal cancer patients whose unfavorable outcome is associated to HSP110.