ATUAÇÃO ESTATAL E DIREITOS FUNDAMENTAIS NO CONTEXTO DA PANDEMIA DO COVID-19

A pandemia da COVID-19, provocada pelo Sars-Cov-2 coronavírus, exsurge-se num mundo globalizado, com técnicas científicas desenvolvidas para monitoramento e prevenção de riscos e com Estados comprometidos em assegurar às pessoas os direitos inerentes ao homem. Em meio à calamidade pública, desafia-se o Estado a atuar sob os moldes de intervenção ou abstenção, a fim de garantir a maior eficácia aos direitos fundamentais positivados, sem incorrer em excesso ou em deficiência de proteção

Correction: Phosphorescent bio-based resin for digital light processing (DLP) 3D-printing

Correction for 'Phosphorescent bio-based resin for digital light processing (DLP) 3D-printing' by Mirko Maturi et al., Green Chem., 2020, 22, 6212–6224, DOI: 10.1039/D0GC01983F

Accessory proteins of the zDHHC family of S-acylation enzymes

Almost two decades have passed since seminal work in Saccharomyces cerevisiae identified zinc finger DHHC domain-containing (zDHHC) enzymes as S-acyltransferases. These enzymes are ubiquitous in the eukarya domain, with 23 distinct zDHHC-encoding genes in the human genome. zDHHC enzymes mediate the bulk of S-acylation (also known as palmitoylation) reactions in cells, transferring acyl chains to cysteine thiolates, and in so-doing affecting the stability, localisation and function of several thousand proteins. Studies using purified components have shown that the minimal requirements for S-acylation are an appropriate zDHHC enzyme-substrate pair and fatty acyl-CoA. However, additional proteins including GCP16 (also known as Golga7), Golga7b, huntingtin and selenoprotein K, have been suggested to regulate the activity, stability and trafficking of certain zDHHC enzymes. In this Review, we discuss the role of these accessory proteins as essential components of the cellular S-acylation system

Identification of key features required for efficient S-acylation and plasma membrane targeting of Sprouty-2

Sprouty-2 is an important regulator of growth factor signalling and a tumour suppressor protein. The defining feature of this protein is a cysteine-rich domain (CRD) that contains twenty-six cysteines and is modified by S-acylation. In this study, we show that the CRD of Sprouty-2 is differentially modified by S-acyltransferase enzymes. The high specificity/low activity zDHHC17 enzyme mediated restricted S-acylation of Sprouty-2, and cysteines-265/268 were identified as key targets of this enzyme. In contrast, the low specificity/high activity zDHHC3/zDHHC7 enzymes mediated more expansive modification of the Sprouty-2 CRD. Nevertheless, S-acylation by all enzymes enhanced Sprouty-2 expression, suggesting that S-acylation stabilises this protein. In addition, we identified two charged residues (aspartate-214 and lysine-223), present on opposite faces of a predicted alpha helix in the CRD, which are essential for S-acylation of Sprouty-2. Interestingly, mutations that perturbed S-acylation also led to a loss of plasma membrane localisation of Sprouty-2 in PC12 cells. This study provides insight into the mechanisms and outcomes of Sprouty-2 S-acylation, and highlights distinct patterns of S-acylation mediated by different classes of zDHHC enzymes

Seleção de microrganismos produtores de enzimas hidrolíticas isolados da região do meio oeste de Santa Catarina, Brasil

A tecnologia enzimática vem sendo utilizada por oferecer vantagens no estabelecimento de um processo tecnologicamente limpo. Enzimas hidrolíticas estão entre os principais alvos da pesquisa biotecnológica pelo grande potencial de aplicação no setor industrial. Com o objetivo de selecionar e caracterizar microrganismos amilolíticos, pectinolíticos e xilanolíticos utilizaram-se solos e resíduos agroindustriais como fonte para o isolamento. Como resultados, dezoito cepas produtoras de amilase foram isoladas. Destas, onze apresentaram Índice Enzimático (IE) superior a 2.0 e foram determinados os perfis de regulação por repressão catabólica, sendo que três apresentaram características desejadas. Estes foram cultivados em meio líquido e tiveram a atividade de amilase sacarificante determinada. Também foram selecionados quatro microrganismos produtores de pectinase, que em cultivo em meio líquido confirmaram produção das enzimas pectina liase (PMGL) e de poligalacturanase (PG). Da mesma forma, a partir de cultivo em meio sólido, foram isolados treze microrganismos produtores de xilanases, sendo onze bactérias e dois fungos filamentosos. Cinco dos microrganismos que se destacaram quanto ao IE foram cultivados em meio líquido e analisados quando à produção da enzima, assim, um fungo filamentoso e uma bactéria, originários de solo, destacaram-se como produtores de xilanase. Apesar de haver a necessidade de aprofundamentos nos estudos quanto à caracterização desses isolados e das enzimas produzidas, os resultados positivos das atividades enzimáticas demonstraram que as metodologias utilizadas e os microrganismos selecionados são promissores na obtenção das respectivas enzimas com potencial para aplicação na indústria.Palavras-chave: Enzimas hidrolíticas. Isolamento de Microrganismos. Microbiologia Aplicada

Cylinder pressure based calibration model for engines using ethanol, hydrogen and natural gas as alternative fuels

This paper proposes a novel virtual engine calibration method for alternative fuels using thermodynamic simulation for in-cylinder pressure prediction. Based on known engine data, including the crank angle of the peak cylinder pressure, the optimization problem is defined for a desired indicated mean effective pressure. The decision variables are the combustion and heat transfer model parameters The method was tested for three different engines of different sizes, operating with ethanol, hydrogen and natural gas, and different equivalence ratios. The Wiebe model and a quasi-dimensional fractal combustion model were compared. The results showed that the method was able to successfully predict the in-cylinder pressure curve, with a coefficient of determination higher than 0.99. Furthermore, the method predicted the peak pressure and the crank angle corresponding to 50% of mass fraction burned with a maximum deviation of 2.5% and 1.5 °CA, respectively

Violência financeira/econômica contra a pessoa idosa: protocolo de revisão de escopo

Objetivo: Mapear na literatura nacional e internacional, sobre o tema central da violência financeira econômica contra a pessoa idosa. Método: O protocolo fundamentou-se nas recomendações metodológicas propostas pelo Joanna Briggs Institute - Manual for Evidence Synthesis e Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) Checklist, com a finalidade de responder à pergunta de pesquisa: “Quais são tipos de estudos e resultados que a literatura científica traz sobre violência financeira/econômica contra a pessoa idosa”? Tendo por base o mnemônico PCC (População; Conceito; Contexto), onde “P” refere-se aos idosos; “C” se refere ao fenômeno violência e, “C” refere-se à violência financeira/econômica. As fontes de dados serão as seguintes bases de dados, nacionais e internacionais: Scientific Electronic Library Online (SciELO.org), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), Medical Literature Analysis and Retrieval System Online (MEDLINE/PubMed), SciVerse Scopus (Scopus), Cummulative Index to Nursing and Allied Health Literature (CINAHL), Base de Dados em Enfermagem (BDENF), Biblioteca Digital Brasileira de Teses e Dissertações (BDTD), Embase (Elsevier), Cochrane Library, Catálogo de Teses e Dissertações (CAPES), Google Acadêmico,  e, ainda,  repositórios, bancos de teses e dissertações e as referências encontradas nos estudos selecionados. Esse protocolo encontra-se registrado no Open Science Framework (OSF), sob o DOI 10.17605/OSF.IO/VZ7G9

S-acylation of Sprouty and SPRED proteins by the S-acyltransferase zDHHC17 involves a novel mode of enzyme-substrate interaction

S-Acylation is an essential post-translational modification, which is mediated by a family of twenty-three zDHHC enzymes in humans. Several thousand proteins are modified by S-acylation; however, we lack a detailed understanding of how enzyme-substrate recognition and specificity is achieved. Previous work showed that the ankyrin repeat domain of zDHHC17 (ANK17) recognizes a short linear motif, known as the zDHHC ANK binding motif (zDABM) in substrate protein SNAP25, as a mechanism of substrate recruitment prior to S-acylation. Here, we investigated the S-acylation of the Sprouty and SPRED family of proteins by zDHHC17. Interestingly, although Sprouty-2 (Spry2) contains a zDABM that interacts with ANK17, this mode of binding is dispensable for S-acylation, and indeed removal of the zDABM does not completely ablate binding to zDHHC17. Furthermore, the related SPRED3 protein interacts with and is efficiently S-acylated by zDHHC17 despite lacking a zDABM. We undertook mutational analysis of SPRED3 to better understand the basis of its zDABM-independent interaction with zDHHC17. This analysis found that the cysteine-rich SPR domain of SPRED3, which is the defining feature of all Sprouty and SPRED proteins, interacts with zDHHC17. Surprisingly, the interaction with SPRED3 was independent of ANK17. Our mutational analysis of Spry2 was consistent with the SPR domain of this protein containing a zDHHC17 binding site, and Srpy2 also showed detectable binding to a zDHHC17 mutant lacking the ANK domain. Thus, zDHHC17 can recognize its substrates through ANK domain and zDABM-dependent and –independent mechanisms, and some substrates display more than one mode of binding to this enzyme

Assessment of the effect of sphingosine kinase inhibitors on apoptosis,unfolded protein response and autophagy of T-cell acute lymphoblastic leukemia cells; indications for novel therapeutics

Sphingosine 1-phosphate (S1P) is a bioactive lipid that is formed by the phosphorylation of sphingosine and catalysed by sphingosine kinase 1 (SK1) or sphingosine kinase 2 (SK2). Sphingosine kinases play a fundamental role in many signaling pathways associated with cancer, suggesting that proteins belonging to this signaling network represent potential therapeutic targets. Over the last years, many improvements have been made in the treatment of T-cell acute lymphoblastic leukemia (T-ALL); however, novel and less toxic therapies are still needed, especially for relapsing and chemo-resistant patients. Here, we analyzed the therapeutic potential of SKi and ROMe, a sphingosine kinase 1 and 2 inhibitor and SK2-selective inhibitor, respectively. While SKi induced apoptosis, ROMe initiated an autophagic cell death in our in vitro cell models. SKi treatment induced an increase in SK1 protein levels in Molt-4 cells, whereas it activated the endoplasmic reticulum (ER) stress/unfolded protein response (UPR) pathway in Jurkat and CEM-R cells as protective mechanisms in a sub-population of T-ALL cells. Interestingly, we observed a synergistic effect of SKi with the classical chemotherapeutic drug vincristine. In addition, we reported that SKi affected signaling cascades implicated in survival, proliferation and stress response of cells. These findings indicate that SK1 or SK2 represent potential targets for treating T-ALL