Structural and thermodynamic characterization of a highly amyloidogenic dimer of transthyretin involved in a severe cardiomyopathy.

Transthyretin (TTR) is a homotetrameric protein involved in the transport of thyroxine. More than 150 different mutations have been described in the TTR gene, several of them associated with familial amyloid cardiomyopathy (FAC). Recently, our group described a new variant of TTR in Brazil, namely A39D-TTR, which causes a severe cardiac condition. Position 39 is in the AB loop, a region of the protein that is located within the thyroxine-binding channels and is involved in tetramer formation. In the present study we solved the structure and characterize the thermodynamic stability of this new variant of TTR using urea and high hydrostatic pressure (HHP). Interestingly, during the process of purification, A39D-TTR turned out to be a dimer and not a tetramer, a variation that might be explained by the close contact of the four aspartic acids at position 39, where they face each other inside the thyroxine channel. In the presence of sub-denaturing concentrations of urea, bis-ANS binding and dynamic light scattering revealed A39D-TTR in the form of a molten-globule dimer. Co-expression of A39D and WT isoforms in the same bacterial cell did not produce heterodimers or heterotetramers, suggesting that somehow a negative charge at the AB loop precludes tetramer formation. A39D-TTR proved to be highly amyloidogenic, even at mildly acidic pH values where WT-TTR does not aggregate. Interestingly, despite being a dimer, aggregation of A39D-TTR was inhibited by diclofenac, which binds to the thyroxine channel in the tetramer, suggesting the existence of other pockets in A39D-TTR able to accommodate this molecule.

Transcriptional profile of ribosome-associated quality control components and their associated phenotypes in mammalian cells.

During protein synthesis, organisms detect translation defects that induce ribosome stalling and result in protein aggregation. The Ribosome-associated Quality Control (RQC) complex, comprising TCF25, LTN1, and NEMF, is responsible for identifying incomplete protein products from unproductive translation events, targeting them for degradation. Although RQC disruption causes adverse effects on vertebrate neurons, data regarding mRNA/protein expression and regulation across tissues are lacking. Employing high-throughput methods, we analyzed public datasets to explore RQC gene expression and phenotypes. Our findings revealed widespread expression of RQC components in human tissues; however, silencing of RQC yielded only mild negative effects on cell growth. Notably, TCF25 exhibited elevated mRNA levels that were not reflected in the protein content. We experimentally demonstrated that this disparity arose from post-translational protein degradation by the proteasome. Additionally, we observed that cellular aging marginally influenced RQC expression, leading to reduced mRNA levels in specific tissues. Our results suggest the necessity of RQC expression in all mammalian tissues. Nevertheless, when RQC falters, alternative mechanisms seem to compensate, ensuring cell survival under nonstress conditions.

Suspicious of Acute Kidney Graft Rejection: Tacrolimus Pharmacokinetics Under Methylprednisolone Therapy.

BACKGROUND: Acute rejection remains one of the main complications in the first months after transplantation and may influence long-term outcomes. Tacrolimus has proven its usefulness in solid organ transplants and its monitoring through the application of pharmacokinetic concepts to optimize individual drug therapy. OBJECTIVE: This research proposes to evaluate the tacrolimus pharmacokinetic parameters in patients suspected of acute kidney graft rejection under methylprednisolone pulse therapy. METHODS: Eleven adult tacrolimus-treated renal recipients were selected from a prospective, single-arm, single-center cohort study, with suspicion of acute rejection although in use of methylprednisolone pulses therapy. They were followed up for three months posttransplantation, being tacrolimus trough serum concentrations determined using a chemiluminescent magnetic immunoassay, and pharmacokinetic parameters were estimated by using a nonlinear mixed-effects model implemented by Monolix 2020R1. A tacrolimus trough serum concentration range of 8 to 12 ng.mL-1 was considered therapeutic. RESULTS: Six patients showed acute cellular rejection, and two of them in addition had an antibody- mediated rejection. Tacrolimus trough serum concentration was below the reference range in eight patients. Most patients showed a high tacrolimus concentration intrapatient and pharmacokinetic parameters variability. CONCLUSION: The obtained pharmacokinetics parameters helped in understanding the kidney recipient patients' tacrolimus behavior, assisting in the improvement of individual drug therapy and reducing the risk of acute rejection episodes.

Impact of pathogenic bacterial communities present in wastewater on aquatic organisms: Application of nanomaterials for the removal of these pathogens.

Contaminated wastewater (WW) can cause severe hazards to numerous delicate ecosystems and associated life forms. In addition, human health is negatively impacted by the presence of microorganisms in water. Multiple pathogenic microorganisms in contaminated water, including bacteria, fungi, yeast, and viruses, are vectors for several contagious diseases. To avoid the negative impact of these pathogens, WW must be free from pathogens before being released into stream water or used for other reasons. In this review article, we have focused on pathogenic bacteria in WW and summarized the impact of the different types of pathogenic bacteria on marine organisms. Moreover, we presented a variety of physical and chemical techniques that have been developed to provide a pathogen-free aquatic environment. Among the techniques, membrane-based techniques for trapping hazardous biological contaminants are gaining popularity around the world. Besides, novel and recent advancements in nanotechnological science and engineering suggest that many waterborne pathogens could be inactivated using nano catalysts, bioactive nanoparticles, nanostructured catalytic membranes, nanosized photocatalytic structures, and electrospun nanofibers and processes have been thoroughly examined.

Gestores municipais de saúde da Paraíba: percepções, abordagens e práticas inerentes ao fenômeno da judicialização / Municipal health managers in the state of Paraíba, Brazil: perceptions, approaches and practices relatedto the phenomenon of judicialization / Gestores de salud municipales en el estado de Paraíba, Brasil: percepciones, enfoques y prácticas asociados conel fenómeno de la judicialización

Objetivo: analisar a percepção de gestores municipais paraibanos sobre o tema da judicialização da saúde. Metodologia: trata-se de uma pesquisa descritiva e analítica, com delimitação transversal, realizada em 2018. Foram aplicados questionários a 31 gestores de saúde municipais, que representam proporcionalmente à 75% da população do estado da Paraíba. Resultados: os resultados apontam a judicialização como fenômeno mais fortemente ligado a fatores externos à gestão do Sistema Único de Saúde (SUS). O processo de discussão da judicialização tem mais ressonância nos espaços das Comissões Intergestores, e os órgãos representativos e o Ministério Público são os maiores aliados no enfrentamento. Identifica-se fragilidades na construção dos instrumentos de gestão e, entre as soluções possíveis, estão o maior conhecimento desse fenômeno pelos órgãos e setores de gestão do SUS, bem como na qualificação das ações de regulação. Conclusão: percebe-se a complexidade da judicialização da saúde, desde a sua compreensão até a forma desafiadora de gerenciá-la. É fundamental estabelecer estratégias que promovam a comunicação e a informação, ampliando o conhecimento dos órgãos de controle e dos setores de gestão do SUS, a fim de lidar de forma mais eficaz com esse fenômeno.

Objective: to analyze the perceptions of municipal leaders in the state of Paraíba, Brazil, on the topic of health judicialization. Methodology: it is a descriptive and analytical study of a cross-sectional nature, carried out in 2018. Questionnaires were administered to 31 municipal health managers, proportionately representing 75% of the population of the state of Paraíba, Brazil. Results: judicialization is a phenomenon more associated with factors outside the management of the Brazilian Unified Health System (SUS). The discussion process about judicialization tends to take place in the Inter-institutional Commissions.The representative bodies and the Public Ministry are the main allies in the dispute. Weaknesses in the construction of administrative instruments have been identified, and solutions include better knowledge of this phenomenon by the SUS administrative bodies and sectors, and in the qualification of regulatory measures. Conclusion: the complexity of health judicializationis perceived, from its understanding to the difficulties of managing it. It is important to develop strategies that promote communication and information and improve the knowledge of regulatory bodies and SUS administrative sectors to better deal with this phenomenon.

Objetivo: analizar la percepción de los gestores municipales del estado de Paraíba, en Brasil, sobre la judicialización de la salud. Metodología: se trata de una investigación descriptiva y analítica, con delimitación transversal, realizada en 2018. Se aplicaron cuestionarios a 31 gestores municipales de salud, que representan proporcionalmente el 75% de la población del estado de Paraíba, en Brasil. Resultados: los resultados apuntan a la judicialización como un fenómeno más fuertemente ligado a factores externos a la gestión del Sistema Único de Salud (SUS) brasileño. Los órganos de representación y el Ministerio Público son los principales aliados in el enfrentamiento, y el proceso de discusión de la judicialización tiene mayor resonancia en los espacios de las Comisiones Interinstitucionales. Fue posible identificar debilidades en la construcción de instrumentos de gestióny, entre las posibles soluciones se encuentran un mayor conocimiento de este fenómeno por parte de los órganos y sectores gestores del SUS, así como en la calificación de las acciones normativas. Conclusión: se reconoce la complejidad de la judicialización de la salud, desde cómo se entiende hasta cómo se maneja. Para enfrentar con mayor eficacia este fenómeno, es necesario establecer estrategias que promuevan la comunicación y la información, ampliando el conocimiento de los órganos de control y de los sectores de gestión del SUS.

Biosynthesis of Nanoparticles Using Plant Extracts and Essential Oils.

Plant extracts and essential oils have a wide variety of molecules with potential application in different fields such as medicine, the food industry, and cosmetics. Furthermore, these plant derivatives are widely interested in human and animal health, including potent antitumor, antifungal, anti-inflammatory, and bactericidal activity. Given this diversity, different methodologies were needed to optimize the extraction, purification, and characterization of each class of biomolecules. In addition, these plant products can still be used in the synthesis of nanomaterials to reduce the undesirable effects of conventional synthesis routes based on hazardous/toxic chemical reagents and associate the properties of nanomaterials with those present in extracts and essential oils. Vegetable oils and extracts are chemically complex, and although they are already used in the synthesis of nanomaterials, limited studies have examined which molecules are effectively acting in the synthesis and stabilization of these nanostructures. Similarly, few studies have investigated whether the molecules coating the nanomaterials derived from these extracts and essential oils would bring benefits or somehow reduce their potential activity. This synergistic effect presents a promising field to be further explored. Thus, in this review article, we conducted a comprehensive review addressing the main groups of molecules present in plant extracts and essential oils, their extraction capacity, and available methodologies for their characterization. Moreover, we highlighted the potential of these plant products in the synthesis of different metallic nanomaterials and their antimicrobial capacity. Furthermore, we correlated the extract's role in antimicrobial activity, considering the potential synergy between molecules from the plant product and the different metallic forms associated with nanomaterials.

Environmental Implications Associated with the Development of Nanotechnology: From Synthesis to Disposal.

Nanotechnology remains under continuous development. The unique, fascinating, and tunable properties of nanomaterials make them interesting for diverse applications in different fields such as medicine, agriculture, and remediation. However, knowledge about the risks associated with nanomaterials is still poorly known and presents variable results. Furthermore, the interaction of nanomaterials with biological systems and the environment still needs to be clarified. Moreover, some issues such as toxicity, bioaccumulation, and physicochemical transformations are found to be dependent on several factors such as size, capping agent, and shape, making the comparisons even more complex. This review presents a comprehensive discussion about the consequences of the use and development of nanomaterials regarding their potential risks to the environment as well as human and animal health. For this purpose, we reviewed the entire production chain from manufacturing, product development, applications, and even product disposal to raise the important implications at each stage. In addition, we present the recent developments in terms of risk management and the recycling of nanomaterials. Furthermore, the advances and limitations in the legislation and characterization of nanomaterials are also discussed.

Forest-Fruticulture Conversion Alters Soil Traits and Soil Organic Matter Compartments.

Fruticulture in the Amazonian Rainforest is one of the main causes of deforestation, biodiversity loss, and soil erosion. Fruticulture plays a key role in the soil traits and soil organic matter (SOM) compartments by altering the soil ecosystem. Our aim was to assess the influence of Forest-Fruticulture conversion on soil traits, and SOM fractions in Brazil's Legal Amazon. The experiment was carried out in field conditions using four land uses as main treatments: Bixa orellana, Theobroma grandiflorum, Paullinia cupana, and the Amazon Rainforest. The soil physicochemical traits were analyzed using samples that were collected from 0-5, 5-10, and 10-20 cm soil depth by using grids (10 × 10 m) with 36 sampling points. Our results showed that the Fruticulture promoted an increase in bulk density, GMD, aggregate diameter, soil porosity, gravimetric moisture, sand, clay, carbon associated with humic acid, and, the sum of bases (K+, Ca2+, and Mg2+), while the Amazon Rainforest showed the highest values of silt, soil P content, SOC, p-SOC, m-SOC, carbon associated with fulvic acid, humine, and soil C stock. Overall, the fruticulture farming systems have negative effects on SOM compartments. The results of our study highlight the importance of considering fruticulture with endemic plant species by promoting soil fertility and soil aggregation.

Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production.

The ferulic acid (FA) represents a high-value molecule with applications in the cosmetic and pharmaceutical industries. This aromatic molecule is derived from lignin and can be enzymatically converted in other commercially interesting molecules, such as vanillin and bioplastics. This process starts with a common step of FA activation via CoA-thioesterification, catalyzed by feruloyl-CoA synthetases. Therefore, here, we report the successfully expression, purification as well as the initial structural and biochemical characterization of a stable, correctly folded, and catalytically active bacterial feruloyl-CoA synthase (here named FCS3) isolated from a lignin-degrading microbial consortium. The purification of recombinant FCS3 to near homogeneity was achieved using affinity chromatography. The FCS3 structure is composed of a mixture of α and ß secondary structures and most likely forms stable homodimers in solution. The FCS3 presented a notable structural stability at alkaline pH values and it was able to convert FA and coenzyme A (CoA) into feruloyl-CoA complex at room temperature. This study should provide a useful basis for future biotechnological applications of FCS3, especially in the field of conversion of lignin-derived FA into high value compounds.

Determinantes da judicialização da saúde: uma análise bibliográfica / Determinants of health judicialization: a bibliographical analysis / Determinantes de la judicialización de la salud: un análisis bibliográfico

Objetivo: a judicialização da saúde é um fenômeno multifacetado e requer a construção de caminhos que possibilitem a análise de fatores que, isolada ou conjuntamente, influenciam em sua constituição; assim, torna-se necessário identificar quais são os fatores apontados pela literatura científica nacional como causas da judicialização no campo da saúde. Metodologia: foi feita uma revisão integrativa de literatura, fundamentada no estudo exploratório-descritivo e bibliográfico de 101 publicações contidas na base de dados da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, usando cinco descritores compatíveis com a pesquisa, a saber: judicialização e saúde e Brasil; judicialização da saúde e Direito; judicialização e saúde e causas; judicialização e saúde e consequências; e judicialização e direito à saúde. Resultados: dos 101 artigos catalogados, 30 foram selecionados; tal amostra foi organizada em três categorias, a partir de sua temática: 14 artigos discutiam a judicialização da assistência farmacêutica; 10 artigos abordavam a judicialização do direito à saúde; e 6 artigos debatiam sobre a judicialização com ênfase nas práticas de gestão. Conclusão: ainda é incipiente a produção intelectual brasileira para a identificação e sistematização dos determinantes da judicialização. Contudo, foi possível identificar caminhos que permitiram uma leitura preliminar sobre a temática, que foram divididos em três categorias: judicialização da assistência farmacêutica; judicialização do direito à saúde; e judicialização e práticas de gestão

Objective: health judicialization is a multifaceted phenomenon; thus, it is necessary to identify what are the factors that cause this phenomenon in Brazilian scientific literature. Methods: it was conducted an integrative literature review, based on an exploratory-descriptive and bibliographic study of 101 publications available in Capes database. The search used 5 descriptors: judicialization and health and Brazil; health judicialization and law; judicialization and health and causes; judicialization and health and consequences; and judicialization and the right to health. Results: of a corpus of 101 articles, the authors selected 30, which were organized into three categories based on its theme: 14 articles discussed the judicialization of pharmaceutical assistance; 10 articles addressed the judicialization of the right to health; and 6 articles discussed judicialization with emphasis on management practices. Conclusion: Brazilian intellectual production is still incipient to identify and systematize the determinants of health judicialization. However, it was possible to have a preliminary understanding of the main themes, which were divided into three categories: the judicialization of pharmaceutical assistance, the judicialization of the right to health and the judicialization and management practices.

Objetivo: la judicialización de la salud es un fenómeno multifacético y requiere la construcción de caminos que permitan analizar los factores que, individualmente o en conjunto, inciden en su constitución; por tanto, se hace necesario identificar cuáles son los factores señalados por la literatura científica nacional como causas de judicialización en el campo de la salud. Metodología: se realizó una revisión integradora de la literatura, a partir de un estudio exploratorio-descriptivo y bibliográfico de 101 publicaciones contenidas en la base de datos de la Coordinación para el Perfeccionamiento del Personal de Educación Superior, utilizando cinco descriptores compatibles con la investigación, a saber: judicialización y salud. y Brasil; judicialización de la salud y el derecho; judicialización y salud y causas; judicialización y salud y consecuencias; y judicialización y derecho a la salud. Resultados: de los 101 artículos catalogados, se seleccionaron 30; esta muestra se organizó en tres categorías, en función de su temática: 14 artículos discutieron la judicialización de la asistencia farmacéutica; 10 artículos abordaron la judicialización del derecho a la salud; y 06 artículos discutieron la judicialización con énfasis en las prácticas de gestión. Conclusión: la producción intelectual brasileña es aún incipiente para la identificación y sistematización de los determinantes de la judicialización. Sin embargo, fue posible identificar caminos que permitieron una lectura preliminar sobre el tema, que se dividió en tres categorías: la primera, judicialización de la asistencia farmacéutica, seguida de la judicialización del derecho a la salud y finalmente, judicialización y prácticas de gestión.

Fire lead to disturbance on organic carbon under sugarcane cultivation but is recovered by amendment with vinasse.

Sugarcane burning has been widely practiced in Brazil and worldwide. In the long term, this farming practice can cause soil erosion, reduction in organic carbon (OC) and consequently, changes in the structure of soil organic matter (SOM). Such changes may be difficult to reverse. This study aimed to assess the medium- and long-term effects of sugarcane burning on SOM characteristics, both in terms of quantity and structural quality and evaluate the application of vinasse as a strategy to attenuate fire-induced changes in burned soil. The experiment was conducted in a 50-year-old sugarcane field on soils classed as Cambissolo Háplico (Inceptisol). Four plots were sampled: a) burning of sugarcane for harvest for 37 years (SCB37); b) renewal of the sugarcane field and burning for harvest for 3 years (SCB3); c) renewal of the sugarcane field without burning for harvest for 3 years (SCWB), and d) renewal of the sugarcane field and burning for harvest with the application of vinasse for 3 years (SCV). Chemical and physical characterization of SOM was performed by solid-state spectroscopy (UV-vis, ATR-FTIR e 13C NMR CP/MAS) and chemometric techniques. The results showed that sugarcane burning drastically impacts SOM content and its chemical structure, however, the application of vinasse preserves and restores the soil from the fire effects. Content of soil OC, particulate OC, mineral-associated OC, humic acid, humin and light fraction OM that were affected by fire, had an increase and recovery of contents by the vinasse application. Solid state spectroscopy showed that labile structures were lost in humic acids (HA) by fire and recalcitrant structures were preserved. The application of vinasse incorporated fragments of lipids and carbohydrates in HA structure. Burning sugar cane straw affects the integrity of soil organic matter but can be restored by applying vinasse.

Dps protein is related to resistance of Mycobacterium abscessus subsp. massiliense against stressful conditions.

Mycobacterium abscessus subsp. massiliense (Mycma) belongs to the Mycobacterium abscessus complex and is a rapidly growing non-tuberculous mycobacterium. The chronic pulmonary, skin, and soft tissue infections that it causes may be difficult to treat due to its intrinsic resistance to the commonly used antimicrobial drugs, making it a serious world public health problem. Iron is an essential nutrient for the growth of microorganisms; nonetheless, it can be toxic when in excess. Thus, bacteria require an iron homeostasis mechanism to succeed in different environments. DNA-binding proteins from starved cells (Dps) are miniferritins with the property to act as additional iron storage proteins but also can bind to DNA, protecting it against hydroxyl radical. Annotation of the Mycma genome revealed the gene mycma_03135 with 79% sequential identity when compared to MSMEG_3242 gene from M. smegmatis mc2 155, which codifies for a known Dps. Recombinant Dps from M. abscessus (rMaDps) was produced in Escherichia coli, purified in soluble form and shown to form high mass oligomers in solution with ferroxidase activity, DNA binding, and protection against damage. The expression of the mycma_03135 gene was induced during Mycma growth in the presence of hydrogen peroxide (H2O2). Additionally, the expression of rMaDps by E. coli conferred greater resistance to H2O2. Thus, this study is the first to identify and characterize a Dps from M. abscessus. KEY POINTS: Mycobacterium abscessus subsp. massiliense express a miniferritin protein (Dps). Mycma Dps binds to DNA and protects against oxidative stress.