Fragment-Based Discovery of Allosteric Inhibitors of SH2 Domain-Containing Protein Tyrosine Phosphatase-2 (SHP2).

The ubiquitously expressed protein tyrosine phosphatase SHP2 is required for signaling downstream of receptor tyrosine kinases (RTKs) and plays a role in regulating many cellular processes. Genetic knockdown and pharmacological inhibition of SHP2 suppresses RAS/MAPK signaling and inhibit the proliferation of RTK-driven cancer cell lines. Here, we describe the first reported fragment-to-lead campaign against SHP2, where X-ray crystallography and biophysical techniques were used to identify fragments binding to multiple sites on SHP2. Structure-guided optimization, including several computational methods, led to the discovery of two structurally distinct series of SHP2 inhibitors binding to the previously reported allosteric tunnel binding site (Tunnel Site). One of these series was advanced to a low-nanomolar lead that inhibited tumor growth when dosed orally to mice bearing HCC827 xenografts. Furthermore, a third series of SHP2 inhibitors was discovered binding to a previously unreported site, lying at the interface of the C-terminal SH2 and catalytic domains.

Role of Cystathionine ß-Synthase and 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics of Murine Breast Cancer Cells.

Cystathionine ß-synthase (CBS), CSE (cystathionine γ-lyase) and 3-mercaptopyruvate sulfurtransferase (3-MST) have emerged as three significant sources of hydrogen sulfide (H2S) in various forms of mammalian cancer. Here, we investigated the functional role of CBS' and 3-MST's catalytic activity in the murine breast cancer cell line EO771. The CBS/CSE inhibitor aminooxyacetic acid (AOAA) and the 3-MST inhibitor 2-[(4-hydroxy-6-methylpyrimidin-2-yl)sulfanyl]-1-(naphthalen-1-yl)ethan-1-one (HMPSNE) were used to assess the role of endogenous H2S in the modulation of breast cancer cell proliferation, migration, bioenergetics and viability in vitro. Methods included measurements of cell viability (MTT and LDH assays), cell proliferation and in vitro wound healing (IncuCyte) and cellular bioenergetics (Seahorse extracellular flux analysis). CBS and 3-MST, as well as expression were detected by Western blotting; H2S production was measured by the fluorescent dye AzMC. The results show that EO771 cells express CBS, CSE and 3-MST protein, as well as several enzymes involved in H2S degradation (SQR, TST, and ETHE1). Pharmacological inhibition of CBS or 3-MST inhibited H2S production, suppressed cellular bioenergetics and attenuated cell proliferation. Cell migration was only inhibited by the 3-MST inhibitor, but not the CBS/CSE inhibitor. Inhibition of CBS/CSE of 3-MST did not significantly affect basal cell viability; inhibition of 3-MST (but not of CBS/CSE) slightly enhanced the cytotoxic effects of oxidative stress (hydrogen peroxide challenge). From these findings, we conclude that endogenous H2S, generated by 3-MST and to a lower degree by CBS/CSE, significantly contributes to the maintenance of bioenergetics, proliferation and migration in murine breast cancer cells and may also exert a minor role as a cytoprotectant.

Validation of a New HPLC-DAD Method to Quantify 3-Deoxyanthocyanidins Based on Carajurin in Medicinal Plants and for Chemical Ecology Studies.

Anthocyanidins are found in nature mainly as pelargonidin, cyanidin, peonidin, delphinidin, petunidin, and malvidin derivatives. These compounds are found free or as glycoside derivatives which are responsible for the color (red, blue, and violet) of some foods and are responsible for attracting seed dispersers. They are grouped into 3-hydroxyanthocyanidins, 3-deoxyanthocyanidins (3D-anth), and O-methylated anthocyanidins. A new method was developed and validated to quantify 3D-anth in plant-rich extracts. To test the new method, Arrabidaea chica Verlot was selected as it is widely used in folk medicine, and it is rich in 3D-anth. The new method was developed by HPLC-DAD and expressed 3D-anth as carajurin content. Carajurin was chosen as the reference standard due to its role as a biological marker for the antileishmanial activity for A. chica. The selected method used a silica-based phenyl column, a mobile phase composed of potassium dihydrogen phosphate buffer, acetonitrile, and methanol, in a gradient elution mode and detection at 480 nm. The method reliability was confirmed by verifying selectivity, linearity, precision, recovery, and robustness. This method contributes to quality control and development of a possible active pharmaceutical ingredient from A. chica as well as it can be used to evaluate 3D-anth in plant extracts with chemical ecology interest.

Fibrx Rocking Chair: Design and Application of Tailored Timber as an Embedded Frame for Natural Fibre-Reinforced Polymer (NFRP) Coreless Winding.

The building industry needs to innovate towards a more sustainable future and can do so through a combination of more renewable material choices and less wasteful fabrication processes. To address these issues, a hybrid material and fabrication system was developed using laminated timber veneer and natural fibre-reinforced composites (NFRPs), two materials that are leveraged for their potential of strategic material placement in additive processes towards programmed material behaviour and performance. The main contribution is in the hybrid fabrication approach, using thin, bent laminated veneer as an embedded frame for coreless filament winding of NFRP, which removes the need for temporary, wasteful formwork that is typically required to achieve structurally performative bent timber or FRP elements. Integrative methods are developed for the design, simulation, and fabrication of a rocking chair prototype that illustrates the architectural potential of the developed fabrication approach.

A commentary on the use of pharmacoenhancers in the pharmaceutical industry and the implication for DMPK drug discovery strategies.

Paxlovid, a drug combining nirmatrelvir and ritonavir, was designed for the treatment of COVID-19 and its rapid development has led to emergency use approval by the FDA to reduce the impact of COVID-19 infection on patients.In order to overcome potentially suboptimal therapeutic exposures, nirmatrelvir is dosed in combination with ritonavir to boost the pharmacokinetics of the active product.Here we consider examples of drugs co-administered with pharmacoenhancers.Pharmacoenhancers have been adopted for multiple purposes such as ensuring therapeutic exposure of the active product, reducing formation of toxic metabolites, changing the route of administration, and increasing the cost-effectiveness of a therapy.We weigh the benefits and risks of this approach, examining the impact of technology developments on drug design and how enhanced integration between cross-discipline teams can improve the outcome of drug discovery.

Efficacy of Clinically Used PARP Inhibitors in a Murine Model of Acute Lung Injury.

Poly(ADP-ribose) polymerase 1 (PARP1), as a potential target for the experimental therapy of acute lung injury (ALI), was identified over 20 years ago. However, clinical translation of this concept was not possible due to the lack of clinically useful PARP inhibitors. With the clinical introduction of several novel, ultrapotent PARP inhibitors, the concept of PARP inhibitor repurposing has re-emerged. Here, we evaluated the effect of 5 clinical-stage PARP inhibitors in oxidatively stressed cultured human epithelial cells and monocytes in vitro and demonstrated that all inhibitors (1-30 µM) provide a comparable degree of cytoprotection. Subsequent in vivo studies using a murine model of ALI compared the efficacy of olaparib and rucaparib. Both inhibitors (1-10 mg/kg) provided beneficial effects against lung extravasation and pro-inflammatory mediator production-both in pre- and post-treatment paradigms. The underlying mechanisms include protection against cell dysfunction/necrosis, inhibition of NF-kB and caspase 3 activation, suppression of the NLRP3 inflammasome, and the modulation of pro-inflammatory mediators. Importantly, the efficacy of PARP inhibitors was demonstrated without any potentiation of DNA damage, at least as assessed by the TUNEL method. These results support the concept that clinically approved PARP inhibitors may be repurposable for the experimental therapy of ALI.

Profile of tobacco growers with green tobacco sickness in Southern Brazil.

Introduction: Green tobacco sickness affects tobacco growers while handling tobacco leaves, regardless of the cultivation stage. Objectives: To characterize the sociodemographic and occupational profiles of tobacco growers with green tobacco sickness, as well as their health habits. Methods: This was a cross-sectional, descriptive study with information from a database obtained from a previous study conducted in a Southern Brazilian municipality. The data were analyzed using absolute and relative frequency. Results: We identified 8 cases of tobacco growers with green tobacco sickness, whose sociodemographic and occupational profiles and health habits were described. Conclusions: The data obtained in this study corroborate the existing literature on tobacco growers. The sociodemographic and occupational profiles and the health habits of our study participants have already been described in other studies, as well as of tobacco growers without green tobacco sickness.

Effects of the PARP Inhibitor Olaparib on the Response of Human Peripheral Blood Leukocytes to Bacterial Challenge or Oxidative Stress.

Prior studies demonstrate the activation of poly-(ADP-ribose) polymerase 1 (PARP1) in various pathophysiological conditions, including sepsis. We have assessed the effect of olaparib, a clinically used PARP1 inhibitor, on the responses of human peripheral blood leukocytes (PBMCs) obtained from healthy volunteers in response to challenging with live bacteria, bacterial lipopolysaccharide (LPS), or oxidative stress (hydrogen peroxide, H2O2). The viability of PBMCs exposed to olaparib or to the earlier generation PARP inhibitor PJ-34 (0.1-1000 µM) was monitored using Annexin V and 7-aminoactinomycin D. To evaluate the effects of olaparib on the expression of PARP1 and its effects on protein PARylation, PBMCs were stimulated with Staphylococcus aureus with or without olaparib (1-10 µM). Changes in cellular levels of nicotinamide adenine dinucleotide (NAD+) and adenosine triphosphate (ATP), as well as changes in mitochondrial membrane potential (MMP), were measured in PBMCs exposed to H2O2. Bacterial killing was evaluated in PBMCs and polymorphonuclear leukocytes (PMNs) incubated with S. aureus. Cytokine production was measured in supernatants using a cytometric bead array. Reactive oxygen species (ROS), nitric oxide (NO) production, and phagocytic activity of monocytes and neutrophils were measured in whole blood. For ROS and NO production, samples were incubated with heat-killed S. aureus; phagocytic activity was assessed using killed Escherichia coli conjugated to FITC. Olaparib (0.1-100 µM) did not adversely affect lymphocyte viability. Olaparib also did not interfere with PARP1 expression but inhibits S. aureus-induced protein PARylation. In cells challenged with H2O2, olaparib prevented NAD+ and ATP depletion and attenuated mitochondrial membrane depolarization. LPS-induced production of TNF-α, MIP-1α, and IL-10 by PBMCs was also reduced by olaparib. Monocytes and neutrophils displayed significant increases in the production of ROS and NO after stimulation with S. aureus and phagocytic (E. coli) and microbicidal activity, and these responses were not suppressed by olaparib. We conclude that, at clinically relevant concentrations, olaparib exerts cytoprotective effects and modulates inflammatory cytokine production without exerting adverse effects on the cells' ability to phagocytose or eradicate pathogens. The current data support the concept of repurposing olaparib as a potential experimental therapy for septic shock.

Vivências de estudantes de enfermagem na preceptoria em saúde / Experiences of nursing students on preceptorship in health / Experiencias de estudiantes de enfermería sobre la preceptoría en salud

Objetivo:conhecer as vivências de estudantes de enfermagem na preceptoria em saúde. Método: estudo exploratório e com abordagem qualitativa, realizado entre o primeiro semestre de 2019 e o último de 2020, junto a 24 alunos do último semestre do curso de enfermagem, em uma universidade pública de Mato Grosso, Brasil. A coleta de dados foi realizada de forma remota. Para a análise dos dados, utilizou-se a análise temática. Resultados: o processo de transição do formato tradicional do estágio supervisionado de enfermagem para uma nova modalidade de ensino-aprendizagem e acompanhamento em campo prático, foi concebido pelos estudantes com certa apreensão, pelo desconhecimento da estratégia, mas remetendo a ideia de maior liberdade e autonomia. Consideram que um preceptor deve ser comprometido com a ética e a formação de qualidade. Quanto a coordenação do estágio, veem como importante interlocução entre alunos e preceptores, estreitando laços e oportunizando momentos de trocas, com direcionamentos e apoio para o melhor aproveitamento. Conclusão: as vivências dos estudantes de enfermagem na preceptoria foram positivas, o que pode impactar na atuação profissional futura.

Objective:to know the experiences of nursing students in health preceptorship. Method:exploratory study with a qualitative approach, carried out between the first semester of 2019 and the last semester of 2020, with 24 students from the last semester of the nursing course, at a public university in Mato Grosso, Brazil. Data collection was performed remotely. For data analysis, thematic analysis was used. Results:the transition process from the traditional format of the supervised nursing internship to a new modality of teaching-learning and monitoring in the practical field, was conceived by the students with some apprehension, due to the lack of knowledge of the strategy, but referring to the idea of greater freedom and autonomy. They consider that a preceptor must be committed to ethics and quality training. As for the coordination of the internship, they see it as an important dialogue between students and preceptors, strengthening ties and providing opportunities for exchanges, with directions and support for the best use. Conclusion:the experiences of nursing students in the preceptorship were positive, which can impact their future professional performance.

Objetivo:conocer las experiencias de los estudiantes de enfermería en la preceptoría en salud. Método:estudio exploratorio con enfoque cualitativo, realizado entre el primer semestre de 2019 y último semestre de 2020, con 24 estudiantes del último semestre de enfermería, en una universidad pública de Mato Grosso, Brasil. La recolección de datos se realizó de forma remota. Para el análisis de los datos se utilizó el análisis temático. Resultados:el proceso de transición del formato tradicional de pasantía de enfermería supervisada a una nueva modalidad de enseñanza-aprendizaje y acompañamiento en el campo práctico, fue concebido por los estudiantes con cierta aprensión, debido al desconocimiento de la estrategia, pero refiriéndose a la ideade mayor libertad y autonomía. Consideran que un preceptor debe estar comprometido con la ética y la formación de calidad. En cuanto a la coordinación de la pasantía, la ven como un importante diálogo entre alumnos y preceptores, estrechando lazos y brindando oportunidades de intercambio, con orientaciones y apoyos para el mejor aprovechamiento. Conclusión:las experiencias de los estudiantes de enfermería en la preceptoría fueron positivas, lo que puede impactar en su futuro desempeño profesional.

A validated liquid chromatography-tandem mass spectroscopy method for the quantification of tolinapant in human plasma.

Tolinapant (ASTX660), a pan-selective inhibitor of apoptosis protein antagonist with dual cIAP/XIAP activity, was identified as a clinical candidate in preclinical efficacy, pharmacokinetic and safety studies. In order to assess tolinapant in first-in-human Phase I/II clinical trials, a validated bioanalytical method was required to determine plasma pharmacokinetics. Tolinapant and d4-tolinapant were extracted from human plasma using liquid-liquid extraction. Separation chromatography was performed on a Acquity BEH C18 1.7 µM, 50 mm × 2.1 mm i.d. column, using a mobile phase of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Mass spectrometry detection was performed by positive turbo ion spray ionisation, in multiple reaction monitoring mode. The method was validated according to the US Food and Drug Administration (FDA) guidelines. The method has a quantifiable linear range of 1-500 ng/mL (r 2 = 0.999). The intra- and inter-day coefficients of variation were < 11.4%. Dilution QC samples agreed with prepared concentrations, with a precision of 1.5% and accuracy of 101%. Tolinapant mean recoveries ranged from 85.1-94.4 % with negligible matrix effects. A highly sensitive and selective LC-MS/MS bioanalytical method was developed and validated. The method was successfully applied in Phase 1/2 clinical trials to determine the human pharmacokinetic profile of tolinapant.

Potentially toxic elements and rare earth elements in sandy soils from the Brazilian Cerrado.

This study aims to determine the concentration of potentially toxic elements (PTEs) and rare earth elements (REEs) in Brazilian sandy soils under the Cerrado at the Parnaíba-São Francisco Basin transition. We also explored the geochemical correlation between these elements and pH, cation exchange capacity (CEC), total organic carbon (TOC), sand, clay, oxides from secondary minerals, and chemical index of alteration for each basin. Mineralogical, physical, and chemical analyses were used to examine PTE and REE geochemistry in six sand soil profiles from the Brazilian Cerrado. The background concentrations of these elements are low, but soils from the Parnaíba Basin have higher concentrations of PTEs than soils from the São Francisco Basin. In soils from the Parnaíba Basin, mainly Al2O3 has relevance in the V and Cr geochemistry, as these elements increase with increasing Al2O3 content. On the other hand, the REEs have CEC as a soil attribute of higher relevance in the geochemistry of those elements is soils from the Parnaíba Basin, and this relevance divides the TOC, Fe2O3, and TiO2 minerals from the clay fraction. In soils from the São Francisco Basin, the geochemistry of PTEs is possibly associated with kaolinite, especially Cu, V, and Zn. In contrast, the Ba concentration was associated with the presence of feldspar. Unlike soils from the Parnaíba Basin, the REEs do not correlate with the studied soil attributes, except for Ho and Lu. Ho had a positive association with Al2O3. Ho and Lu are negatively related to the presence of iron oxides.

Discovery of ASTX029, A Clinical Candidate Which Modulates the Phosphorylation and Catalytic Activity of ERK1/2.

Aberrant activation of the mitogen-activated protein kinase pathway frequently drives tumor growth, and the ERK1/2 kinases are positioned at a key node in this pathway, making them important targets for therapeutic intervention. Recently, a number of ERK1/2 inhibitors have been advanced to investigational clinical trials in patients with activating mutations in B-Raf proto-oncogene or Ras. Here, we describe the discovery of the clinical candidate ASTX029 (15) through structure-guided optimization of our previously published isoindolinone lead (7). The medicinal chemistry campaign focused on addressing CYP3A4-mediated metabolism and maintaining favorable physicochemical properties. These efforts led to the identification of ASTX029, which showed the desired pharmacological profile combining ERK1/2 inhibition with suppression of phospho-ERK1/2 (pERK) levels, and in addition, it possesses suitable preclinical pharmacokinetic properties predictive of once daily dosing in humans. ASTX029 is currently in a phase I-II clinical trial in patients with advanced solid tumors.

Clinical Positioning of the IAP Antagonist Tolinapant (ASTX660) in Colorectal Cancer.

Inhibitors of apoptosis proteins (IAPs) are intracellular proteins, with important roles in regulating cell death, inflammation, and immunity. Here, we examined the clinical and therapeutic relevance of IAPs in colorectal cancer. We found that elevated expression of cIAP1 and cIAP2 (but not XIAP) significantly correlated with poor prognosis in patients with microsatellite stable (MSS) stage III colorectal cancer treated with 5-fluorouracil (5FU)-based adjuvant chemotherapy, suggesting their involvement in promoting chemoresistance. A novel IAP antagonist tolinapant (ASTX660) potently and rapidly downregulated cIAP1 in colorectal cancer models, demonstrating its robust on-target efficacy. In cells co-cultured with TNFα to mimic an inflammatory tumor microenvironment, tolinapant induced caspase-8-dependent apoptosis in colorectal cancer cell line models; however, the extent of apoptosis was limited because of inhibition by the caspase-8 paralogs FLIP and, unexpectedly, caspase-10. Importantly, tolinapant-induced apoptosis was augmented by FOLFOX in human colorectal cancer and murine organoid models in vitro and in vivo, due (at least in part) to FOLFOX-induced downregulation of class I histone deacetylases (HDAC), leading to acetylation of the FLIP-binding partner Ku70 and downregulation of FLIP. Moreover, the effects of FOLFOX could be phenocopied using the clinically relevant class I HDAC inhibitor, entinostat, which also induced acetylation of Ku70 and FLIP downregulation. Further analyses revealed that caspase-8 knockout RIPK3-positive colorectal cancer models were sensitive to tolinapant-induced necroptosis, an effect that could be exploited in caspase-8-proficient models using the clinically relevant caspase inhibitor emricasan. Our study provides evidence for immediate clinical exploration of tolinapant in combination with FOLFOX in poor prognosis MSS colorectal cancer with elevated cIAP1/2 expression.

ASTX029, a Novel Dual-mechanism ERK Inhibitor, Modulates Both the Phosphorylation and Catalytic Activity of ERK.

The MAPK signaling pathway is commonly upregulated in human cancers. As the primary downstream effector of the MAPK pathway, ERK is an attractive therapeutic target for the treatment of MAPK-activated cancers and for overcoming resistance to upstream inhibition. ASTX029 is a highly potent and selective dual-mechanism ERK inhibitor, discovered using fragment-based drug design. Because of its distinctive ERK-binding mode, ASTX029 inhibits both ERK catalytic activity and the phosphorylation of ERK itself by MEK, despite not directly inhibiting MEK activity. This dual mechanism was demonstrated in cell-free systems, as well as cell lines and xenograft tumor tissue, where the phosphorylation of both ERK and its substrate, ribosomal S6 kinase (RSK), were modulated on treatment with ASTX029. Markers of sensitivity were highlighted in a large cell panel, where ASTX029 preferentially inhibited the proliferation of MAPK-activated cell lines, including those with BRAF or RAS mutations. In vivo, significant antitumor activity was observed in MAPK-activated tumor xenograft models following oral treatment. ASTX029 also demonstrated activity in both in vitro and in vivo models of acquired resistance to MAPK pathway inhibitors. Overall, these findings highlight the therapeutic potential of a dual-mechanism ERK inhibitor such as ASTX029 for the treatment of MAPK-activated cancers, including those which have acquired resistance to inhibitors of upstream components of the MAPK pathway. ASTX029 is currently being evaluated in a first in human phase I-II clinical trial in patients with advanced solid tumors (NCT03520075).

Questionário de Estratégias Específicas a Fissura (USSQ): adaptação e propriedades psicométricas iniciais para o Brasil / Cuestionario de Estrategias Específicas de Antojo (USSQ): adaptación y propiedades psicométricas iniciales para Brasil / Brazilian Adaptation of the Urge-Specific Strategies Questionnaire (USSQ): Adaptation and Initial Psychometric Properties for Brazil

Resumo Habilidades de enfrentamento são uma das variáveis mais importantes para a prevenção de recaída ao álcool e outras drogas. O Questionário de Estratégias Específicas à Fissura (USSQ) é um instrumento desenvolvido nos EE. UU. para a avaliação do repertório de enfrentamento de um indivíduo diante da fissura. Como não foram encontradas informações sobre suas qualidades psicométricas no Brasil, o objetivo deste estudo foi adaptar o instrumento para o país. Participaram do estudo 378 pessoas em tratamento por transtorno de uso de substâncias em Centros de Atenção Psicossocial - Álcool e outras Drogas (CAPS-AD), sendo a maioria homens, de baixa escolaridade e usuários de crack. Os instrumentos utilizados foram um questionário de caracterização geral, o USSQ e o Inventário de Depressão Maior. Foram realizados procedimentos de adaptação transcultural e investigações de evidências de validade e precisão. Os resultados da validade de conteúdo indicaram que os 20 itens representavam adequadamente a definição teórica de enfrentamento, eram adequados para a população brasileira e relevantes para a avaliação do enfrentamento imediato. Considerando aspectos teóricos e psicométricos, uma estrutura interna unifatorial de 12 itens foi considerada a melhor opção para a versão brasileira do USSQ, com índices de ajuste satisfatórios, invariância de medida para usuários de álcool e crack e para pessoas em desintoxicação e abstinentes, bem como relação significativa com as variáveis externas tempo de abstinência e nível de depressão. Conclui-se que a adaptação brasileira do USSQ possui qualidades psicométricas iniciais para mensurar o repertório de habilidades de enfrentamento de usuários de substâncias.

Abstract Coping skills are one of the most important variables of alcohol and drug relapse prevention. The Urge-Specific Strategies Questionnaire (USSQ) is an instrument developed in the USA that aims to evaluate the coping skills repertoire of an individual who is facing craving. As no information about its psychometric properties was found in Brazil, the aim of this study was to adapt the USSQ to the Brazilian context. Participants were 378 addicts in treatment for substance use disorder at Psychosocial Care Centers-Alcohol and Other Drugs (CAPS-AD). Most were men with low education and crack cocaine addicts. The instruments applied were a questionnaire of general characterization, the USSQ, and the Major Depression Inventory. Procedures performed were cross-cultural adaptation and validity and reliability assessments. Results of the content validity indicated that all 20 items adequately represented the theoretical definition and were suitable to assess the immediate coping skills in Brazilian culture. Considering theoretical and psychometric issues, a single-factor internal structure with 12 items was the best choice for the Brazilian version of the USSQ. It showed satisfactory fit indexes, measurement invariance for alcohol and crack cocaine users, and for people in detoxing and abstinence, as well as a significant correlation with external variables such as length of abstinence and depression level. The conclusion of the study is that the Brazilian USSQ has initial psychometric qualities to assess the coping skills repertoire of substance addicts.

Resumen Las habilidades de afrontamiento son una de las variables más importantes para prevenir la recaída en el alcohol y otras drogas. El Cuestionario de Estrategias Específicas de Antojo (USSQ) es un instrumento desarrollado en los EE. UU. para la evaluación del repertorio de afrontamiento de un individuo frente a un antojo de droga. Como no se encontró información sobre sus cualidades psicométricas en Brasil, el objetivo de este estudio fue adaptar el instrumento para el país. Participaron 378 personas en el tratamiento por trastorno de uso de sustancias en los Centros de Atención Psico-social-Alcohol y otras Drogas (CAPS-AD), la mayoría hombres, con bajo nivel educativo y consumidores de crack. Los instrumentos utilizados fueron un cuestionario de caracterización general, el USSQ y el Inventario de Depresión Mayor. Se llevaron a cabo procedimientos de adaptación transcultural y evaluaciones de la validez y fiabilidad. Los resultados de la validez de contenido indicaron que los 20 ítems representaban adecuadamente la definición teórica de afrontamiento, eran adecuados para la población brasileña y relevantes, para la evaluación del afrontamiento inmediato. Considerando aspectos teóricos y psicométricos, una estructura interna unifactorial de 12 ítems fue considerada la mejor opción para la versión brasileña del USSQ, con tasas de ajuste satisfactorias, invariancia de medición para consumidores de alcohol y crack y para personas en desintoxicación y abstinentes, así como relación significativa con las variables externas tiempo de abstinencia y nivel de depresión. Se concluye que la adaptación brasileña del USSQ tiene cualidades psicométricas iniciales para medir el repertorio de habilidades de afrontamiento de los consumidores de sustancias.

Pathological pulmonary vascular remodeling is induced by type V collagen in a model of scleroderma.

OBJECTIVE: The pulmonary vascular remodeling in systemic sclerosis (SSc) is poorly understood and animal models are lacking. Type V collagen (COLV) is elevated in SSc and is implicated in the pathogenesis, and immunization with human COLV induces SSc-like skin and lung changes in rabbits and mice. Here we tested the hypothesis that COLV immunization will induce pathological and functional changes that phenocopy SSc-associated pulmonary vascular disease. METHODS: Pulmonary vascular changes in rabbits immunized with human COLV were extensively characterized by a combination of histology, electron microscopy and immunohistochemistry. Physiologic changes induced by COLV in explanted pulmonary artery rings were evaluated. The pattern of histopathologic alterations and gene expression induced in immunized rabbits were compared to those in SSc patients. RESULTS: COLV immunization was accompanied by striking pulmonary vascular abnormalities, characterized by reduced capillary density, perivascular inflammation, endothelial cell injury and collagen accumulation, that closely phenocopy changes seen in SSc patients. Moreover, pulmonary arteries from immunized rabbits showed impaired ex vivo vascular relaxation. Expression of COL5A2 was significantly increased in the lungs from immunized rabbits (p = 0.02), as well as in patients with SSc (P = 0.02). CONCLUSION: COLV immunity in rabbits is associated with marked vascular remodeling in the lung that phenocopies early-stage human SSc-associated pulmonary vascular disease. COLV immunization therefore represents a novel approach to model SSc pulmonary vascular pathology. Moreover, our findings suggest that COLV might represent a novel pathogenic autoantigen in SSc and future studies with the present model should be developed for possible association with PAH.

In situ evidence of collagen V and signaling pathway of found inflammatory zone 1 (FIZZ1) is associated with silicotic granuloma in lung mice.

Inhalation of silica particles causes silicosis: an occupational lung disease characterized by persistent inflammation with granuloma formation that leads to tissue remodeling and impairment of lung function. Although silicosis has been studied intensely, little is known about the crucial cellular mechanisms that initiate and drive the process of inflammation and fibrosis. Recently, found in inflammatory zone 1 (FIZZ1) protein, produced by alveolar macrophages and fibroblasts have been shown to induce the proliferation of myofibroblasts and their transdifferentiation, causing tissue fibrosis. Moreover, autoimmunogenic collagen V, produced by alveolar epithelial cells and fibroblasts, is involved in the pathophysiology of interstitial pulmonary fibrosis and bleomycin-induced lung fibrosis. Based on the aforementioned we hypothesized that FIZZ1 and collagen V may be involved in the silicotic granuloma process in mice lungs. Male C57BL/6 mice (N = 20) received intratracheal administration of silica particles (Silica; 20 mg in 50 µL saline) or saline (Control; 50 µL). After 15 days, the lung histology was performed through immunohistochemistry and morphometric analysis. Within silicotic granulomas, collagen V and FIZZ1 increased, while peroxisome proliferator-activated receptor gamma (PPARγ) positive cells decreased. In addition, the expression of proteins Notch-1, alpha smooth muscle actin (α-SMA) and macrophages163 (CD163) were higher in silicotic granulomas than control lungs. A significant positive correlation was found between collagen V and FIZZ1 (r = 0.70; p < 0.05), collagen V and Notch-1 (r = 0.72; p < 0.05), whereas Collagen V was inversely associated with peroxisome proliferator-activated receptor gamma (r=-0.69; p < 0.05). These findings suggested that collagen V association with FIZZ1, Notch-1 and PPARγ might be a key pathogenic mechanism for silicotic granulomas in mice lungs.

Poly(ADP-Ribose) Polymerase Inhibition in Acute Lung Injury. A Reemerging Concept.

PARP1, the major isoform of a family of ADP-ribosylating enzymes, has been implicated in the regulation of various biological processes including DNA repair, gene transcription, and cell death. The concept that PARP1 becomes activated in acute lung injury (ALI) and that pharmacological inhibition or genetic deletion of this enzyme can provide therapeutic benefits emerged over 20 years ago. The current article provides an overview of the cellular mechanisms involved in the pathogenetic roles of PARP1 in ALI and provides an overview of the preclinical data supporting the efficacy of PARP (poly[ADP-ribose] polymerase) inhibitors. In recent years, several ultrapotent PARP inhibitors have been approved for clinical use (for the therapy of various oncological diseases): these newly-approved PARP inhibitors were recently reported to show efficacy in animal models of ALI. These observations offer the possibility of therapeutic repurposing of these inhibitors for patients with ALI. The current article lays out a potential roadmap for such repurposing efforts. In addition, the article also overviews the scientific basis of potentially applying PARP inhibitors for the experimental therapy of viral ALI, such as coronavirus disease (COVID-19)-associated ALI.

C and P pool restoration by a no-tillage system on Brazilian Cerrado Oxisol in Piauí State.

The Cerrado soil is under constant modification, especially because of the use of agricultural systems, which affect soil carbon (C) and phosphorus (P) functioning. Thus, the objective of this study was to determine the C and P dynamics in Brazilian Cerrado Oxisol in Piauí State under natural and anthropic conditions, considering that conservational agricultural management and no-tillage systems can restore the C and P pools in that soil. Four soil samples with distinct characteristics (native Cerrado, NC; burned native Cerrado, BNC; conventional tillage agricultural system, CTS; and no-tillage agricultural system, NTS) were collected in the study area for chemical and physical laboratory analysis. The total organic carbon (TOC) concentrations found were 33 g kg-1, 27 g kg-1, 26 g kg-1, and 20 g kg-1 for CTS, NTS, NC, and BNC, respectively. The NTS had a total nitrogen (TN) concentration of 2.0 g kg-1. The CTS had 33.4 g kg-1 of soil-oxidizable C, followed by the NTS with 27.2 g kg-1. In both studied layers, the NTS had an organic P concentration > 200 mg kg-1. The higher TOC concentration in the CTS was because of the higher content of clay in comparison with that in the NTS. The organic P in the NTS was associated with a less labile fraction of C. Thus, despite the disturbance caused by agricultural systems, the adoption of the NTS could be an influential strategy in agricultural systems to restore soil organic functioning in the Brazilian Cerrado Oxisol in Piauí State.

Adipose-derived stem cells and adipose-derived stem cell-conditioned medium modulate in situ imbalance between collagen I- and collagen V-mediated IL-17 immune response recovering bleomycin pulmonary fibrosis.

The immunogenic collagen V (Col V) and the proinflammatory cytokine interleukin (IL)-17 have been implicated in the pathogenesis of multiple autoimmune diseases. Col V is also up-regulated during adipogenesis and can stimulate adipocyte differentiation in vitro. Conditioned medium (CM) generated from adipose-derived mesenchymal stem cells (MSCs) reduces bleomycin (BLM)-induced lung injury in rats, suggesting a crucial role in situ of immunomodulatory factors secreted by MSCs in these beneficial effects. In the present work, we investigated this hypothesis, analyzing levels of plasma inflammatory mediators and inflammatory and fibrotic mediators in the lung tissue of BLM-injured rats after treatment with MSCs and CM. Pulmonary fibrosis was intratracheally induced by BLM. After 10 days, BLM animals were further randomized into subgroups receiving saline, MSCs, or CM intravenously. On days 14 and 21, the animals were euthanized, and the lungs were examined through protein expression of nitric oxide synthase (NOS), IL-17, transforming growth factor-ß (TGF-ß), vascular endothelial growth factor, endothelin-1, and the immunogenic Col V through histological quantitative evaluation and plasma levels of fibrinogen, Von Willebrand factor, and platelet-derived growth factor (PDGF). Rats that had been injected with MSCs and CM showed a significant increase in weight and significant improvements at 14 and 21 days after intravenous injection at both time points of analysis of plasma fibrinogen, PDGF, and Von Willebrand factor and NOS-2 expression, supporting an early anti-inflammatory action, thus reducing TGF-ß and collagen I fibers. In contrast, intravenous injection of CM was able to significantly increase the deposition of Col V fibers and IL-17 on both day 14 and day 21 as compared with the amount observed in rats from the BLM group and MSC groups. In conclusion, this study reinforces previous observations on the therapeutic properties of MSCs and CM and is the first report to demonstrate the association of its actions with immunomodulatory biomarkers on lung tissue. We concluded that adipose-derived stem cells and adipose-derived stem cells-CM modulate an in situ imbalance between collagen I- and Col V-mediated IL-17 immune response, emerging as a promising therapeutic option for recovering from BLM pulmonary fibrosis.