Nonsense-mediated RNA decay and its bipolar function in cancer

ReviewNonsense-mediated decay (NMD) was first described as a quality-control mechanism that targets and rapidly degrades aberrant mRNAs carrying premature termination codons (PTCs). However, it was found that NMD also degrades a significant number of normal transcripts, thus arising as a mechanism of gene expression regulation. Based on these important functions, NMD regulates several biological processes and is involved in the pathophysiology of a plethora of human genetic diseases, including cancer. The present review aims to discuss the paradoxical, pro- and anti-tumorigenic roles of NMD, and how cancer cells have exploited both functions to potentiate the disease. Considering recent genetic and bioinformatic studies, we also provide a comprehensive overview of the present knowledge of the advantages and disadvantages of different NMD modulation-based approaches in cancer therapy, reflecting on the challenges imposed by the complexity of this disease. Furthermore, we discuss significant advances in the recent years providing new perspectives on the implications of aberrant NMD-escaping frameshifted transcripts in personalized immunotherapy design and predictive biomarker optimization. A better understanding of how NMD differentially impacts tumor cells according to their own genetic identity will certainly allow for the application of novel and more effective personalized treatments in the near future.Gonçalo Nogueira, Rafael Fernandes, and Juan Fernandez García-Moreno are recipients of a fellowship from BioSys PhD programme PD65-2012 (SFRH/PD/BD/130959/2017, SFRH/BD/114392/2016 and SFRH/PD/BD/142898/2018, respectively) from FCT. This work was partially supported by UID/MULTI/04046/2019 Research Unit grant (to BioISI).info:eu-repo/semantics/publishedVersio

Tandem Thio-Michael Addition/Remote Lactone Activation of 5-Hydroxymethylfurfural-Derived δ-Lactone-Fused Cyclopentenones

Funding Information: We thank the Fundação para a Ciência e a Tecnologia (SFRH/BD/120829/2016, SFRH/BD/148211/2019, UIDB/04138/2020, UIDP/04138/2020, PTDC/QUI‐QOR/32008/2017 and GHTM‐UID/04413/2020). The project leading to this application has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No 951996. J. A. S. C. thanks the Fundação para a Ciência e a Tecnologia (FCT) for Scientific Employment Stimulus 2020/02383/CEECIND. Publisher Copyright: © 2022 The Authors. ChemSusChem published by Wiley-VCH GmbH.The creation of structurally diverse chemical entities from fairly simple biorefinery products remains a challenge. In this work 5-hydroxymethylfurfural (HMF) was identified as a key synthon for preparing highly complex cyclopentenones (CP) via tandem 1,4-addition/elimination/remote lactone activation to external O- and N-nucleophiles in δ-lactone-fused-CPs hotspots. This scaffold was also reactive enough to be incorporated into model cysteine-peptides in low concentrations, paving the way to a potential translation generating complexity in the synthesis of small peptides. The new enones also exhibited activity against intraerythrocytic Plasmodium falciparum (IC50=1.32 μm).publishersversionpublishe

Effect of polymerization cycles on gloss, roughness, hardness and impact strength of acrylic resins

The aim of this study was to evaluate the conventional and boiled polymerization cycles on gloss, roughness, hardness and impact strength of acrylic resins. Samples were made for each Classico and QC-20 materials (n=10) in dental stone molds obtained from rectangular metallic matrices embedded in metallic flasks. The powder-liquid ratio and manipulation of the acrylic resins' were accomplished according to manufacturers' instructions and the resins were conventionally packed in metallic flasks. After polymerization by (1) conventional: 74 °C for 9 h (Classico) and (2) boiled: 20 min (QC-20) cycles, the samples were deflasked after cooling at room temperature and conventionally finished and polished. The properties were evaluated after storage in water at 37 °C for 24 h. Gloss was verified with Multi Gloss 268 meter (Konica Minolta), surface roughness was measured with Surfcorder SE 1700 rugosimeter (Kosaka), Knoop hardness number was obtained with HMV-200 microdurometer, and impact strength was measured in an Otto Wolpert-Werke device by Charpy system (40 kpcm). Data were subjected to Student's t-test (at α=0.05). The results were: Gloss: 67.7 and 62.2 for Classico and QC-20 resins, respectively; Surface roughness: 0.874 and 1.469 Ra-µm for Classico and QC-20, respectively; Knoop hardness: 27.4 and 26.9 for Classico and QC-20, respectively; and Impact strength: 37.6 and 33.6 kgf/cm2 for Classico and QC-20, respectively. No statistically significant difference (p>0.05)were found between the resins for the evaluated properties. In conclusion, conventional and boiled polymerization cycles had similar effects on gloss, roughness, hardness and impact strength of both Classico and QC-20 resins272176180CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQSem informaçãoO propósito neste estudo foi avaliar os ciclos de polimerização convencional e por fervura sobre o brilho, rugosidade, dureza e resistência ao impacto de resinas acrílicas. Amostras foram confeccionadas para cada resina Clássico ou QC-20 (n=20) em moldes de gesso obtidos de matrizes metálicas retangulares incluídas em muflas metálicas. A proporção monômero/polímero das resinas e manipulação foram de acordo com as recomendações dos fabricantes e a massa convencionalmente incluída em muflas metálicas. Após polimerização nos ciclos (1) convencional: 74 °C por 9 horas (Clássico) e (2) fervura: 20 min (QC-20), as amostras foram demufladas após esfriadas em temperatura ambiente e convencionalmente acabadas e polidas. As propriedades foram avaliadas após armazenagem das amostras em água a 37 °C por 24 h. O brilho foi verificado com medidor Multi Gloss 268 (Konica Minolta), a rugosidade avaliada com rugosímetro Surfcorder SE 1700 (Kosaka), a dureza Knoop foi obtida com microdurômetro HMV-200 (Shimadzu) e a resistência ao impacto determinada pelo sistema Charpy (Otto Wolpert Werke). Os dados submetidos ao teste t de Student (α=0.05) mostraram que Os resultados foram: brilho: 67,7 e 62,2 para Clássico e QC-20, respectivamente; rugosidade: 0,874 e 1,469 Ra-µm para Clássico e QC-20, respectivamente; dureza: 27,4 e 26,9 para Clássico e QC-20, respectivamente; e resistência ao impacto: 37,6 e 33,6 kgf/cm2 para Clássico e QC-20, respectivamente. Não houve diference estatisticamente significante entre as resinas para as propriedades avaliadas. Conclui-se que os ciclos de polimerização convencional e por fervura promoveram similares efeitos sobre o brilho, rugosidade, dureza e resistência ao impacto para ambas as resinas Clássico e QC-2

Avaliação in vivo da interação leucócito-endotélio mesentérico após ligadura e punção cecal e remoção cirúrgica do foco sééptico

PURPOSE: Cecal ligation and puncture (CLP) has been used as a useful model for the induction of polymicrobial sepsis. Necrotic tissue resection and peritoneal lavage (REL) are the surgical procedures for controlling perforated appendicitis. The aim of this study was to evaluate leukocyte-endothelial interactions in the rat mesentery in vivo after CLP and REL. METHODS: Thirty-seven male Wistar rats (250-300 g) underwent laparotomy and were randomly assigned to the following groups: 1) SHAM; 2) CLP: animals submitted to CLP, 3) CLP+REL: animals submitted to CLP and REL. Mesenteric leukocyte-endothelial interactions were studied by intravital microscopy assessed once in each animal (3-5 postcapillary venules, 15-25 µm diameter) 24 hours after intervention. Follow-up was performed in all animals; this included analysis of glycemia, lactate, hematocrit, white blood cell count as well as a functional score that was the sum of scoring on the following parameters: alertness, mobility, piloerection, diarrhea, encrusted eyes, and dirty nose and tail. RESULTS: None of the animals showed significant changes in body weight (265 ± 20 g) or in hematocrit levels (46% ± 2%) during the experimental protocol. Compared to SHAM animals, CLP animals showed an increased number of rolling (2x), adherent, and migrating leukocytes (7x) in the mesenteric microcirculation, an increase in blood glucose (136 ± 8 mg/dL), lactate (3.58 ± 0.94 mmol/L), white cell count (23,570 ± 4,991 cells/mm³) and functional alterations (score 11 ± 1), characterized by impaired alertness and mobility, and presence of piloerection, diarrhea, encrusted eyes, and dirty nose and tail. The REL procedure normalized the number of rolling, adherent, and migrated leukocytes in the mesentery; glycemia; lactate; and white blood cell count. The REL procedure also improved the functional score (7 ± 1). CONCLUSION: Local and systemic inflammation was induced by CLP, while REL completely overcame the inflammatory process.OBJETIVO: O procedimento de ligadura cecal e perfuração (CLP) tem sido usado como um modelo útil de indução de sepse polimicrobiana. A ressecção do tecido necrosado e lavagem peritoneal (REL) são procedimentos cirúrgicos freqüentemente utilizados para controlar uma apendicite perfurada. O objetivo desse estudo foi avaliar in vivo as interações leucócito-endotélio no mesentério de ratos após a CLP e REL. MÉTODOS: Trinta e sete ratos Wistar machos (250-300 g) foram submetidos à laparotomia e aleatoriamente divididos em grupos: 1) SHAM, 2) CLP: ratos submetidos à CLP, 3) CLP+REL: animais submetidos à CLP e REL. As interações leucócito-endotélio no mesentério foram estudadas através de microscopia intravital somente uma vez em cada animal (3-5 vênulas pós-capilares, 15-25 µm diâmetro), 24-horas após as intervenções. A evolução clínica foi realizada em todos os animais, incluindo glicemia, lactato, hematócrito, número total de células brancas e um escore funcional, o qual foi considerado como a somatória dos seguintes parâmetros: estado de alerta, mobilidade, piloereção, diarréia, olhos encrustados, e nariz e cauda sujos. RESULTADOS: Os animais não apresentaram alterações significantes no peso (265 ± 20 g) e hematócrito (46 ± 2%) ao longo do estudo. Comparados ao SHAM, os animais CLP apresentaram aumento no número de leucócitos em rolamento (2x), aderidos (7x) e migrados (7x) na microcirculação mesentérica, aumentos da glicemia (136 ± 8 mg/dL), lactato (3,58 ± 0,94 mmol/L), leucocitose (23.570 ± 4.991 células/mm³) e alterações clínicas (escore 11±1), caracterizadas por comprometimento do estado de alerta e mobilidade, e presença de piloereção, diarréia, olhos encrustados, nariz e cauda sujos. REL normalizou o número de leucócitos em rolamento, aderidos e migrados no mesentério, a glicemia, o lactato e o número de leucócitos circulantes. REL também melhorou o escore clínico (7 ± 1). CONCLUSÃO: A CLP induziu inflamação local e sistêmica. A REL resolveu, por completo, o processo inflamatório

Relationship between the nursing practice environment and the therapeutic relationship in acute mental health units: a cross-sectional study

The therapeutic relationship constitutes the central axis of mental health nursing. The clinical practice environment has been empirically related to the quality of care. However, the relationship between the two constructs is unknown in the setting of mental health units. We aimed to examine whether the practice environment and nurses' characteristics influence the therapeutic relationship in mental health units. Through a cross-sectional design, data were collected via an online form completed by nurses in 18 mental health units. Linear regression was used to examine the relationship between the clinical practice environment and the therapeutic relationship. Questionnaires were completed by 198 participants. The mean age was 33.8 (SD 9.1) years, 71.7% were women, and only 20.2% had a specialist qualification in mental health. The therapeutic relationship was better when there was a more favourable practice environment (B: 3.111; 95% CI: 1.46-4.75). The most influential environment-related factor was the nursing foundations for quality of care (B: 2.124; 95% CI: 0.17-4.07). The factors associated with a high-quality therapeutic relationship were a more favourable practice environment and the presence of more foundations for quality nursing care, coupled with higher academic attainment and longer nursing experience. Institutions should take into account the importance of the nursing practice environment in mental health units. Aspects related to the quality of nursing foundations, such as training, the use of nursing language and taxonomy, and the existence of a common nursing philosophy, are influential for a high-quality therapeutic relationship

Polymerization cycles on hardness and surface gloss of denture bases

The aim of this study was to evaluate different polymerization cycles on the hardness and surface gloss of acrylic resins for denture bases. Classico and Vip Cril Plus acrylic resins samples were made in dental stone molds. Powder-liquid ratio and resin manipulation were according to the manufacturers’ instructions, and the resulting mass pressed in metallic fl asks. The polymerization cycles were A - hot water bath at 74°C for 9 hours; B - hot water bath at 74°C for 8 hours + 100°C for 1 hour, and C - hot water bath at 74°C for 2 hours + 100°C for 1 hour. After polymerization, the samples were defl asked and submitted to finishing and polishing procedures, and stored in water at 37°C for 24 hours. A hardness indenter with load of 25 gf for 10 s evaluated the Knoop hardness values. A gloss meter evaluated the surface gloss using a light incidence of 60°. Data were submitted to ANOVA and Tukey’s test (α = 0.05). Hardness: There were signifi cant differences between resins regardless of polymerization cycles (Classico = 22.28 and Vip Cril Plus = 25.83). Significant differences occurred among polymerization cycles regardless of resins (A = 25.83, B = 24.64 and C = 21.73). There was similarity for the resin and cycle interaction (Classico: A = 24.51, B = 22.68 and C = 19.65; Vip Cril Plus: A = 27.15, B = 26.53 and C = 23.81). Surface gloss. Significant differences were shown between resins regardless of polymerization cycle (Classico = 57.26 and Vip Cril Plus = 49.38) and between polymerization cycles regardless of resin (A = 48.82, B = 53.46 and C = 57.68). Statistical diff erences were also found for the resin and cycle interaction (Classico: A = 52.32, B = 63.79 and C = 55.67; Vip Cril Plus: A = 45.32, B = 43.14 and C = 59.69). Diff erent polymerization cycles showed similar eff ects on hardness and different effects on the surface gloss of denture base acrylic resins

Absence of the caspases 1/11 modulates liver global lipid profile and gut microbiota in high-fat-diet-induced obese mice

Obesity is a chronic disease with rising worldwide prevalence and largely associated with several other comorbidities, such as cancer, non-alcoholic fatty liver disease (NAFLD), and metabolic syndrome. Hepatic steatosis, a hallmark of NAFLD, is strongly correlated with obesity and has been correlated with changes in the gut microbiota, which can promote its development through the production of short-chain fatty acids (SCFAs) that regulate insulin resistance, bile acid, choline metabolism, and inflammation. Recent studies have suggested a controversial role for the inflammasome/caspase-1 in the development of obesity and non-alcoholic steatohepatitis (NASH). Here, we evaluated the role of inflammasome NLRP3 and caspases 1/11 in the establishment of obesity and hepatic steatosis in diet-induced obese mice, correlating them with the global lipid profile of the liver and gut microbiota diversity. After feeding wild-type, caspases 1/11, and NLRP3 knockout mice with a standard fat diet (SFD) or a high-fat diet (HFD), we found that the caspases 1/11 knockout mice, but not NLRP3 knockout mice, were more susceptible to HFD-induced obesity, and developed enhanced hepatic steatosis even under SFD conditions. Lipidomics analysis of the liver, assessed by MALDI-MS analysis, revealed that the HFD triggered a significant change in global lipid profile in the liver of WT mice compared to those fed an SFD, and this profile was modified by the lack of caspases 1/11 and NLRP3. The absence of caspases 1/11 was also correlated with an increased presence of triacylglycerol in the liver. Gut microbial diversity analysis, using 16S rRNA gene sequencing, showed that there was also an increase of Proteobacteria and a higher Firmicutes/Bacteroidetes ratio in the gut of caspases 1/11 knockout mice fed an HFD. Overall, mice without caspases 1/11 harbored gut bacterial phyla involved with weight gain, obesity, and hepatic steatosis. Taken together, our data suggest an important role for caspases 1/11 in the lipid composition of the liver and in the modulation of the gut microbial community composition. Our results further suggest that HFD-induced obesity and the absence of caspases 1/11 may regulate both lipid metabolism and gut microbial diversity, and therefore may be associated with NAFLD and obesity10CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP312359/2016-02016/22577-6This research was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq -#312359/2016-0). CM was funded by the Canada Research Chair Program, the Canadian Foundation for Innovation, McGill University, and the Canadian Institutes for Health Research (PJT-149098). ME was funded by the São Paulo Research Foundation (FAPESP) (2016/22577-6)

Characterization of Systemic Disease Development and Paw Inflammation in a Susceptible Mouse Model of Mayaro Virus Infection and Validation Using X-ray Synchrotron Microtomography

Mayaro virus (MAYV) is an emerging arthropod-borne virus endemic in Latin America and the causative agent of arthritogenic febrile disease. Mayaro fever is poorly understood; thus, we established an in vivo model of infection in susceptible type-I interferon receptor-deficient mice (IFNAR−/−) to characterize the disease. MAYV inoculations in the hind paws of IFNAR−/− mice result in visible paw inflammation, evolve into a disseminated infection and involve the activation of immune responses and inflammation. The histological analysis of inflamed paws indicated edema at the dermis and between muscle fibers and ligaments. Paw edema affected multiple tissues and was associated with MAYV replication, the local production of CXCL1 and the recruitment of granulocytes and mononuclear leukocytes to muscle. We developed a semi-automated X-ray microtomography method to visualize both soft tissue and bone, allowing for the quantification of MAYV-induced paw edema in 3D with a voxel size of 69 µm3. The results confirmed early edema onset and spreading through multiple tissues in inoculated paws. In conclusion, we detailed features of MAYV-induced systemic disease and the manifestation of paw edema in a mouse model extensively used to study infection with alphaviruses. The participation of lymphocytes and neutrophils and expression of CXCL1 are key features in both systemic and local manifestations of MAYV disease

Lipophosphoglycans from \u3cem\u3eLeishmania amazonensis\u3c/em\u3e Strains Display Immunomodulatory Properties via TLR4 and Do Not Affect Sand Fly Infection

The immunomodulatory properties of lipophosphoglycans (LPG) from New World species of Leishmania have been assessed in Leishmania infantum and Leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. This glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved Gal(β1,4)Man(α1)-PO4 backbone of repeat units. Here, the immunomodulatory activity of LPGs from Leishmania amazonensis, the causative agent of diffuse cutaneous leishmaniasis, was evaluated in two strains from Brazil. One strain (PH8) was originally isolated from the sand fly and the other (Josefa) was isolated from a human case. The ability of purified LPGs from both strains was investigated during in vitro interaction with peritoneal murine macrophages and CHO cells and in vivo infection with Lutzomyia migonei. In peritoneal murine macrophages, the LPGs from both strains activated TLR4. Both LPGs equally activate MAPKs and the NF-κB inhibitor p-IκBα, but were not able to translocate NF-κB. In vivo experiments with sand flies showed that both stains were able to sustain infection in L. migonei. A preliminary biochemical analysis indicates intraspecies variation in the LPG sugar moieties. However, they did not result in different activation profiles of the innate immune system. Also those polymorphisms did not affect infectivity to the sand fly

Pleiotropic antifibrotic actions of aspirin-triggered resolvin D1 in the lungs

Introduction: Pulmonary fibrosis is a destructive, progressive disease that dramatically reduces life quality of patients, ultimately leading to death. Therapeutic regimens for pulmonary fibrosis have shown limited benefits, hence justifying the efforts to evaluate the outcome of alternative treatments. Methods: Using a mouse model of bleomycin (BLM)-induced lung fibrosis, in the current work we asked whether treatment with pro-resolution molecules, such as pro-resolving lipid mediators (SPMs) could ameliorate pulmonary fibrosis. To this end, we injected aspirin-triggered resolvin D1 (7S,8R,17R-trihydroxy-4Z,9E,11E,13Z,15E19Z-docosahexaenoic acid; ATRvD1; i.v.) 7 and 10 days after BLM (intratracheal) challenge and samples were two weeks later. Results and discussion: Assessment of outcome in the lung tissues revealed that ATRvD1 partially restored lung architecture, reduced leukocyte infiltration, and inhibited formation of interstitial edema. In addition, lung tissues from BLM-induced mice treated with ATRvD1 displayed reduced levels of TNF-α, MCP-1, IL-1-β, and TGF-β. Of further interest, ATRvD1 decreased lung tissue expression of MMP-9, without affecting TIMP-1. Highlighting the beneficial effects of ATRvD1, we found reduced deposition of collagen and fibronectin in the lung tissues. Congruent with the anti-fibrotic effects that ATRvD1 exerted in lung tissues, α-SMA expression was decreased, suggesting that myofibroblast differentiation was inhibited by ATRvD1. Turning to culture systems, we next showed that ATRvD1 impaired TGF-β-induced fibroblast differentiation into myofibroblast. After showing that ATRvD1 hampered extracellular vesicles (EVs) release in the supernatants from TGF-β-stimulated cultures of mouse macrophages, we verified that ATRvD1 also inhibited the release of EVs in the bronco-alveolar lavage (BAL) fluid of BLM-induced mice. Motivated by studies showing that BLM-induced lung fibrosis is linked to angiogenesis, we asked whether ATRvD1 could blunt BLM-induced angiogenesis in the hamster cheek pouch model (HCP). Indeed, our intravital microscopy studies confirmed that ATRvD1 abrogates BLM-induced angiogenesis. Collectively, our findings suggest that treatment of pulmonary fibrosis patients with ATRvD1 deserves to be explored as a therapeutic option in the clinical setting.Fil: Guilherme, Rafael F.. Universidade Federal do Rio de Janeiro; BrasilFil: Silva, José Bruno N.F.. Universidade Federal do Rio de Janeiro; Brasil. Universidade Federal do Tocantins; BrasilFil: Waclawiack, Ingrid. Universidade Federal do Rio de Janeiro; BrasilFil: Fraga Junior, Vanderlei S.. Universidade Federal do Rio de Janeiro; BrasilFil: Nogueira, Thaís O.. Universidade Federal do Rio de Janeiro; BrasilFil: Pecli, Cyntia. Universidade Federal do Rio de Janeiro; BrasilFil: Araújo Silva, Carlla A.. Universidade Federal do Rio de Janeiro; BrasilFil: Magalhães, Nathalia S.. Ministerio de Salud de Brasil. Fundación Oswaldo Cruz. Instituto Oswaldo Cruz;Fil: Lemos, Felipe S.. Ministerio de Salud de Brasil. Fundación Oswaldo Cruz. Instituto Oswaldo Cruz;Fil: Bulant, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil; ArgentinaFil: Blanco, Pablo Javier. Laboratório Nacional para Computação Científica; BrasilFil: Serra, Rafaela. Universidade Federal do Rio de Janeiro; BrasilFil: Svensjö, Erik. Universidade Federal do Rio de Janeiro; BrasilFil: Scharfstein, Júlio. Universidade Federal do Rio de Janeiro; BrasilFil: Moraes, João A.. Universidade Federal do Rio de Janeiro; BrasilFil: Canetti, Claudio. Universidade Federal do Rio de Janeiro; BrasilFil: Benjamim, Claudia F.. Universidade Federal do Rio de Janeiro; Brasi