Gasdermin-D activation by SARS-CoV-2 triggers NET and mediate COVID-19 immunopathology.

BACKGROUND: The release of neutrophil extracellular traps (NETs) is associated with inflammation, coagulopathy, and organ damage found in severe cases of COVID-19. However, the molecular mechanisms underlying the release of NETs in COVID-19 remain unclear. OBJECTIVES: We aim to investigate the role of the Gasdermin-D (GSDMD) pathway on NETs release and the development of organ damage during COVID-19. METHODS: We performed a single-cell transcriptome analysis in public data of bronchoalveolar lavage. Then, we enrolled 63 hospitalized patients with moderate and severe COVID-19. We analyze in blood and lung tissue samples the expression of GSDMD, presence of NETs, and signaling pathways upstreaming. Furthermore, we analyzed the treatment with disulfiram in a mouse model of SARS-CoV-2 infection. RESULTS: We found that the SARS-CoV-2 virus directly activates the pore-forming protein GSDMD that triggers NET production and organ damage in COVID-19. Single-cell transcriptome analysis revealed that the expression of GSDMD and inflammasome-related genes were increased in COVID-19 patients. High expression of active GSDMD associated with NETs structures was found in the lung tissue of COVID-19 patients. Furthermore, we showed that activation of GSDMD in neutrophils requires active caspase1/4 and live SARS-CoV-2, which infects neutrophils. In a mouse model of SARS-CoV-2 infection, the treatment with disulfiram inhibited NETs release and reduced organ damage. CONCLUSION: These results demonstrated that GSDMD-dependent NETosis plays a critical role in COVID-19 immunopathology and suggests GSDMD as a novel potential target for improving the COVID-19 therapeutic strategy.

Effect of Nd:YAG laser irradiation, used as a desensitizing strategy, on bond strength to simulated hypersensitive dentin.

OBJECTIVES: To evaluate the effect of Nd:YAG laser irradiation as a prior desensitizing strategy on immediate and medium-term microtensile bond strength (µTBS) to simulated hypersensitive dentin. MATERIALS AND METHODS: Flat mid-coronal dentin was obtained from third molars and submitted to a 600-grit SiC paper (1 min; N: normal dentin) or subsequently challenged with citric acid (6%, 1 min; H: simulated hypersensitive dentin). Afterwards, dentin was or was not (C: control; HC, NC - each n = 7) irradiated with Nd:YAG laser (L: laser; 1.0 W/10 Hz/100 mJ/4 irradiations of 50-60 s; HL, NL - each n = 7). A 2-step self-etch adhesive (Clearfil SE Bond) was applied and composite (Filtek Z350) buildups were constructed. After 24-h (distilled water/37 °C) storage, specimens were sectioned into beams and tested (µTBS; 0.5 mm/min) immediately or after 6-month aging. Three-way ANOVA and Tukey tests were applied (α = 0.05). Qualitative evaluation of the adhesive interface (n = 1 extra tooth per group) was performed by Confocal Laser Scanning Microscopy. RESULTS: Substrate condition (p < 0.001), laser irradiation (p < 0.001), and aging (p = 0.002) influenced the results. Furthermore, there was interaction between substrate and irradiation (p < 0.001). Laser irradiation favored µTBS exclusively to hypersensitive dentin immediately and after aging. µTBS to hypersensitive dentin was higher than that to the normal substrate only when it was laser-irradiated. In any case, immediate µTBS was always higher than that after aging. CLSM revealed longer and more numerous resin tags for simulated hypersensitive dentin, and shorter and fewer resin tags for laser-irradiated dentin. No differences were observed in the hybrid layer itself. CONCLUSION: Nd:YAG laser irradiation prior to restoration favored the µTBS of a self-etch adhesive and resin composite to hypersensitive dentin. CLINICAL RELEVANCE: Desensitizing strategies are usually tried before performing restorative treatments in hypersensitive dentin; therefore, they may influence behavior of the adhesive interface established. However, instead of causing concern, Nd:YAG laser irradiation revealed a favorable effect on the aforementioned interface.

Transcriptionally active LTR retrotransposons in Eucalyptus genus are differentially expressed and insertionally polymorphic.

BACKGROUND: In Eucalyptus genus, studies on genome composition and transposable elements (TEs) are particularly scarce. Nearly half of the recently released Eucalyptus grandis genome is composed by retrotransposons and this data provides an important opportunity to understand TE dynamics in Eucalyptus genome and transcriptome. RESULTS: We characterized nine families of transcriptionally active LTR retrotransposons from Copia and Gypsy superfamilies in Eucalyptus grandis genome and we depicted genomic distribution and copy number in two Eucalyptus species. We also evaluated genomic polymorphism and transcriptional profile in three organs of five Eucalyptus species. We observed contrasting genomic and transcriptional behavior in the same family among different species. RLC_egMax_1 was the most prevalent family and RLC_egAngela_1 was the family with the lowest copy number. Most families of both superfamilies have their insertions occurring <3 million years, except one Copia family, RLC_egBianca_1. Protein theoretical models suggest different properties between Copia and Gypsy domains. IRAP and REMAP markers suggested genomic polymorphisms among Eucalyptus species. Using EST analysis and qRT-PCRs, we observed transcriptional activity in several tissues and in all evaluated species. In some families, osmotic stress increases transcript values. CONCLUSION: Our strategy was successful in isolating transcriptionally active retrotransposons in Eucalyptus, and each family has a particular genomic and transcriptional pattern. Overall, our results show that retrotransposon activity have differentially affected genome and transcriptome among Eucalyptus species.

[The experience of forced sexual intercourse among undergraduate students: associated factors and possible consequences]. / Experiência de relação sexual forçada entre estudantes de graduação: fatores associados e possíveis consequências.

The scope of this article is to investigate the prevalence of the experience of forced sexual intercourse on life and its possible consequences on mental and sexual health among undergraduate students at a public university in Brazil. It is a quantitative cross-sectional study with 996 undergraduate students. Sampling was carried out systematically by single-stage clusters. Descriptive, bivariate and multivariate analyses were performed using the Poisson Regression model, and the etiological fractions of the consequences of forced sexual intercourse were calculated. The lifetime prevalence of forced sexual intercourse was 12.1%. Female sex, non-heterosexual sexual orientation, first sexual intercourse before the age of 14, child food insecurity and domestic violence were associated with a higher prevalence of the outcome. Individuals who experienced forced sexual intercourse were more likely to be diagnosed with sexually transmitted infections and to represent a potential suicide risk. The study revealed the problems of sexual violence in the university context.

O objetivo deste artigo é investigar a prevalência de relação sexual forçada na vida e suas possíveis consequências na saúde mental e sexual em acadêmicos de cursos de graduação de uma universidade pública no Brasil. Estudo transversal quantitativo com 996 estudantes de graduação. A amostragem realizada de forma sistemática por conglomerados. Foi usado o modelo de Regressão de Poisson, e foram calculadas as frações etiológicas das consequências de relação sexual forçada. A prevalência de relação sexual forçada ao longo da vida foi de 12,1%. Sexo feminino, orientação sexual não heterossexual, primeira relação sexual antes de 14 anos de idade, insegurança alimentar e violência no contexto doméstico na infância foram associados à maior prevalência do desfecho. Indivíduos que sofreram relação sexual forçada foram mais propensos a apresentar diagnóstico de infecções sexualmente transmissíveis e de ter risco de suicídio. O estudo evidenciou a problemática da violência sexual no contexto universitário.

Experiência de relação sexual forçada entre estudantes de graduação: fatores associados e possíveis consequências / The experience of forced sexual intercourse among undergraduate students: associated factors and possible consequences

Resumo O objetivo deste artigo é investigar a prevalência de relação sexual forçada na vida e suas possíveis consequências na saúde mental e sexual em acadêmicos de cursos de graduação de uma universidade pública no Brasil. Estudo transversal quantitativo com 996 estudantes de graduação. A amostragem realizada de forma sistemática por conglomerados. Foi usado o modelo de Regressão de Poisson, e foram calculadas as frações etiológicas das consequências de relação sexual forçada. A prevalência de relação sexual forçada ao longo da vida foi de 12,1%. Sexo feminino, orientação sexual não heterossexual, primeira relação sexual antes de 14 anos de idade, insegurança alimentar e violência no contexto doméstico na infância foram associados à maior prevalência do desfecho. Indivíduos que sofreram relação sexual forçada foram mais propensos a apresentar diagnóstico de infecções sexualmente transmissíveis e de ter risco de suicídio. O estudo evidenciou a problemática da violência sexual no contexto universitário.

Abstract The scope of this article is to investigate the prevalence of the experience of forced sexual intercourse on life and its possible consequences on mental and sexual health among undergraduate students at a public university in Brazil. It is a quantitative cross-sectional study with 996 undergraduate students. Sampling was carried out systematically by single-stage clusters. Descriptive, bivariate and multivariate analyses were performed using the Poisson Regression model, and the etiological fractions of the consequences of forced sexual intercourse were calculated. The lifetime prevalence of forced sexual intercourse was 12.1%. Female sex, non-heterosexual sexual orientation, first sexual intercourse before the age of 14, child food insecurity and domestic violence were associated with a higher prevalence of the outcome. Individuals who experienced forced sexual intercourse were more likely to be diagnosed with sexually transmitted infections and to represent a potential suicide risk. The study revealed the problems of sexual violence in the university context.