Increased Alveolar Epithelial Damage Markers and Inflammasome-Regulated Cytokines Are Associated with Pulmonary Superinfection in ARDS.

Acute respiratory distress syndrome (ARDS) is a life-threatening form of respiratory failure defined by dysregulated immune homeostasis and alveolar epithelial and endothelial damage. Up to 40% of ARDS patients develop pulmonary superinfections, contributing to poor prognosis and increasing mortality. Understanding what renders ARDS patients highly susceptible to pulmonary superinfections is therefore essential. We hypothesized that ARDS patients who develop pulmonary superinfections display a distinct pulmonary injury and pro-inflammatory response pattern. Serum and BALF samples from 52 patients were collected simultaneously within 24 h of ARDS onset. The incidence of pulmonary superinfections was determined retrospectively, and the patients were classified accordingly. Serum concentrations of the epithelial markers soluble receptor for advanced glycation end-products (sRAGE) and surfactant protein D (SP-D) and the endothelial markers vascular endothelial growth factor (VEGF) and angiopoetin-2 (Ang-2) as well as bronchoalveolar lavage fluid concentrations of the pro-inflammatory cytokines interleukin 1ß (IL-1ß), interleukin 18 (IL-18), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-a) were analyzed via multiplex immunoassay. Inflammasome-regulated cytokine IL-18 and the epithelial damage markers SP-D and sRAGE were significantly increased in ARDS patients who developed pulmonary superinfections. In contrast, endothelial markers and inflammasome-independent cytokines did not differ between the groups. The current findings reveal a distinct biomarker pattern that indicates inflammasome activation and alveolar epithelial injury. This pattern may potentially be used in future studies to identify high-risk patients, enabling targeted preventive strategies and personalized treatment approaches.

Impaired ketogenesis ties metabolism to T cell dysfunction in COVID-19.

Anorexia and fasting are host adaptations to acute infection, and induce a metabolic switch towards ketogenesis and the production of ketone bodies, including ß-hydroxybutyrate (BHB)1-6. However, whether ketogenesis metabolically influences the immune response in pulmonary infections remains unclear. Here we show that the production of BHB is impaired in individuals with SARS-CoV-2-induced acute respiratory distress syndrome (ARDS) but not in those with  influenza-induced ARDS. We found that BHB promotes both the survival of and the production of interferon-γ by CD4+ T cells. Applying a metabolic-tracing analysis, we established that BHB provides an alternative carbon source to fuel oxidative phosphorylation (OXPHOS) and the production of bioenergetic amino acids and glutathione, which is important for maintaining the redox balance. T cells from patients with SARS-CoV-2-induced ARDS were exhausted and skewed towards glycolysis, but could be metabolically reprogrammed by BHB to perform OXPHOS, thereby increasing their functionality. Finally, we show in mice that a ketogenic diet and the delivery of BHB as a ketone ester drink restores CD4+ T cell metabolism and function in severe respiratory infections, ultimately reducing the mortality of mice infected with SARS-CoV-2. Altogether, our data reveal that BHB is an alternative source of carbon that promotes T cell responses in pulmonary viral infections, and highlight impaired ketogenesis as a potential confounding factor in severe COVID-19.

Cytosolic d-type CpG-oligonucleotides induce a type I interferon response by activating the cGAS-STING signaling pathway.

Cytosolic DNA receptor cyclic GMP-AMP (cGAMP) synthase (cGAS) has been shown to be critically involved in the detection of cytosolic, self- and non-self-DNA, initiating a type I IFN response through the adaptor protein Stimulator of Interferon Genes (STING) and interferon regulatory factor 3 (IRF3). Current studies propose that canonical binding of dsDNA by cGAS depends on DNA length, but not on base sequence. In contrast, activation of TLR9 is sequence dependent. It requires unmethylated CpG dinucleotides in microbial DNA, which is mimicked by synthetic oligodeoxynucleotides (ODN). Here, we provide evidence that d-type ODN (D-ODN), but not K-type ODN (K-ODN), bind to human cGAS and activate downstream signaling. Transfection of D-ODN into a TLR9-deficient, human monocytic cell line (THP-1) induced phosphorylation of IRF3 and secretion of IFN. This response was absent in cells with CRISPR/Cas9-mediated cGAS- or STING-deficiency. Utilizing a protein pulldown approach, we further demonstrate direct binding of D-ODN to cGAS. Induction of a type I IFN response by D-ODN was confirmed in human primary monocytes and monocyte-derived macrophages. These results are relevant to our understanding of self-nonself-discrimination by cGAS and to the pharmacologic effects of ODN, which currently are investigated in clinical studies.

Diseño y Evaluación de un micro viscosímetro de bajo costo utilizando un resonador de cristal de cuarzo y Arduino / Designing and evaluating a low-cost micro-viscometer using a quartz crystal resonator (QCR) and an Arduino DUE microcontroller board / Desenho e Avaliação de um micro viscosímetro de baixo custo usando um ressonador de cristal de quartzo e Arduino

Resumen El objetivo principal de este artículo es proponer el diseño de un micro viscosímetro de bajo coste utilizando un resonador de cristal de cuarzo (QCR) y una tarjeta Arduino DUE. En el artículo se explican los pasos del diseño del sensor y también su evaluación con dos líquidos de diferentes características: diluciones de glicerol en agua (comportamiento Newtoniano) y diluciones de albúmina de huevo extraída de huevos frescos en agua (comportamiento No-Newtoniano). Este estudio está relacionado con el interés de desarrollar nuevas herramientas para el diagnóstico de enfermedades cardiovasculares y artríticas.

Abstract This article outlines a design for a low-cost micro-viscometer, using a quartz crystal resonator (QCR) and an Arduino DUE programmable microcontroller board. The article explains the steps involved in designing the sensor and also how it was evaluated regarding two liquids having different characteristics: dilute aqueous glycerol (Newtonian behaviour) and dilutions of egg-white extracted from fresh eggs in water (non-Newtonian behaviour). This study was related to interest in developing new tools for diagnosing cardiovascular and arthritic diseases.

Resumo O principal objetivo deste trabalho é propor o projeto de um micro viscosímetro de baixo custo usando um ressonador de cristal de quartzo (QCR) e um cartão Arduino DUE. O artigo explica as etapas do desenho do sensor e também sua avaliação com dois líquidos de diferentes características: diluições de glicerol em água (comportamento newtoniano) e diluições de albumina de ovo extraídas de ovos frescos em água (comportamento não newtoniano). Este estudo está relacionado ao interesse de desenvolver novas ferramentas para o diagnóstico de doenças cardiovasculares e artríticas.