Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) appeared just over 7 months ago in Wuhan, China. Early reports from China indicated that although some cases are asymptomatic, 20% of COVID-19 cases follow a severe course, necessitating hospitalization, with a quarter of hospitalized patients requiring intensive care unit (ICU) facilities [1]. Later reports from China and other countries substantiated these data, although ICU admission rates, proportion of patients receiving invasive mechanical ventilation (IMV), and mortality rates differ considerably between studies [2]. The life-threatening form of respiratory failure, acute respiratory distress syndrome (ARDS) is a frequent complication in COVID-19 [3]. The severity of ARDS is classified into categories of mild, moderate, and severe, depending on the degree of hypoxemia [4]. Patients with moderate-tosevere ARDS require invasive mechanical ventilation (IMV) and have a poor prognosis [4]. The incidence of ARDS and specifically, moderate-to-severe ARDS, among COVID-19 patients is currently unknown [5]. We describe here the results of a survey of clinical studies reporting COVID-19-associated ARDS in hospitalized patients since the beginning of the COVID-19 pandemic in January until the end of July 2020. Our aim was to obtain a clearer picture of the incidence of COVID-19-associated ARDS in hospitalized patients on a global level, to better define the burden to healthcare

Expression of recombinant proteins in Escherichia coli often results in the formation of insoluble inclusion bodies, In case of expression of eukaryotic proteins containing cysteine, which may form disulfide bonds in the native active protein, often nonnative inter- and intramolecular disulfide bonds exist in the inclusion bodies. Hence, several methods have been developed to isolate recombinant eukaryotic polypeptides from inclusion bodies, and to generate native disulfide bonds, to get active proteins. This article summarizes the different steps and methods of isolation and renaturation of eukaryotic proteins containing disulfide bonds, which have been expressed in E. coli as inclusion bodies, and shows which methods originally developed for studying the folding mechanism of naturally occurring proteins have been successfully adapted for reactivation of recombinant eukaryotic proteins.

Patients with chronic pancreatitis experience substantial impairments in health-related quality of life. The severity of chronic pancreatitis-related symptoms is directly associated with patient function and well-being. These data offer further insight into the impact of chronic pancreatitis on patient health status and may serve as the basis for the development of disease-specific instruments, which are needed to measure the effect of therapeutic interventions on patient-derived health outcomes.

Rationale: Alveolar liquid clearance is regulated by Na 1 uptake through the apically expressed epithelial sodium channel (ENaC) and basolaterally localized Na 1 -K 1 -ATPase in type II alveolar epithelial cells. Dysfunction of these Na 1 transporters during pulmonary inflammation can contribute to pulmonary edema.Objectives: In this study, we sought to determine the precise mechanism by which the TIP peptide, mimicking the lectin-like domain of tumor necrosis factor (TNF), stimulates Na 1 uptake in a homologous cell system in the presence or absence of the bacterial toxin pneumolysin (PLY).Methods: We used a combined biochemical, electrophysiological, and molecular biological in vitro approach and assessed the physiological relevance of the lectin-like domain of TNF in alveolar liquid clearance in vivo by generating triple-mutant TNF knock-in mice that express a mutant TNF with deficient Na 1 uptake stimulatory activity.Measurements and Main Results: TIP peptide directly activates ENaC, but not the Na 1 -K 1 -ATPase, upon binding to the carboxyterminal domain of the a subunit of the channel. In the presence of PLY, a mediator of pneumococcal-induced pulmonary edema, this binding stabilizes the ENaC-PIP2-MARCKS complex, which is necessary for the open probability conformation of the channel and preserves ENaC-a protein expression, by means of blunting the protein kinase C-a pathway. Triple-mutant TNF knock-in mice are more prone than wild-type mice to develop edema with low-dose intratracheal PLY, correlating with reduced pulmonary ENaC-a subunit expression.Conclusions: These results demonstrate a novel TNF-mediated mechanism of direct ENaC activation and indicate a physiological role for the lectin-like domain of TNF in the resolution of alveolar edema during inflammation.

Antibiotics-induced release of the pore-forming virulence factor pneumolysin (PLY) in patients with pneumococcal pneumonia results in its presence days after lungs are sterile and is a major factor responsible for the induction of permeability edema. Here we sought to identify major mechanisms mediating PLY-induced endothelial dysfunction. We evaluated PLY-induced endothelial hyperpermeability in human lung microvascular endothelial cells (HL-MVECs) and human lung pulmonary artery endothelial cells in vitro and in mice instilled intratracheally with PLY. PLY increases permeability in endothelial monolayers by reducing stable and dynamic microtubule content and modulating VE-cadherin expression. These events, dependent upon an increased calcium influx, are preceded by protein kinase C (PKC)-a activation, perturbation of the RhoA/Rac1 balance, and an increase in myosin light chain phosphorylation. At later time points, PLY treatment increases the expression and activity of arginase in HL-MVECs. Arginase inhibition abrogates and suppresses PLY-induced endothelial barrier dysfunction by restoring NO generation. Consequently, a specific PKC-a inhibitor and the TNF-derived tonoplast intrinsic protein peptide, which blunts PLY-induced PKC-a activation, are able to prevent activation of arginase in HL-MVECs and to reduce PLY-induced endothelial hyperpermeability in mice. Arginase I (AI) 1/2 /arginase II (AII) 2/2 C57BL/6 mice, displaying a significantly reduced arginase I expression in the lungs, are significantly less sensitive to PLY-induced capillary leak than their wild-type or AI 1/1 /AII 2/2 counterparts, indicating an important role for arginase I in PLYinduced endothelial hyperpermeability. These results identify PKC-a and arginase I as potential upstream and downstream therapeutic targets in PLY-induced pulmonary endothelial dysfunction.Keywords: PKC; arginase; pneumococcus; pneumolysin; TNF Although severe pneumonia remains the leading cause of mortality worldwide in children aged less than 5 years (1), community-acquired pneumonia represents a major cause of morbidity and mortality mainly in elderly patients (2). Despite the use of potent antibiotics and aggressive intensive care support, the fatality rate associated with Streptococcus pneumoniae, accounting for 45% of all cases of community-acquired pneumonia, is approximately 20% (1, 2). Pulmonary permeability edema, a major complication of severe pneumonia characterized by endothelial hyperpermeability, can occur days after initiation of antibiotics therapy when tissues are sterile and the pneumonia is clearing and correlates with the presence of the bacterial virulence factor pneumolysin (PLY) (3, 4). This cytoplasmic hemolytic protein is released during bacterial lysis, as occurs after treatment with b-lactam antibiotics (5). PLYinduced lung injury was suggested to result from direct pneumotoxic effects on the alveolar-capillary barrier rather than from resident or recruited phagocytic cells (6).Upon binding of PLY to cholesterol in cell membranes, oligo...

Patients with chronic pancreatitis experience substantial deteriorations in health-related quality of life compared with the general population. The Short Form-36 proved to be a feasible, reliable and valid measure for descriptive studies of patients with chronic pancreatitis, but ceiling effects may limit its usefulness as an outcome measure in the assessment of treatment effects.

This letter introduces a method based on electroluminescence imaging to determine mappings of the local series resistance of large area semiconductor devices such as solar cells. The method combines the local electroluminescence emission Φi(U) and its derivative Φi′(U) with respect to the applied voltage U. The combined analysis of these two quantities yields the local series resistance Rise and proves the physical validity of the used current transport model and thus the physical relevance of the determined Rise value. The method is verified on a monocrystalline silicon solar cell with local shunts and local series resistance problems.

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