Host and microbiome features of secondary infections in lethal covid-19

Secondary infections contribute significantly to covid-19 mortality but driving factors remain poorly understood. Autopsies of 20 covid-19 cases and 14 controls from the first pandemic wave complemented with microbial cultivation and RNA-seq from lung tissues enabled description of major organ pathologies and specification of secondary infections. Lethal covid-19 segregated into two main death causes with either dominant diffuse alveolar damage (DAD) or secondary pneumonias. The lung microbiome in covid-19 showed a reduced biodiversity and increased prototypical bacterial and fungal pathogens in cases of secondary pneumonias. RNA-seq distinctly mirrored death causes and stratified DAD cases into subgroups with differing cellular compositions identifying myeloid cells, macrophages and complement C1q as strong separating factors suggesting a pathophysiological link. Together with a prominent induction of inhibitory immune-checkpoints our study highlights profound alterations of the lung immunity in covid-19 wherein a reduced antimicrobial defense likely drives development of secondary infections on top of SARS-CoV-2 infection

Potentiometric detection of the metabolic activity of human tumor cells

Monitoring of cellular viability is a key part of toxicological assays in vitro. On-line monitoring of metabolic activity would be particularly useful for evaluation of responses to potential therapeutic compounds. Current assays are mostly based on fluorescent dyes and optical detection methods. These methods offer high sensitivity and specificity, however are not suitable for long-term on-line observations. Electrochemical methods can be an alternative for current protocols. Electrochemical detection is low cost and label-free, therefore suitable for long-term cell culture monitoring. In this work investigations on human cancer cells viability will be presented. Cells were cultured as two-dimensional monolayer or three-dimensional spheroids. Different cell culture media were examined. Potentiometric detection was used for continuous monitoring of cell culture as well as end-point investigations. Different growth phases were identified using applied method. Finally, response to an anticancer drug was successfully observed

Consumption of sweetened beverages as a risk factor of colonization of oral cavity by fungi - eating habits of university students

Foods rich in sugar are an excellent substrate for the microorganisms that inhabit the initial sections of the gastrointestinal tract, and one of the most commonly available sources of sugar is the sweetened drink. Students represent an interesting sub-population; the large number of classes and associated stress levels promote fixing of unhealthy behaviors, e.g. tendency to consume a lot of sweetened drinks, for example cola-type or energetic drinks. Aim of this study was to determine the relationship between the amount of sugar consumed in beverages and the prevalence of fungi in the oral cavity. The investigated material consisted of oral washings. Participants completed original questionnaire regarding beverages consumed. The relationship between the consumption of sweetened beverages and risk of the presence of fungi in the oral cavity was determined. Fungi were isolated from 68.1% of examined subjects. Seven species of the genus Candida were observed. Higher prevalence of fungi was seen in the oral cavity of subjects who declared consumption of beverages containing sugar. 37.8% of respondents were found to consume with beverages doses of sugar exceeding the recommended daily requirement. Significantly greater prevalence of oral cavity fungi was noted in those exceeding the recommended GDA (76.3%), compared to of those who were not (68.7%). There were positive correlations between occurrence of fungi and consumption of sweetened carbonated drinks or adding sugar to coffee and tea. The addition of sugar to coffee/tea and sugar consumption above the recommended daily amount significantly increases the risk of colonization of the oral cavity by fungi. Students, due to invalid nutritional habits especially excessive consumption of beverages containing large amounts of sugar, belong to a group with a predisposition to the occurrence of fungi in the oral cavity

Loss of LRP1 promotes acquisition of contractile-myofibroblast phenotype and release of active TGF-beta 1 from ECM stores

International audienceIn healing tissue, fibroblasts differentiate to α-smooth muscle actin (SMA)-expressing contractile-myofibroblasts, which pull the wound edges together ensuring proper tissue repair. Uncontrolled expansion of the myofibroblast population may, however, lead to excessive tissue scarring and finally to organ dysfunction. Here, we demonstrate that the loss of low-density lipoprotein receptor-related protein (LRP) 1 overactivates the JNK1/2-c-Jun-Fra-2 signaling pathway leading to the induction of α-SMA and periostin expression in human lung fibroblasts (hLF). These changes are accompanied by increased contractility of the cells and the integrin- and protease-dependent release of active transforming growth factor (TGF)-β1 from the extracellular matrix (ECM) stores. Liberation of active TGF-β1 from the ECM further enhances α-SMA and periostin expression thus accelerating the phenotypic switch of hLF. Global gene expression profiling of LRP1-depleted hLF revealed that the loss of LRP1 affects cytoskeleton reorganization, cell-ECM contacts, and ECM production. In line with these findings, fibrotic changes in the skin and lung of Fra-2 transgenic mice were associated with LRP1 depletion and c-Jun overexpression. Altogether, our results suggest that dysregulation of LRP1 expression in fibroblasts in healing tissue may lead to the unrestrained expansion of contractile myofibroblasts and thereby to fibrosis development. Further studies identifying molecules, which regulate LRP1 expression, may provide new therapeutic options for largely untreatable human fibrotic diseases

BDNF/TrkB signaling augments smooth muscle cell proliferation in pulmonary hypertension.

Pulmonary hypertension (PH) is a life-threatening disorder that is characterized by pulmonary arterial smooth muscle cell (PASMC) hyperplasia. Until now, little was been known about early changes that underlie the manifestation of PH. To characterize these early changes, we performed whole-genome microarray analysis of lungs from mice exposed to either 24 hours hypoxia or normoxia. TrkB, a member of the tyrosine kinase receptor family, and its ligand, brain-derived neurotrophic factor (BDNF), were strongly up-regulated in hypoxic mouse lungs, as well as in arteries of patients suffering from idiopathic pulmonary arterial hypertension (IPAH). BDNF stimulation of PASMC in vitro resulted in increased proliferation, TrkB and ERK1/2 phosphorylation, and nuclear translocation of the transcription factor early growth response factor 1 (Egr-1). In addition, increased Egr-1 expression was observed in idiopathic PAH lungs. The pro-proliferative effect of BDNF was attenuated by TrkB kinase inhibitor (K252a) or ERK1/2 inhibitor (U0126) pretreatment, and by knocking down Egr-1. Consequently, we have identified the BDNF-TrkB-ERK1/2 pathway as a proproliferative signaling pathway for PASMC in PH. Interference with this pathway may thus serve as an attractive reverse remodeling approach

Pathobiology, pathology and genetics of pulmonary hypertension: Update from the Cologne Consensus Conference 2018

The European guidelines, which focus on clinical aspects of pulmonary hypertension (PH), provide only minimal information about the pathophysiological concepts of PH. Here, we review this topic in greater detail, focusing on specific aspects in the pathobiology, pathology and genetics, which include mechanisms of vascular inflammation, the role of transcription factors, ion channels/ion channel diseases, hypoxic pulmonary vasoconstriction, genetics/epigenetics, metabolic dysfunction, and the potential future role of histopathology of PH in the modern era of PH therapy. In addition to new insights in the pathobiology of this disease, this working group of the Cologne Consensus Conference also highlights novel concepts and potential new therapeutic targets to further improve the treatment options in PAH. (c) 2018 Published by Elsevier B.V

Pathobiology, pathology and genetics of pulmonary hypertension: Recommendations of the Cologne Consensus Conference 2016

The 2015 European Guidelines on Diagnosis and Treatment of Pulmonary Hypertension (PH) are also valid for Germany. While the guidelines contain detailed recommendations regarding clinical aspects of pulmonary arterial hypertension (PAH) and other forms of PH, they contain only a relatively short paragraph on novel findings on the pathobiology, pathology, and genetics. However, these are of great importance for our understanding of this complex disease both from a clinical and scientific point of view, and they are essential for the development of novel treatment strategies. To this end, a number of current data are relevant, prompting a detailed commentary to the guidelines, and the consideration of new scientific data. In June 2016, a Consensus Conference organized by the PH working groups of the German Society of Cardiology (DGK), the German Society of Respiratory Medicine (DGP) and the German Society of Pediatric Cardiology (DGPK) was held in Cologne, Germany. This conference aimed to solve practical and controversial issues surrounding the implementation of the European Guidelines in Germany. To this end, a number of working groups was initiated, one of which was specifically dedicated to the pathobiology, pathology and genetics of PH. This article summarizes the results and recommendations of this working group