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Abstract Influenza A virus (IAV) infection mobilizes bone marrow-derived macrophages (BMDM) that gradually undergo transition to tissue-resident alveolar macrophages (TR-AM) in the inflamed lung. Combining high-dimensional single-cell transcriptomics with complex lung organoid modeling, in vivo adoptive cell transfer, and BMDM-specific gene targeting, we found that transitioning (“regenerative”) BMDM and TR-AM highly express Placenta-expressed transcript 1 (Plet1). We reveal that Plet1 is released from alveolar macrophages, and acts as important mediator of macrophage-epithelial cross-talk during lung repair by inducing proliferation of alveolar epithelial cells and re-sealing of the epithelial barrier. Intratracheal administration of recombinant Plet1 early in the disease course attenuated viral lung injury and rescued mice from otherwise fatal disease, highlighting its therapeutic potential.

IntroductionProtein kinases are indispensable reversible molecular switches that adapt and control protein functions during cellular processes requiring rapid responses to internal and external events. Bacterial infections can affect kinase-mediated phosphorylation events, with consequences for both innate and adaptive immunity, through regulation of antigen presentation, pathogen recognition, cell invasiveness and phagocytosis. Streptococcus pneumoniae (Spn), a human respiratory tract pathogen and a major cause of community-acquired pneumoniae, affects phosphorylation-based signalling of several kinases, but the pneumococcal mediator(s) involved in this process remain elusive. In this study, we investigated the influence of pneumococcal H2O2 on the protein kinase activity of the human lung epithelial H441 cell line, a generally accepted model of alveolar epithelial cells.MethodsWe performed kinome analysis using PamGene microarray chips and protein analysis in Western blotting in H441 lung cells infected with Spn wild type (SpnWT) or with SpnΔlctOΔspxB -a deletion mutant strongly attenuated in H2O2 production- to assess the impact of pneumococcal hydrogen peroxide (H2O2) on global protein kinase activity profiles.ResultsOur kinome analysis provides direct evidence that kinase activity profiles in infected H441 cells significantly vary according to the levels of pneumococcal H2O2. A large number of kinases in H441 cells infected with SpnWT are significantly downregulated, whereas this no longer occurs in cells infected with the mutant SpnΔlctOΔspxB strain, which lacks H2O2. In particular, we describe for the first time H2O2-mediated downregulation of Protein kinase B (Akt1) and activation of lymphocyte-specific tyrosine protein kinase (Lck) via H2O2-mediated phosphorylation.

Similar to human–human interaction (HHI), gaze is an important modality in conversational human–robot interaction (HRI) settings. Previously, human-inspired gaze parameters have been used to implement gaze behavior for humanoid robots in conversational settings and improve user experience (UX). Other robotic gaze implementations disregard social aspects of gaze behavior and pursue a technical goal (e.g., face tracking). However, it is unclear how deviating from human-inspired gaze parameters affects the UX. In this study, we use eye-tracking, interaction duration, and self-reported attitudinal measures to study the impact of non-human inspired gaze timings on the UX of the participants in a conversational setting. We show the results for systematically varying the gaze aversion ratio (GAR) of a humanoid robot over a broad parameter range from almost always gazing at the human conversation partner to almost always averting the gaze. The main results reveal that on a behavioral level, a low GAR leads to shorter interaction durations and that human participants change their GAR to mimic the robot. However, they do not copy the robotic gaze behavior strictly. Additionally, in the lowest gaze aversion setting, participants do not gaze back as much as expected, which indicates a user aversion to the robot gaze behavior. However, participants do not report different attitudes toward the robot for different GARs during the interaction. In summary, the urge of humans in conversational settings with a humanoid robot to adapt to the perceived GAR is stronger than the urge of intimacy regulation through gaze aversion, and a high mutual gaze is not always a sign of high comfort, as suggested earlier. This result can be used as a justification to deviate from human-inspired gaze parameters when necessary for specific robot behavior implementations.

Social robots are often envisioned as companions for older adults and chronically ill people to support them in everyday life (taking medication, staying physically active, etc.). For all these tasks a high-level of personalization is required, in order to achieve satisfying support as well as a high degree of engagement to motivate people to continue a healthy lifestyle. In the Austrian project RoboGen, we wanted to achieve exactly that. Using an off-the-shelf low-cost robotic platform (Q.bo One), we extended it with a learning agent and designed the interaction in a gender-sensitive way. In this brief communication we present the evaluation results derived from a case study with potential users and semi-structured interviews with experts. Our results show the challenges that come with off-the-shelf robotic platforms and usability (speech recognition, text-to-speech output etc.), also highlight the perception of the potential users as well as how experts rated the concept.

Idiopathic pulmonary fibrosis is associated with myofibroblast activation in the lungs and metabolic alterations. Here, the authors show that the antidiabetic drug metformin has antifibrotic effects in human-derived samples and mouse models, by modulating a number of metabolic pathways to induce lipogenic transdifferentiation of myofibroblasts.

Cells of the pulmonary vasculature show a hyperproliferative phenotype in pulmonary arterial hypertension (PAH), thus contributing to the disease pathogenesis. Here the authors show that cyclin-dependent kinases are overactivated in PAH, and that their pharmacological inhibition attenuates the disease in two independent rodent models

Endurance training induces several adaptations in substrate metabolism, especially in relation to glycogen conservation. The study aimed to investigate differences in the metabolism of lipids, lipid-like substances, and amino acids between highly trained and untrained subjects using targeted metabolomics. Depending on their maximum relative oxygen uptake (VO2max), subjects were categorized as either endurance-trained (ET) or untrained (UT). Resting blood was taken and plasma isolated. It was screened for changes of 345 metabolites, including amino acids and biogenic amines, acylcarnitines, glycerophosphocholines (GPCs), sphingolipids, hexoses, bile acids, and polyunsaturated fatty acids (PUFAs) by using liquid chromatography coupled to tandem mass spectrometry. Acylcarnitine (C14:1, down in ET) and five GPCs (lysoPC a C18:2, up in ET; PC aa C42:0, up in ET; PC ae C38:2, up in ET; PC aa C38:5, down in ET; lysoPC a C26:0, down in ET) were differently regulated in ET compared to UT. TCDCA was down-regulated in athletes, while for three ratios of bile acids CA/CDCA, CA/(GCA+TCA), and DCA/(GDCA+TDCA) an up-regulation was found. TXB2 and 5,6-EET were down-regulated in the ET group and 18S-HEPE, a PUFA, showed higher levels in 18S-HEPE in endurance-trained subjects. For PC ae C38:2, TCDCA, and the ratio of cholic acid to chenodeoxycholic acid, an association with VO2max was found. Numerous phospholipids, acylcarnitines, glycerophosphocholines, bile acids, and PUFAs are present in varying concentrations at rest in ET. These results might represent an adaptation of lipid metabolism and account for the lowered cardiovascular risk profile of endurance athletes.

Pulmonary arterial hypertension is a severe respiratory disease characterized by pulmonary artery remodeling. RV dysfunction and dysregulated circulating metabolomics are associated with adverse outcomes in pulmonary arterial hypertension. We investigated effects of tadalafil and macitentan alone or in combination on the RV and plasma metabolomics in SuHx and PAB models. For SuHx model, rats were injected with SU5416 and exposed to hypoxia for three weeks and then were returned to normoxia and treated with either tadalafil (10 mg/kg in chow) or macitentan (10 mg/kg in chow) or their combination (both 10 mg/kg in chow) for two weeks. For PAB model, rats were subjected to either sham or PAB surgery for three weeks and treated with above-mentioned drugs from week 1 to week 3. Following terminal echocardiographic and hemodynamic measurements, tissue samples were collected for metabolomic, histological and gene expression analysis. Both SuHx and PAB rats developed RV remodeling/dysfunction with severe and mild plasma metabolomic alterations, respectively. In SuHx rats, tadalafil and macitentan alone or in combination improved RV remodeling/function with the effects of macitentan and combination therapy being superior to tadalafil. All therapies similarly attenuated SuHx-induced changes in plasma metabolomics. In PAB rats, only macitentan improved RV remodeling/function, while only tadalafil attenuated PAB-induced changes in plasma metabolomics.

Escherichia (E.) coli is the main causative pathogen of neonatal and post-weaning diarrhea and edema disease in swine production. There is a significant health concern due to an increasing number of human infections associated with food and/or environmental-borne pathogenic and multidrug-resistant E. coli worldwide. Monitoring the presence of pathogenic and antimicrobial-resistant E. coli isolates is essential for sustainable disease management in livestock and human medicine. A total of 102 E. coli isolates of diseased pigs were characterized by antimicrobial and biocide susceptibility testing. Antimicrobial resistance genes, including mobile colistin resistance genes, were analyzed by PCR and DNA sequencing. The quinolone resistance-determining regions of gyrA and parC in ciprofloxacin-resistant isolates were analyzed. Clonal relatedness was investigated by two-locus sequence typing (CH clonotyping). Phylotyping was performed by the Clermont multiplex PCR method. Virulence determinants were analyzed by customized DNA-based microarray technology developed in this study for fast and economic molecular multiplex typing. Thirty-five isolates were selected for whole-genome sequence-based analysis. Most isolates were resistant to ampicillin and tetracycline. Twenty-one isolates displayed an ESBL phenotype and one isolate an AmpC β-lactamase-producing phenotype. Three isolates had elevated colistin minimal inhibitory concentrations and carried the mcr-1 gene. Thirty-seven isolates displayed a multi-drug resistance phenotype. The most predominant β-lactamase gene classes were blaTEM-1 (56%) and blaCTX-M-1 (13.71%). Mutations in QRDR were observed in 14 ciprofloxacin-resistant isolates. CH clonotyping divided all isolates into 51 CH clonotypes. The majority of isolates belonged to phylogroup A. Sixty-four isolates could be assigned to defined pathotypes wherefrom UPEC was predominant. WGS revealed that the most predominant sequence type was ST100, followed by ST10. ST131 was detected twice in our analysis. This study highlights the importance of monitoring antimicrobial resistance and virulence properties of porcine E. coli isolates. This can be achieved by applying reliable, fast, economic and easy to perform technologies such as DNA-based microarray typing. The presence of high-risk pathogenic multi-drug resistant zoonotic clones, as well as those that are resistant to critically important antibiotics for humans, can pose a risk to public health. Improved protocols may be developed in swine farms for preventing infections, as well as the maintenance and distribution of the causative isolates.

It has been shown previously that increased circulating endothelial cells-derived extracellular vesicles represent an important pathological attribute of pulmonary hypertension. Although it is a well-known fact that inflammatory cells may also release extracellular vesicles, and pulmonary hypertension is a disease associated with abnormal inflammation, there is no profound knowledge with regard to the role of inflammatory cells-derived extracellular vesicles. Therefore, our study demonstrated that circulating levels of extracellular vesicles derived from T-cells are enhanced in various pulmonary hypertension forms and that endothelial cells-derived extracellular vesicles may have distinctive profiles in different clinical subgroups of pulmonary hypertension, which still remains as a poorly treatable and life-threatening disorder.

We present the robot developed within the Hobbit project, a socially assistive service robot aiming at the challenge of enabling prolonged independent living of elderly people in their own homes. We present the second prototype (Hobbit PT2) in terms of hardware and functionality improvements following first user studies. Our main contribution lies within the description of all components developed within the Hobbit project, leading to autonomous operation of 371 days during field trials in Austria, Greece, and Sweden. In these field trials, we studied how 18 elderly users (aged 75 years and older) lived with the autonomously interacting service robot over multiple weeks. To the best of our knowledge, this is the first time a multifunctional, low-cost service robot equipped with a manipulator was studied and evaluated for several weeks under real-world conditions. We show that Hobbit’s adaptive approach towards the user increasingly eased the interaction between the users and Hobbit. We provide lessons learned regarding the need for adaptive behavior coordination, support during emergency situations, and clear communication of robotic actions and their consequences for fellow researchers who are developing an autonomous, low-cost service robot designed to interact with their users in domestic contexts. Our trials show the necessity to move out into actual user homes, as only there can we encounter issues such as misinterpretation of actions during unscripted human-robot interaction.

This article reports three case studies on the usability and acceptance of an industrial robotic prototype in the context of human-robot cooperation. The three case studies were conducted in the framework of a two-year project named AssistMe, which aims at developing different means of interaction for programming and using collaborative robots in a user-centered manner. Together with two industrial partners and a technological partner, two different application scenarios were implemented and studied with an off-the-shelf robotic system. The operators worked with the robotic prototype in laboratory conditions (two days), in a factory context (one day) and in an automotive assembly line (three weeks). In the article, the project and procedures are described in detail, including the quantitative and qualitative methodology. Our results show that close human-robot cooperation in the industrial context needs adaptive pacing mechanisms in order to avoid a change of working routines for the operators and that an off-the-shelf robotic system is still limited in terms of usability and acceptance. The touch panel, which is needed for controlling the robot, had a negative impact on the overall user experience. It creates a further intermediate layer between the user, the robot and the work piece and potentially leads to a decrease in productivity. Finally, the fear of the worker of being replaced by an improved robotic system was regularly expressed and adds an additional anthropocentric dimension to the discussion of human-robot cooperation, smart factories and the upcoming Industry 4.0.