Vector-borne Trypanosoma brucei parasites develop in artificial human skin and persist as skin tissue forms.

Transmission of Trypanosoma brucei by tsetse flies involves the deposition of the cell cycle-arrested metacyclic life cycle stage into mammalian skin at the site of the fly's bite. We introduce an advanced human skin equivalent and use tsetse flies to naturally infect the skin with trypanosomes. We detail the chronological order of the parasites' development in the skin by single-cell RNA sequencing and find a rapid activation of metacyclic trypanosomes and differentiation to proliferative parasites. Here we show that after the establishment of a proliferative population, the parasites enter a reversible quiescent state characterized by slow replication and a strongly reduced metabolism. We term these quiescent trypanosomes skin tissue forms, a parasite population that may play an important role in maintaining the infection over long time periods and in asymptomatic infected individuals.

Data augmentation in natural language processing: a novel text generation approach for long and short text classifiers.

In many cases of machine learning, research suggests that the development of training data might have a higher relevance than the choice and modelling of classifiers themselves. Thus, data augmentation methods have been developed to improve classifiers by artificially created training data. In NLP, there is the challenge of establishing universal rules for text transformations which provide new linguistic patterns. In this paper, we present and evaluate a text generation method suitable to increase the performance of classifiers for long and short texts. We achieved promising improvements when evaluating short as well as long text tasks with the enhancement by our text generation method. Especially with regard to small data analytics, additive accuracy gains of up to 15.53% and 3.56% are achieved within a constructed low data regime, compared to the no augmentation baseline and another data augmentation technique. As the current track of these constructed regimes is not universally applicable, we also show major improvements in several real world low data tasks (up to +4.84 F1-score). Since we are evaluating the method from many perspectives (in total 11 datasets), we also observe situations where the method might not be suitable. We discuss implications and patterns for the successful application of our approach on different types of datasets.

Digital Resilience in Dealing with Misinformation on Social Media during COVID-19: A Web Application to Assist Users in Crises.

In crises such as the COVID-19 pandemic, it is crucial to support users when dealing with social media content. Considering digital resilience, we propose a web app based on Social Network Analysis (SNA) to provide an overview of potentially misleading vs. non-misleading content on Twitter, which can be explored by users and enable foundational learning. The latter aims at systematically identifying thematic patterns which may be associated with misleading information. Additionally, it entails reflecting on indicators of misleading tweets which are proposed to approach classification of tweets. Paying special attention to non-expert users of social media, we conducted a two-step Think Aloud study for evaluation. While participants valued the opportunity to generate new knowledge and the diversity of the application, qualities such as equality and rapidity may be further improved. However, learning effects outweighed individual costs as all users were able to shift focus onto relevant features, such as hashtags, while readily pointing out content characteristics. Our design artifact connects to learning-oriented interventions regarding the spread of misleading information and tackles information overload by a SNA-based plug-in.

Comparison of the Newborn Infant Parasympathetic Evaluation (NIPE™) index to changes in heart rate to detect intraoperative nociceptive stimuli in healthy and critically ill children below 2 years: An observational study.

BACKGROUND: The validity of current tools for intraoperative objective assessment of nociception/antinociception balance during anesthesia in young and very young surgery children is unknown. AIM: Primary aim of the study was to test the hypothesis that the Newborn Infant Parasympathetic Evaluation (NIPE) index performs better in indicating nociception in anesthetized children below 2 years than changes in heart rate. Secondary aims were to evaluate associations between intraoperative changes in NIPE index values and postoperative pain and emergence delirium. METHODS: Fifty-one children aged <2 years who underwent surgery were included in this prospective observational study. Patients were assigned to either group 1 (healthy children, n = 31) or group 2 (critically ill, ventilated premature infants and neonates, n = 20). The NIPE index and heart rate in response to three defined nociceptive stimuli were continuously recorded. Two different scales, Kindliche Unbehagens- und Schmerzskala (KUS) and Pediatric Anesthesia Emergence Delirium (PAED) as well as a Pain Questionnaire were used to assess postoperative pain levels and emergence delirium. RESULTS: In total, 110 nociceptive events were evaluated. The analysis revealed a statistically significant association between a decrease in the NIPE index and all nociceptive stimuli, with a sensitivity of 92% and a specificity of 96%. The mean percentage decrease ranged from approx. 15%-30% and was highly statistically significant in both groups and for each of the nociceptive events except for venous puncture (p = .004). In contrast, no consistent change in heart rate was demonstrated. The KUS and PAED scale scores were significantly associated with the duration of anesthesia (p = .04), but not with intraoperative NIPE depression. CONCLUSION: The NIPE index was reliable for assessing intraoperative nociception in children aged <2 years and was more reproducible for detecting specific nociceptive stimuli during general anesthesia than heart rate. An effect on postoperative outcome is still elusive.

Dual-Use and Trustworthy? A Mixed Methods Analysis of AI Diffusion Between Civilian and Defense R&D.

Artificial Intelligence (AI) seems to be impacting all industry sectors, while becoming a motor for innovation. The diffusion of AI from the civilian sector to the defense sector, and AI's dual-use potential has drawn attention from security and ethics scholars. With the publication of the ethical guideline Trustworthy AI by the European Union (EU), normative questions on the application of AI have been further evaluated. In order to draw conclusions on Trustworthy AI as a point of reference for responsible research and development (R&D), we approach the diffusion of AI across both civilian and military spheres in the EU. We capture the extent of technological diffusion and derive European and German patent citation networks. Both networks indicate a low degree of diffusion of AI between civilian and defense sectors. A qualitative investigation of project descriptions of a research institute's work in both civilian and military fields shows that military AI applications stress accuracy or robustness, while civilian AI reflects a focus on human-centric values. Our work represents a first approach by linking processes of technology diffusion with normative evaluations of R&D.

Unexpected plasticity in the life cycle of Trypanosoma brucei.

African trypanosomes cause sleeping sickness in humans and nagana in cattle. These unicellular parasites are transmitted by the bloodsucking tsetse fly. In the mammalian host's circulation, proliferating slender stage cells differentiate into cell cycle-arrested stumpy stage cells when they reach high population densities. This stage transition is thought to fulfil two main functions: first, it auto-regulates the parasite load in the host; second, the stumpy stage is regarded as the only stage capable of successful vector transmission. Here, we show that proliferating slender stage trypanosomes express the mRNA and protein of a known stumpy stage marker, complete the complex life cycle in the fly as successfully as the stumpy stage, and require only a single parasite for productive infection. These findings suggest a reassessment of the traditional view of the trypanosome life cycle. They may also provide a solution to a long-lasting paradox, namely the successful transmission of parasites in chronic infections, despite low parasitemia.

Differences and similarities in comfort and discomfort experience in nine countries in Asia, the Americas and Europe.

In order to investigate differences in comfort and discomfort experiences amongst different regions of the world (America, Asia and Europe), a cross cultural study was performed. A questionnaire was sent to participants out in nine countries (Brazil, Canada, the USA, China, Indonesia, Thailand, Germany, Italy and the Netherlands). In total 795 participants completed the questionnaires. All countries score the comfort of a luxurious bed higher than a simple bed, first-class seats higher than economy class and all countries rate the comfort lower when the duration of sitting increases. The study suggests that in the USA and Canada softer beds, hammocks, more luxurious seats and softer pillows are scored as more comfortable compared with the other countries. There are indications that China and Germany prefer a harder mattress than in the other countries. For pillows, the differences between countries are large, which might show that much is influenced by habitude or hesitation to use something new. The Asian countries score the comfort of a brace neck pillow higher, which might be because these participants better realise the benefits better or feel less concerned to wear something that might give the appearance of an orthotic device. Further studies are needed to confirm these suggestions. The study shows that obvious differences are seen in all countries, which makes the construct of comfort internationally comparable. Practitioner summary: In designing and manufacturing globally, it is important to know how different parts of the world experience (dis)comfort. This study did not show large cultural differences amongst nine countries. Some differences emerge regarding pillows, perhaps as differences in sleeping habits play a role. Abbreviations: MANOVA: multivariate analysis of variance; VDA: Vargha and Delaney's A statistic; USA: United States of America.

Low pressure gas electron diffraction: An experimental setup and case studies.

Principles of low pressure gas electron diffraction are introduced. An experimental setup has been constructed for measuring the electron diffraction patterns of gaseous samples at pressures below 10-3 mbar. Test measurements have been performed for benzoic acid at T = 287 K corresponding to a vapor pressure of the substance P = 2 × 10-4 mbar, for iodoform CHI3 at T = 288 K (P = 4 × 10-4 mbar), and for carbon tetraiodide CI4 at T = 290 K (P = 1 × 10-4 mbar). Due to the low experimental temperature, thermal decomposition of CI4 has been prevented, which was unavoidable in previous classical measurements at higher temperatures. From the obtained data, the molecular structures have been successfully refined. The most important semi-empirical equilibrium molecular parameters are re(Car-Car)av = 1.387(5) Å in benzoic acid, re(C-I) = 2.123(3) Å in iodoform, and re(C-I) = 2.133(7) Å in carbon tetraiodide. The determined parameters showed consistency with the theoretically predicted values. A critical comparison with the results of the earlier investigations has also been done.

Combined gas electron diffraction and mass spectrometric experimental setup at Bielefeld University.

We have designed and constructed a combined experimental setup for synchronous measurements of electron diffraction patterns and mass-spectra of gas samples. Test measurements have been performed for acetic acid at two temperatures, 296 K and 457 K. Electron diffraction data have been analyzed taking into account mass spectra measured in the same experiments. From the diffraction intensities, molecular structures and mole fractions of the acetic acid monomer and dimer have been refined. The obtained results demonstrate the importance of measuring mass spectra in gas electron diffraction experiments. In particular, it is possible to detect the sample decomposition, which can be used for the optimization of experimental conditions and for the data interpretation. The length of the hydrogen bond in the acetic acid dimer determined in this work, re(O⋯H) = 1.657(9) Å, is in good agreement with modern theoretical predictions. We recommend measuring the diffraction patterns of acetic acid for the calibration of the sample pressure in the diffraction volume.

Recombinant Adenosine Deaminase Ameliorates Inflammation, Vascular Disease, and Fibrosis in Preclinical Models of Systemic Sclerosis.

OBJECTIVE: Systemic sclerosis (SSc) is characterized by fibrosis, vascular disease, and inflammation. Adenosine signaling plays a central role in fibroblast activation. We undertook this study to evaluate the therapeutic effects of adenosine depletion with PEGylated adenosine deaminase (PEG-ADA) in preclinical models of SSc. METHODS: The effects of PEG-ADA on inflammation, vascular remodeling, and tissue fibrosis were analyzed in Fra-2 mice and in a B10.D2→BALB/c (H-2d ) model of sclerodermatous chronic graft-versus-host disease (GVHD). The effects of PEG-ADA were confirmed in vitro in a human full-thickness skin model. RESULTS: PEG-ADA effectively inhibited myofibroblast differentiation and reduced pulmonary fibrosis by 34.3% (with decreased collagen expression) (P = 0.0079; n = 6), dermal fibrosis by 51.8% (P = 0.0006; n = 6), and intestinal fibrosis by 17.7% (P = 0.0228; n = 6) in Fra-2 mice. Antifibrotic effects of PEG-ADA were also demonstrated in sclerodermatous chronic GVHD (reduced by 38.4%) (P = 0.0063; n = 8), and in a human full-thickness skin model. PEG-ADA treatment decreased inflammation and corrected the M2/Th2/group 2 innate lymphoid cell 2 bias. Moreover, PEG-ADA inhibited proliferation of pulmonary vascular smooth muscle cells (reduced by 40.5%) (P < 0.0001; n = 6), and prevented thickening of the vessel walls (reduced by 39.6%) (P = 0.0028; n = 6) and occlusions of pulmonary arteries (reduced by 63.9%) (P = 0.0147; n = 6). Treatment with PEG-ADA inhibited apoptosis of microvascular endothelial cells (reduced by 65.4%) (P = 0.0001; n = 6) and blunted the capillary rarefication (reduced by 32.5%) (P = 0.0199; n = 6). RNA sequencing demonstrated that treatment with PEG-ADA normalized multiple pathways related to fibrosis, vasculopathy, and inflammation in Fra-2 mice. CONCLUSION: Treatment with PEG-ADA ameliorates the 3 cardinal features of SSc in pharmacologically relevant and well-tolerated doses. These findings may have direct translational implications, as PEG-ADA has already been approved by the Food and Drug Administration for the treatment of patients with ADA-deficient severe combined immunodeficiency disease.

Development of a Classifier to Determine Factors Causing Cybersickness in Virtual Reality Environments.

Objective: The goal of this contribution is to develop a classifier able to determine if cybersickness (CS) has occurred after immersion in a virtual reality (VR) scenario, based on a combination of biosignals and game parameters. Methods: We collected electrocardiographic, electrooculographic, respiratory, and skin conductivity data from a total of 66 participants. In addition, we also captured relevant game parameters such as avatar linear and angular speed as well as acceleration, head movements, and on-screen collisions. The data were collected while the participants were in a 10-minute VR experience, which was developed in Unity. The experience forced rotation and lateral movements upon the participants to provoke CS. A baseline was captured during a first simple scenario. The data were then split in per-level, per-60-second, and per-30-second windows. Furthermore, participants filled a pre- and postimmersion simulator sickness questionnaire. Simulator sickness scores were then used as a reference for binary (CS vs. no CS) and ternary (no CS-mild CS-severe CS) classification patterns. Several classification methods (support vector machines, K-nearest neighbors, and neural networks) were tested. Results: A maximum classification accuracy of 82% was achieved for binary classification and 56% for ternary classification. Conclusion: Given the sample size and the variety of movement patterns presented in the demonstration, we conclude that a combination of biosignals and game parameters suffice to determine the occurrence of CS. However, substantial further research is required to improve binary classification accuracy to adequate values for real-life scenarios and to determine better approaches to classify its severity.

Icosahedral Carbaboranes with Peripheral Hydrogen-Chalcogenide Groups: Structures from Gas Electron Diffraction and Chemical Shielding in Solution.

Carbaboranes 1,2-(EH)2 -closo-1,2-C2 B10 H10 (E=S, Se) were prepared, in the case of E=Se for the first time. Their semi-experimental equilibrium molecular structures were established by the concerted use of quantum-chemical calculations and gas electron diffraction. A method was developed and implemented to quantify the contribution of experimental data to each refined structural parameter. The accuracy of the experimental structures and those calculated at the MP2 level of theory were gauged by comparison of experimental 11 B NMR chemical shifts with quantum-chemically computed values; the inclusion of electron correlation (GIAO-MP2) provided superior results. For the purpose of geometrical prediction, the remaining group 16 elements were considered, and the icosahedral structures for E=O and Te were also computed; for E=O the same theoretical approach was used as for E=S, and for E=Te a description similar to that for E=Se was employed.

Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA.

The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the vulnerable epithelium is primarily achieved by mucins that form a gel-like structure adherent to the apical cell surface. This mucus layer constitutes a physical and chemical barrier between the microbial flora and the underlying epithelium. Mucus is critical to the maintenance of a homeostatic relationship between the microbiota and its host. Subtle deviations from this dynamic interaction may result in major implications for health. The protocol in this article describes the procedures to grow low mucus-producing HT29 and high mucus-producing HT29-MTX-E12 cells, maintain cells and use them for mucus quantification by ELISA. Additionally, it is described how to assess the amount of secreted adherent mucus. This system can be used to study the protective effect of mucus, e.g., against bacterial toxins, to test the effect of different culture conditions on mucus production or to analyze diffusion of molecules through the mucus layer. Since the ELISA used in this protocol is available for different species and mucus proteins, also other cell types can be used.

An adherent mucus layer attenuates the genotoxic effect of colibactin.

The human gastrointestinal tract is a complex ecosystem in which epithelial cells and microorganisms of the intestinal microbiota live in symbiosis. Certain members of the microbiota, in particular Escherichia coli strains of the B2 phylotype, carry the polyketide synthase-island encoding the genotoxin colibactin. Colibactin is a nonribosomal peptide or polyketide-nonribosomal peptide hybrid of still unsolved structure, which induces DNA double strand breaks (DSBs) in eukaryotic cells. However, direct contact between live bacteria and host cell is required in order to elicit these genotoxic effects. In this study, we used a variety of cell culture models, among them, a 3D cell culture approach based on decellularised small intestinal submucosa, to investigate whether the intestinal mucus layer has the potential to interfere with colibactin activity. We demonstrate that the expression of mucins and the formation of an adherent mucus layer significantly increased with increasing complexity of cell culture. Moreover, we show that the presence of an adherent mucus layer on epithelial cells attenuates the genotoxic activity of colibactin, by preventing the induction of DNA-DSBs. Removal of the adherent mucus layer restored the occurrence of DNA-DSBs.

Preparation of a Three-Dimensional Full Thickness Skin Equivalent.

In vitro test systems are a promising alternative to animal models. Due to the use of human cells in a three-dimensional arrangement that allows cell-cell or cell-matrix interactions these models may be more predictive for the human situation compared to animal models or two-dimensional cell culture systems. Especially for dermatological research, skin models such as epidermal or full-thickness skin equivalents (FTSE) are used for different applications. Although epidermal models provide highly standardized conditions for risk assessment, FTSE facilitate a cellular crosstalk between the dermal and epidermal layer and thus can be used as more complex models for the investigation of processes such as wound healing, skin development, or infectious diseases. In this chapter, we describe the generation and culture of an FTSE, based on a collagen type I matrix and provide troubleshooting tips for commonly encountered technical problems.

Tris(perfluorotolyl)borane-A Boron Lewis Superacid.

Tris[tetrafluoro-4-(trifluoromethyl)phenyl]borane (BTolF) was prepared by treating boron tribromide with tetrameric F3 CC6 F4 -CuI . The F3 CC6 F4 -CuI was generated from F3 CC6 F4 MgBr and copper(I) bromide. Lewis acidities of BTolF evaluated by the Gutmann-Beckett method and calculated fluoride-ion affinities are 9 and 10 %, respectively, higher than that of tris(pentafluorophenyl)borane (BCF) and even higher than that of SbF5 . The molecular structures of BTolF and BCF were determined by gas-phase electron diffraction, that of BTolF also by single-crystal X-ray diffraction.

Osteopontin is elevated in patients with mitral annulus calcification independent from classic cardiovascular risk factors.

BACKGROUND: Osteopontin (OPN) regulates the Ca(++)-deposition in bone and coronary arteries. Elevated OPN were also associated with (aortic) valve calcification in healthy individuals. This study aimed to investigate the association between OPN levels and mitral annulus calcification (MAC) in patients with coronary artery disease (CAD). METHODS: In this cross-sectional study OPN-levels were measured in 223 non-or ex-smoking patients (160 male, mean age: 61,09 ± 11,02 years; 63 female: mean age: 67,49 ± 7,87 years) with CAD. Plasma OPN levels were measured by ELISA and MAC was evaluated by echocardiography. RESULTS: Forward stepwise logistic regression analysis (likelihood quotient) showed significantly higher OPN-levels in patients with MAC compared to patient without calcified mitral annulus independent from the classic risk factors age and severity of coronary artery disease (CAD). In addition to age and the severity of CAD, the circulating OPN amount was a significant predictor for MAC. CONCLUSIONS: This is the first clinical trial which observed increased circulating OPN levels in MAC, suggesting a distinct role of OPN in the process of MAC. Considering the current knowledge about OPN it is more likely that OPN does not promote but counteracts valve calcification and therefore is elevated in course of a calcification processes.

Structural Analysis of Perfluoropropanoyl Fluoride in the Gas, Liquid, and Solid Phases.

The coexistence of two conformers in perfluoropropanoyl fluoride, CF3CF2C(O)F, differing in the CC-CF dihedral angle (gauche 85(10)% and anti 15(10)%), has been determined by means of gas-phase electron diffraction (GED). Quantum-chemical calculations performed at the MP2 and B3LYP approximations and cc-pVTZ basis sets reproduce the experimental values with confidence. By contrast, FTIR spectra give no clear evidence for the anti-conformer in the gas phase. Information on this less abundant but stable rotamer is obtained from matrix-isolation/FTIR spectroscopy and liquid Raman spectroscopy. In situ crystallization and single-crystal X-ray diffraction (XRD) data reveal the presence of solely the gauche-conformation in the solid state. A set of intermolecular interactions including C═O···C═O, C-F···F-C, and F···C═O is detected. The nature of bonding and the relative stabilities of gauche- and anti-conformers are explored using natural bond orbitals.

Structures of Trichloromethyl Thiocyanate, CCl3 SCN, in Gaseous and Crystalline State.

Trichloromethyl thiocyanate, CCl3 SCN, was structurally studied in both the gas and crystal phases by means of gas electron diffraction (GED) and single-crystal X-ray diffraction (XRD), respectively. Both experimental studies and quantum chemical calculations indicate a staggered orientation of the CCl3 group relative to the SCN group. This conclusion is supported by the similarity of the C-SCN bond length to that of the anti-structure of CH2 ClSCN (Berrueta Martínez et al. Phys. Chem. Chem. Phys. 2015, 17, 15805-15812). Bond lengths and angles are similar for gas and crystal CCl3 SCN structures; however, the crystal structure presents different intermolecular interactions. These include halogen and chalcogen type interactions, the geometry of which was studied. Characteristic C-Y⋅⋅⋅N angles (Y=Cl or S) close to 180° provide evidence for typical σ-hole interactions along the halogen/chalcogen-carbon bond in N⋅⋅⋅Cl and N⋅⋅⋅S, intermolecular units.

Gas-phase structure of 2,2,2-trichloroethyl chloroformate studied by electron diffraction and quantum-chemical calculations.

The molecular structure and conformational properties of 2,2,2-trichloroethyl chloroformate, ClC(O)OCH2CCl3 were determined experimentally using gas-phase electron diffraction (GED) and theoretically based on quantum-chemical calculations at the MP2 and DFT levels of theory. Further experimental measurements such as UV-visible, IR and Raman spectroscopy were complemented with the corresponding theoretical studies. All experimental results and calculations confirm the presence of two conformers namely anti-gauche (C1 symmetry) and anti-anti (Cs symmetry). The conformational preference was rationalised by NBO and AIM analyses. Molecular properties such as ionisation potential, electronegativity, chemical potential, chemical hardness and softness were deduced from HOMO-LUMO analyses. The TD-DFT approach was applied to assign the electronic transitions observed in the UV-visible spectrum. A detailed interpretation of the infrared and Raman spectra of the title compound are reported. Using calculated frequencies as a guide, IR and Raman spectra also provide evidence for the presence of both C1 and Cs conformers.