Introduction: Facial expression scoring has proven to be useful for pain evaluation in humans. In the last decade, equivalent scales have been developed for various animal species, including large domestic animals. The research question of this systematic review was as follows: is facial expression scoring (intervention) a valid method to evaluate pain (the outcome) in large domestic animals (population)?

Method: We searched two databases for relevant articles using the search string: “grimace scale” OR “facial expression” AND animal OR “farm animal” NOT “mouse” NOT “rat” NOT “laboratory animal.” The risk of bias was estimated by adapting the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) checklist.

Results: The search strategy extracted 30 articles, with the major share on equids and a considerable number on cows, pigs, and sheep. Most studies evaluated facial action units (FAUs), including the eye region, the orbital region, the cheek or the chewing muscles, the lips, the mouth, and the position of the ears. Interobserver reliability was tested in 21 studies. Overall FAU reliability was substantial, but there were differences for individual FAUs. The position of the ear had almost perfect interobserver reliability (interclass coefficient (ICC): 0.73–0.97). Validity was tested in five studies with the reported accuracy values ranging from 68.2 to 80.0%.

Discussion: This systematic review revealed that facial expression scores provide an easy method for learning and reliable test results to identify whether an animal is in pain or distress. Many studies lack a reference standard and a true control group. Further research is warranted to evaluate the test accuracy of facial expression scoring as a live pen side test.

A detailed investigation of Valsartan`s light-induced degradation is reported. Based on UPLC-HRMS studies of stressed solutions several degradation pathways are proposed. Some of the proposed structures were obtained by forced degradation experiments. Examination of analytical data including crystal structures allowed for structural revision of one literature-known degradant. In addition, irradiation under aerobic conditions revealed further degradative pathways leading to two previously unknown decomposition products. Mechanistic considerations regarding their origin helped to identify structural weak points of Valsartan under the influence of light.

Plant fungal pathogens are an increasing emerging threat as climate change progresses. Sphaeropsis sapinea (syn. Diplodia sapinea), the causal fungal agent of Diplodia tip blight, is a major pathogen of pines of forestry and ornamental relevance in Europe and worldwide. Here, we combined molecular-based field surveys in a common-garden setting and across an elevation gradient with historical records, cultivation-based growth experiments and microscopy to report on host and abiotic constraints on the distribution of S. sapinea. Using the arboretum at the Botanical Garden Berlin, Germany, to control for environmental variability, S. sapinea was detected on all seven host Pinus species we studied. However, P. sylvestris is the only species in which the fungus was detected in symptomless needles at the arboretum, and the most frequently recorded host for over a century, suggesting that it is the main, and perhaps, potential original host. In addition, sampling symptomatic needles in four out of the seven same species across a gradient from 200 to 2,100 m of elevation in the French Alps, S. sapinea was not detected at elevation higher than 800 m. Abiotic constraints were also supported by reduced growth of isolates of S. sapinea at low temperature under controlled conditions, but a 35°C prior stress exposure increased the subsequent growth of S. sapinea within its optimal temperature range (20-30°C). Altogether, our study thus not only suggests that S. sapinea is more likely to cause tip blight in P. sylvestris compared to the other species we studied, but also that in the current context of global climate change with predicted temperature increases, the fungus could infect a wider range of pine hosts and locations worldwide.

Background

Vector-borne diseases are of increasing importance in Germany. Since 2015, autochthonous cases have been increasingly documented in Berlin/Brandenburg.

Objectives

Describe autochthonous Babesia canis infection in the Berlin/Brandenburg region.

Animals

Forty-nine dogs with autochthonous B. canis infection.

Methods

Evaluation of history, clinical signs, laboratory abnormalities, treatment, and outcome.

Results

Dogs were presented between March and August (9) and September and January (40) in the years 2015-2021. Historical and clinical findings were lethargy (100%), pale mucous membranes (63%), fever (50%), and pigmenturia (52%). Common clinicopathological findings were thrombocytopenia (100%), anemia (85%), intravascular hemolysis (52%), pancytopenia (41%), and systemic inflammatory response syndrome (SIRS; 37%). Babesia detection was based on blood smear evaluation (n = 40) and PCR targeting the 18S rRNA gene of piroplasms (n = 49). Sequencing indicated 99.47% to 100% identity to B. canis sequences from GenBank. All dogs were treated with imidocarb (2.4-6.3 mg/kg; median, 5 mg/kg); 8 dogs received 1, 35 received 2, and 1 dog each received 3, 4, or 5 injections, respectively. Continued PCR-positive results were detected in 7 dogs after the 1st, in 5 after the 2nd, in 2 after the 3rd, and in 1 28 days after the 4th injection. Four dogs were euthanized and 3 dogs died.

Conclusions and Clinical Importance

Autochthonous B. canis infections in Berlin/Brandenburg were associated with severe clinicopathological changes, SIRS, and multiorgan involvement. Testing by PCR during and after treatment is advisable to monitor treatment success. Screening of blood donors in high-risk areas and year-round tick protection is strongly recommended.

We studied the effects of the quantum delocalization in space of the hydrogen atoms of water in the aggregation process of two fullerene molecules. We considered a case using a purely repulsive water–fullerene interaction, as such a situation has shown that water-mediated effects play a key role in the aggregation process. This study becomes feasible, at a reduced computational price, by combining the path integral (PI) molecular dynamics (MD) method with a recently developed open-system MD technique. Specifically, only the mandatory solvation shell of the two fullerene molecules was considered at full quantum resolution, while the rest of the system was represented as a mean-field macroscopic reservoir of particles and energy. Our results showed that the quantum nature of the hydrogen atoms leads to a sizable difference in the curve of the free energy of aggregation; that is, that nuclear quantum effects play a relevant role.

When described by a one-dimensional reaction coordinate, pair-reaction rates in a solvent depend, in addition to the potential barrier height and the friction coefficient, on the potential shape, the effective mass, and the friction relaxation spectrum, but a rate theory that accurately accounts for all of these effects does not exist. After a review of classical reaction-rate theories, we show how to extract all parameters of the generalized Langevin equation (GLE) and, in particular, the friction memory function from molecular dynamics (MD) simulations of two prototypical pair reactions in water, the dissociation of NaCl and of two methane molecules. The memory exhibits multiple time scales and, for NaCl, pronounced oscillatory components. Simulations of the GLE by Markovian embedding techniques accurately reproduce the pair-reaction kinetics from MD simulations without any fitting parameters, which confirms the accuracy of the approximative form of the GLE and of the parameter extraction techniques. By modification of the GLE parameters, we investigate the relative importance of memory, mass, and potential shape effects. Neglect of memory slows down NaCl and methane dissociation by roughly a factor of 2; neglect of mass accelerates reactions by a similar factor, and the harmonic approximation of the potential shape gives rise to slight acceleration. This partial error cancellation explains why Kramers’ theory, which neglects memory effects and treats the potential shape in harmonic approximation, describes reaction rates better than more sophisticated theories. In essence, all three effects, friction memory, inertia, and the potential shape nonharmonicity, are important to quantitatively describe pair-reaction kinetics in water.

We consider a generalized Langevin equation (GLE) in which the deterministic force, the mass and the friction kernel are configuration-dependent, i.e. general nonlinear functions of the reaction coordinate. We introduce a projection operator that allows for a self-consistent Markovian embedding of such GLEs. Self-consistency means that trajectories generated by the Markovian embedding are described by a GLE with the same configuration-dependent deterministic force, mass and friction kernel. Using the projection operator, we derive a closed-form relation between the parameters of the Markovian embedding Langevin equations and the parameters of the GLE. This is accomplished by applying the projection operator formalism to the system of Markovian embedding stochastic equations.

Cryo-soft X-ray tomography (cryo-SXT) is a powerful method to investigate the ultrastructure of cells, offering resolution in the tens of nm range and strong contrast for membranous structures without requirement for labeling or chemical fixation. The short acquisition time and the relatively large volumes acquired allow for fast acquisition of large amounts of tomographic image data. Segmentation of these data into accessible features is a necessary step in gaining biologically relevant information from cryo-soft X-ray tomograms. However, manual image segmentation still requires several orders of magnitude more time than data acquisition. To address this challenge, we have here developed an end-to-end automated 3D-segmentation pipeline based on semi-supervised deep learning. Our approach is suitable for high-throughput analysis of large amounts of tomographic data, while being robust when faced with limited manual annotations and variations in the tomographic conditions. We validate our approach by extracting three-dimensional information on cellular ultrastructure and by quantifying nanoscopic morphological parameters of filopodia in mammalian cells.

Background

Porcine cytomegalovirus (PCMV) is a porcine roseolovirus (PCMV/PRV) which is widely distributed in pigs. Transmission of PCMV/PRV in preclinical xenotransplantations was shown to significantly reduce the survival time of the pig transplants in non-human primates. PCMV/PRV was also transmitted in the first transplantation of a pig heart into a human patient. To analyze how PCMV/PRV could be introduced into pig breeds, especially considering cloned transgenic pigs, and subsequently spread in breeding facilities, we screened ovaries and derived materials which are used to perform somatic cell nuclear transfer (SCNT).

Methods

DNA was isolated from ovarian tissues, follicular fluids, oocytes with cumulus cells, denuded oocytes and parthenotes. A real-time PCR with PCMV/PRV-specific primers and a probe was performed to detect PCMV/PRV. Furthermore, a Western blot assay using a recombinant fragment of the gB protein of PCMV/PRV was performed to screen for virus-specific antibodies in the follicular fluids.

Results

PCMV/PRV was found by real-time PCR in ovarian tissues, in the follicular fluid and in oocytes. In parthenotes the virus could not be detected, most-likely due to the low amount of DNA used. By Western blot assay specific antibodies against PCMV/PRV were found in 19 of 20 analyzed follicular fluids.

Conclusion

PCMV/PRV was found in ovarian tissues, in the follicular fluids and also in denuded oocytes, indicating that the virus is present in the animals of which the oocytes were taken from. Despite several washing steps of the denuded oocytes, which are subsequently used for microinjection or SCNT, the virus could still be detected. Therefore, the virus could infect oocytes during genetic modifications or stay attached to the surface of the oocytes, potentially infecting SCNT recipient animals.

Obtaining accurate ground and low-lying excited states of electronic systems is crucial in a multitude of important applications. One ab initio method for solving the Schrödinger equation that scales favorably for large systems is variational quantum Monte Carlo (QMC). The recently introduced deep QMC approach uses ansatzes represented by deep neural networks and generates nearly exact ground-state solutions for molecules containing up to a few dozen electrons, with the potential to scale to much larger systems where other highly accurate methods are not feasible. In this paper, we extend one such ansatz (PauliNet) to compute electronic excited states. We demonstrate our method on various small atoms and molecules and consistently achieve high accuracy for low-lying states. To highlight the method’s potential, we compute the first excited state of the much larger benzene molecule, as well as the conical intersection of ethylene, with PauliNet matching results of more expensive high-level methods.

Porcine cytomegalovirus (PCMV), a porcine roseolovirus (PRV) that is closely related to human herpesviruses 6 and 7, is commonly found in commercial pigs. PCMV/PRV is important in xenotransplantation, because in preclinical trials in which pig organs were transplanted into non-human primates, transmission of PCMV/PRV was shown to be associated with significantly reduced survival of the xenotransplants. PCMV/PRV was also transmitted in the first transplantation of a pig heart into a human patient worldwide and apparently contributed to the death of the patient. The prevalence of PCMV/PRV in wild boars is largely unknown. In this study, we screened wild boars from several areas of northern Italy and Germany to test for the presence of PCMV/PRV using PCR-based and Western blot assays. By Western blot analysis, 54% and 82% of Italian and German wild boars, respectively, were found to be PCMV/PRV positive, while 36% and 60%, respectively, tested positive by real-time polymerase chain reaction (PCR). These data indicate that the virus is common in German and Italian wild boars and that the Western blot assay detected a PCMV/PRV infection more often than did real-time PCR. The data also indicate that pigs raised for xenotransplantation should be protected from contact with materials from wild boars and commercial pigs.

Batrachochytrium salamandrivorans (Bsal), a species related to the destructive pathogen Batrachochytrium dendrobatidis (Bd), was found and identified in Europe in 2013. Now, a decade later, a large amount of information is available. This includes data from studies in the field, reports of infection in captive amphibians, laboratory studies testing host susceptibility, and data from prospective studies that test for Bsal’s presence in a location. We conducted a systematic review of the published literature and compiled a dataset of Bsal tests. We identified 67 species that have been reported positive for Bsal, 20 of which have a threatened conservation status. The distribution of species that have been found with infection encompasses 69 countries, highlighting the potential threat that Bsal poses. We point out where surveillance to detect Bsal have taken place and highlight areas that have not been well monitored. The large number of host species belonging to the families Plethodontidae and Salamandridae suggests a taxonomic pattern of susceptibility. Our results provide insight into the risk posed by Bsal and identifies vulnerable species and areas where surveillance is needed to fill existing knowledge gaps.

TGFβs, BMPs and Activins regulate numerous developmental and homeostatic processes and signal through hetero-tetrameric receptor complexes composed of two types of serine/threonine kinase receptors. Each of the 33 different ligands possesses unique affinities towards specific receptor types. However, the lack of specific tools hampered simultaneous testing of ligand binding towards all BMP/TGFβ receptors. Here we present a N-terminally Halo- and SNAP-tagged TGFβ/BMP receptor library to visualize receptor complexes in dual color. In combination with fluorescently labeled ligands, we established a Ligand Surface Binding Assay (LSBA) for optical quantification of receptor-dependent ligand binding in a cellular context. We highlight that LSBA is generally applicable to test (i) binding of different ligands such as Activin A, TGFβ1 and BMP9, (ii) for mutant screens and (iii) evolutionary comparisons. This experimental set-up opens opportunities for visualizing ligand-receptor binding dynamics, essential to determine signaling specificity and is easily adaptable for other receptor signaling pathways.

Citizen Science (CS) is a research approach that has become popular in recent years and offers innovative potential for dialect research in ornithology. As the scepticism about CS data is still widespread, we analysed the development of a 3-year CS project based on the song of the Common Nightingale (Luscinia megarhynchos) to share best practices and lessons learned. We focused on the data scope, individual engagement, spatial distribution and species misidentifications from recordings generated before (2018, 2019) and during the COVID-19 outbreak (2020) with a smartphone using the ‘Naturblick’ app. The number of nightingale song recordings and individual engagement increased steadily and peaked in the season during the pandemic. 13,991 nightingale song recordings were generated by anonymous (64%) and non-anonymous participants (36%). As the project developed, the spatial distribution of recordings expanded (from Berlin based to nationwide). The rates of species misidentifications were low, decreased in the course of the project (10–1%) and were mainly affected by vocal similarities with other bird species. This study further showed that community engagement and data quality were not directly affected by dissemination activities, but that the former was influenced by external factors and the latter benefited from the app. We conclude that CS projects using smartphone apps with an integrated pattern recognition algorithm are well suited to support bioacoustic research in ornithology. Based on our findings, we recommend setting up CS projects over the long term to build an engaged community which generates high data quality for robust scientific conclusions.

Class imbalance poses a major challenge for machine learning as most supervised learning models might exhibit bias towards the majority class and under-perform in the minority class. Cost-sensitive learning tackles this problem by treating the classes differently, formulated typically via a user-defined fixed misclassification cost matrix provided as input to the learner. Such parameter tuning is a challenging task that requires domain knowledge and moreover, wrong adjustments might lead to overall predictive performance deterioration. In this work, we propose a novel cost-sensitive boosting approach for imbalanced data that dynamically adjusts the misclassification costs over the boosting rounds in response to model’s performance instead of using a fixed misclassification cost matrix. Our method, called AdaCC, is parameter-free as it relies on the cumulative behavior of the boosting model in order to adjust the misclassification costs for the next boosting round and comes with theoretical guarantees regarding the training error. Experiments on 27 real-world datasets from different domains with high class imbalance demonstrate the superiority of our method over 12 state-of-the-art cost-sensitive boosting approaches exhibiting consistent improvements in different measures, for instance, in the range of [0.3–28.56%] for AUC, [3.4–21.4%] for balanced accuracy, [4.8–45%] for gmean and [7.4–85.5%] for recall.

Splicing is one key mechanism determining the state of any eukaryotic cell. Apart from linear splice variants, circular splice variants (circRNAs) can arise via non-canonical splicing involving a back-splice junction (BSJ). Most existing methods only identify circRNAs via the corresponding BSJ, but do not aim to estimate their full sequence identity or to identify different, alternatively spliced circular isoforms arising from the same BSJ. We here present CYCLeR, the first computational method for identifying the full sequence identity of new and alternatively spliced circRNAs and their abundances while simultaneously co-estimating the abundances of known linear splicing isoforms. We show that CYCLeR significantly outperforms existing methods in terms of F score and quantification of transcripts in simulated data. In a in a comparative study with long-read data, we also show the advantages of CYCLeR compared to existing methods. When analysing Drosophila melanogaster data, CYCLeR uncovers biological patterns of circRNA expression that other methods fail to observe.

Introduction

Handling sexual interactions in a competent manner is a key skill for young adults, which is linked to positive aspects of sexual and general well-being. Several research conceptualizations of sexual competence have been proposed in the literature, but little is known about how young adults define sexual competence and what consequences they consider low sexual competence may have.

Methods

In this qualitative study conducted in 2019, 571 university students (365 women, 206 men) from Germany with a mean age of 22.6 years provided open-ended answers to two questions: (Q1) What do you consider to be sexual competence? (Q2) What consequences can low sexual competence have? Combining thematic analysis and qualitative content analysis, responses were coded into 264 categories that were then condensed into 30 latent themes, with 14 themes referring to Q1 and 16 themes referring to Q2. All categories showed strong inter-coder agreement.

Result

Participants defined sexual competence in a multi-faceted way and in partial overlap with research definitions. Gender differences emerged in four themes (needs/desires, communication, skills/abilities, and setting boundaries/limits). Participants’ statements about the consequences of low sexual competence corresponded closely with their definitions of sexual competence. Gender differences emerged in five themes (risk of sexual victimization and sexual aggression, problems in sexual communication, problematic [sexual] risk behavior, negative influence on [sexual] satisfaction, and lack of skills).

Conclusions and Policy Implications

The implications of the findings for research conceptualizations of sexual competence, for designing interventions to promote sexual competence, and for policy measures designed to reduce sexual aggression are discussed.

According to the United Nations, an estimated 26.6 million people worldwide were refugees in 2021. Experiences before, during, and after flight increase psychological distress and contribute to a high prevalence of mental disorders. The resulting high need for mental health care is generally not reflected in the actual mental health care provision for refugees. A possible strategy to close this gap might be to offer smartphone-delivered mental health care. This systematic review summarizes the current state of research on smartphone-delivered interventions for refugees, answering the following research questions: (1) Which smartphone-delivered interventions are available for refugees? (2) What do we know about their clinical (efficacy) and (3) nonclinical outcomes (e.g., feasibility, appropriateness, acceptance, and barriers)? (4) What are their dropout rates and dropout reasons? (5) To what extent do smartphone-delivered interventions consider data security? Relevant databases were systematically searched for published studies, gray literature, and unpublished information. In total, 456 data points were screened. Twelve interventions were included (nine interventions from 11 peer-reviewed articles and three interventions without published study reports), comprising nine interventions for adult refugees and three for adolescent and young refugees. Study participants were mostly satisfied with the interventions, indicating adequate acceptability. Only one randomized controlled trial (RCT; from two RCTs and two pilot RCTs) found a significant reduction in the primary clinical outcome compared to the control group. Dropout rates ranged from 2.9 to 80%. In the discussion, the heterogeneous findings are integrated into the current state of literature.

Various drug samples (N = 249; drug substances, tablets, capsules, solutions, crèmes, and more) from the European pharmaceutical market were collected since 2019 and analyzed for 16 nitrosamines (NAs). In 2.0% of the cases, NAs were detected. These findings included four active pharmaceutical ingredients already known for potential NA contamination: losartan (N-nitrosodimethylamine [NDMA] and N-nitrosodiethylamine, simultaneously), valsartan (NDMA), metformin (NDMA) and ranitidine (NDMA). The fifth new finding, which has not been reported yet, discovered contamination of a molsidomine tablet sample with N-nitrosomorpholine (NMor). The tablet contained 144% of the toxicological allowable intake for NMor. NMor was included in our screening from the beginning and is currently the focus of regulatory authorities, but was added to the guidelines only last year. Thus, it may not have been the focus of regulatory investigations for too long. Our results indicate that the majority of drug products in the market are nonhazardous in terms of patient safety and drug purity. Unfortunately, the list of individual affected products keeps growing constantly and new NA cases, such as molsidomine or nitrosated drug substances (nitrosamine drug substance-related impurities [NDSRI]), continue to emerge. We therefore expect nitrosamine screenings to remain a high priority.