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Abstract In vertebrates and plants, dsRNA plays crucial roles as PAMP and as a mediator of RNAi. How higher fungi respond to dsRNA is not known. We demonstrate that Magnaporthe oryzae (Mo), a globally significant crop pathogen, internalizes dsRNA across a broad size range of 21 to about 3000 bp. Incubation of fungal conidia with 10 ng/µL dsRNA, regardless of size or sequence, induced aberrant germ tube elongation, revealing a strong sequence-unspecific effect of dsRNA in this fungus. Accordingly, the synthetic dsRNA analogue poly(I:C) and dsRNA of various sizes and sequences elicited canonical fungal stress pathways, including nuclear accumulation of the stress marker mitogen-activated protein kinase Hog1p and production of ROS. Leaf application of dsRNA to the cereal model species Brachypodium distachyon suppressed the progression of leaf blast disease. Notably, the sequence-unspecific effect of dsRNA depends on higher doses, while pure sequence-specific effects were observed at low concentrations of dsRNA (

Adaptive expertise is a highly sought after, but difficult to achieve, outcome of mathematics education. Many teaching methods appear to support the development of adaptive expertise only in a small proportion of students. Game-based learning environments may be useful for supporting adaptive expertise. Therefore, we carried out a quasi-experimental classroom intervention to examine the possibility of using a game-based learning environment to promote adaptive rational number knowledge, a potential indicator of adaptive expertise in the domain of rational numbers. The Number Line Elaboration and Exploration learning environment relied on the increasing elaboration of the number line analogy as a means for students to explore connections between multiple aspects of rational number knowledge. Our results show the game-based learning environment was successful in promoting adaptive rational number knowledge. These results provide directions for the development and examination of how learning environments may be able to support adaptive expertise.

Background: Pediatric patients with intestinal failure are critically dependent on central venous access for nutrition and fluids. Long-term central venous access can be complicated by recurrent infections, catheter malfunction, and venous stricture and thrombosis. Prior studies have discussed hybrid procedures to salvage suboptimal central venous access sites; however, data is very limited. Methods: This is a retrospective review of six pediatric patients with intestinal failure (IF) and long-term total parenteral nutrition (TPN) dependence who underwent one or more hybrid procedures for achieving complex vascular access, vascular mapping, and/or salvage of vascular access sites. Results: Median age at the time of intervention was 1.4 years (range 2.5 weeks – 2.6 years) with a median weight of 10 kg (range 3.3–13.8 kg). The median number of lifetime central lines was 4 (range 2–6). Indications for hybrid intervention included line fractures, occlusions, dislodgement, recurrent infections, and refractory central line infection. The most common procedures included vascular access, vein mapping, and balloon angioplasty of occluded central veins. The median procedure time was 4.6 h (range 1.3–5.9 h) with a median procedural radiation dose of 2.2 Gycm2 (range 0.1–6.7 Gycm2). All patients who underwent hybrid procedures had successful exchange and/or rehabilitation of the at-risk access site. Conclusions: These cases highlight the importance of vascular mapping for identifying potential access sites, as well as techniques for successful vascular rehabilitation for maintenance or salvage of existing central venous access. A multidisciplinary hybrid approach is a feasible and effective means of maintaining central venous access.

Mathematics anxiety hinders students' mathematical achievement already in primary school, but research on its effects beyond whole number knowledge is limited. The main aim of the current study is to examine how state and trait mathematics anxiety relate to performance across five tasks that are relevant for the development of mathematics in primary school, including a measure of adaptive expertise with school mathematics. These include mathematical tasks with non-symbolic quantities, whole numbers, and rational numbers. The participants were 406 primary school students attending the 5th grade (N = 188) and 6th grade (N = 218). Our results showed that state anxiety varies across task type. Furthermore, students' self-evaluated state and trait mathematics anxiety had varying negative relations with performance depending on the task type. In particular, we found that mathematics anxiety may limit students' adaptive expertise with rational numbers, even after controlling for other relevant mathematical skills. Overall, our results indicate that existing accounts on the role mathematics anxiety plays in school mathematics should expand to consider differences across task type and measures of anxiety.

After 4 years of the COVID-19 pandemic, SARS-CoV-2 continues to circulate with epidemic waves caused by evolving new variants. Although the rapid development of vaccines and approved antiviral drugs has reduced virus transmission and mitigated the symptoms of infection, the continuous emergence of new variants and the lack of simple-use (non-hospitalized, easy timing, local delivery, direct acting, and host-targeting) treatment modalities have limited the effectiveness of COVID-19 vaccines and drugs. Therefore, novel therapeutic approaches against SARS-CoV-2 infection are still urgently needed. As a positive-sense single-stranded RNA virus, SARS-CoV-2 is highly susceptible to RNA interference (RNAi). Accordingly, small interfering (si)RNAs targeting different regions of SARS-CoV-2 genome can effectively block the expression and replication of the virus. However, the rapid emergence of new SARS-CoV-2 variants with different genomic mutations has led to the problem of viral escape from the targets of RNAi strategy, which has increased the potential of off-target effects by siRNA and decreased the efficacy of long-term use of siRNA treatment. In our study, we enzymatically generated a set of Dicer-substrate (D)siRNA swarms containing DsiRNAs targeting single or multiple conserved sequences of SARS-CoV-2 genome by using in vitro transcription, replication and Dicer digestion system. Pre-transfection of these DsiRNA swarms into Vero E6-TMPRSS2 cells inhibited the replication of several SARS-CoV-2 variants, including the recent Omicron subvariants BQ.1.1 and XBB.1.5. This in vitro investigation of novel DsiRNA swarms provides solid evidence for the feasibility of this new RNAi strategy in the prevention and treatment of SARS-CoV-2 infection.

A universal taxonomy of viruses is essential for a comprehensive view of the virus world and for communicating the complicated evolutionary relationships among viruses. However, there are major differences in the conceptualisation and approaches to virus classification and nomenclature among virologists, clinicians, agronomists, and other interested parties. Here, we provide recommendations to guide the construction of a coherent and comprehensive virus taxonomy, based on expert scientific consensus. Firstly, assignments of viruses should be congruent with the best attainable reconstruction of their evolutionary histories, i.e., taxa should be monophyletic. This fundamental principle for classification of viruses is currently included in the International Committee on Taxonomy of Viruses (ICTV) code only for the rank of species. Secondly, phenotypic and ecological properties of viruses may inform, but not override, evolutionary relatedness in the placement of ranks. Thirdly, alternative classifications that consider phenotypic attributes, such as being vector-borne (e.g., "arboviruses"), infecting a certain type of host (e.g., "mycoviruses," "bacteriophages") or displaying specific pathogenicity (e.g., "human immunodeficiency viruses"), may serve important clinical and regulatory purposes but often create polyphyletic categories that do not reflect evolutionary relationships. Nevertheless, such classifications ought to be maintained if they serve the needs of specific communities or play a practical clinical or regulatory role. However, they should not be considered or called taxonomies. Finally, while an evolution-based framework enables viruses discovered by metagenomics to be incorporated into the ICTV taxonomy, there are essential requirements for quality control of the sequence data used for these assignments. Combined, these four principles will enable future development and expansion of virus taxonomy as the true evolutionary diversity of viruses becomes apparent.

Abstract Background Brachycephalic dogs display sleep‐disordered breathing (SDB). The risk factors for SDB remain unknown. Objectives To identify risk factors for SDB. We hypothesized that brachycephaly, increasing severity of brachycephalic obstructive airway syndrome (BOAS), excess weight, and aging predispose to SDB. Animals Sixty‐three privately owned pet dogs were prospectively recruited: 28 brachycephalic and 35 normocephalic (mesaticephalic or dolicocephalic) dogs. Methods Prospective observational cross‐sectional study with convenience sampling. Recording with the neckband was done over 1 night at each dog's home. The primary outcome measure was the obstructive respiratory event index (OREI). Body condition score (BCS) was assessed, and BOAS severity was graded for brachycephalic dogs. Results Brachycephaly was a significant risk factor for high OREI value (ratio of the geometric means 5.6, 95% confidence interval [CI] 3.2‐9.9; P

Abstract Background High-flow nasal oxygen (HFNO) therapy is a non-invasive respiratory support method that provides oxygen-enriched, warmed, and humidified air to respiratory-compromised patients. It is widely used in human medical care, but in veterinary medicine it is still a relatively new method. No practical guidelines exist for its use in canine pneumonia patients, although they could potentially benefit from HFNO therapy. This study aims to provide a new, safe, non-invasive, and effective treatment protocol for oxygen supplementation of non-sedated dogs with pneumonia. Methods Twenty privately owned dogs with pneumonia will receive HFNO therapy at a flow rate of 1–2 L/kg, and the fraction of inspired oxygen will be determined individually (ranging from 21% to 100%). HFNO therapy will continue as long as oxygen support is needed based on clinical evaluation. Patients will be assessed thrice daily during their hospitalisation, with measured primary outcomes including partial pressure of oxygen, oxygen saturation, respiratory rate and type, days in hospital, and survival to discharge. Discussion The proposed protocol aims to provide a practical guideline for applying HFNO to dogs hospitalised due to pneumonia. The protocol could enable more efficient and well-tolerated oxygenation than traditional methods, thus hastening recovery and improving survival of pneumonia patients.

The canonical lytic-lysogenic binary has been challenged in recent years, as more evidence has emerged on alternative bacteriophage infection strategies. These infection modes are little studied, and yet they appear to be more abundant and ubiquitous in nature than previously recognized, and can play a significant role in the ecology and evolution of their bacterial hosts. In this review, we discuss the extent, causes and consequences of alternative phage lifestyles, and clarify conceptual and terminological confusion to facilitate research progress. We propose distinct definitions for the terms 'pseudolysogeny' and 'productive or non-productive chronic infection', and distinguish them from the carrier state life cycle, which describes a population-level phenomenon. Our review also finds that phages may change their infection modes in response to environmental conditions or the physiological state of the host cell. We outline known molecular mechanisms underlying the alternative phage-host interactions, including specific genetic pathways and their considerable biotechnological potential. Moreover, we discuss potential implications of the alternative phage lifestyles for microbial biology and ecosystem functioning, as well as applied topics such as phage therapy.

Background: Adaptive expertise is a highly valued outcome of mathematics curricula. One aspect of adaptive expertise with rational numbers is adaptive rational number knowledge, which refers to the ability to integrate knowledge of numerical characteristics and relations in solving novel tasks. Even among students with strong conceptual and procedural knowledge of rational numbers, there are substantial individual differences in adaptive rational number knowledge. Aims: We aimed to examine how a wide range of domain-general and mathematically specific skills and knowledge predicted different aspects of rational number knowledge, including procedural, conceptual, and adaptive rational number knowledge. Sample: 173 6th and 7th grade students from a school in the southeastern US (51% female) participated in the study. Methods: At three time points across 1.5 years, we measured students' domain-general and domain-specific skills and knowledge. We used multiple hierarchal regression analysis to examine how these predictors related to rational number knowledge at the third time point. Result: Prior knowledge of rational numbers, general mathematical calculation knowledge, and spontaneous focusing on multiplicative relations (SFOR) tendency uniquely predicted adaptive rational number knowledge, after taking into account domain-general and mathematically specific skills and knowledge. Although conceptual knowledge of rational numbers and general mathematical achievement also predicted later conceptual and procedural knowledge of rational numbers, SFOR tendency did not. Conclusion: Results suggest expanding investigations of mathematical development to also explore different features of adaptive expertise as well as spontaneous mathematical focusing tendencies.

Abstract Background Sleep‐disordered breathing (SDB), defined as any difficulty in breathing during sleep, occurs in brachycephalic dogs. Diagnostic methods for SDB in dogs require extensive equipment and laboratory assessment. Objectives To evaluate the usability of a portable neckband system for detection of SDB in dogs. We hypothesized that the neckband is a feasible method for evaluation of SDB and that brachycephaly predisposes to SDB. Animals Twenty‐four prospectively recruited client‐owned dogs: 12 brachycephalic dogs and 12 control dogs of mesocephalic or dolicocephalic breeds. Methods Prospective observational cross‐sectional study with convenience sampling. Recording was done over 1 night at each dog's home. The primary outcome measure was the obstructive Respiratory Event Index (OREI), which summarized the rate of obstructive SDB events per hour. Additionally, usability, duration of recording, and snore percentage were documented. Results Brachycephalic dogs had a significantly higher OREI value (Hodges‐Lehmann estimator for median difference = 3.5, 95% confidence interval [CI] 2.2‐6.8; P

Abstract Background Salivary bile acids are used to diagnose extraesophageal reflux (EER) and to evaluate the risk of reflux aspiration that is associated with respiratory diseases in dogs. Objectives To study total bile acid (TBA) concentrations in saliva and in bronchoalveolar lavage fluid (BALF) to investigate EER and reflux aspiration in dogs with respiratory diseases and in healthy dogs. Animals Thirty‐one West Highland White Terriers (WHWTs) with idiopathic pulmonary fibrosis (IPF), 12 dogs with inflammatory airway disease (IAD), 6 dogs with recurrent pneumonia (RP), 26 brachycephalic dogs (BD), 27 healthy WHWTs (HW), 52 healthy dogs (HD). All privately‐owned dogs. Methods Saliva and BALF were collected from dogs in each group. Results Salivary TBA concentrations were higher in IPF (median 0.1692 μM, interquartile range [IQR] 0.1115‐0.2925 μM, Cohen's d 3.4, 95% confidence interval [CI] 2.2‐4.0, P

Coxsackievirus B (CVB) enteroviruses are common pathogens that can cause acute and chronic myocarditis, dilated cardiomyopathy, aseptic meningitis, and they are hypothesized to be a causal factor in type 1 diabetes. The licensed enterovirus vaccines and those currently in clinical development are traditional inactivated or live attenuated vaccines. Even though these vaccines work well in the prevention of enterovirus diseases, new vaccine technologies, like virus-like particles (VLPs), can offer important advantages in the manufacturing and epitope engineering. We have previously produced VLPs for CVB3 and CVB1 in insect cells. Here, we describe the production of CVB3-VLPs with enhanced production yield and purity using an improved purification method consisting of tangential flow filtration and ion exchange chromatography, which is compatible with industrial scale production. We also resolved the CVB3-VLP structure by Cryo-Electron Microscopy imaging and single particle reconstruction. The VLP diameter is 30.9 nm on average, and it is similar to Coxsackievirus A VLPs and the expanded enterovirus cell-entry intermediate (the 135s particle), which is ~2 nm larger than the mature virion. High neutralizing and total IgG antibody levels, the latter being a predominantly Th2 type (IgG1) phenotype, were detected in C57BL/6J mice immunized with non-adjuvanted CVB3-VLP vaccine. The structural and immunogenic data presented here indicate the potential of this improved methodology to produce highly immunogenic enterovirus VLP-vaccines in the future.

Mathematics anxiety hinders students' mathematical achievement already in primary school, but research on its effects beyond whole number knowledge is limited. The main aim of the current study is to examine how state and trait mathematics anxiety relate to performance across five tasks that are relevant for the development of mathematics in primary school, including a measure of adaptive expertise with school mathematics. These include mathematical tasks with non-symbolic quantities, whole numbers, and rational numbers. The participants were 406 primary school students attending the 5th grade (N = 188) and 6th grade (N = 218). Our results showed that state anxiety varies across task type. Furthermore, students' self-evaluated state and trait mathematics anxiety had varying negative relations with performance depending on the task type. In particular, we found that mathematics anxiety may limit students' adaptive expertise with rational numbers, even after controlling for other relevant mathematical skills. Overall, our results indicate that existing accounts on the role mathematics anxiety plays in school mathematics should expand to consider differences across task type and measures of anxiety.

IntroductionVirus-like particles (VLPs) are similar in size and shape to their respective viruses, but free of viral genetic material. This makes VLP-based vaccines incapable of causing infection, but still effective in mounting immune responses. Noro-VLPs consist of 180 copies of the VP1 capsid protein. The particle tolerates C-terminal fusion partners, and VP1 fused with a C-terminal SpyTag self-assembles into a VLP with SpyTag protruding from its surface, enabling conjugation of antigens via SpyCatcher.MethodsTo compare SpyCatcher-mediated coupling and direct peptide fusion in experimental vaccination, we genetically fused the ectodomain of influenza matrix-2 protein (M2e) directly on the C-terminus of norovirus VP1 capsid protein. VLPs decorated with SpyCatcher-M2e and VLPs with direct M2 efusion were used to immunize mice.Results and discussionWe found that direct genetic fusion of M2e on noro-VLP raised few M2e antibodies in the mouse model, presumably because the short linker positions the peptide between the protruding domains of noro-VLP, limiting its accessibility. On the other hand, adding aluminum hydroxide adjuvant to the previously described SpyCatcher-M2e-decorated noro-VLP vaccine gave a strong response against M2e. Surprisingly, simple SpyCatcher-fused M2e without VLP display also functioned as a potent immunogen, which suggests that the commonly used protein linker SpyCatcher-SpyTag may serve a second role as an activator of the immune system in vaccine preparations. Based on the measured anti-M2e antibodies and cellular responses, both SpyCatcher-M2e as well as M2e presented on the noro-VLP via SpyTag/Catcher show potential for the development of universal influenza vaccines.

Abstract Background Prolonged tissue hypoxia caused by chronic pulmonary disease is commonly regarded as an important mechanism in the development of secondary polycythemia, but little clinical data are available to support this hypothesis. Objective To study the prevalence and severity of erythrocytosis accompanying chronic hypoxic pulmonary disease in dogs. Animals Forty‐seven dogs with hypoxic chronic pulmonary disease, 27 dogs with nonhypoxic chronic pulmonary disease, and 60 healthy controls. Methods Dogs with chronic pulmonary disease and chronic hypoxemia (partial pressure of arterial oxygen [PaO2] .05 for all). Conclusions and Clinical Importance Polycythemia is uncommon, and usually mild if present, in dogs with chronic hypoxia caused by pulmonary disease.

In vertebrates and plants, dsRNA plays crucial roles as PAMP and as a mediator of RNAi. How higher fungi respond to dsRNA is not known. We demonstrate that Magnaporthe oryzae (Mo), a globally significant crop pathogen, internalizes dsRNA across a broad size range of 21 to about 3000 bp. Incubation of fungal conidia with 10 ng/µL dsRNA, regardless of size or sequence, induced aberrant germ tube elongation, revealing a strong sequence-unspecific effect of dsRNA in this fungus. Accordingly, the synthetic dsRNA analogue poly(I:C) and dsRNA of various sizes and sequences elicited canonical fungal stress pathways, including nuclear accumulation of the stress marker mitogen-activated protein kinase Hog1p and production of ROS. Leaf application of dsRNA to the cereal model species Brachypodium distachyon suppressed the progression of leaf blast disease. Notably, the sequence-unspecific effect of dsRNA depends on higher doses, while pure sequence-specific effects were observed at low concentrations of dsRNA (< 0.03 ng/µL). The protective effects of dsRNA were further enhanced by maintaining a gap of at least seven days between dsRNA application and inoculation, and by stabilising the dsRNA in alginate-chitosan nanoparticles. Overall, our study opens up additional possibilities for the development and use of dsRNA pesticides in agriculture. Exogenous dsRNA controls the rice blast fungus Magnaporthe oryzae via stress responses and RNAi. Plant protection improves with dsRNA delivery in chitosan-alginate nanoparticles, offering new possibilities for dsRNA-based pesticides. [ABSTRACT FROM AUTHOR]

Double-stranded RNA (dsRNA) molecules are mediators of RNA interference (RNAi) in eukaryotic cells. RNAi is a conserved mechanism of post-transcriptional silencing of genes cognate to the sequences of the applied dsRNA. RNAi-based therapeutics for the treatment of rare hereditary diseases have recently emerged, and the first sprayable dsRNA biopesticide has been proposed for registration. The range of applications of dsRNA molecules will likely expand in the future. Therefore, cost-effective methods for the efficient large-scale production of high-quality dsRNA are in demand. Conventional approaches to dsRNA production rely on the chemical or enzymatic synthesis of single-stranded (ss)RNA molecules with a subsequent hybridization of complementary strands. However, the yield of properly annealed biologically active dsRNA molecules is low. As an alternative approach, we have developed methods based on components derived from bacteriophage phi6, a dsRNA virus encoding RNA-dependent RNA polymerase (RdRp). Phi6 RdRp can be harnessed for the enzymatic production of high-quality dsRNA molecules. The isolated RdRp efficiently synthesizes dsRNA in vitro on a heterologous ssRNA template of any length and sequence. To scale up dsRNA production, we have developed an in vivo system where phi6 polymerase complexes produce target dsRNA molecules inside Pseudomonas cells.

Serious games for learning have received increased attention in recent years. However, empirical studies on students' gaming experiences throughout the developmental process of serious games and discussions regarding game design are missing. The aims of the present study were to analyze students' gaming experiences while playing four consecutive versions of the Number Navigation Game (NNG)--a mathematical game-based learning environment focusing on flexibility and adaptivity with whole-number arithmetic; and to provide an extensive review of the NNG developmental and design process over 3 years with focus on how and why the design decisions were made, and how those choices affected students' gaming experiences. The study employed a mixed-methods design of quantitative and qualitative research. The Game Experience Questionnaire about eight core game experience dimensions was answered by different groups of students at primary schools in Finland in three different experiments after students played four versions of the NNG from 2014 to 2016. Six semi structured interviews related to students' game experiences, preferences and game features of the latest version of NNG were conducted. Overall, results indicate that improvement in game's usability and clarity in the user interface has positive impacts on students' game experiences. Furthermore, there seems to be a clear advantage in having better aesthetics and value in improving extrinsic elements that could contribute to maintain players' enthusiasm and situational interest in serious games.

Contrasting theories exist regarding how Norway spruce recolonized Fennoscandia after the last glaciation. Here, the authors provide evidences from sedimentary ancient DNA and modern population genomics to support that Norway spruce was present in southern Fennoscandia shortly after deglaciation and the early Holocene migration from the east.

Abstract Background Canine idiopathic pulmonary fibrosis (CIPF) is a chronic, progressive, interstitial fibrosing lung disease, manifesting as cough, exercise intolerance and ultimately, dyspnea and respiratory failure. It mainly affects West Highland white terriers (WHWTs), lacks curable treatment and has a poor prognosis. Aspiration of gastroesophageal refluxate may play a role in the development of CIPF. In the first part of this study, we completed label-free quantitative proteomic analysis of bronchoalveolar lavage fluid (BALF) from CIPF and healthy WHWTs. In the second part, we evaluated potential protein markers of reflux aspiration from canine gastric juice and vomitus and whether these were present in BALF from the two groups. Results Across all BALF samples, 417 proteins were identified, and of these, 265 proteins were identified by two or more unique tryptic peptides. Using the 265 high confidence assignments, the quantitative proteome profiles were very similar in the two cohorts, but they could be readily resolved by principal component analysis on the basis of differential protein expression. Of the proteins that were differentially abundant in the two groups, several (including inflammatory and fibrotic markers) were elevated in CIPF, and a smaller, more diverse group of proteins were diminished in CIPF. No protein markers indicative of reflux aspiration were identified. Conclusions Label-free proteomics allowed discrimination between CIPF and healthy WHWTs, consistent with fibrotic process but did not provide clear evidence for gastrointestinal aspiration. The measurement of proteins may provide a proteomics signature of CIPF that could be used to evaluate treatment options.

Abstract Herpes simplex virus type 1 (HSV‐1) is a human pathogen that causes recurrent infections. Acyclovir‐resistant strains exist and can cause severe complications, which are potentially untreatable with current therapies. We have developed siRNA swarms that target a 653 base pair long region of the essential HSV gene UL29. As per our previous results, the anti‐UL29 siRNA swarm effectively inhibits the replication of circulating HSV strains and acyclovir‐resistant HSV strains in vitro, while displaying a good safety profile. We investigated a single intranasal therapeutic dose of a siRNA swarm in mice, which were first inoculated intranasally with HSV‐1 and given treatment 4 h later. We utilized a luciferase‐expressing HSV‐1 strain, which enabled daily follow‐up of infection with in vivo imaging. Our results show that a single dose of a UL29‐targeted siRNA swarm can inhibit the replication of HSV‐1 in orofacial tissue, which was reflected in ex vivo HSV titers and HSV DNA copy numbers as well as by a decrease in a luciferase‐derived signal. Furthermore, the treatment had a tendency to protect mice from severe clinical symptoms and delay the onset of the symptoms. These results support the development of antiviral siRNA swarms as a novel treatment for HSV‐1 infections.

BackgroundGastroesophageal reflux and microaspiration (MA) of gastric juice are associated with various human respiratory diseases but not in dogs.ObjectiveTo detect the presence of bile acids in bronchoalveolar lavage fluid (BALF) of dogs with various respiratory diseases.AnimalsTwenty-seven West Highland White Terriers (WHWTs) with canine idiopathic pulmonary fibrosis (CIPF), 11 dogs with bacterial pneumonia (BP), 13 with chronic bronchitis (CB), 9 with eosinophilic bronchopneumopathy (EBP), 19 with laryngeal dysfunction (LD), 8 Irish Wolfhounds (IWHs) with previous BPs, 13 healthy WHWTs, all privately owned dogs, and 6 healthy research colony Beagles METHODS: Prospective cross-sectional observational study with convenience sampling of dogs. Bile acids were measured by mass spectrometry in BALF samples. Total bile acid (TBA) concentration was calculated as a sum of 17 different bile acids.ResultsConcentrations of TBA were above the limit of quantification in 78% of CIPF, 45% of BP, 62% of CB, 44% of EBP, 68% of LD, and 13% of IWH dogs. In healthy dogs, bile acids were detected less commonly in Beagles (0/6) than in healthy WHWTs (10/13). Concentrations of TBA were significantly higher in CIPF (median 0.013 μM, range not quantifiable [n.q.]-0.14 μM, P

A universal taxonomy of viruses is essential for a comprehensive view of the virus world and for communicating the complicated evolutionary relationships among viruses. However, there are major differences in the conceptualisation and approaches to virus classification and nomenclature among virologists, clinicians, agronomists, and other interested parties. Here, we provide recommendations to guide the construction of a coherent and comprehensive virus taxonomy, based on expert scientific consensus. Firstly, assignments of viruses should be congruent with the best attainable reconstruction of their evolutionary histories, i.e., taxa should be monophyletic. This fundamental principle for classification of viruses is currently included in the International Committee on Taxonomy of Viruses (ICTV) code only for the rank of species. Secondly, phenotypic and ecological properties of viruses may inform, but not override, evolutionary relatedness in the placement of ranks. Thirdly, alternative classifications that consider phenotypic attributes, such as being vector-borne (e.g., “arboviruses”), infecting a certain type of host (e.g., “mycoviruses,” “bacteriophages”) or displaying specific pathogenicity (e.g., “human immunodeficiency viruses”), may serve important clinical and regulatory purposes but often create polyphyletic categories that do not reflect evolutionary relationships. Nevertheless, such classifications ought to be maintained if they serve the needs of specific communities or play a practical clinical or regulatory role. However, they should not be considered or called taxonomies. Finally, while an evolution-based framework enables viruses discovered by metagenomics to be incorporated into the ICTV taxonomy, there are essential requirements for quality control of the sequence data used for these assignments. Combined, these four principles will enable future development and expansion of virus taxonomy as the true evolutionary diversity of viruses becomes apparent. Transforming an existing phenotypic classification of viruses into one based on evolutionary relationships that can accommodate the vast number of viruses characterized in metagenomics and environmental studies is an ongoing challenge. This Consensus View explains how such a taxonomy can be expanded to encapsulate viral diversity and to recognize independent biological origins of different virus groups.