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Abstract Equine asthma (EA) is an inflammatory disease of the lower airways driven by mediators released from cells. Extracellular vesicles (EVs) are vehicles for lipid mediators, which possess either pro-inflammatory or dual anti-inflammatory and pro-resolving functions. In this study, we investigated how the respiratory fatty acid (FA) profile reflects airway inflammatory status. The FA composition of bronchoalveolar lavage fluid (BALF), BALF supernatant, and bronchoalveolar EVs of healthy horses (n = 15) and horses with mild/moderate EA (n = 10) or severe EA (SEA, n = 5) was determined with gas chromatography and mass spectrometry. The FA profiles distinguished samples with different diagnoses in all sample types, yet they were insufficient to predict the health status of uncategorized samples. Different individual FAs were responsible for the discrimination of the diagnoses in different sample types. Particularly, in the EVs of SEA horses the proportions of palmitic acid (16:0) decreased and those of eicosapentaenoic acid (20:5n-3) increased, and all sample types of asthmatic horses had elevated dihomo-γ-linolenic acid (20:3n-6) proportions. The results suggest simultaneous pro-inflammatory and resolving actions of FAs and a potential role for EVs as vehicles for lipid mediators in asthma pathogenesis. EV lipid manifestations of EA can offer translational targets to study asthma pathophysiology and treatment options.

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

Super-resolution microscopy techniques can be challenging for live cells and thick samples. Here, the authors propose a method to reduce beam intensity and remove out-of-focus fluorescence background in image-scanning microscopy (ISM) and its combination with stimulated emission depletion (STED).

The authors developed an open-source, low-cost, multi-channel time-tagging module for fluorescence lifetime image scanning microscopy and correlation spectroscopy that can tag in parallel multiple single-photon events with 30 ps precision.

This study aims to determine the effect of social media marketing activities on brand trust, brand equity and brand loyalty in social media. The study uses the Structural Equation Modeling (SEM) method with SPSS 3.3.3 software with a sample of 450 respondents determined by the simple random sampling method who had experience of using social media for at least six months. Data was obtained by distributing online questionnaires using google form. The results show that social media marketing has a positive effect on brand trust, social media marketing has a positive influence on brand equity, and social media marketing has a positive influence on brand loyalty. Brand trust has a positive influence on SMEs Performance, Brand equity has a positive influence on SMEs Performance and finally brand loyalty has a positive influence on SMEs Performance.

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

Although the RDF query language SPARQL has a reputation for being opaque and difficult for traditional humanists to learn, it holds great potential for opening up vast amounts of Linked Open Data to researchers willing to take on its challenges. This is especially true in the field of premodern manuscripts studies as more and more datasets relating to the study of manuscript culture are made available online. This paper explores the results of a two-year long process of collaborative learning and knowledge transfer between the computer scientists and humanities researchers from the Mapping Manuscript Migrations (MMM) project to learn and apply SPARQL to the MMM dataset. The process developed into a wider investigation of the use of SPARQL to analyse the data, refine research questions, and assess the research potential of the MMM aggregated dataset and its Knowledge Graph. Through an examination of a series of six SPARQL query case studies, this paper will demonstrate how the process of learning and applying SPARQL to query the MMM dataset returned three important and unexpected results: 1) a better understanding of a complex and imperfect dataset in a Linked Open Data environment, 2) a better understanding of how manuscript description and associated data involving the people and institutions involved in the production, reception, and trade of premodern manuscripts needs to be presented to better facilitate computational research, and 3) an awareness of need to further develop data literacy skills among researchers in order to take full advantage of the wealth of unexplored data now available to them in the Semantic Web.

Extracellular vesicles (EVs) are membrane-bound particles that engage in inflammatory reactions by mediating cell–cell interactions. Previously, EVs have been isolated from the bronchoalveolar lavage fluid (BALF) of humans and rodents. The aim of this study was to investigate the number and size distribution of EVs in the BALF of asthmatic horses (EA, n = 35) and healthy horses (n = 19). Saline was injected during bronchoscopy to the right lung followed by manual aspiration. The retrieved BALF was centrifuged twice to remove cells and biological debris. The supernatant was concentrated and EVs were isolated using size-exclusion chromatography. Sample fractions were measured with nanoparticle tracking analysis (NTA) for particle number and size, and transmission electron microscopy and confocal laser scanning microscopy were used to visualize EVs. The described method was able to isolate and preserve EVs. The mean EV size was 247 ± 35 nm (SD) in the EA horses and 261 ± 47 nm in the controls by NTA. The mean concentration of EVs was 1.38 × 1012 ± 1.42 × 1012 particles/mL in the EA horses and 1.33 × 1012 ± 1.07 × 1012 particles/mL in the controls with no statistically significant differences between the groups. With Western blotting and microscopy, these particles were documented to associate with EV protein markers (CD63, TSG101, HSP70, EMMPRIN, and actin) and hyaluronan. Equine BALF is rich in EVs of various sizes, and the described protocol is usable for isolating EVs. In the future, the role of EVs can be studied in horses with airway inflammation.

Abstract The combination of confocal laser-scanning microscopy (CLSM) and fluorescence fluctuation spectroscopy (FFS) is a powerful tool in studying fast, sub-resolution biomolecular processes in living cells. A detector array can further enhance CLSM-based FFS techniques, as it allows the simultaneous acquisition of several samples–essentially images—of the CLSM detection volume. However, the detector arrays that have previously been proposed for this purpose require tedious data corrections and preclude the combination of FFS with single-photon techniques, such as fluorescence lifetime imaging. Here, we solve these limitations by integrating a novel single-photon-avalanche-diode (SPAD) array detector in a CLSM system. We validate this new implementation on a series of FFS analyses: spot-variation fluorescence correlation spectroscopy, pair-correlation function analysis, and image-derived mean squared displacement analysis. We predict that the unique combination of spatial and temporal information provided by our detector will make the proposed architecture the method of choice for CLSM-based FFS.

BackgroundResistance to antibiotics is an increasing and major threat to global health. While the large majority of antimicrobial use occurs in the community where antibiotics are available without prescription, we did not find any studies investigating community-level factors influencing the inappropriate use of antibiotics in the Democratic Republic of Congo (DRC), where non-prescription antibiotic use is prevalent.MethodsThis qualitative study was conducted from April 1st 2019 to May 5th 2019 and consisting of in-depth semi-structured interviews, utilizing purposive and snowball sampling schemes to recruit adult heads of households in the Pakadjuma slum, in Kinshasa, DRC. Participants with differing medical and educational backgrounds were selected. We employed a thematic analysis approach to explore community knowledge and use of antibiotics in the sampled population.ResultsA total of 18 participants with a median age of 35 years were interviewed. The majority was female (77.7%), had at least a secondary education (83.4%), and unemployed (61.1%). We found that participants were familiar with the term "antibiotics", but had limited knowledge of the indications and risks of antibiotics, including the risk of antibiotic resistance. Inappropriate use of antibiotics was common and there was frequent self-medication of non-prescribed medicines for a range of non-indicated conditions such as menstruation. Having limited income was the most commonly reported reason for not visiting a health facility for appropriate health care.ConclusionInappropriate use of antibiotics is a widespread practice and is influenced by lack of adequate knowledge of antibiotic use, indications and risks, prevalent self-medication, and financial barriers to accessing appropriate health care. There is need for both community education as well as structural interventions addressing poverty in order to reduce the inappropriate use of antibiotics in the Pakadjuma slum in Kinshasa.

Semanttinen parlamentti -hankkeessa 2020–2022 luodaan eduskunnan tietokannoista ja niihin liittyvistä muista aineistoista uudenlainen linkitetyn avoimen datan (Linked Open Data, LOD) palvelu, tietoinfrastruktuuri ja semanttinen portaali Parlamenttisampo – eduskunta semant­tisessa webissä, joiden avulla tutkitaan poliittista kulttuuria ja kieltä. Dataa linkittämällä voi-daan rikastaa eduskuntadataa muilla tietolähteillä kuten biografisella tiedolla, terminologioilla ja lainsäädännön dokumenteilla. Parlamenttisampo on kieli- ja semanttisen webin teknologioihin perustuva palvelukokonaisuus tutkijoita, kansalaisia, mediaa ja valtionhallintoa varten. Artik­kelissa esitellään hankkeen visio, ensimmäisiä tuloksia ja niiden hyödyntämismahdollisuuksia: Eduskunnan kaikkien täysistuntojen 1907–2021 yli 900 000 puheesta on valmistunut linkitetyn datan tietämysgraafi (knowledge graph); data on myös saatavilla XML-muodossa, jossa hyö­dynnetään uutta kansainvälistä Parla-CLARIN-formaattia. Ensimmäistä kertaa eduskunnan puheiden koko aikasarja on muunnettu dataksi ja datapalveluksi yhtenäisessä muodossa. Lisäksi puheet on yhdistetty eduskunnan kansanedustajien tietokannasta luotuun ja muista tietolähteistä rikastettuun toiseen tietämysgraafiin laajemmaksi ontologiaperustaiseksi datapalveluksi Fin- Parla. Datapalvelua voidaan käyttää eduskuntatutkimukseen parlamentaarisesta ja edustuksel-lisesta kulttuurista sekä poliittisen kielen käytöstä analysoimalla kansanedustajien täysistunnois­sa pitämiä puheita ja poliitikkojen verkostoja data-analyysin keinoin. Palvelun rajapinnan avulla voidaan myös kehittää eri käyttäjäryhmille sovelluksia, kuten hankkeessa valmistuva Parlament­tisampo.fi-portaali.

Metabarcoding analyses of bacterial and eukaryotic communities have been proposed as efficient tools for environmental impact assessment. It has been unclear, however, to which extent these analyses can provide similar or differing information on the ecological status of the environment. Here, we used 16S and 18S rRNA gene metabarcoding to compare eutrophication-induced shifts in sediment bacterial and eukaryotic community structure in relation to a range of porewater, sediment and bottom-water geochemical variables, using data obtained from six stations near a former rainbow trout farm in the Archipelago Sea (Baltic Sea). Shifts in the structure of both community types were correlated with a shared set of variables, including porewater ammonium concentrations and the sediment depth-integrated oxygen consumption rate. Distance-based redundancy analyses showed that variables typically employed in impact assessments, such as bottom water nutrient concentrations, explained less of the variance in community structure than alternative variables (e.g., porewater NH4+ inventories and sediment depth-integrated O2 consumption rates) selected due to their low collinearity (up to 40 vs. 58% of the variance explained, respectively). In monitoring surveys where analyses of both bacterial and eukaryotic communities may be impossible, either 16S or 18S rRNA gene metabarcoding can serve as reliable indicators of wider ecological impacts of eutrophication.

Foraminifera are unicellular eukaryotes that are an integral part of benthic fauna in many marine ecosystems, including the deep sea, with direct impacts on benthic biogeochemical cycles. In these systems, different foraminiferal species are known to have a distinct vertical distribution, i.e., microhabitat preference, which is tightly linked to the physico-chemical zonation of the sediment. Hence, foraminifera are well-adapted to thrive in various conditions, even under anoxia. However, despite the ecological and biogeochemical significance of foraminifera, their ecology remains poorly understood. This is especially true in terms of the composition and diversity of their microbiome, although foraminifera are known to harbor diverse endobionts, which may have a significant meaning to each species’ survival strategy. In this study, we used 16S rRNA gene metabarcoding to investigate the microbiomes of five different deep-sea benthic foraminiferal species representing differing microhabitat preferences. The microbiomes of these species were compared intra- and inter-specifically, as well as with the surrounding sediment bacterial community. Our analysis indicated that each species was characterized with a distinct, statistically different microbiome that also differed from the surrounding sediment community in terms of diversity and dominant bacterial groups. We were also able to distinguish specific bacterial groups that seemed to be strongly associated with particular foraminiferal species, such as the family Marinilabiliaceae for Chilostomella ovoidea and the family Hyphomicrobiaceae for Bulimina subornata and Bulimina striata. The presence of bacterial groups that are tightly associated to a certain foraminiferal species implies that there may exist unique, potentially symbiotic relationships between foraminifera and bacteria that have been previously overlooked. Furthermore, the foraminifera contained chloroplast reads originating from different sources, likely reflecting trophic preferences and ecological characteristics of the different species. This study demonstrates the potential of 16S rRNA gene metabarcoding in resolving the microbiome composition and diversity of eukaryotic unicellular organisms, providing unique in situ insights into enigmatic deep-sea ecosystems.

Fourier ring correlation (FRC) analysis is commonly used in fluorescence microscopy to measure effective image resolution. Here, the authors demonstrate that FRC can also be leveraged in blind image restoration methods, such as image deconvolution.

Abstract Background The World Health Organization recommends the use of artemisinin-based combination therapy (ACT) to treat uncomplicated malaria for the control of malaria across the world. There are several types of ACT used across malaria-endemic countries, yet there is little information about preferences and adherence practices regarding different types of ACT. The objective of this study was to evaluate levels of adherence to two types of ACT, artemether–lumefantrine (AL) and artesunate + amodiaquine (ASAQ), for the treatment of uncomplicated malaria among prescribers and patients in Guinea in 2016. Methods The study included a review of records of malaria patients and three health-facility, cross-sectional surveys. Patients diagnosed with uncomplicated malaria and prescribed ACT (n = 1830) were recruited and visited in their home after receiving the medication and administered a questionnaire regarding ACT adherence. Prescribers (n = 115) and drug dispensers (n = 43) were recruited at the same public health facilities and administered questionnaires regarding prescribing practices and opinions regarding the national treatment policies and protocols. Results According to the registry review, 35.8% of all-cause consultations were recorded as malaria. Of these, 26.6% were diagnosed clinically without documentation of laboratory confirmation. The diagnosis of uncomplicated malaria represented 64.1% of malaria cases among children under 5 years and 74.9% of those 5 years of age and older. An ACT was prescribed for 83.5% of cases of uncomplicated malaria. Among participants in the study, ACT adherence was 95.4% (95% CI 94.4, 96.3). Overall, about one in four patients (23.4%; 95% CI 21.5, 25.3) reported experiencing adverse events. While patients prescribed ASAQ were significantly more likely to report experiencing adverse effects than patients on AL (p

The Mapping Manuscript Migrations (MMM) project transformed three separate datasets relating to the history and provenance of medieval and Renaissance manuscripts into a unified knowledge graph. The source databases are: Schoenberg Database of Manuscripts, from the Schoenberg Institute for Manuscript Studies, University of Pennsylvania; Bibale, from the Institut de recherche et d’histoire des textes (IRHT-CNRS, Paris); and Medieval Manuscripts in Oxford Libraries, from the Bodleian Libraries, University of Oxford. The data consist of more than 20 million RDF triples which have been mapped to the MMM Data Model. The model combines classes and properties from CIDOC-CRM and FRBR, together with some specific MMM elements. The Knowledge Graph was created using the MMM data transformation pipeline. The MMM dataset is available from the Zenodo repository, and can be directly deployed on a SPARQL endpoint using a docker recipe. To test and demonstrate its usefulness, the MMM Knowledge Graph is in use in the MMM Semantic Portal: https://mappingmanuscriptmigrations.org.

Abstract Background The Finnish neurological function testing battery for dogs (FINFUN) was developed to meet the increasing demand for objective outcome measures in veterinary physiotherapy. The testing battery should provide consistent, reproducible results and have established face and content validity. Internal consistency and intra- and inter-rater reliability of the FINFUN were also investigated. Results The FINFUN comprised 11 tasks: lying, standing up from lying, sitting, standing up from sitting, standing, proprioceptive positioning, starting to walk, walking, trotting, walking turns and walking stairs. A score from 0 to 4, (0: unable to perform task, 4: performing task with normal motor function) was given for each task, the maximum score being 44. Twenty-six dogs were filmed when performing the FINFUN. Seven observers scored the performances from the video recordings. The FINFUN was considered to have appropriate face and content validity based on a pilot study, clinical experience and critical reflection of the development process. Its internal consistency was excellent, with no Cronbach’s alpha values below 0.922. The intra-rater reliability for total score of experienced observers was almost perfect: 0.999 (observer 1) and 0.994 (observer 2). The inter-rater reliability for both experienced and novice observers’ total scores was also almost perfect (0.919–0.993). Analysis of each individual task showed substantial intra-rater and inter-rater agreement for the tasks “lying” and “sitting”. Conclusions The FINFUN is an objective, valid and reliable tool with standardized scoring criteria for evaluation of motor function in dogs recovering from spinal cord injury.

The Mn / Ca of calcium carbonate tests of living (rose-Bengal-stained) benthic foraminifera (Elphidium batialis, Uvigerina spp., Bolivina spissa, Nonionellina labradorica and Chilostomellina fimbriata) were determined in relation to pore water manganese (Mn) concentrations for the first time along a bottom water oxygen gradient across the continental slope along the NE Japan margin (western Pacific). The local bottom water oxygen (BWO) gradient differs from previous field study sites focusing on foraminiferal Mn / Ca and redox chemistry, therefore allowing further resolution of previously observed trends. The Mn / Ca ratios were analysed using laser ablation inductively coupled plasma-mass spectrometer (ICP-MS), allowing single-chamber determination of Mn / Ca. The incorporation of Mn into the carbonate tests reflects environmental conditions and is not influenced by ontogeny. The inter-species variability in Mn / Ca reflected foraminiferal in-sediment habitat preferences and associated pore water chemistry but also showed large interspecific differences in Mn partitioning. At each station, Mn / Ca ratios were always lower in the shallow infaunal E. batialis, occupying relatively oxygenated sediments, compared to intermediate infaunal species, Uvigerina spp. and B. spissa, which were typically found at greater depth, under more reducing conditions. The highest Mn / Ca was always recorded by the deep infaunal species N. labradorica and C. fimbriata. Our results suggest that although partitioning differs, Mn / Ca ratios in the intermediate infaunal taxa are promising tools for palaeoceanographic reconstructions as their microhabitat exposes them to higher variability in pore water Mn, thereby making them relatively sensitive recorders of redox conditions and/or bottom water oxygenation.

Foraminifera are ubiquitous marine protists with an important role in the benthic carbon cycle. However, morphological observations often fail to resolve their exact taxonomic placement and there is a lack of field studies on their particular trophic preferences. Here, we propose the application of metabarcoding as a tool for the elucidation of the in situ feeding behavior of benthic foraminifera, while also allowing the correct taxonomic assignment of the feeder, using the V9 region of the 18S (small subunit; SSU) rRNA gene. Living foraminiferal specimens were collected from two intertidal mudflats of the Wadden Sea and DNA was extracted from foraminiferal individuals and from the surrounding sediments. Molecular analysis allowed us to confirm that our foraminiferal specimens belong to three genetic types: Ammonia sp. T6, Elphidium sp. S5 and Haynesina sp. S16. Foraminiferal intracellular eukaryote communities reflected to an extent those of the surrounding sediments but at different relative abundances. Unlike sediment eukaryote communities, which were largely determined by the sampling site, foraminiferal intracellular eukaryote communities were driven by foraminiferal species, followed by sediment depth. Our data suggests that Ammonia sp. T6 can predate on metazoan classes, whereas Elphidium sp. S5 and Haynesina sp. S16 are more likely to ingest diatoms. These observations, alongside the use of metabarcoding in similar ecological studies, significantly contribute to our overall understanding of the ecological roles of these protists in intertidal benthic environments and their position and function in the benthic food webs.

Quantitative variation of nine flavonoid compounds and two related phenolic acids in several parts of three garden varieties of Cosmos bipinnatus Cav. were examined by; means of paper chromatography followed by a spectrophotometric procedure.