Isolation of extracellular vesicles from the bronchoalveolar lavage fluid of healthy and asthmatic horses

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.Peer reviewe

Kuvapatteristosta virtaa vuorovaikutukseen : työvälineen kehittäminen varhaiskasvatuksen ammattilaisille monikulttuuriseen asiakastyöhön

Tämän kehittämishankkeen tehtävänä oli tuottaa työväline maahanmuuttajalasten vanhempien ja päivähoidon työntekijöiden vuorovaikutuksen tueksi. Työväline konkretisoitui mahdollisimman toimivaksi, yksinkertaiseksi ja informatiiviseksi kuvapatteristoksi, jonka käytön tuli olla helppoa ja vaivatonta. Kuvapatteriston tavoitteena oli luoda mahdollisuus vuorovaikutukseen ilman yhteistä kieltä, toimia puheen tukena, parantaa maahanmuuttajavanhempien osallisuutta lastensa päiväkodin arjessa sekä kehittää kasvatuskumppanuuden laatua. Työvälineen tuottamisen lähtökohdat kumpusivat työelämästä; osa maahanmuuttajalasten vanhemmista ei kielitaitonsa puutteellisuudesta johtuen kyennyt ymmärrettävään kommunikointiin päiväkodin henkilöstön kanssa. Kehittämisen menetelminä käytettiin tiimityöskentelyä, parityöskentelyä, keskustelua ja neuvottelevaa arviointia. Kehittämishanketta pohjaava teoria koostuu varhaiskasvatuksesta ja kasvatuskumppanuudesta, monikulttuurisuudesta varhaiskasvatuksen näkökulmasta sekä vuorovaikutuksen eri muodoista. Kehittämishankkeen prosessi eteni ideoinnin, suunnittelun ja tuottamisen kautta valmiiseen tuotokseen ja raporttiin. Näiden vaiheiden lomassa toteutettiin parhaan tuloksen aikaansaamiseksi arviointia ja palautteen kautta tuotetun materiaalin muokkausta. Tuotoksena syntyi kattava, mutta vain oleellisia asioita sisältävä kuvapatteristo, joka koostui tilannekuvista ja niiden rinnalle luoduista erilaisia tunnetiloja ilmentävistä kasvo-, sekä ”kyllä” ja ”ei”-korteista. Valmiin tuotoksen arvioi toimeksiantaja, jonka palautteen pohjalta se vielä muokattiin viimeisteltyyn ja lopulliseen muotoonsa. Kehittämishankkeen tekijät olivat tyytyväisiä lopputulokseen niin tuotoksen, kuin raportoinninkin osalta ja he uskovat kuvapatteriston monikäyttöisyyden antavan jokaiselle käyttäjälle mahdollisuuden toimia sen kanssa itselleen parhaalta tuntuvalla tavalla. Tuotoksen koettiin vastaavan ennalta määrättyihin tavoitteisiin ja kehittämishankkeen etenemisprosessi kokonaisuudessaan toimi tekijöiden ammatillisen kasvun ja kehityksen vahvistajana.This development project was about producing a tool to support immigrant parents’ and day care professionals’ interaction. The tool was concretised with an image composition, which was made to be as functional, simple and informative as possible. Its usage had to be easy and effortless. Objectives of the image composition was to create a possibility for interaction without a common language, support speech, enhance the inclusion of immigrant parents in everyday life of their children’s day care center and improve the quality of educational partnership. The base of this development project emerged from working life; some of the immigrant parents weren’t able to understandably communicate with daycare workers since their language skills were inadequate. Development methods such as teamwork, pair work, discussion and negotiative evaluation was used in this project. The theory, on which the development project is based, consists of early childhood education, educational partnership, multiculturalism in point of view of early childhood education and different forms of interaction. Process of the development project progressed through different phases, such as generating ideas, planning and production. At the side of these phases, evaluation and modifying of the produced material was executed in order to achieve the best results. As an output, arose a comprehensive image composition, which included only relevant matter. The image composition consisted of situation images, facial, different emotion expressing images, as well as “yes” and “no” images. The finished output was evaluated by the principal. Based on the feedback that was given, the output was modified into a finished and final form. Makers of the development projects were pleased with the results and they believe that the versatility of the image composition will offer a possibility for each user to function with it in the best way they see fit. The output was experienced to meet the objectives made in the beginning and the process as a hole was felt to strengthen the makers’ professional growth and advancement

Fatty acid fingerprints in bronchoalveolar lavage fluid and its extracellular vesicles reflect equine asthma severity

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.Peer reviewe