Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

ObjectiveNatural killer (NK) cells are a part of the innate immune system and first-line defense against cancer. Since they possess natural mechanisms to recognize and kill tumor cells, NK cells are considered as a potential option for an off-the-shelf allogeneic cell-based immunotherapy. Here, our objective was to identify the optimal cytokine-based, feeder-free, activation and expansion protocol for cytotoxic NK cells against glioblastoma in vitro.MethodsNK cells were enriched from human peripheral blood and expanded for 16 days with different activation and cytokine combinations. The expansion conditions were evaluated based on NK cell viability, functionality, expansion rate and purity. The cytotoxicity and degranulation of the expanded NK cells were measured in vitro from co‑cultures with the glioma cell lines U‑87 MG, U‑87 MG EGFR vIII, LN-229, U-118 and DK-MG. The best expansion protocols were selected from ultimately 39 different conditions: three magnetic cell‑selection steps (Depletion of CD3+ cells, enrichment of CD56+ cells, and depletion of CD3+ cells followed by enrichment of CD56+ cells); four activation protocols (continuous, pre-activation, re-activation, and boost); and four cytokine combinations (IL-2/15, IL‑21/15, IL‑27/18/15 and IL-12/18/15).ResultsThe expansion rates varied between 2-50-fold, depending on the donor and the expansion conditions. The best expansion rate and purity were gained with sequential selection (Depletion of CD3+ cells and enrichment of CD56+ cells) from the starting material and pre-activation with IL‑12/18/15 cytokines, which are known to produce cytokine-induced memory-like NK cells. The cytotoxicity of these memory-like NK cells was enhanced with re-activation, diminishing the donor variation. The most cytotoxic NK cells were produced when cells were boosted at the end of the expansion with IL-12/18/15 or IL-21/15.ConclusionAccording to our findings the ex vivo proliferation capacity and functionality of NK cells is affected by multiple factors, such as the donor, composition of starting material, cytokine combination and the activation protocol. The cytokines modified the NK cells' phenotype and functionality, which was evident in their reactivity against the glioma cell lines. To our knowledge, this is the first comprehensive comparative study performed to this extent, and these findings could be used for upscaling clinical NK cell manufacturing

Developing a Feasible and Credible Method for Analyzing Healthcare Documents as Written Data

Healthcare provides a rich, and constantly increasing, number of written documents, which are underutilized in research data for health and nursing sciences, but previous literature has only provided limited guidance on the process of document analysis. The aim of this paper is to provide a methodological framework for analyzing health care documents as written data, based on a systematic methodological review and the research team's experience of the method. Based on the results, the methods consist of seven phases: (i) identify the purpose, (ii) determine the document selection strategy, (iii) select or design an extraction matrix, (iv) carry out pilot testing, (v) collect and analyze the data, (vi) consider the credibility, and (vii) ethics of the study. The framework that has been developed can be used to carry out document analysis studies that are both feasible and credible

Työntekijöiden ja esimiesten näkemyksiä etäjohtamisesta

Opinnäytetyön tarkoituksena oli tutkia etäjohtamista esimiesten ja työntekijöiden näkökulmista. Tutkimusmenetelmänä oli kvalitatiivinen tutkimus johon tietoa kerättiin teemahaastatteluilla. Tutkimukseen osallistui kolme yritystä jotka kaikki toimivat IT-alalla. Yksi yritys tarjosi palveluita IT-infrastruktuurin kehittämiseen ja hallintaan, toinen oli tiedolla johtamiseen erikoistunut palveluyritys ja kolmannen liiketoimintaa olivat palvelu- ja projektijohtamisen käytännöt. Jokaisesta yrityksestä haastatteluun osallistui vähintään yksi esimies ja kaksi asiantuntijaa. Kaikki haastatellut henkilöt tekivät töitä myös etätyönä ja heitä myös etäjohdettiin. Haastatteluista nousi selkeästi esille neljä teemaa: ihminen, organisaatio, tekniikka ja kehittäminen. Opinnäytetyössä keskitytään etätyön ja etäjohtamisen inhimillisiin piirteisiin eikä etätyössä käytettyjä tekniikoita ja sovelluksia esitellä tarkemmin. Opinnäytetyössä haettiin vastauksia siihen miten työntekijöiden ja esimiesten näkemykset etäjohtamisesta ja sen tarpeista eroavat toisistaan ja siihen miten etäjohtamista voitaisiin kehittää. Haastatteluista voitiin nähdä, että näkemykset olivat melko samankaltaisia. Suurimmat erot syntyivät yritysten organisaatiokulttuurista ja -rakenteista. Kehityskohteet liittyivät myös yritysten organisaatiorakenteisiin joissa on tärkeää ottaa myös etätyön ja etäjohtamiseen tuoma ulottuvuus huomioon. Etäjohtamista voi myös kehittää muokkaamalla yrityksen viestintää ja kommunikointia etätyömalliin sopivammaksi.The purpose of this thesis was to study distance management from the perspectives of employees and managers. Qualitative study was used as the research method, to which data was collected by theme interviews. Three companies, all from the information technology industry, took part in the study. One of the companies offered services for IT-infrastructure development and management, the other company specialised in management by knowledge, and the third company’s business area were service and project management practices. From each company at least one manager and two experts participated in the interview. All of the people interviewed worked and were magaged from the distance. Four themes clearly stood out during the interviews: people, organization, technology and development. The thesis concentrates on the humane qualities of distance work and management. It does not present the techniques or applications used in detail. The thesis sought out the answers to how the employees’ and managers’ views on distance management differ and how distance management could be developed. Based on the interviews a conclusion can be made that the views do not differ very much from each other. The biggest differences arise from the culture and structures of the companies. Distance management can be developed by modifying the communication for the needs of distance work. The organization culture and operating models should be rethought from the perspective of distance work

Effects of Phosphate and Light on Growth of and Bioactive Peptide Production by the Cyanobacterium Anabaena Strain 90 and Its Anabaenopeptilide Mutant

Cyanobacteria synthesize several types of bioactive secondary metabolites. Anabaena strain 90 produces three types of bioactive peptides, microcystins (inhibitors of protein phosphatases 1 and 2A), anabaenopeptilides, and anabaenopeptins (serine protease inhibitors). To investigate the role of the anabaenopeptilides in Anabaena, wild-type strain 90 (WT) and its anabaenopeptilide deficient mutant (MU) were cultured with various light and phosphate levels to evaluate the effects and coeffects of these growth factors on the concentrations of the three classes of peptides and the growth characteristics. WT and MU grew in comparable ways under the different growth conditions. The total peptide concentration in WT was significantly higher than that in MU (2.5 and 1.4 μg/mg [dry weight], respectively). Interestingly, the average concentration of anabaenopeptins was significantly higher in MU than in WT (0.59 and 0.24 μg/mg [dry weight], respectively). The concentration of microcystins was slightly but not statistically significantly higher in MU than in WT (1.0 and 0.86 μg/mg [dry weight], respectively). In WT, the highest peptide concentrations were usually found after 13 days in cultures grown at medium light intensities (23 μmol m(−2) s(−1)) and with the highest phosphate concentrations (2,600 μg liter(−1)). In MU, the highest peptide concentrations were found in 13-day-old cultures grown at medium light intensities (23 μmol m(−2) s(−1)) and with phosphate concentrations greater than 100 μg liter(−1). The higher concentrations of anabaenopeptins in MU may compensate for the absence of anabaenopeptilides. These findings clearly indicate that these compounds may have some linked function in the producer organism, the nature of which remains to be discovered

DataSheet_1_Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro.docx

ObjectiveNatural killer (NK) cells are a part of the innate immune system and first-line defense against cancer. Since they possess natural mechanisms to recognize and kill tumor cells, NK cells are considered as a potential option for an off-the-shelf allogeneic cell-based immunotherapy. Here, our objective was to identify the optimal cytokine-based, feeder-free, activation and expansion protocol for cytotoxic NK cells against glioblastoma in vitro.MethodsNK cells were enriched from human peripheral blood and expanded for 16 days with different activation and cytokine combinations. The expansion conditions were evaluated based on NK cell viability, functionality, expansion rate and purity. The cytotoxicity and degranulation of the expanded NK cells were measured in vitro from co‑cultures with the glioma cell lines U‑87 MG, U‑87 MG EGFR vIII, LN-229, U-118 and DK-MG. The best expansion protocols were selected from ultimately 39 different conditions: three magnetic cell‑selection steps (Depletion of CD3+ cells, enrichment of CD56+ cells, and depletion of CD3+ cells followed by enrichment of CD56+ cells); four activation protocols (continuous, pre-activation, re-activation, and boost); and four cytokine combinations (IL-2/15, IL‑21/15, IL‑27/18/15 and IL-12/18/15).ResultsThe expansion rates varied between 2-50-fold, depending on the donor and the expansion conditions. The best expansion rate and purity were gained with sequential selection (Depletion of CD3+ cells and enrichment of CD56+ cells) from the starting material and pre-activation with IL‑12/18/15 cytokines, which are known to produce cytokine-induced memory-like NK cells. The cytotoxicity of these memory-like NK cells was enhanced with re-activation, diminishing the donor variation. The most cytotoxic NK cells were produced when cells were boosted at the end of the expansion with IL-12/18/15 or IL-21/15.ConclusionAccording to our findings the ex vivo proliferation capacity and functionality of NK cells is affected by multiple factors, such as the donor, composition of starting material, cytokine combination and the activation protocol. The cytokines modified the NK cells' phenotype and functionality, which was evident in their reactivity against the glioma cell lines. To our knowledge, this is the first comprehensive comparative study performed to this extent, and these findings could be used for upscaling clinical NK cell manufacturing.</p

Jatkoväylällä : Yhteistyöllä ammatillisesta koulutuksesta ammattikorkeakouluun

Tässä kirjassa kuvataan Jatkoväylä-hankkeen tuloksina ammatillisen koulutuksen ja ammattikorkeakoulujen yhteistyönä syntyneitä rakenteita, kokemuksia ja toimintamalleja jatkoväyläopinnoista ja -ohjauksesta. Nämä tiivistyvät kirjan lopussa valtakunnallisiksi suosituksiksi jatkoväyläopintojen ja -ohjauksen järjestämisestä ammatillisen toisen asteen koulutuksen ja ammattikorkeakoulun yhteistyönä. Valtakunnallisesti jatkoväyläopinnoilla ja -ohjauksella tarkoitetaan ammatillisen toisen asteen oppilaitoksen ja ammattikorkeakoulun välisiä siirtymävaihetta tukevia opintoja ja ohjausta. Käytössä on myös väyläopinnot ja -ohjaus -käsite, jota käytetään paikallisesti eri koulutusorganisaatioissa kuvaamaan jatkoväyläopintoja ja -ohjausta. Kirjan artikkeleissa käytetään molempia käsitteitä koulutusorganisaatiosta riippuen. Jatkoväyläopinnot ovat avoimen ammattikorkeakoulun opintoja, joita suorittavat ammatillisen toisen asteen opiskelijat. Jatkoväyläopintoja ja -ohjausta suunnitellaan ja toteutetaan ammatillisen toisen asteen oppilaitoksen ja ammattikorkeakoulun yhteistyönä