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Sökning: WFRF:(Manneberg Otto)

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1.
  • Manneberg, Otto, 1981-, et al. (författare)
  • En struktur för ökad funktionell kunskap hos studenten från räkneövningar
  • 2008
  • Konferensbidrag (refereegranskat)abstract
    • En struktur för räkneövningar med syftet att ge studenten ökade funktionella kunskaper och färdigheter i ämnet presenteras. Strukturen är avsedd att aktivera studenten både innan och under en räkneövning. Genom kamraträttning av frivilliga hemuppgifter frikopplade från bonussystem, student-student-diskussioner samt diskussion i helklass antas studenten uppmuntras till djupare lärstrategier och nå en högre taxonominivå inom ämnet.Strukturen utprovas på två studentgrupper; andraårsstudenter på ett civilingenjörsprogram på KTH, samt förstårsstudenter på optikerutbildningen på Karolinska Institutet som läser optikkurser vid KTH. Utvärdering sker under pågående kurs genom enkäter till alla studenterna samt intervjuer av optikerstudenter, samt efter fullgången kurs genom intervjuer av assistenter och ingenjörsstudenter. Resultaten visar att studenterna ställer sig positiva till det nya systemet och anser att det ger dem en djupare nivå av förståelse, att det skapar en mer transparent bild av rättningsprocessen, samt att det ökar interaktionen mellan studenter och skapar en bättre klassrumsmiljö. Deltagandet är stort bland optikerstudenterna, medan ingenjörsstudenterna anser att de har betydligt svårare att hitta tid till hemuppgifterna.
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  • Agostinho, Ana, et al. (författare)
  • High density of REC8 constrains sister chromatid axes and prevents illegitimate synaptonemal complex formation
  • 2016
  • Ingår i: EMBO Reports. - Stockholm : Wiley-VCH Verlagsgesellschaft. - 1469-221X .- 1469-3178. ; 17:6, s. 901-913
  • Tidskriftsartikel (refereegranskat)abstract
    • During meiosis, cohesin complexes mediate sister chromatid cohesion (SCC), synaptonemal complex (SC) assembly and synapsis. Here, using super-resolution microscopy, we imaged sister chromatid axes in mouse meiocytes that have normal or reduced levels of cohesin complexes, assessing the relationship between localization of cohesin complexes, SCC and SC formation. We show that REC8 foci are separated from each other by a distance smaller than 15% of the total chromosome axis length in wild-type meiocytes. Reduced levels of cohesin complexes result in a local separation of sister chromatid axial elements (LSAEs), as well as illegitimate SC formation at these sites. REC8 but not RAD21 or RAD21L cohesin complexes flank sites of LSAEs, whereas RAD21 and RAD21L appear predominantly along the separated sister-chromatid axes. Based on these observations and a quantitative distribution analysis of REC8 along sister chromatid axes, we propose that the high density of randomly distributed REC8 cohesin complexes promotes SCC and prevents illegitimate SC formation.
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4.
  • Grondin, Yohann, et al. (författare)
  • Pulmonary delivery of d-methionine is associated with an increase in ALCAR and glutathione in cochlear fluids.
  • 2013
  • Ingår i: Hearing Research. - : Elsevier BV. - 0378-5955 .- 1878-5891. ; 298, s. 93-103
  • Tidskriftsartikel (refereegranskat)abstract
    • In animals, hearing loss resulting from cochlear mechanosensory cell damage can be mitigated by antioxidants such as d-methionine (d-met) and acetyl-l-carnitine (ALCAR). The systemic routes of administration of these compounds, that must of necessity transit trough the cochlear fluids, may affect the antioxidant levels in the cochlea and the resulting oto-protective effect. In this study, we analyzed the pharmacokinetics of [C]d-met in the cochlea and four other tissues after intratracheal (IT), intranasal (IN), and oral by gavage (OG) administration and compared it to intravenous administration (IV). We then analyzed the effect of these four routes on the antioxidant content of the cochlear fluids after d-met or ALCAR administration, by liquid chromatography/mass spectrometry. Our results showed that the concentration of methionine and ALCAR in cochlear fluids significantly increased after their respective systemic administration. Interestingly, d-met administration also contributed to an increase of ALCAR. Our results also showed that the delivery routes differently affected the bioavailability of administered [C]d-met as well as the concentrations of methionine, ALCAR and the ratio of oxidized to reduced glutathione. Overall, pulmonary delivery via IT administration achieved high concentrations of methionine, ALCAR, and oxidative-related metabolites in cochlear fluids, in some cases surpassing IV administration, while IN route appeared to be the least efficacious. To our knowledge, this is the first report of the direct measurements of antioxidant levels in cochlear fluids after their systemic administration. This report also demonstrates the validity of the pulmonary administration of antioxidants and highlights the different contributions of d-met and ALCAR allowing to further investigate their impact on oxidative stress in the cochlear microenvironment.
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  • Guldevall, Karolin, et al. (författare)
  • Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
  • 2010
  • Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 5:11, s. e15453-
  • Tidskriftsartikel (refereegranskat)abstract
    • New markers are constantly emerging that identify smaller and smaller subpopulations of immune cells. However, there is a growing awareness that even within very small populations, there is a marked functional heterogeneity and that measurements at the population level only gives an average estimate of the behaviour of that pool of cells. New techniques to analyze single immune cells over time are needed to overcome this limitation. For that purpose, we have designed and evaluated microwell array systems made from two materials, polydimethylsiloxane (PDMS) and silicon, for high-resolution imaging of individual natural killer (NK) cell responses. Both materials were suitable for short-term studies (<4 hours) but only silicon wells allowed long-term studies (several days). Time-lapse imaging of NK cell cytotoxicity in these microwell arrays revealed that roughly 30% of the target cells died much more rapidly than the rest upon NK cell encounter. This unexpected heterogeneity may reflect either separate mechanisms of killing or different killing efficiency by individual NK cells. Furthermore, we show that high-resolution imaging of inhibitory synapse formation, defined by clustering of MHC class I at the interface between NK and target cells, is possible in these microwells. We conclude that live cell imaging of NK-target cell interactions in multi-well microstructures are possible. The technique enables novel types of assays and allow data collection at a level of resolution not previously obtained. Furthermore, due to the large number of wells that can be simultaneously imaged, new statistical information is obtained that will lead to a better understanding of the function and regulation of the immune system at the single cell level.
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7.
  • Guldevall, Karolin, 1981-, et al. (författare)
  • Microchip screening platform for assessment of cytotoxic effector cells
  • Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
    • Here we report a screening platform for assessment of the cytotoxic potential of individual natural killer (NK) or T cells within larger populations. Human primary NK cells or human Epstein-Barr virus (EBV)- specific T cells were distributed across a silicon-glass microchip containing 32 400 individual microwells loaded with target cells. Through fluorescence screening and automated image analysis the numbers of effector and live or dead target cells in each well could be assessed at different time-points after initial mixing. Cytotoxicity was also studied by time-lapse live-cell imaging in microwells quantifying the killing potential of individual NK cells. Although most resting NK cells (≈75%) were non-cytotoxic to the leukemia cell line K562, some NK cells were able to kill several (≥3) target cells within the 12 hours long experiment. We demonstrate that this assay can be used to enumerate and characterize cytotoxic cells, something that could find clinical applications, e.g. in the selection of donors for stem cell transplantation or generation of highly specific and cytotoxic cells for adoptive immunotherapy.
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8.
  • Guldevall, Karolin, et al. (författare)
  • Microchip screening Platform for single cell assessment of NK cell cytotoxicity
  • 2016
  • Ingår i: Frontiers in Immunology. - : FRONTIERS MEDIA SA. - 1664-3224. ; 7
  • Tidskriftsartikel (refereegranskat)abstract
    • Here, we report a screening platform for assessment of the cytotoxic potential of individual natural killer (NK) cells within larger populations. Human primary NK cells were distributed across a silicon-glass microchip containing 32,400 individual microwells loaded with target cells. Through fluorescence screening and automated image analysis, the numbers of NK and live or dead target cells in each well could be assessed at different time points after initial mixing. Cytotoxicity was also studied by time-lapse live-cell imaging in microwells quantifying the killing potential of individual NK cells. Although most resting NK cells (approximate to 75%) were non-cytotoxic against the leukemia cell line K562, some NK cells were able to kill several (>= 3) target cells within the 12-h long experiment. In addition, the screening approach was adapted to increase the chance to find and evaluate serial killing NK cells. Even if the cytotoxic potential varied between donors, it was evident that a small fraction of highly cytotoxic NK cells were responsible for a substantial portion of the killing. We demonstrate multiple assays where our platform can be used to enumerate and characterize cytotoxic cells, such as NK or T cells. This approach could find use in clinical applications, e.g., in the selection of donors for stem cell transplantation or generation of highly specific and cytotoxic cells for adoptive immunotherapy.
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9.
  • Hultström, Jessica, et al. (författare)
  • Proliferation and viability of adherent cells manipulated by standing-wave ultrasound in a microfluidic chip
  • 2007
  • Ingår i: Ultrasound in Medicine and Biology. - : Elsevier BV. - 0301-5629 .- 1879-291X. ; 33, s. 145-151
  • Tidskriftsartikel (refereegranskat)abstract
    • Ultrasonic-standing-wave (USW) technology has potential to become a standard method for gentle and contactless cell handling in microfluidic chips. We investigate the viability of adherent cells exposed to USWs by studying the proliferation rate of recultured cells following ultrasonic trapping and aggregation of low cell numbers in a microfluidic chip. The cells form 2-D aggregates inside the chip and the aggregates are held against a continuous flow of cell culture medium perpendicular to the propagation direction of the standing wave. No deviations in the doubling time from expected values (24 to 48 h) were observed for COS-7 cells held in the trap at acoustic pressure amplitudes up to 0.85 MPa and for times ranging between 30 and 75 min. Thus, the results demonstrate the potential of ultrasonic standing waves as a tool for gentle manipulation of low cell numbers in microfluidic systems.
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10.
  • Hultström, Jessica, et al. (författare)
  • Proliferation and viability of COS-7 cells trapped by standing-wave ultrasound in a microfluidic chip
  • 2006
  • Ingår i: Micro Total Analysis Systems - Proceedings of MicroTAS 2006 Conference. - : Japan Academic Association Inc. ; , s. 449-451
  • Konferensbidrag (refereegranskat)abstract
    • We study cell viability after ultrasonic-standing-wave trapping of low cell numbers in a microfluidic chip by recultivation of the trapped cells. The cell proliferation rate is estimated by counting of initial and final cell numbers and shows normal cell growth. The results demonstrate the potential of ultrasonic standing waves as a tool for gentle and long-term manipulation of low cell numbers in microfluidic systems.
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