| 1. |
- Al-Jubair, Tamim, et al.
(author)
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Characterization of human aquaporin protein-protein interactions using microscale thermophoresis (MST)
- 2022
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In: STAR Protocols. - : Elsevier BV. - 2666-1667. ; 3:2
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Journal article (peer-reviewed)abstract
- Aquaporin water channels (AQPs) are membrane proteins that maintain cellular water homeostasis. The interactions between human AQPs and other proteins play crucial roles in AQP regulation by both gating and trafficking. Here, we describe a protocol for characterizing the interaction between a human AQP and a soluble interaction partner using microscale thermophoresis (MST). MST has the advantage of low sample consumption and high detergent compatibility enabling AQP protein-protein interaction investigation with a high level of control of components and environment. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Roche et al. (2017).
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| 2. |
- Al-Jubair, Tamim, et al.
(author)
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High-yield overproduction and purification of human aquaporins from Pichia pastoris
- 2022
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In: STAR Protocols. - : Elsevier BV. - 2666-1667. ; 3:2
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Journal article (peer-reviewed)abstract
- Aquaporins (AQPs) are membrane-bound water channels that play crucial roles in maintaining the water homeostasis of the human body. Here, we present a protocol for high-yield recombinant expression of human AQPs in the methylotropic yeast Pichia pastoris and subsequent AQP purification. The protocol typically yields 1–5 mg AQP per g of yeast cell at >95% purity and is compatible with any membrane protein cloned into Pichia pastoris, although expression levels may vary. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Frick et al. (2014).
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| 3. |
- Bill, Susanna, et al.
(author)
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Litteratur om tillitsbegreppet
- 2014
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In: DigiTrust: Tillit i det digitala. Tvärvetenskapliga perspektiv från ett forskningsprojekt. - 9789197989367 ; , s. 15-20
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Book chapter (pop. science, debate, etc.)abstract
- Detta kapitel visar på hur forskning om tillit är spridd över en rad ämnesområden och forskardiscipliner, och tar upp några av de tydligaste områdena.
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| 4. |
- Bill, Susanna, et al.
(author)
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Rethinking User Involvement in the Front End of Healthcare Innovation
- 2011
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Conference paper (peer-reviewed)abstract
- The purpose of this paper is to share insights from an empirical study within the Swedish healthcare system, based on a case study of the largest business unit of a Swedish medical technology company and its key customer segment. The study involving a Swedish medical technology company and three Swedish hospitals indicates need to move away from an archetypical, largely homogeneous view of the user, towards a more flexible, heterogeneous user perspective. Following from that insight the paper outlines a preliminary model aimed at improving the conditions of the firm for user involvement in the front end of healthcare innovation. In the proposed model, different users are brought to the foreground in different parts of the front-end innovation process. The research performed aimed at putting the business unit in a more beneficial position to involve users in the front end of innovation by increasing their understanding of a) which user types they actually have access to, b) which roles that different user types may take in the various stages of the innovation front-end.
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| 5. |
- Bolling, Hans, 1966-
(author)
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Sin egen hälsas smed : Idéer, initiativ och organisationer inom svensk motionsidrott 1945–1981
- 2005
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Doctoral thesis (other academic/artistic)abstract
- This dissertation studies the spread of sports for all in Sweden during the years 1945 to 1981. The purposes of the dissertation are twofold: in part to survey the forms of physical activities which were launched as sports for all after 1945, in part to answer the question: Why have almost all voluntarily organized sports in Sweden been part of one organization since the 1970s? In order to handle the diversities of activities that can fall within the concept of sport, two principal abstractions of the concept are used: one rigorous and one flexible. Which definition one uses influences how physical activities are organized in a society. Earlier research into the history of the Swedish sports movement has concluded that it has had a relatively high degree of autonomy in relation to the state. This finding is questioned in this dissertation. Sveriges Riksidrottsförbund (RF) was the largest Swedish sports organization throughout the 20th century and at same time the organization the government relied on to develop sports policies and distribute the financial contribution from the state to the sports movement. This means that RF has played two roles, as an umbrella organization within the Swedish sports movement and as leader of the organizations within the Swedish sports movement, popular movement and semi-public authority. The dissertation shows that the two roles, that RF played, have caused conflicts of interest within the organization. That is made plain when one studies the spread of sports for all. Most members of the organization just wanted to practise different sports and were not interested in the leading organization’s desire to promote a great many different kinds of physical acitivites according to a flexible concept of sport. These members were not interested in strengthening the organization’s leading position within sports. There are not many conceptions that are so universally and uncritically accepted as the conception of the connection between physical activity and health. Sports for all came to age in a society where more and more people were told to use part of their leisure time to take part in physical activities. A societal consensus prevailed that the population’s health was on the decline due to the increased standard of living, which was creating an inactive and unhealthy population. This has meant that sports for all have been an asset of power for the sports organizations and that they have fought for authority and control over sports for all; a struggle fought over the language and thoughts as much as over sport activities. Since 1945 large campaigns to get the population to become more physically active irrespective of physical ability have been common.
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| 6. |
- Kitchen, Philip, et al.
(author)
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Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells
- 2020
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In: STAR Protocols. - : Elsevier BV. - 2666-1667. ; 1:3
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Journal article (peer-reviewed)abstract
- Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).
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| 7. |
- Kitchen, Philip, et al.
(author)
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Plasma Membrane Abundance of Human Aquaporin 5 Is Dynamically Regulated by Multiple Pathways.
- 2015
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In: PloS one. - : Public Library of Science (PLoS). - 1932-6203. ; 10:11
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Journal article (peer-reviewed)abstract
- Aquaporin membrane protein channels mediate cellular water flow. Human aquaporin 5 (AQP5) is highly expressed in the respiratory system and secretory glands where it facilitates the osmotically-driven generation of pulmonary secretions, saliva, sweat and tears. Dysfunctional trafficking of AQP5 has been implicated in several human disease states, including Sjögren's syndrome, bronchitis and cystic fibrosis. In order to investigate how the plasma membrane expression levels of AQP5 are regulated, we studied real-time translocation of GFP-tagged AQP5 in HEK293 cells. We show that AQP5 plasma membrane abundance in transfected HEK293 cells is rapidly and reversibly regulated by at least three independent mechanisms involving phosphorylation at Ser156, protein kinase A activity and extracellular tonicity. The crystal structure of a Ser156 phosphomimetic mutant indicates that its involvement in regulating AQP5 membrane abundance is not mediated by a conformational change of the carboxy-terminus. We suggest that together these pathways regulate cellular water flow.
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| 8. |
- Kitchen, Philip, et al.
(author)
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Targeting Aquaporin-4 Subcellular Localization to Treat Central Nervous System Edema
- 2020
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In: Cell. - : Elsevier BV. - 0092-8674. ; 181:4, s. 19-799
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Journal article (peer-reviewed)abstract
- Swelling of the brain or spinal cord (CNS edema) affects millions of people every year. All potential pharmacological interventions have failed in clinical trials, meaning that symptom management is the only treatment option. The water channel protein aquaporin-4 (AQP4) is expressed in astrocytes and mediates water flux across the blood-brain and blood-spinal cord barriers. Here we show that AQP4 cell-surface abundance increases in response to hypoxia-induced cell swelling in a calmodulin-dependent manner. Calmodulin directly binds the AQP4 carboxyl terminus, causing a specific conformational change and driving AQP4 cell-surface localization. Inhibition of calmodulin in a rat spinal cord injury model with the licensed drug trifluoperazine inhibited AQP4 localization to the blood-spinal cord barrier, ablated CNS edema, and led to accelerated functional recovery compared with untreated animals. We propose that targeting the mechanism of calmodulin-mediated cell-surface localization of AQP4 is a viable strategy for development of CNS edema therapies.
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| 9. |
- Kitchen, Philip, et al.
(author)
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Water channel pore size determines exclusion properties but not solute selectivity
- 2019
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In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 9:1
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Journal article (peer-reviewed)abstract
- Aquaporins (AQPs) are a ubiquitous family of transmembrane water channel proteins. A subgroup of AQP water channels also facilitates transmembrane diffusion of small, polar solutes. A constriction within the pore, the aromatic/arginine (ar/R) selectivity filter, is thought to control solute permeability: previous studies on single representative water channel proteins suggest narrow channels conduct water, whilst wider channels permit passage of solutes. To assess this model of selectivity, we used mutagenesis, permeability measurements and in silico comparisons of water-specific as well as glycerol-permeable human AQPs. Our studies show that single amino acid substitutions in the selectivity filters of AQP1, AQP4 and AQP3 differentially affect glycerol and urea permeability in an AQP-specific manner. Comparison between in silico-calculated channel cross-sectional areas and in vitro permeability measurements suggests that selectivity filter cross-sectional area predicts urea but not glycerol permeability. Our data show that substrate discrimination in water channels depends on a complex interplay between the solute, pore size, and polarity, and that using single water channel proteins as representative models has led to an underestimation of this complexity.
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| 10. |
- Larsson, Andreas, et al.
(author)
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Collaborative Healthcare Innovation in Sweden
- 2012
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In: Sustainable Innovation. - New York : Springer. - 9781461420767 - 9781461420774 ; , s. 49-62
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Book chapter (peer-reviewed)abstract
- Abstract in UndeterminedThe medical technology (medtech) industry in Sweden is situated withina complex innovation ecosystem, in which various stakeholders from the public,private and academic sectors need to collaborate to meet demands on effective andeffi cient healthcare. Demographics are changing and those in need of healthcare arenot only larger in numbers than ever but they are also more knowledgeable anddemanding. Increasing innovative performance is crucial in both the private andpublic healthcare sectors, but bold steps forward need to be taken in light of stricterrules and regulations for how healthcare stakeholders should manage both theirinternal processes and the ways in which they interact with other stakeholders in thelarger innovation system. The traditional way in which medtech companies gainaccess to user needs, primarily working through a sales–purchasing relationshipwith the public healthcare sector, is outdated and needs to be replaced with anincreasingly collaborative and cocreative model of healthcare innovation.This chapter describes experiences and lessons learned from InnoPlant, a 3-year(2008–2011) action learning project involving three companies from the Swedishmedtech industry, two county/regional councils responsible for public healthcare,and four academic institutions—carried out within the framework of the SwedishProduct Innovation Engineering program (PIEp). The purpose of the project was toadvance the capability of stakeholders from the public, private, and academic sectorsto collaborate in the cocreation of healthcare innovations.
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