| 1. |
- Lönn, Peter, et al.
(författare)
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High-throughput in situ mapping of phosphorylated protein complexes across the cell cycle and in response to drugs
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Interactions and posttranslational modifications (PTMs) of proteins orchestrate cellular responses to cytokines, drugs or other agents, but it has been difficult to monitor and characterize these dynamic events at high-throughput. Here, we have established a semi-automated system for large-scale in situ proximity ligation assays (isPLA). The protocol combines isPLA in microtiter wells with automated microscopy and computer-based image analysis whereby specific protein phosphorylations and interactions are digitally recorded in cells, along with measurements of morphological features. We demonstrate how this platform can improve analysis of cellular signaling by investigating TGF-b responsive Smad2 linker phosphorylations and complex formations over time and across millions of individual cells. We depict single cell responses in relation to e.g. local cell crowding and cell cycle progression via measurements of DNA content and nuclear size. Finally, we illustrate the application of the protocol for demonstrating drug effects by screening a library of phosphatase inhibitors. In summary, our approach expands the scope for image-based single cell analyses by combining observations of protein interactions and modifications with morphological details of individual cells at high throughput.
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| 2. |
- Lönn, Peter, et al.
(författare)
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Image-based high-throughput mapping of TGF-beta-induced phosphocomplexes at a single-cell level
- 2021
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Ingår i: Communications Biology. - : Springer Nature. - 2399-3642. ; 4
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Tidskriftsartikel (refereegranskat)abstract
- To improve our ability to monitor cellular responses to e.g. cytokines or drugs, Lonn et al have developed a semi-automated system for large-scale in situ proximity ligation assays (isPLA) in HaCAT keratinocyte cells. Their approach expands the scope for image-based single cell analyses by combining observations of protein interactions and modifications with morphological details of individual cells at high throughput. Protein interactions and posttranslational modifications orchestrate cellular responses to e.g. cytokines and drugs, but it has been difficult to monitor these dynamic events in high-throughput. Here, we describe a semi-automated system for large-scale in situ proximity ligation assays (isPLA), combining isPLA in microtiter wells with automated microscopy and computer-based image analysis. Phosphorylations and interactions are digitally recorded along with subcellular morphological features. We investigated TGF-beta-responsive Smad2 linker phosphorylations and complex formations over time and across millions of individual cells, and we relate these events to cell cycle progression and local cell crowding via measurements of DNA content and nuclear size of individual cells, and of their relative positions. We illustrate the suitability of this protocol to screen for drug effects using phosphatase inhibitors. Our approach expands the scope for image-based single cell analyses by combining observations of protein interactions and modifications with morphological details of individual cells at high throughput.
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| 3. |
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| 4. |
- Backman Lönn, Beatrice, et al.
(författare)
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Clarifying the role of clinical research nurses working in Sweden, using the clinical trial nursing questionnaire : swedish version
- 2022
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Ingår i: Nursing Open. - : John Wiley & Sons. - 2054-1058. ; 9:5, s. 2434-2443
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Tidskriftsartikel (refereegranskat)abstract
- Aim: To explore the role of CRNs in Sweden and differences in competences and tasks, using the Clinical Trial Nursing Questionnaire - Swedish version (CTNQ-SWE).Design: A cross-sectional survey.Methods: Participants were identified through strategic sampling. Data were analysed by descriptive and comparative statistics.Results: The respondents were experienced nurses who felt proficient in their role, they felt more acceptance by the principal investigators than by nursing colleagues. A majority of CRNs are involved in all procedures specified in the CTNQ-SWE. The most often performed tasks, also rated as the most important by the CRNs, concerned informed consent and management of investigational products. The education was often informal: with a lack of job descriptions and professional development plans. Need of formal specialist education was expressed.Conclusions: Knowledge about the role description can be used by clinical research enterprise internationally and healthcare organizations aiming to support CRNs in their role.
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| 5. |
- Backman Lönn, Beatrice, 1974-, et al.
(författare)
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Clinical research nurses perceive their role as being like the hub of a wheel without real power : Empirical qualitative research
- 2024
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Ingår i: Nursing Open. - : John Wiley & Sons. - 2054-1058. ; 11:5
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Tidskriftsartikel (refereegranskat)abstract
- Aim: This study aimed to explore the experiences of being a clinical research nurse (CRN), in Sweden.Design: A qualitative study analysing individual interview data.Methods: Interviews with 10 participants were conducted in April 2017 and repeated with five participants in May 2022. A semi-structured interview guide was used to cover topics such as experiences of working in a new role and professional challenges related to the role. The transcribed interviews were analysed inductively using qualitative content analysis.Results: The main theme revealed that the CRNs experienced their work role as being like a hub in a wheel, using an ethical compass, but without real power. The six themes identified showed that CRNs worked independently and relied on clinical experiences as nurses but needed more education. They not only had a sense of duty but also too large responsibilities. Furthermore, they viewed their work as valuable and important. However, they needed an accentuated ethical compass and were also affected by power relations that negatively impacted work.Conclusion: Working as a CRN means being in a central position and working independently, which requires diverse skills and competencies. CRNs, however, face and manage complex ethical and practical challenges without real power. They experience huge responsibilities but need education and acknowledgement, indicating a need for improvement. This is an important message to stakeholders and managers about the necessity of taking adequate action to support CRNs who are crucial resources in clinical research.Patient or Public Contribution: No patient or public contribution.
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| 6. |
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| 7. |
- Backman Lönn, Beatrice, 1974-
(författare)
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The registered nurse as a clinical research nurse
- 2023
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Clinical research studies are important for the developmentof new treatments in healthcare. The quality of clinical research relies on the competence, skills, and knowledge of the research team. Clinical research nurses (CRNs) are important members of clinical research teams as they are responsible for various tasks specified in study procedures. Internationally, nurses have been engaged as CRNs in in many areas of medicine, such as oncological research, for several decades. However, there is a lack of consensus concerning their professional role and a clear work description is lacking. In Sweden, nurses are becoming increasingly involved in research as CRNs, yet the CRN role is undefined as there is no Swedish national competence description that could guide CRNs’ work tasks, requirements for education, and management of ethical issues. Furthermore, the processof becoming a CRN is not understood. Increasing international research into the CRN role has shown that the role differs between countries.There is a lack of Swedish studies, including studies of the process of transitioning from being a registered nurse (RN) to a CRN. Since consensus is lacking concerning the CRN role in Sweden, we need to explore it further and examine how nursing perspectives are influencing it. The overall aim of this thesis was therefore to explore the professional role of CRNs in Sweden and the transition of RNs to being CRNs.Methods: Both quantitative (studies I and II) and qualitative (studies III and IV) methods were used. The analyses are based on data from questionnaires and individual interviews of CRNs in Sweden. In study I, descriptive statistics and test–retest analyses were used to analyze the validity and reliability of the translated CTNQ-SWE. In study II, descriptive statistics and one-way ANOVA were used to analyze reported work tasks, perceptions, and differences between groups of CRNs. In studies III and IV, qualitative content analysis according to Graneheimet al., (2004) was used to analyze the transition, i.e., becoming and being a CRN.Results: CRNs perform new and diverse work tasks and are often involved throughout the study process, with the greatest activity in data management and the actual conducting and evaluation of clinical studies (e.g., scheduling and performing procedures and tests according to the research protocols). There is seldom any work description or competence framework to guide nurses through the transition to the CRN role, placing them in an uncertain position where they must struggle to adaptiv to their new role and work tasks. To perform these new work activities, CRNs need more support as well as education in research procedures, regulations, and ethics. During the transition, CRNs improve their knowledge of research, regulations, and ethics and increase their experience of clinical research practice and collaboration in networks. They also learn and develop from dealing with challenging situations, such as the informed consent process. Furthermore, problem solving, study requirements, and ethical reasoning are emphasized as challenging. When passing through the different phases of transition to the CRN role, the nurses achieve expanded competence, change didentity, and growing confidence as informal leaders. CRNs also advocate for patient rights and mentor others involved in research, but they simultaneously lack acknowledgement and a formal leadership role.Conclusions and clinical implications: This thesis provides important insights into the CRN role in Sweden. It highlights the transition of RNs to becoming CRNs as well as their experiences of working as and becoming CRNs. The thesis reports that CRNs, overall, seem to be satisfied with their work, but some of their experiences highlight that skills, education, and organizational structures require improvement; forexample, appropriate introduction and support are required during the transition to the CRN role. Ethical challenges call for ongoing discussion in research teams. One conclusion is that CRNs’ competences and tasks need to be clarified. Developing clear competence pathways for nurses to become CRNs, including mentorship and support, could be one way of acknowledging CRNs’ important work, thereby creating a better outlook for high-quality clinical research procedures. Making the CRN professional title more homogeneous nationally and internationally would make comparisons easier. This would clarify CRNs’ work tasks and role in Sweden and extend the nursing perspective on ethics. This thesis adds to a slowly growing body of literature on the CRN role and is relevant to both practitioners and policymakers. Additionally, it illustrates a need to develop both educational and mentoring support aswell as career pathways for RNs to become CRNs, which could improve the quality of clinical research in Sweden.
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| 8. |
- Backman Lönn, Beatrice, et al.
(författare)
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Transitioning to the clinical research nurse role : a qualitative descriptive study
- 2022
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Ingår i: Journal of Advanced Nursing. - : John Wiley & Sons. - 0309-2402 .- 1365-2648. ; 78:11, s. 3817-3829
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Tidskriftsartikel (refereegranskat)abstract
- Background: Studies have reported on the important role of the clinical research nurse in clinical studies. Yet, there is no international consensus about the role's competencies and tasks. Furthermore, the literature offers a little description of the career pathway from a ward-based registered nurse to a clinical research nurse. More knowledge about this specific role could benefit the nursing profession as well as increase the quality of clinical research.Aim: The aim of the study was to explore Swedish registered nurses' experiences transitioning into the clinical research nurse role.Design: The study had a qualitative design. Data were collected via semi-structured interviews. Inductive qualitative content analysis was employed.Methods: Ten participants (i.e., clinical research nurses) were interviewed in the spring of 2017. A semi-structured interview guide was used to address the transition into the clinical research nurse role, experience working in a new role, experience of ethical dilemmas and experience of organizational and professional issues related to the role. The interviews were analysed inductively using qualitative content analysis.Results: The registered nurses described experiencing reality shock when they became clinical research nurses; that is, it was a challenging and transforming experience. The main theme, a challenging transition, was developed from the four subthemes highlighting that it defied their previous nursing role. They experienced an unclear professional identity, extended professional mandate, increased professional status and growing ethical consciousness in their new role.Conclusion: The results highlight that registered nurses who became clinical research nurses had needs that were both distinct from and overlapped with those of their former professional role as registered nurses. To avoid reality shocks, the development of clear competence pathways for nurses to become clinical research nurses, including introduction, mentorship and continued support, is necessary. Making their professional title more homogeneous, nationally and internationally, would facilitate role identification and comparisons in research.
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| 9. |
- Berntsson, Lars-Olof, et al.
(författare)
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EAST-ADL 2.0 Specification
- 2008
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Rapport (refereegranskat)abstract
- This specification of the EAST ADL 2.0 is based on the EAST-ADL developed in the EAST EEA projectand has been further refined and harmonized with on-going modelling appraches in the automotiveindustry. It presents the modeling infrastructure, i.e. how the modeling elements should be represented inthe language and the UML representation. For each package a usage example is provided.The EAST-ADL 2.0 is harmonized with AUTOSAR.The metamodel and UML profile of EAST ADL 2.0 is defined in two steps: A domain (automotive)metamodel is defined, capturing only the domain specific needs of the language, without adding the UML2details. The basic concepts of UML are used for this purpose, such as classes, compositions andassociations. Based on the domain metamodel, a UML2 profile for the domain metamodel is defined,specifying stereotypes with properties and constraints.Comments on the content of this document are welcomed, and should be directed to .Please download the latest available specification and the XMI file ready for use in UML2 tools from the website.
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| 10. |
- Björkesten, Johan, et al.
(författare)
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A multiplex platform for digital measurement of circular DNA reaction products
- 2020
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Ingår i: Nucleic Acids Research. - : OXFORD UNIV PRESS. - 0305-1048 .- 1362-4962. ; 48:13
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Tidskriftsartikel (refereegranskat)abstract
- Digital PCR provides high sensitivity and unprecedented accuracy in DNA quantification, but current approaches require dedicated instrumentation and have limited opportunities for multiplexing. Here, we present an isothermal platform for digital enumeration of DNA reaction products in multiplex via standard fluorescence microscopy. Circular DNA strands, which may result from a wide range of molecular detection reactions, are captured on streptavidin-coated surfaces via hybridized biotinylated primers, followed by rolling circle amplification (RCA). The addition of 15% polyethylene glycol 4000 during RCA resulted in uniform, easily recorded reaction products. Immobilized DNA circles were visualized as RCA products with 100% efficiency, as determined by droplet digital PCR. We confirmed previous reports about the influence on RCA by sequence composition and size of RCA templates, and we developed an efficient one-step restaining procedure for sequential multiplexing using toehold-triggered DNA strand displacement. Finally, we exemplify applications of this digital readout platform by demonstrating more than three orders of magnitude improved sensitivity by digital measurement of prostate specific antigen (PSA) (detection threshold similar to 100 pg/l), compared to a commercial enzyme-linked immunosorbent assay (ELISA) with analogue readout (detection threshold similar to 500 ng/l), using the same antibody pair.
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