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- de Oliveira, Felipe Marques Souza
(författare)
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Development and Application of Proximity Assays for Proteome Analysis in Medicine
- 2018
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Along with proteins, a myriad of different molecular biomarkers, such as post-translational modifications and autoantibodies, could be used in an attempt to improve disease detection and progression. In this thesis, I build on several iterations of the proximity ligation assay to develop and apply new adaptable methods to facilitate detection of proteins, autoantibodies and post-translational modifications.In paper I, we present an adaptation of the solid-phase proximity ligation assay (SP-PLA) for the detection of post-translational modification of proteins (PTMs). The assay was adapted for the detection of two of the most commons PTMs present in proteins, glycosylation and phosphorylation, offering the encouraging prospect of using detection of PTMs in a diagnostic or prognostic capacity. In paper II, we developed a variant of the proximity ligation assay using micro titer plate for detection and quantification of protein using optical density as readout in the fluorometer, termed PLARCA. With a detection limit considerably lower than ELISA, PLARCA detected femtomolar levels of these proteins in patient samples.In paper III, we aim to compare detection values of samples collected from earlobe capillary, venous plasma, as well as capillary plasma stored in dried plasma spots (DPS) assessed with a 92-plex inflammation panel using multiplex proximity extension assay (PEA). Despite the high variability in protein measurements between the three sample sources, we were able to conclude that earlobe capillary sampling is a suitable less invasive alternative, to venipuncture.In paper IV, we describe the application of PLARCA and proximity extension assay (PEA) for the detection of GAD65 autoantibodies (GADA). Thus, offering highly sensitive and specific autoimmunity detection.
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| 2. |
- Gullberg, Mats, 1953-
(författare)
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Health care professionals' self description
- 1996
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- The aim of the present thesis was to survey and analyse professional self of health care professionals, focusing on perceptions of self, and on their attitudes towards aspects of sense of responsibility.Studies were accomplished with quantitative techniques using questionnaires and/or instruments. A total of 803 respondents was included, and the first study (I) presented the development of the PSDF and comprised 30 registered nurses. The next study (II) examined results from 332 medical laboratory technologists who answered the PSDF. In the third study (III) 68 occupational therapists responded to the PSDF. The fourth study (IV) was a six-year follow-up of 43 registered nurses from the beginning of their training to three years after graduation, and they responded to the PSDF, the SRF and to the Form for ranked sources of encouragement. The fifth study (V) comprised four professional groups from a university hospital in Sweden (physicians, physiotherapists, registered nurses and enrolled nurses), and 312 responded to the set of forms (the PSDF, the SRF, and the Form for ranked sources of encouragement).The main theoretical approach was based on symbolic interactionism, where the self is a significant notion. Concepts on attitude, sense of responsibility and encouragement were important.The results showed that there are components of self description that is 'specific to one professional group, but also components that uncover similarities. Three important components of professional self were outlined; professional conduct, work ability and empathy. Important issues were also encouragement for work done, which influences the professional self, and that the positive attitudes to sense of responsibility showed similarities with characteristics of professional self.In conclusion, two propositions were addressed: that professionals' self description is composed of three meaningful dimensions; professional conduct, work ability and empathy, and that the professional self can be improved throughdevelopment of intrinsic attitudes and behaviours.
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| 3. |
- Gullberg, Mats, 1971-
(författare)
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Proximity Ligation as a Universal Protein Detection Tool
- 2003
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Among the great challenges in biology are the precise quantification of specific sets of proteins and analyses of their patterns of interaction on a much larger scale than is possible today.This thesis presents a novel protein detection technique - proximity ligation - and reports the development and application of a nucleic acid amplification technique, RCA. Proximity ligation converts information about the presence or co-localization of specific proteins to unique sets of nucleic acid sequences. For detection of target proteins or protein complexes the coincident binding by pairs or triplets of specific protein-binding reagents are required. Oligonucleotide-extensions attached to those binding reagents are joined by a DNA ligase and subsequently analyzed by standard molecular genetic techniques. The technique is shown to sensitively detect an assortment of proteins using different types of binders converted to proximity probes, including SELEX aptamers and mono- and polyclonal antibodies. I discuss factors important for using the technique to analyze many proteins simultaneously.Quantification of target molecules requires precise amplification and detection. I show how rolling circle amplification, RCA, can be used for precise quantification of circular templates using modified molecular beacons with real-time detection. The combination of proximity-probe templated circularization and RCA results in a sensitive method with high selectivity, capable of visualizing individual immobilized proteins. This technique is used for localized detection of a set of individual proteins and protein complexes at sub-cellular resolution.
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| 4. |
- Lundin, Per, 1971-
(författare)
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Bilsamhället : Ideologi, expertis och regelskapande i efterkrigstidens Sverige
- 2008
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- During the 1950s the number of cars in Sweden increased almost fivefold and the country attained the highest level of car ownership per capita in Europe. The rise and establishment of mass motoring was dramatically illustrated by the violent encounter between the car and the city. Despite the fact that congestion as well as road accidents were well-known, everyday occurrences, they reached previously unimagined heights through the growth of mass motoring. In this doctoral thesis Per Lundin focuses on the emergence of a group of planning experts as the key advocates of the idea of the “car society” as the solution to these problems. By fully adapting society to the car it would be possible to eliminate congestion and road accidents, thus affirming the continuing advance of the car. This ideal, which originated in the United States, became the goal and the dream of these experts.The general question addressed by Lundin is to what extent the actions and the ideologies of the experts interacted with the advent of mass motoring and the extensive urban building during the post-war period. In order to answer this question Lundin analyzes, firstly how the planning experts laid claim to the problems of congestion and road accidents, thereby restating them as exclusive planning problems, secondly how guidelines and standards for a car-conscious planning of cities and communities were developed based on the formulation of this problem and thirdly how, and to what extent, the guidelines and standards concerned were implemented in the town planning process.In the thesis Lundin argues that the dreams about a post-war Swedish society entirely adapted to the car by and large were realized. One important explanation of the fact that the physical adaptation of cities and societies to the car could proceed so quickly, on such a large scale and in similar forms all over the country, is found in the planning rules developed by the experts. The rules were the embodiment of the untroubled and unreflecting dreams nourished by the planning experts of the 1950s and the 1960s. Through the rules these ideological conceptions were reinforced and disseminated in a manner almost unable to stop. As the rules quickly were integrated with the planning instruments of administrative bodies locally, regionally and nationally, they set the tone for the extensive urban renewal of the following decades.
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