| 11. |
- Almqvist, Helena, et al.
(author)
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CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil
- 2016
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In: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 7
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Journal article (peer-reviewed)abstract
- Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular target binding, exemplified by human thymidylate synthase (TS). The screen selected accurately for all the tested known drugs acting on TS. We also identified TS inhibitors with novel chemistry and marketed drugs that were not previously known to target TS, including the DNA methyltransferase inhibitor decitabine. By following the cellular uptake and enzymatic conversion of known drugs we correlated the appearance of active metabolites over time with intracellular target engagement. These data distinguished a much slower activation of 5-fluorouracil when compared with nucleoside-based drugs. The approach establishes efficient means to associate drug uptake and activation with target binding during drug discovery.
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| 12. |
- Backman, Helena, et al.
(author)
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Severe asthma : A population study perspective
- 2019
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In: Clinical and Experimental Allergy. - : John Wiley & Sons. - 0954-7894 .- 1365-2222. ; 49:6, s. 819-828
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Journal article (peer-reviewed)abstract
- BackgroundSevere asthma is a considerable challenge for patients, health care professionals and society. Few studies have estimated the prevalence of severe asthma according to modern definitions of which none based on a population study.ObjectiveTo describe characteristics and estimate the prevalence of severe asthma in a large adult population‐based asthma cohort followed for 10‐28 years.MethodsN=1006 subjects with asthma participated in a follow‐up during 2012‐14, when 830 (mean age 59y, 56% women) still had current asthma. Severe asthma was defined according to three internationally well‐known criteria: the ATS workshop definition from 2000 used in the US Severe Asthma Research Program (SARP), the 2014 ATS/ERS Task force definition and the GINA 2017. All subjects with severe asthma according to any of these criteria were undergoing respiratory specialist care, and were also contacted by telephone to verify treatment adherence.ResultsThe prevalence of severe asthma according to the three definitions was 3.6% (US SARP), 4.8% (ERS/ATS Taskforce), and 6.1% (GINA) among subjects with current asthma. Although all were using high ICS doses and other maintenance treatment, >40% had uncontrolled asthma according to the asthma control test. Severe asthma was related to age >50 years, nasal polyposis, impaired lung function, sensitization to aspergillus, and tended to be more common in women. Further, neutrophils in blood significantly discriminated severe asthma from other asthma.Conclusions and clinical relevanceSevere asthma differed significantly from other asthma in terms of demographic, clinical and inflammatory characteristics, results suggesting possibilities for improved treatment regimens of severe asthma. The prevalence of severe asthma in this asthma cohort was 4‐6%, corresponding to approximately 0.5% of the general population.
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| 13. |
- Kalsum, Sadaf, 1977-
(author)
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Characterizing phenotypes of Mycobacterium tuberculosis and exploring anti-mycobacterial compounds through high content screening
- 2018
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Doctoral thesis (other academic/artistic)abstract
- Tuberculosis (TB), an airborne disease and one of the top 10 causes of death globally, is caused by Mycobacterium tuberculosis (Mtb). Current standard therapy for TB treatment includes multiple drugs for a period of at least 6 months. The long therapy duration is to sterilize a small sub-population of drug-tolerant bacteria, a characteristic related to biofilm formation, which otherwise responsible for disease relapse. On the other hand, because of such a long treatment period, patient adherence to therapy becomes difficult, which results in the emergence of multidrug-resistant (MDR) or, in worst cases, extensively drug-resistant (XDR)-TB. TB is primarily a disease of lungs and alveolar macrophages are one of the first host cell types to encounter Mtb following aerosol transmission. A well-established role of macrophages in immune defense is phagocytosis, but recent studies also demonstrated that upon interaction with large aggregates of microbes or cord-forming mycobacterial species, macrophages could produce extracellular traps known as macrophage extracellular traps (METs). METs have a DNA backbone with embeds histones and could trap a wide range of microorganisms, but may or may not be able to kill them. Natural products are always a promising starting point for drug discovery because of their wide range of activity. A large number of world’s population is still using extracts from different parts of plants as the primary source of medicines against diseases including TB. Today much effort is being invested by academia in screening campaigns that allows for fast discovery of new active compounds. Thanks to the use of automated technology such as automated microscopy or automated image analysis (known as high content screening, HCS) phenotypic drug discovery has become easier to perform. Therefore, the identification of highly effective compounds to combat infectious diseases like TB can be facilitated by the use of host-pathogen assays at the early stages of drug screening studies.This thesis describes the characterization and antibiotic sensitivity of different phenotypes of Mtb namely planktonic, cord-forming and biofilm-producing phenotypes that arise due to different culture conditions. The culture of Mtb with a high percentage of a detergent (Tween-80) and standing condition promoted planktonic phenotype while a culture with a low amount of Tween-80 and more aeration due to shaking promoted cording and biofilm phenotypes. Primary human macrophages upon interaction with the shaken culture of wild-type Mtb died by releasing METs. Whereas, the shaken cultures of early secreted antigenic target-6 (ESAT-6), an important virulence factor of Mtb, deletion mutant strain could not induce MET formation showing that the cord formation is related to virulence. Moreover, the biofilm phenotype of Mtb is more tolerant to two first-line antibiotics isoniazid (INH) and rifampicin (RIF) as compared to cording and planktonic phenotypes which demand a search of more effective TB therapy. A screening campaign based on a whole-cell assay using different ethanolic crude extracts of many African plants lead to the discovery of a hit, i.e., a chloroform fraction of Khaya senegalensis bark, which showed non-significant inhibition of intracellular growth of a virulent strain of Mtb was selected for further purification and evaluation. Lastly, we have also developed and validated an HCS assay to explore new compounds against intracellular Mtb in human macrophages. INH and RIF, which were found most effective in our system were used in a combination as a positive control to calculate a Z’ factor value, which confirmed our assay to be suitable for HCS.In conclusion, this thesis not only highlights the biology of TB infection, but also discusses the development of a pathophysiologically relevant assay that can be used in the identification of novel compound(s) that has either direct anti-mycobacterial activity (antibiotic), acts by stimulating the host cell immune mechanisms (immunomodulator) or acts by counteracting virulence factors (virulence blocker).
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| 14. |
- Lundbäck, Andreas, et al.
(author)
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Modeling and Experimental Measurement with Synchrotron Radiation of Residual Stresses in Laser Metal Deposited Ti-6Al-4V
- 2016
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In: Proceedings of the 13th World Conference on Titanium. - Hoboken, NJ, USA : John Wiley & Sons, Inc.. - 9781119293668 - 9781119296126 ; , s. 1279-1282
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Conference paper (peer-reviewed)abstract
- There are many challenges in producing aerospace components by additive manufacturing (AM). One of them is to keep the residual stresses and deformations to a minimum. Another one is to achieve the desired material properties in the final component. A computer model can be of great assistance when trying to reduce the negative effects of the manufacturing process. In this work a finite element model is used to predict the thermo-mechanical response during the AM-process. This work features a physically based plasticity model coupled with a microstructure evolution model for the titanium alloy Ti -6Al-4V. Residual stresses in AM components were measured non-destructively using high-energy synchrotron X-ray diffraction on beam line ID15A at the ESRF, Grenoble. The results are compared with FE model predictions of residual stresses. During the process, temperatures and deformations was continuously measured. The measured and computed thermal history agrees well. The result with respect to the deformations agrees well qualitatively. Meaning that the change in deformation in each sequence is well predicted but there is a systematic error that is summing so that the quantitative agreement is lost.
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| 15. |
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| 16. |
- Lundbäck, Andreas, et al.
(author)
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Modelling and Simulation of Metal Deposition on a Ti-6al-4v Plate
- 2015
-
Conference paper (other academic/artistic)abstract
- There are many challenges in producing aerospace components by metal deposition (MD). One of them is to keep the residual stresses and deformations to a minimum. Anotherone is to achieve the desired material properties in the final component. A computer model can be of great assistance when trying to reduce the negative effects of the manufacturing process. In this work a finite element model is used to predict the thermo-mechanical response during the MD-process. This work features a pysically based plasticity model coupled with a microstructure evolution model for the titanium alloy Ti-6Al-4V. A thermally driven microstructure model is used to derive the evolution of the non-equilibrium compositions of α-phases and β-phase. Addition of material is done by activation of elements. The method is taking large deformations into consideration and adjusts the shape and position of the activated elements. This is particularilly important when adding material onto thin and flexible structures. The FE-model can be used to evaluate the effect of different welding sequenses. Validation of the model is performed by comparing measured deformations, strains, residual stresses and temperatures with the computed result. The deformations, strains and temepratures are measured during the process. The deformations are measured with a LVDT-gauge at one location. The strains are measured with a strain gauge at the same location as the deformations. The temperature is measured at five locations, close to the weld and with an increasing distance of one millimeter between each thermo couple. The residual stresses in MD component were measured non-destructively using high-energy synchrotron X-ray diffraction on beam line ID15A at the ESRF, Grenoble.
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| 17. |
- Mateus, André, 1986-, et al.
(author)
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Prediction of intracellular exposure bridges the gap between target- and cell-based drug discovery
- 2017
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In: Proceedings of the National Academy of Sciences of the United States of America. - : NATL ACAD SCIENCES. - 0027-8424 .- 1091-6490. ; 114:30, s. E6231-E6239
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Journal article (peer-reviewed)abstract
- Inadequate target exposure is a major cause of high attrition in drug discovery. Here, we show that a label-free method for quantifying the intracellular bioavailability (F-ic) of drug molecules predicts drug access to intracellular targets and hence, pharmacological effect. We determined F-ic in multiple cellular assays and cell types representing different targets from a number of therapeutic areas, including cancer, inflammation, and dementia. Both cytosolic targets and targets localized in subcellular compartments were investigated. F-ic gives insights on membrane-permeable compounds in terms of cellular potency and intracellular target engagement, compared with biochemical potency measurements alone. Knowledge of the amount of drug that is locally available to bind intracellular targets provides a powerful tool for compound selection in early drug discovery.
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| 18. |
- Mubeen, Saad, et al.
(author)
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Developing predictable vehicular distributed embedded systems on multi-core
- 2016
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In: Advances in Intelligent Systems and Computing, Volume 448. - Cham : Springer International Publishing. - 9783319324661 ; , s. 1273-1277
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Conference paper (peer-reviewed)abstract
- In this paper we address the challenges related to supporting modeland component-based development of predictable software for vehicular distributed embedded systems, utilizing multi-core platforms. We present a research plan for the development of new methods and techniques to deal with these challenges. The techniques will support various aspects such as modeling of the software architecture; supporting multiple criticality levels; verifying predictability of the system using end-to-end timing analysis; code generation; and providing a predictable runtime support on multi-core platforms by virtualizing a certified single-core real-time operating system. As a proof of concept, we will implement the newly developed techniques in a commercial tool chain (Rubus-ICE). The efficacy of the newly developed techniques and the extended tool chain will be demonstrated on the industrial case studies.
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| 19. |
- Perzanowski, Matthew S, et al.
(author)
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Relevance of specific IgE antibody titer to the prevalence, severity, and persistence of asthma among 19-year-olds in northern Sweden
- 2016
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In: Journal of Allergy and Clinical Immunology. - : Elsevier BV. - 0091-6749 .- 1097-6825. ; 138:6, s. 1582-1590
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Journal article (peer-reviewed)abstract
- BACKGROUND: Although sensitization to indoor allergens is strongly associated with asthma, there are questions as to how this relates to asthma symptoms.OBJECTIVE: We sought to study the relevance of IgE antibodies to cat and dog allergens in an area in which (1) the climate discourages cockroach, fungal, and mite growth and (2) dander allergens are known to be present in schools and houses without animals.METHODS: IgE to 8 allergens was tested in 963 sera from a population-based study on 19-year-olds, and associations with asthma symptoms, diagnosis, and treatment were examined. In positive sera IgE to specific cat and dog allergens was also assayed.RESULTS: IgE specific for animal dander had the highest prevalence and strongest relationship to asthma diagnosis. Furthermore, asthma severity, as judged by the frequency of symptoms and use of treatment, was directly associated with the titer of IgE antibodies to animal dander. Among the 103 subjects who had current asthma at age 19 years, 50 had asthma before age 12 years. Among those 50, the odds ratios for asthma related to any IgE antibodies to animal dander or high-titer IgE antibodies (≥17.5 IU/mL) were 9.2 (95% CI, 4.9-17) and 13 (95% CI, 6.9-25), respectively. In multivariable analysis IgE antibodies to Fel d 1 and Can f 5 were each associated with current asthma.CONCLUSION: High-titer IgE antibodies to cat and dog allergens were strongly associated with the diagnosis, severity, and persistence of asthma; however, a large proportion of patients with current asthma did not live in a house with a cat or dog.
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| 20. |
- Porsbjerg, C., et al.
(author)
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Nordic consensus statement on the systematic assessment and management of possible severe asthma in adults
- 2018
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In: European Clinical Respiratory Journal. - : Informa UK Limited. - 2001-8525. ; 5:1
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Journal article (peer-reviewed)abstract
- Although a minority of asthma patients suffer from severe asthma, they represent a major clinical challenge in terms of poor symptom control despite high-dose treatment, risk of exacerbations, and side effects. Novel biological treatments may benefit patients with severe asthma, but are expensive, and are only effective in appropriately targeted patients. In some patients, symptoms are driven by other factors than asthma, and all patients with suspected severe asthma ('difficult asthma') should undergo systematic assessment, in order to differentiate between true severe asthma, and 'difficult-to-treat' patients, in whom poor control is related to factors such as poor adherence or co-morbidities. The Nordic Consensus Statement on severe asthma was developed by the Nordic Severe Asthma Network, consisting of members from Norway, Sweden, Finland, Denmark, Iceland and Estonia, including representatives from the respective national respiratory scientific societies with the aim to provide an overview and recommendations regarding the diagnosis, systematic assessment and management of severe asthma. Furthermore, the Consensus Statement proposes recommendations for the organization of severe asthma management in primary, secondary, and tertiary care.
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