| 1. |
- Wagner, Samuel, et al.
(författare)
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Tuning Escherichia coli for membrane protein overexpression
- 2008
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 105:38, s. 14371-17376
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Tidskriftsartikel (refereegranskat)abstract
- A simple generic method for optimizing membrane protein overexpression in Escherichia coli is still lacking. We have studied the physiological response of the widely used “Walker strains” C41(DE3) and C43(DE3), which are derived from BL21(DE3), to membrane protein overexpression. For unknown reasons, overexpression of many membrane proteins in these strains is hardly toxic, often resulting in high overexpression yields. By using a combination of physiological, proteomic, and genetic techniques we have shown that mutations in the lacUV5 promoter governing expression of T7 RNA polymerase are key to the improved membrane protein overexpression characteristics of the Walker strains. Based on this observation, we have engineered a derivative strain of E. coli BL21(DE3), termed Lemo21(DE3), in which the activity of the T7 RNA polymerase can be precisely controlled by its natural inhibitor T7 lysozyme (T7Lys). Lemo21(DE3) is tunable for membrane protein overexpression and conveniently allows optimizing overexpression of any given membrane protein by using only a single strain rather than a multitude of different strains. The generality and simplicity of our approach make it ideal for high-throughput applications.
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| 2. |
- Apweiler, Rolf, et al.
(författare)
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Approaching clinical proteomics : current state and future fields of application in cellular proteomics
- 2009
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Ingår i: Cytometry. Part A : the journal of the International Society for Analytical Cytology. - : Wiley. - 1552-4922. ; 75A:10, s. 816-832
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Forskningsöversikt (refereegranskat)abstract
- Recent developments in proteomics technology offer new opportunities for clinical applications in hospital or specialized laboratories including the identification of novel biomarkers, monitoring of disease, detecting adverse effects of drugs, and environmental hazards. Advanced spectrometry technologies and the development of new protein array formats have brought these analyses to a standard, which now has the potential to be used in clinical diagnostics. Besides standardization of methodologies and distribution of proteomic data into public databases, the nature of the human body fluid proteome with its high dynamic range in protein concentrations, its quantitation problems, and its extreme complexity present enormous challenges. Molecular cell biology (cytomics) with its link to proteomics is a new fast moving scientific field, which addresses functional cell analysis and bioinformatic approaches to search for novel cellular proteomic biomarkers or their release products into body fluids that provide better insight into the enormous biocomplexity of disease processes and are suitable for patient stratification, therapeutic monitoring, and prediction of prognosis. Experience from studies of in vitro diagnostics and especially in clinical chemistry showed that the majority of errors occurs in the preanalytical phase and the setup of the diagnostic strategy. This is also true for clinical proteomics where similar preanalytical variables such as inter- and intra-assay variability due to biological variations or proteolytical activities in the sample will most likely also influence the results of proteomics studies. However, before complex proteomic analysis can be introduced at a broader level into the clinic, standardization of the preanalytical phase including patient preparation, sample collection, sample preparation, sample storage, measurement, and data analysis is another issue which has to be improved. In this report, we discuss the recent advances and applications that fulfill the criteria for clinical proteomics with the focus on cellular proteomics (cytoproteomics) as related to preanalytical and analytical standardization and to quality control measures required for effective implementation of these technologies and analytes into routine laboratory testing to generate novel actionable health information. It will then be crucial to design and carry out clinical studies that can eventually identify novel clinical diagnostic strategies based on these techniques and validate their impact on clinical decision making.
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| 3. |
- Apweiler, Rolf, et al.
(författare)
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Approaching clinical proteomics : current state and future fields of application in fluid proteomics
- 2009
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Ingår i: Clinical Chemistry and Laboratory Medicine. - 1434-6621 .- 1437-4331. ; 47:6, s. 724-744
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Forskningsöversikt (refereegranskat)abstract
- The field of clinical proteomics offers opportunities to identify new disease biomarkers in body fluids, cells and tissues. These biomarkers can be used in clinical applications for diagnosis, stratification of patients for specific treatment, or therapy monitoring. New protein array formats and improved spectrometry technologies have brought these analyses to a level with potential for use in clinical diagnostics. The nature of the human body fluid proteome with its large dynamic range of protein concentrations presents problems with quantitation. The extreme complexity of the proteome in body fluids presents enormous challenges and requires the establishment of standard operating procedures for handling of specimens, increasing sensitivity for detection and bioinformatical tools for distribution of proteomic data into the public domain. From studies of in vitro diagnostics, especially in clinical chemistry, it is evident that most errors occur in the preanalytical phase and during implementation of the diagnostic strategy. This is also true for clinical proteomics, and especially for fluid proteomics because of the multiple pretreatment processes. These processes include depletion of high-abundance proteins from plasma or enrichment processes for urine where biological variation or differences in proteolytic activities in the sample along with preanalytical variables such as inter- and intra-assay variability will likely influence the results of proteomics studies. However, before proteomic analysis can be introduced at a broader level into the clinical setting, standardization of the preanalytical phase including patient preparation, sample collection, sample preparation, sample storage, measurement and data analysis needs to be improved. In this review, we discuss the recent technological advances and applications that fulfil the criteria for clinical proteomics, with the focus on fluid proteomics. These advances relate to preanalytical factors, analytical standardization and quality-control measures required for effective implementation into routine laboratory testing in order to generate clinically useful information. With new disease biomarker candidates, it will be crucial to design and perform clinical studies that can identify novel diagnostic strategies based on these techniques, and to validate their impact on clinical decision-making.
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| 4. |
- Bienert, Gern, 2008, et al.
(författare)
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A subgroup of plant aquaporins facilitate the bi-directional diffusion of As(OH)3 and Sb(OH)3 across membranes
- 2008
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Ingår i: BMC Biology. - : Springer Science and Business Media LLC. - 1741-7007. ; 6:26
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Tidskriftsartikel (refereegranskat)abstract
- Background Arsenic is a toxic and highly abundant metalloid that endangers human health through drinking water and the food chain. The most common forms of arsenic in the environment are arsenate (As(V)) and arsenite (As(III)). As(V) is a non-functional phosphate analog that enters the food chain via plant phosphate transporters. Inside cells, As(V) becomes reduced to As(III) for subsequent extrusion or compartmentation. Although much is known about As(III) transport and handling in microbes and mammals, the transport systems for As(III) have not yet been characterized in plants. Results Here we show that the Nodulin26-like Intrinsic Proteins (NIPs) AtNIP5;1 and AtNIP6;1 from Arabidopsis thaliana, OsNIP2;1 and OsNIP3;2 from Oryza sativa, and LjNIP5;1 and LjNIP6;1 from Lotus japonicus are bi-directional As(III) channels. Expression of these NIPs sensitized yeast cells to As(III) and antimonite (Sb(III)), and direct transport assays confirmed their ability to facilitate As(III) transport across cell membranes. On medium containing As(V), expression of the same NIPs improved yeast growth, probably due to increased As(III) efflux. Our data furthermore provide evidence that NIPs can discriminate between highly similar substrates and that they may have differential preferences in the direction of transport. A subgroup of As(III) permeable channels that group together in a phylogenetic tree required N-terminal truncation for functional expression in yeast. Conclusion This is the first molecular identification of plant As(III) transport systems and we propose that metalloid transport through NIPs is a conserved and ancient feature. Our observations are potentially of great importance for improved remediation and tolerance of plants, and may provide a key to the development of low arsenic crops for food production.
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| 5. |
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| 6. |
- Boman, Johan, 1955, et al.
(författare)
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Elemental characterization of aerosols in urban and rural locations in Bangladesh
- 2005
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Ingår i: X-Ray Spectrometry. - : Wiley. - 0049-8246 .- 1097-4539. ; 34:5, s. 460-467
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Tidskriftsartikel (refereegranskat)abstract
- Bangladesh became the first country in South Asia to ban the use of lead in gasoline in 1999. In 2003, two-stroke engine auto rickshaws were banned in the city centre of Dhaka, the capital of Bangladesh and one of the Asian mega-cities, in an attempt to improve the air quality. In July and August 2003, fine aerosol particles (PM2.5) were collected in Dhaka and in Rangpur in the northern part of the country. The aim was to study the effect of the action and to characterize PM2.5 particles in urban and rural areas of Bangladesh. Sampling was done with a cyclone of Dewell-Higgins type equipped with Teflon filters with a pore size of 3 mu m. Day- and night-time filters were collected in an attempt to understand the possible difference in air pollution between the two periods. An energy-dispersive x-ray fluorescence (EDXRF) spectrometer with three-axial geometry was utilized for the analysis of the elemental content of the particles. Large variations of concentrations for most elements in both Dhaka and in Rangpur were observed. Most of the elemental concentrations in Dhaka in daytime were higher than at night whereas the opposite was the case for Rangpur. The concentrations showed a significant influence of human activities and weather. Considerable concentrations of Pb were found despite the official ban of leaded gasoline. Motor transport, especially the two-stroke engine taxis and elements remobilized by burning, were the major sources identified. This is the first report on elemental concentrations in PM2.5 particles in both Dhaka and Rangpur. Copyright (c) 2005 John Wiley & Sons, Ltd.
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| 7. |
- Boman, Johan, 1955, et al.
(författare)
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Elemental content of PM2.5 aerosol particles collected in Göteborg during the Göte-2005 campaign in February 2005
- 2008
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Ingår i: Atmos. Chem. Phys. Discuss. ; 8:2, s. 7703-7724
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Tidskriftsartikel (refereegranskat)abstract
- Göte–2005 was a measurement campaign in the city of Göteborg with the aim of studying the influence of the winter thermal inversions on urban air pollution. Elemental speciation of PM2.5 aerosol particles, collected on Teflon filters at three urban sites and one rural site in the Göteborg region, was a major part of the study. Trace element analysis was done by Energy Dispersive X-Ray Fluorescence (EDXRF) spectrometry and the concentrations of S, Cl, K, Ca, Ti, V, Mn, Fe, Ni, Cu, Zn, Br and Pb were determined. The elemental content of the particles in combination with information of local wind speed and direction and also backward trajectories were used to estimate the source areas of the pollutants. We can conclude that S, V, Ni, Br, and Pb have their main sources outside the Göteborg area, since we cannot see elevated concentrations of these elements during an inversion episode. Sea traffic and harbour activities were also identified, primarily by the S and V content of the particles. This study proves that the elemental analysis by EDXRF presents valuable information for tracing the origin of air masses arriving at a measurement site.
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| 8. |
- Boman, Johan, 1955, et al.
(författare)
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Elemental content of PM2.5 aerosol particles collected in Göteborg during the Göte-2005 campaign in February 2005
- 2009
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Ingår i: Atmospheric Chemistry and Physics. - : Copernicus GmbH. - 1680-7316 .- 1680-7324. ; 9, s. 2597-2606
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Tidskriftsartikel (refereegranskat)abstract
- The Göte-2005 measurement campaign aimed at studying the influence of the winter thermal inversions on urban air pollution. Elemental speciation of PM2.5 aerosol particles, collected on Teflon filters at three urban sites and one rural site in the Göteborg region, was a major part of the study. Trace element analysis was done by Energy Dispersive X-Ray Fluorescence (EDXRF) spectrometry and the concentrations of S, Cl, K, Ca, Ti, V, Mn, Fe, Ni, Cu, Zn, Br and Pb were determined. The elemental content of the particles, local wind speed and direction, and backward trajectories were used to investigate possible sources for the pollutants. We concluded that S, V, Ni, Br, and Pb had their main sources outside the central Göteborg area, since elevated concentrations of these elements were not observed during an inversion episode. Sea traffic and harbour activities were identified, primarily by the S and V content of the particles. This study showed that the elemental analysis by EDXRF presents valuable information for tracing the origin of air masses arriving at a measurement site.
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| 9. |
- Gatari, Michael J, et al.
(författare)
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Assessment of inorganic content of PM(2.5) particles sampled in a rural area north-east of Hanoi, Vietnam.
- 2006
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Ingår i: The Science of the total environment. - : Elsevier BV. - 0048-9697 .- 1879-1026. ; 368:2-3, s. 675-85
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Tidskriftsartikel (refereegranskat)abstract
- Atmospheric aerosols from seven rural sites in northern Vietnam, east of Hanoi, were sampled and analyzed. The aim of the study was to evaluate trace elemental and black carbon (BC) concentrations in fine particles (PM(2.5)) and to investigate the influence of the Pha Lai power plant and other pollution sources on regional air quality. Seven measurement stations were set up at selected rural sites and a campaign consisting of 12 two-day measurement periods was conducted from the end of May until the end of October 2000. At each location a pair of samplers was installed consisting of a cyclone loaded with Teflon filters and a modified Millipore air monitoring cartridge loaded with glass fibre filters. The obtained samples were analyzed for trace elements by Energy Dispersive X-ray Fluorescence (EDXRF), while a black smoke detector was used for BC analysis. Seventeen trace elements were analyzed; Br, Ca, Cl, Cr, Cu, Fe, K, Mn, Ni, Pb, Rb, S, Se, Sr, Ti, V and Zn and their concentrations evaluated. The results showed that BC, Ca, Cl, Fe, K and S dominated in the sampled atmospheric aerosols. The measured concentrations of the potentially hazardous trace elements Cr, Mn, Ni and Pb were all below the limits defined by Vietnamese standards of ambient air quality. Statistical evaluations indicated that coal and heavy fuel oil combustion were major sources of atmospheric pollutants in the area and that biomass burning and road transport had a marked influence on regional air quality. It was concluded that the Pha Lai power plant was the major source of coal combustion emissions. Trace element emissions originating from river transportation were suggested as another major source of atmospheric pollutants. The results indicate that the elemental concentrations in PM(2.5) are strongly influenced by seasonal variations. Further measurements are required to identify the impact of long-range transported continental air masses on the air quality of the investigated rural area.
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| 10. |
- Gatari, Michael J, et al.
(författare)
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Elemental composition of tropospheric aerosols in Hanoi, Vietnam and Nairobi, Kenya.
- 2005
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Ingår i: The Science of the total environment. - : Elsevier BV. - 0048-9697 .- 1879-1026. ; 341:1-3, s. 241-9
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Tidskriftsartikel (refereegranskat)abstract
- Air pollution problems in major cities within the developing countries need to be studied. There are scanty measurements from the developing countries on airborne particles despite their adverse implications to human health, visibility and climate. One of the major sources of anthropogenic air pollution is energy production. Energy demand is bound to increase as population increases, especially in major cities of the world. Fine particles, particles with aerodynamic diameter < or = 2.5 microm, are mainly anthropogenic and these particles were collected in the capital cities of Vietnam and Kenya. A cyclone airborne particle collector was used to sample in Hanoi during the months of May to October 2000 and a dichotomous virtual impactor in Nairobi in February 2000. The samples were analysed for elemental content by an energy dispersive X-ray fluorescence (EDXRF) spectrometer. S, Cl, K and Fe exceeded atmospheric concentrations of 100 ng m(-3) at both cities. Atmospheric elemental concentrations in both Hanoi and Nairobi were orders of magnitude higher than their respective rural towns. Traffic, biomass and waste burning emissions were implicated as the main sources of air pollution in Nairobi, while coal combustion and road transport were the major sources in Hanoi. Regional air pollution had a major impact over Hanoi, whereas an influence of that kind was not identified in Nairobi. Pb and other toxic elements had concentration levels below WHO guideline, however, the two cities are threatened by future high levels of air pollution due to the high rate of population growth. Long-term measurements are required in both areas to evaluate if the alarming situation is deteriorating.
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