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- Chilkova, Olga, et al.
(författare)
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The eukaryotic leading and lagging strand DNA polymerases are loaded onto primer-ends via separate mechanisms but have comparable processivity in the presence of PCNA.
- 2007
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Ingår i: Nucleic Acids Research. - : Oxford University Press (OUP). - 1362-4962 .- 0305-1048. ; 35:19, s. 6588-6597
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Tidskriftsartikel (refereegranskat)abstract
- Saccharomyces cerevisiae DNA polymerase delta (Pol delta) and DNA polymerase epsilon (Pol epsilon) are replicative DNA polymerases at the replication fork. Both enzymes are stimulated by PCNA, although to different levels. To understand why and to explore the interaction with PCNA, we compared Pol delta and Pol epsilon in physical interactions with PCNA and nucleic acids (with or without RPA), and in functional assays measuring activity and processivity. Using surface plasmon resonance technique, we show that Pol epsilon has a high affinity for DNA, but a low affinity for PCNA. In contrast, Pol delta has a low affinity for DNA and a high affinity for PCNA. The true processivity of Pol delta and Pol epsilon was measured for the first time in the presence of RPA, PCNA and RFC on single-stranded DNA. Remarkably, in the presence of PCNA, the processivity of Pol delta and Pol epsilon on RPA-coated DNA is comparable. Finally, more PCNA molecules were found on the template after it was replicated by Pol epsilon when compared to Pol delta. We conclude that Pol epsilon and Pol delta exhibit comparable processivity, but are loaded on the primer-end via different mechanisms.
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2. |
- Fuxe, K., et al.
(författare)
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Intramembrane receptor-receptor interactions: a novel principle in molecular medicine
- 2007
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Ingår i: Journal of neural transmission. - : Springer Science and Business Media LLC. - 0300-9564 .- 1435-1463. ; 114:1, s. 49-75
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Tidskriftsartikel (refereegranskat)abstract
- In 1980/81 Agnati and Fuxe introduced the concept of intramembrane receptor-receptor interactions and presented the first experimental observations for their existence in crude membrane preparations. The second step was their introduction of the receptor mosaic hypothesis of the engram in 1982. The third step was their proposal that the existence of intramembrane receptor-receptor interactions made possible the integration of synaptic (WT) and extrasynaptic (VT) signals. With the discovery of the intramembrane receptor-receptor interactions with the likely formation of receptor aggregates of multiple receptors, so called receptor mosaics, the entire decoding process becomes a branched process already at the receptor level in the surface membrane. Recent developments indicate the relevance of cooperativity in intramembrane receptor-receptor interactions namely the presence of regulated cooperativity via receptor-receptor interactions in receptor mosaics (RM) built up of the same type of receptor (homo-oligomers) or of subtypes of the same receptor (RM type1). The receptor-receptor interactions will to a large extent determine the various conformational states of the receptors and their operation will be dependent on the receptor composition (stoichiometry), the spatial organization (topography) and order of receptor activation in the RM. The biochemical and functional integrative implications of the receptor-receptor interactions are outlined and long-lived heteromeric receptor complexes with frozen RM in various nerve cell systems may play an essential role in learning, memory and retrieval processes. Intramembrane receptor-receptor interactions in the brain have given rise to novel strategies for treatment of Parkinson's disease (A2A and mGluR5 receptor antagonists), schizophrenia (A2A and mGluR5 agonists) and depression (galanin receptor antagonists). The A2A/D2, A2A/D3 and A2A/mGluR5 heteromers and heteromeric complexes with their possible participation in different types of RM are described in detail, especially in the cortico-striatal glutamate synapse and its extrasynaptic components, together with a postulated existence of A2A/D4 heteromers. Finally, the impact of intramembrane receptor-receptor interactions in molecular medicine is discussed outside the brain with focus on the endocrine, the cardiovascular and the immune systems.
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3. |
- Paolesse, R., et al.
(författare)
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Insights on the chemistry of a,c-biladienes from a CSPT investigation
- 2008
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Ingår i: New Journal of Chemistry. - : Royal Society of Chemistry (RSC). - 1144-0546 .- 1369-9261. ; 32:7, s. 1162-1166
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Tidskriftsartikel (refereegranskat)abstract
- Computer Screen Photoassisted Technique (CSPT) is a recent interesting approach for the development of optical sensor platforms using ubiquitous devices. We demonstrate here that the information obtained by CSPT measurements can be useful for other than sensing purposes and a computer set and a web camera can also be exploited as spectroscopic technique for the characterization of the compounds used as indicators. CSPT measurement data have been used to characterize novel features of the a,c-biladiene chemistry, which have been later confirmed by photophysical characterization carried out using classical instrumentation. The demonstrated analytical capabilities of a regular computer-web camera set reach a degree of complexity approaching that of fluorescence excitation-emission spectroscopy, something remarkable considering the exploitation of familiar and already well distributed devices. © The Royal Society of Chemistry and the Centre National de la Recherche Scientifique.
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4. |
- Riepl, M., et al.
(författare)
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Molecular gradients : An efficient approach for optimizing the surface properties of biomaterials and biochips
- 2005
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Ingår i: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 21:3, s. 1042-1050
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Tidskriftsartikel (refereegranskat)abstract
- A variety of molecular gradients of alkanethiols with the structure HS-(CH2)m-X (m = 15, X = COOH, CH2NH 2, or CH3) and oligo(ethylene glycol)-terminated alkanethiols with the structures HS-(CH2)15-CO-NH-Eg n (n = 2, 4, or 6), HS-(CH2)15-CO-NH-Eg 2-(CH2)2-NH-CO-(CH2) 4-biotin, and HS-(CH2)15-CO-NH-Eg 6-CH2-COOH were prepared on polycrystalline gold films. These gradients were designed to serve as model surfaces for fundamental studies of protein adsorption and immobilization phenomena. Ellipsometry, infrared spectroscopy, and X-ray photoelectron spectroscopy, operating in scanning mode, were used to monitor the layer composition, gradient profiles, tail group conformation, and overall structural quality of the gradient assemblies. The gradient profiles were found to be 4-10 mm wide, and they increased in width with increasing difference in molecular complexity between the thiols used to form the gradient. The oligo(ethylene glycol) thiols are particularly interesting because they can be used to prepare so-called conformational gradients, that is, gradients that display a variation in oligo(ethylene glycol) chain conformation from all trans on the extreme Eg 2,4 sides, via an amorphous-like phase in the mixing regimes, to helical at the extreme Eg6 sides. We demonstrate herein a series of experiments where the above gradients are used to evaluate nonspecific binding of the plasma protein fibrinogen, and in agreement with previous studies, the highest amounts of nonspecifically bound fibrinogen were observed on all-trans monolayers, that is, on the extreme Eg2,4 sides. Moreover, gradients between Eg2 and a biotinylated analogue have been prepared to optimize the conditions for the immobilization of streptavidin. Ellipsometry and infrared spectroscopy reveal high levels of immobilization over a fairly broad range of compositions in the gradient regime, with a maximum between 50 and 60% of the biotinylated analogue in the monolayer. A pi gradient composed of (NH3+/COO-)-terminated thiols was also prepared and evaluated with respect to its ability to separate differently charged proteins, pepsin, and lysozyme, on a solid surface.
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5. |
- Smits, Mark M, et al.
(författare)
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The fungal-mineral interface: Challanges and considerations of micro-analytical developments
- 2009
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Ingår i: Fungal Biology Reviews. - : Elsevier BV. - 1749-4613 .- 1878-0253. ; 23:4, s. 122-131
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Tidskriftsartikel (refereegranskat)abstract
- Over recent years, the role of fungi, especially mycorrhizal fungi, in the weathering of rock-forming minerals has been increasingly recognised. Much of our understanding of the effects of fungi on mineral weathering is based on macroscopic studies. However, the ability of fungi to translocate materials, including organic acids and siderophores, to specific areas of a mineral surface leads to significant spatial heterogeneity in the weathering process. Thus, geomycologists are confronted with unique challenges of how to comprehend and quantify such a high degree of diversity and complicated arrays of interactions. Recent advances in experimental and analytical techniques have increased our ability to probe the fungal-mineral interface at the resolution necessary to decouple significant biogeochemical processes. Modern microscopy, spectroscopy, mass spectrometry, wet chemistry, and scattering techniques allow for the selective extraction of physical, chemical, and structural data at the micro- to nano-scale. These techniques offer exciting possibilities to study fungal-mineral interactions at the scale of individual hyphae. In this review, we give an overview of some of these techniques with their characteristics, advantages and limitations, and how they can be used to further our understanding of biotic mineral weathering.
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6. |
- Trey, Stacy. M., et al.
(författare)
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Effects of dual cure and surface treatments on coating adhesion to different SMC substrates
- 2009
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Ingår i: Plastics, rubber and composites. - : Informa UK Limited. - 1465-8011 .- 1743-2898. ; 38:2-4, s. 131-137
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Tidskriftsartikel (refereegranskat)abstract
- Sheet moulded compound (SMC) is a highly filled, glass fibre reinforced, thermoset material used in trim and body panel automotive parts. When SMC substrates are coated with conventional thermally cured paints, inherent porosity and entrapped volatiles of the substrate result in popping ('paint pops') defects. UV curable primers (UVP) provide an order of magnitude reduction of paint defects in SMC coatings, but typically have poor adhesion. The present study investigates a series of UVPs, showing the effect of resin functionality and isocyanate additive content on the adhesion of the coatings to SMC substrates. The SMC formulation is also considered regarding how variables such as surface chemistry, morphology, surface area and degree of post-mould emissions affect UVP adhesion. The present study reveals the crucial factors involved in achieving adhesion including the importance of low post-mould emissions, high surface areas, glass fibre and other oxygen moieties on the surface, and improved wetting of the surface.
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8. |
- Adén, Daniel, et al.
(författare)
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A photometric and spectroscopic study of the new dwarf spheroidal galaxy in Hercules. Metallicity, velocities and a clean list of RGB members
- 2009
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Ingår i: Astronomy & Astrophysics. - : EDP Sciences. - 0004-6361 .- 1432-0746. ; 506:3, s. 1147-1168
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Tidskriftsartikel (refereegranskat)abstract
- Our aim is to provide as clean and as complete a sample as possible of red giant branch stars that are members of the Hercules dSph galaxy. With this sample we explore the velocity dispersion and the metallicity of the system. Stromgren photometry and multi-fibre spectroscopy are combined to provide information about the evolutionary state of the stars (via the Stromgren c_1 index) and their radial velocities. Based on this information we have selected a clean sample of red giant branch stars, and show that foreground contamination by Milky Way dwarf stars can greatly distort the results. Our final sample consists of 28 red giant branch stars in the Hercules dSph galaxy. Based on these stars we find a mean photometric metallicity of -2.35 dex which is consistent with previous studies. We find evidence for an abundance spread. Using those stars for which we have determined radial velocities we find a systemic velocity of 45.2 km/s with a dispersion of 3.72 km/s, this is lower than values found in the literature. Furthermore we identify the horizontal branch and estimate the mean magnitude of the horizontal branch of the Hercules dSph galaxy to be V_0=21.17, which corresponds to a distance of 147 kpc. We have shown that a proper cleaning of the sample results in a smaller value for the velocity dispersion of the system. This has implications for galaxy properties derived from such velocity dispersions.
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