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- Almén, Oscar, et al.
(författare)
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Local Governance Diversity in the Unitary Authoritarian State: NGO-State Relations in Guangzhou and Hangzhou
- 2024
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Ingår i: Journal of Contemporary China. - : Routledge. - 1067-0564 .- 1469-9400.
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Tidskriftsartikel (refereegranskat)abstract
- This study investigates how the different political opportunity structures (POS) are related to NGO mobilization in two Chinese cities, Guangzhou and Hangzhou. Based on 48 interviews from 2016–2019, the study finds that variance in NGO mobilization is related to differences such as rules for NGO registration, more or less open-minded local leaders, and a relatively more independent media. NGO governance in Hangzhou is characterized as coopted participation. A few NGOs are allowed some influence in policy making, but in order to be allowed to mobilize, NGOs must accept a certain degree of cooptation. NGO governance in Guangzhou is characterized as constrained autonomy as the government plays a less active role in mobilizing NGOs, and more initiative for policy influence comes from the NGOs themselves.
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| 4. |
- Benkestock, Kurt, et al.
(författare)
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Influence of droplet size, capillary-cone distance and selected instrumental parameters for the analysis of noncovalent protein-ligand complexes by nano-electrospray ionization mass spectrometry
- 2004
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Ingår i: Journal of Mass Spectrometry. - : Wiley. - 1076-5174 .- 1096-9888. ; 39:9, s. 1059-1067
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Tidskriftsartikel (refereegranskat)abstract
- It has been suggested in the literature that nano-electrospray ionization (nano-ESI) mass spectrometry better reflects the equilibrium between complex and free protein in solution than pneumatically assisted electrospray ionization (ESI) in noncovalent interaction studies. However, no systematic studies of the effects of ionization conditions have been performed to support this statement. In the present work, different instrumental and sample-derived parameters affecting the stability of noncovalent complexes during analysis by nano-ESI were investigated. In general, increased values of parameters such as drying gas flow-rate, ion-source temperature, capillary tip voltage and buffer concentration lead to a dissociation of ribonuclease A (RNAse)-cytidine 2'-monophosphate (CMP) and cytidine 5'-triphosphate (CTP) complexes. The size of the electrosprayed droplets was shown to be an important issue. Increasing the capillary to cone distance yielded an increased complex to free protein ratio when a hydrophilic ligand was present and the reverse effect was obtained with a hydrophobic ligand. Important in this regard is the degree of sampling of ions originating from late-generation residue droplets, that is, ions present in the droplet bulk. Sampling of these ions increases with longer capillary-cone distance (flight time). Furthermore, when the sample flow-rate was increased by increasing the capillary internal tip i.d. from 4 to 30 mum, a decreased complex to free protein ratio for the RNAse-CTP system was observed. This behavior was consistent with the change in surface to volume ratio for flow-rates between 2 and 100 nl min(-1). Finally, polarity switching between positive and negative ion modes gave a higher complex to free protein ratio when the ligand and the protein had the same polarity.
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| 5. |
- Eneyskaya, Elena V., et al.
(författare)
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Transglycosylating and hydrolytic activities of the beta-mannosidase from Trichoderma reesei
- 2009
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Ingår i: Biochimie. - : Elsevier BV. - 0300-9084 .- 1638-6183. ; 91:5, s. 632-638
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Tidskriftsartikel (refereegranskat)abstract
- A purified beta-mannosidase (EC 3.2.1.25) from the fungus Trichoderma reesei has been identified as a member of glycoside hydrolase family 2 through mass spectrometry analysis of tryptic peptides. In addition to hydrolysis, the enzyme catalyzes substrate transglycosylation with p-nitrophenyl beta-mannopyranoside. Structures of the major and minor products of this reaction were identified by NMR analysis as p-nitrophenyl mannobiosides and p-nitrophenyl mannotriosides containing beta-(1 -> 4) and beta-(1 -> 3) linkages. The rate of donor substrate hydrolysis increased in presence of acetonitrile and dimethylformamide, while transglycosylation was weakly suppressed by these organic solvents. Differential ultraviolet spectra of the protein indicate that a rearrangement of the hydrophobic environment of the active site following the addition of the organic solvents may be responsible for this hydrolytic activation.
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| 6. |
- Fugelstad, Johanna, et al.
(författare)
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Functional characterization of the pleckstrin homology domain of a cellulose synthase from the Oomycete Saprolegnia monoica
- 2012
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Ingår i: Biochemical and Biophysical Research Communications - BBRC. - : Academic Press. - 0006-291X .- 1090-2104. ; 417:4, s. 1248-1253
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Tidskriftsartikel (refereegranskat)abstract
- Some oomycetes, for instance Saprolegnia parasitica, are severe fish pathogens that cause important economic losses worldwide. Cellulose biosynthesis is a vital process for this class of microorganisms, but the corresponding molecular mechanisms are poorly understood. Of all cellulose synthesizing enzymes known, only some oomycete cellulose synthases contain a pleckstrin homology (PH) domain. Some human PH domains bind specifically to phosphoinositides, but most PH domains bind phospholipids in a non-specific manner. In addition, some PH domains interact with various proteins. Here we have investigated the function of the PH domain of cellulose synthase 2 from the oomycete Saprolegnia monoica (SmCesA2), a species closely related to S. parasitica. The SmCesA2 PH domain is similar to the C-terminal PH domain of the human protein TAPP1. It binds in vitro to phosphoinositides, F-actin and microtubules, and co-localizes with F-actin in vivo. Our results suggest a role of the SmCesA2 PH domain in the regulation, trafficking and/or targeting of the cell wall synthesizing enzyme.
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| 7. |
- Gullfot, Fredrika, 1967-, et al.
(författare)
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Functional Characterization of Xyloglucan Glycosynthases from GH7, GH12, and GH16 Scaffolds
- 2009
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Ingår i: Biomacromolecules. - : American Chemical Society (ACS). - 1525-7797 .- 1526-4602. ; 10:7, s. 1782-1788
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Tidskriftsartikel (refereegranskat)abstract
- Glycosynthases, hydrolytically inactive mutant glycosidases that catalyze glycosylation reactions using glycosyl fluoride donor substrates, are emerging as useful tools for the synthesis of large, complex polysaccharides [Faijes, M.; Planas, A. Carbohydr. Res. 2007, 342, 1581-1594]. Guided by wild-type xyloglucanase activity, we have produced and characterized new glycosynthases for the synthesis of xyloglucan oligo- and polysaccharides, based on family GH7, GH12, and GH16 scaffolds. The Humicola insolens GH7 glycosynthase, HiCel7B E197S, is capable of synthesizing nongalactosylated, XXXG-based homoxyloglucan up to Mw 60000 [G = Glcβ(1→4); X = Xylα(1→6)Glcβ(1→4); L = Galβ(1→2)Xylα(1→6)Glcβ(1→4)], which is among the largest products so far obtained with glycosynthase technology. Novel glycosynthases based on the GH16 xyloglucan hydrolase from Tropaeolum majus (nasturtium), TmNXG1, are capable of synthesizing XLLG-based xyloglucan oligosaccharides at rates feasible for preparative synthesis, thus providing an essential expansion of product range. Finally, a new glycosynthase based on the recently characterized GH12 xyloglucanase from Bacillus licheniformis, BlXG12 E155A, can perform the condensation of xyloglucosyl fluorides, albeit at poor rates. Altogether, the high catalytic efficiency demonstrated by HiCel7B E197S and the extended product range provided by TmNXG1 E94A are key achievements toward a robust and versatile method for the preparative synthesis of homogeneous xyloglucans with regular substitution patterns not available in nature. Such compounds enable in vitro experimental studies regarding the role of particular structural elements for xyloglucan properties and its interaction with cellulose.
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| 8. |
- Malm, Erik, 1981-, et al.
(författare)
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APP : An Automated Proteomics Pipeline for the analysis of mass spectrometry data based on multiple open access tools
- 2014
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Ingår i: BMC Bioinformatics. - : Springer Science and Business Media LLC. - 1471-2105. ; 15:1
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Tidskriftsartikel (refereegranskat)abstract
- Background: Mass spectrometry analyses of complex protein samples yield large amounts of data and specific expertise is needed for data analysis, in addition to a dedicated computer infrastructure. Furthermore, the identification of proteins and their specific properties require the use of multiple independent bioinformatics tools and several database search algorithms to process the same datasets. In order to facilitate and increase the speed of data analysis, there is a need for an integrated platform that would allow a comprehensive profiling of thousands of peptides and proteins in a single process through the simultaneous exploitation of multiple complementary algorithms. Results: We have established a new proteomics pipeline designated as APP that fulfills these objectives using a complete series of tools freely available from open sources. APP automates the processing of proteomics tasks such as peptide identification, validation and quantitation from LC-MS/MS data and allows easy integration of many separate proteomics tools. Distributed processing is at the core of APP, allowing the processing of very large datasets using any combination of Windows/Linux physical or virtual computing resources. Conclusions: APP provides distributed computing nodes that are simple to set up, greatly relieving the need for separate IT competence when handling large datasets. The modular nature of APP allows complex workflows to be managed and distributed, speeding up throughput and setup. Additionally, APP logs execution information on all executed tasks and generated results, simplifying information management and validation.
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| 9. |
- Martins, António, et al.
(författare)
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Ssy5 is a signaling serine protease that exhibits atypical biogenesis and marked S1 specificity
- 2018
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Ingår i: Journal of Biological Chemistry. - : AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC. - 0021-9258 .- 1083-351X. ; 293:22, s. 8362-8378
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Tidskriftsartikel (refereegranskat)abstract
- Ssy5 is a signaling endoprotease that plays a key role in regulating central metabolism, cellular aging, and morphological transitions important for growth and survival of yeast (Saccharomyces cerevisiae) cells. In response to extracellular amino acids, Ssy5 proteolytically activates the transcription factors Stp1 and Stp2, leading to enhanced Ssy1-Ptr3-Ssy5 (SPS) sensor-regulated gene expression. Ssy5 comprises a catalytic (Cat) domain and an extensive regulatory prodomain. Ssy5 is refractory to both broad-spectrum and serine protease-specific inhibitors, confounding its classification as a protease, and no information about Ssy5's cleavage-site preferences and its mechanism of substrate selection is available. Here, using mutational and inhibition experiments, we investigated the biogenesis and catalytic properties of Ssy5 and conclusively show that it is a serine protease. Atypical for the majority of serine proteases, Ssy5's prodomain was obligatorily required in cis during biogenesis for the maturation of the proteolytic activity of the Cat domain. Autolysis and Stp1 and Stp2 cleavage occurred between a cysteine (at the P1 site) and a serine or alanine (at the P1 site) and required residues with short side chains at the P1 site. Substitutions in the Cat domain affecting substrate specificity revealed that residues Phe-634, His-661, and Gly-671 in the S1-binding pocket of this domain are important for Ssy5 catalytic function. This study confirms that the signaling protease Ssy5 is a serine protease and provides a detailed understanding of the biogenesis and intrinsic properties of this key enzyme in yeast.
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| 10. |
- Nordin, Gustav, et al.
(författare)
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Somatic symptoms in sleep disturbance
- 2023
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Ingår i: Psychology, Health & Medicine. - : Taylor & Francis Group. - 1354-8506 .- 1465-3966. ; 28:4, s. 884-894
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Tidskriftsartikel (refereegranskat)abstract
- Despite sleep disturbance and somatic symptoms being common health complaints, the relationship between these disturbances and single somatic symptoms is not well documented. The objectives of this study were to (i) identify somatic symptoms that are particularly associated with sleep disturbance, here referred to as somatic symptoms related to sleep disturbance (SS-SD), (ii) determine increased risk of sleep disturbance for each SS-SD and for a certain number of SS-SD, with and without controlling for anxiety and depression, and (iii) determine sensitivity and specificity for identifying sleep disturbance based on number of SS-SD in a general Swedish sample. Population-based, cross-sectional data based on validated questionnaire instruments were used from participants who constituted a sleep disturbance (n = 864) or a reference (n = 2340) group. Among 15 common somatic symptoms, stomach pain, back pain nausea/gas/indigestion, dizziness, and constipation/loose bowels/diarrhea were identified as SS-SD, with odds ratios of increased risk of sleep disturbance that ranged from 1.93 to 2.44 (1.36–1.79 and 1.54–1.91 when controlled for anxiety and depression, respectively). The risk of sleep disturbance increased by 1.44 times for each SS-SD (1.25 and 1.30 when controlled for anxiety and depression, respectively). A cutoff of two/three or more SS-SD had a sensitivity of 72.5/54.2% and a specificity of 50.0/69.7% for identifying sleep disturbances. When patients present with these somatic symptoms with or without a pathophysiological explanation, primary care clinicians may consider screening for sleep disturbance.
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