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231.
F Nachon L Ehret-Sabatier D Loew C Colas A van Dorsselaer M Goeldner 《Canadian Metallurgical Quarterly》1998,37(29):10507-10513
Purified butyrylcholinesterase (BuChE) was photolabeled by [3H]-p-N, N-dimethylamino benzene diazonium ([3H]DDF) to identify the quaternary ammonium binding sites on this protein [Ehret-Sabatier, L. , Schalk, I., Goeldner, M., and Hirth, C. (1992) Eur. J. Biochem. 203, 475-481]. The covalent photoincorporation occurs with a stoichiometry of one mole of probe per mole of inactivated site and could be fully prevented by several cholinergic inhibitors such as tacrine or tetramethylammonium. After complete deglycosylation of the enzyme using N-glycosidase F, the alkylated protein was trypsinolyzed and the digests were analyzed by HPLC coupled to ES-MS. A direct comparison of tryptic fragments from labeled and unlabeled BuChE allowed us to identify the tryptic peptide Tyr61-Lys103 as carrying the probe. Purification of the labeled peptides by anion-exchange chromatography gave a major radioactive peak which was further fractionated by reversed-phase HPLC leading to three, well-resolved, radioactive peaks. Microsequencing revealed that two of these peaks contained an overlapping sequence starting at Tyr61, while the third peak contained a sequence extending from Thr315. Radioactive signals could be unambiguously attributed to positions corresponding to residues Trp82 and Tyr332. This labeling study establishes the existence of two different binding domains for quaternary ammonium in BuChE and exemplifies additional cation/pi interactions in cholinergic proteins. This work strongly supports the existence of a peripheral anionic site in BuChE, implying residue Tyr332 as a key element. 相似文献
232.
CW van Haselen PV Diederen J Toonstra SC van der Putte LH Rademakers JA Faber WA van Vloten 《Canadian Metallurgical Quarterly》1998,290(11):583-590
Small-cell variants of Sézary syndrome and mycosis fungoides (MF) have been described. However, in these studies the nuclear area of the small-cell variant of MF (SC-MF) as compared to histological classical MF (CL-MF) was not characterized objectively by quantitative electron microscopy. In a 14-year follow-up period, of a total of 76 patch/plaque stage MF patients seen in the Department of Dermatology of the University Hospital Utrecht, 14 (18%) had an infiltrate composed of atypical lymphocytes characterized by a distinctly smaller cell diameter and smaller, hyperchromatic, deeply indented nuclei as compared to the usual cell type of MF. The aim of the investigation was to confirm this observation objectively using quantitative electron microscopy (morphometry) and to define SC-MF as compared to CL-MF. The study was performed on the 14 patients with SC-MF, and 10 patients with clinical and histological CL-MF and 4 patients with chronic eczema. Electron micrographs of sections obtained from each biopsy were analysed by computer to produce the following data: a nuclear contour index (NCI), the mean nuclear area (MNA), the mean nuclear area of the cells above the 75th percentile (P75NA) and the percentage of cells larger than 30 microm2. The values of MNA differed significantly between patients with SC-MF and those with CL-MF (17.6 vs 23.2 microm2; P = 0.02), as did the values of P75NA (20.7 vs 27.9 microm2; P = 0.01). The NCI of the SC-MF and CL-MF patients were similar. These results are consistent with our observations that SC-MF does indeed exist. 相似文献
233.
AH Kolk LF Kox J van Leeuwen S Kuijper HM Jansen 《Canadian Metallurgical Quarterly》1998,11(6):1222-1226
This study examines the diagnostic utility of the polymerase chain reaction (PCR) in 156 patients (five human immunodeficiency virus (HIV) seropositive) suspected of extrapulmonary tuberculosis. The results of PCR in 226 samples from 11 different sites were compared with the results of microscopy and culture. Positive culture results were predicted in 86% of samples by PCR but in only 31% by microscopy. Specificity of PCR was 92%. In cases with culture-proven tuberculosis, PCR identified all 11 microscopy positive cases and 19 of 24 (79%) of the microscopy-negative cases. In four patients, PCR excluded the diagnosis of tuberculosis in microscopy-positive samples, which were later shown to contain mycobacteria other than Mycobacterium tuberculosis or laboratory contaminants. In 20 patients (microscopy, PCR and culture negative) a trial of antituberculous drugs was given, but patients showed no improvement and treatment was stopped. In 17 patients, all culture negative (in nine PCR was positive, three of whom also had positive microscopy) the diagnosis was probable tuberculosis based on clinical findings and response to treatment. This polymerase chain reaction has a much higher sensitivity than microscopy and can facilitate therapeutic decisions for those with suspected extrapulmonary tuberculosis. 相似文献
234.
235.
LS Melchers M Apotheker-de Groot JA van der Knaap AS Ponstein MB Sela-Buurlage JF Bol BJ Cornelissen PJ van den Elzen HJ Linthorst 《Canadian Metallurgical Quarterly》1994,5(4):469-480
A novel chitinase gene of tobacco was isolated and characterized by DNA sequence analysis of a genomic clone and a cDNA clone. Comparative sequence analysis of both clones showed an identity of 94%. The proteins encoded by these sequences do not correspond to any of the previously characterized plant chitinases of classes I-IV and are designated as class V chitinases. Comparison of the chitinase class V peptide sequence with sequences in the Swiss Protein databank revealed significant sequence similarity with bacterial exo-chitinases from Bacillus circulans, Serratia marcescens and Streptomyces plicatus. It was demonstrated that class V chitinase gene expression is induced after treatment of tobacco with different forms of stress, like TMV-infection, ethylene treatment, wounding or ultraviolet irradiation. Two related chitinase class V proteins of 41 and 43 kDa were purified from Samsun NN tobacco leaves inoculated with tobacco mosaic virus. The proteins were purified by Chelating Superose chromatography and gel filtration. In vitro assays demonstrated that class V chitinases have endo-chitinase activity and exhibit antifungal activity toward Trichoderma viride and Alternaria radicina. In addition, it was shown that class V chitinase acts synergistically with tobacco class I beta-1,3-glucanase against Fusarium solani germlings. 相似文献
236.
MA Feijge EC van Pampus C Lacabaratz-Porret K Hamulyàk S Levy-Toledano J Enouf JW Heemskerk 《Canadian Metallurgical Quarterly》1998,102(3):850-859
Increased Ca2+ signal generation may lead to hyperactivity of platelets and contribute to thrombotic complications. Using fura-2-loaded platelets from 51 healthy volunteers, high variability was detected in the Ca2+ responses evoked by the receptor agonists, thrombin and collagen, and the inhibitor of sarco/endoplasmic reticulum Ca2+-ATPases (SERCA), thapsigargin (Tg). Oral intake of 500mg aspirin reduced the magnitude of the Ca2+ responses, and lowered the intra-individual coefficients of variance of the responses by 50%. However, the corresponding inter-individual variance coefficients were only a little influenced by aspirin intake, pointing to subject-dependent factors in Ca2+ handling that are unrelated to thromboxane formation. With each agonist, 6-9% of the subjects had platelets with relatively high Ca2+ responses (> mean + SD) both before and after aspirin intake. In 90% (9/10) of these cases the high responsiveness was confirmed in platelets obtained 6-12 months later. The Tg- but not thrombin-induced Ca2+ responses correlated inversely with the expression levels of SERCA PL/IM 430 (SERCA-3b) in platelets. After aspirin intake, the Ca2+ responses with collagen but not thrombin correlated inversely with SERCA-2b expression. These results suggest that, in the absence of potentiating effects of thromboxane, (i) the amount of PL/IM 430-recognizable SERCA may control the Ca2+ signal when SERCA-2b is specifically inhibited (with Tg), and (ii) the expression of SERCA-2b determine the collagen- but not the thrombin-evoked Ca2+ signal. Accordingly, limited Ca2+-pumping activity by low expression of one of the SERCA isoforms is likely to be one of the factors resulting in increased platelet activity towards collagen or thapsigargin but not thrombin. 相似文献
237.
The medical management of Crohn's disease has changed in recent years, but the mainstay of treatment is still prednisone. A substantial fraction of steroid-treated patients are refractory to therapy and addition of azathioprine or methotrexate has a corticosteroid-sparing effect and increases duration of remission. Controlled ileal release budesonide (9 mg daily) induces clinical remission in 60-70% of patients with Crohn's ileitis or right-sided colitis, and continued budesonide treatment has a finite effect on the duration of remission. The efficacy of mesalazine in active Crohn's disease is limited and high doses are required (4000 mg/day). The role of mesalazine in Crohn's disease in remission is disputed, and there is no evidence of a corticosteroid-sparing effect. 相似文献
238.
Bert Slof Anouschka van Leeuwen Jeroen Janssen Paul A. Kirschner 《Journal of Computer Assisted Learning》2021,37(1):39-50
In computer-supported collaborative learning research, studies examining the combined effects of individual level, group level and within-group differences level measures on individual achievement are scarce. The current study addressed this by examining whether individual, group and within-group differences regarding engagement and prior knowledge predict individual achievement. Engagement was operationalised as group members' exhibited activities in the task space (i.e., discussing domain-content) and social space (i.e., regulating ideas, actions and socioemotional processes). Prior knowledge and achievement were operationalised as group members' performance on a domain-related pre-test and post-test, respectively. Data was collected for 95 triads of secondary education students collaborating on a complex business-economics problem. Subsequently, three different multilevel models were tested to examine the combined effect. First a model with the individual level measures (model 1) was tested and in subsequent models the group level measures (model 2) and within-group levels measures (model 3) were added. Findings indicate model 2 showed the best fit; group members' individual engagement in the social space activities as well as the groups' average prior knowledge positively predicts individual achievement. No effects were found for either group members' or groups' engagement in the task space and for the within-group differences. 相似文献
239.
Huihua Liu Eva Bracht Xin‐an Zhang Bret Bradley Rolf van Dick 《Creativity & Innovation Management》2021,30(1):129-143
An increasing number of individuals work in jobs with little standardization and repetition, that is, with high levels of job non‐routinization. At the same time, demands for creativity are high, which raises the question of how employees can use job non‐routinization to develop creativity. Acknowledging the importance of social processes for creativity, we propose that transformational leaders raise feelings of organizational identification in followers and that this form of identification then helps individuals to develop creativity in jobs with little routinization. This is because organizational members evaluate and promote those ideas as more creative, which are in line with a shared understanding of creativity within the organization. To investigate these relationships, we calculated a mediated moderation model with 173 leader–follower dyads from China. Results confirm our hypotheses that transformational leadership moderates the relationship between job non‐routinization on employee creativity through organizational identification. We conclude that raising feelings of social identity is a key task for leaders today, especially when working in uncertain and fast developing environments with little repetition and the constant need to develop creative ideas. 相似文献
240.
Flow experience, the degree to which a person feels involved in a particular activity, is an important influence on human–computer interaction. Building on Guo and Poole’s (2009) model of flow experience in Web navigation, and van Schaik and Ling's (in press) cognitive-experiential approach to modelling interaction experience, this research demonstrates the crucial role of the preconditions of flow experience in human–computer interaction. In an experiment, the preconditions of flow experience – but not flow experience proper – mediated the effects of artefact complexity, task complexity and intrinsic motivation (as a situation-specific trait) on both flow and task outcome. However, preconditions did not predict overall artefact evaluation. Within a staged model of flow experience, the broader implications of this work for human–computer interaction are explored. 相似文献