Особенности школьной организационной культуры

Рассматривается основное содержание понятия «школьная организационная культура», функции и потенциал использования организационной культуры в общеобразовательных учреждениях. Выявляются особенности школьной организационной культуры. Обосновывается взаимосвязь организационной культуры и социально-психологического климата общеобразовательного учреждени

Substrate Specificity of Clostridial Glucosylating Toxins and Their Function on Colonocytes Analyzed by Proteomics Techniques

<i>Clostridium difficile</i> is the major cause of intestinal infections in hospitals. The major virulence factors are toxin A (TcdA) and toxin B (TcdB), which belong to the group of clostridial glucosylating toxins (CGT) that inactivate small GTPases. After a 24 h incubation period with TcdA or a glucosyltransferase-deficient mutant TcdA (gdTcdA), quantitative changes in the proteome of colonic cells (Caco-2) were analyzed using high-resolution LC–MS/MS and the SILAC technique. The changes in abundance of more than 5100 proteins were quantified. Nearly 800 toxin-responsive proteins were identified that were involved in cell cycle, cell structure, and adhesion as well as metabolic processes. Several proteins localized to mitochondria or involved in lipid metabolism were consistently of higher abundance after TcdA treatment. All changes of protein abundance depended on the glucosyltransferase activity of TcdA. Glucosylation of the known targets of TcdA such as RhoA, RhoC, RhoG was detected by LC–MS/MS. In addition, an almost complete glucosylation of Rap1­(A/B), Rap2­(A/B/C) and a partial glucosylation of Ral­(A/B) and (H/K/N)­Ras were detected. The glucosylation pattern of TcdA was compared to that of other CGT like TcdB, the variant TcdB from <i>C. difficile</i> strain VPI 1470 (TcdBF), and lethal toxin from <i>C. sordellii</i> (TcsL)

Contextual Dynamics of Immigration Attitudes: Regional Differences in Southern Europe

215 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2005.Research in the area of attitudes towards immigration will benefit from a more thorough discussion of the relationship between degrees of political engagement and the variance of political and social tolerance towards immigrants. Drawing upon institutional theory and realistic conflict theory, I further refine a theory of ethnic competition and prejudice in the Southern European context. I argue that popular attitudes towards immigration are correlated with a set of individual level factors (e.g. perceptions of personal and collective threat, as well as measures of political socialization), which are shaped and determined by the contextual characteristics (e.g. economic conditions and demographic characteristics) as well as the type of institutional environment (e.g. the presence or absence of support towards civic institutions) in which inter-group relations are embedded. The characterization of these environments determines the type of in-group/out-group social relations. I first, empirically characterize the type of "civic communities" existing in 50 Southern European regions and then, empirically test its significance in preventing inter-group hostility and the fostering of tolerance towards minority groups. Results show that there is strong significant effect between trust in institutions (such as NGOs and voluntary organizations) and decreased levels of anti-immigrant sentiment and intergroup conflict in Southern Europe. This dissertation provides evidence for the widespread effects that local minority group size and types of institutional trust have on political and social tolerance towards immigrants. Furthermore, evidence is provided that anti-immigrant sentiment has an extensive impact on Southern Europeans' policy opinions.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD

Proteome Alterations of Hippocampal Cells Caused by <i>Clostridium botulinum</i> C3 Exoenzyme

C3bot from <i>Clostridium botulinum</i> is a bacterial mono-ADP-ribosylating enzyme, which transfers an ADP-ribose moiety onto the small GTPases Rho A/B/C. C3bot and the catalytic inactive mutant (C3E174Q) cause axonal and dendritic growth as well as branching in primary hippocampal neurons. In cultured murine hippocampal HT22 cells, protein abundances were analyzed in response to C3bot or C3E174Q treatment using a shotgun proteomics approach. Proteome analyses were performed at four time points over 6 days. More than 4000 protein groups were identified at each time point and quantified in triplicate analyses. On day one, 46 proteins showed an altered abundance, and after 6 days, more than 700 proteins responded to C3bot with an up- or down-regulation. In contrast, C3E174Q had no provable impact on protein abundance. Protein quantification was verified for several proteins by multiple reaction monitoring. Data analysis of altered proteins revealed different cellular processes that were affected by C3bot. They are particularly involved in mitochondrial and lysosomal processes, adhesion, carbohydrate and glucose metabolism, signal transduction, and nuclear proteins of translation and ribosome biogenesis. The results of this study gain novel insights into the function of C3bot in hippocampal cells

Substrate Specificity of Clostridial Glucosylating Toxins and Their Function on Colonocytes Analyzed by Proteomics Techniques

<i>Clostridium difficile</i> is the major cause of intestinal infections in hospitals. The major virulence factors are toxin A (TcdA) and toxin B (TcdB), which belong to the group of clostridial glucosylating toxins (CGT) that inactivate small GTPases. After a 24 h incubation period with TcdA or a glucosyltransferase-deficient mutant TcdA (gdTcdA), quantitative changes in the proteome of colonic cells (Caco-2) were analyzed using high-resolution LC–MS/MS and the SILAC technique. The changes in abundance of more than 5100 proteins were quantified. Nearly 800 toxin-responsive proteins were identified that were involved in cell cycle, cell structure, and adhesion as well as metabolic processes. Several proteins localized to mitochondria or involved in lipid metabolism were consistently of higher abundance after TcdA treatment. All changes of protein abundance depended on the glucosyltransferase activity of TcdA. Glucosylation of the known targets of TcdA such as RhoA, RhoC, RhoG was detected by LC–MS/MS. In addition, an almost complete glucosylation of Rap1­(A/B), Rap2­(A/B/C) and a partial glucosylation of Ral­(A/B) and (H/K/N)­Ras were detected. The glucosylation pattern of TcdA was compared to that of other CGT like TcdB, the variant TcdB from <i>C. difficile</i> strain VPI 1470 (TcdBF), and lethal toxin from <i>C. sordellii</i> (TcsL)

Substrate Specificity of Clostridial Glucosylating Toxins and Their Function on Colonocytes Analyzed by Proteomics Techniques

<i>Clostridium difficile</i> is the major cause of intestinal infections in hospitals. The major virulence factors are toxin A (TcdA) and toxin B (TcdB), which belong to the group of clostridial glucosylating toxins (CGT) that inactivate small GTPases. After a 24 h incubation period with TcdA or a glucosyltransferase-deficient mutant TcdA (gdTcdA), quantitative changes in the proteome of colonic cells (Caco-2) were analyzed using high-resolution LC–MS/MS and the SILAC technique. The changes in abundance of more than 5100 proteins were quantified. Nearly 800 toxin-responsive proteins were identified that were involved in cell cycle, cell structure, and adhesion as well as metabolic processes. Several proteins localized to mitochondria or involved in lipid metabolism were consistently of higher abundance after TcdA treatment. All changes of protein abundance depended on the glucosyltransferase activity of TcdA. Glucosylation of the known targets of TcdA such as RhoA, RhoC, RhoG was detected by LC–MS/MS. In addition, an almost complete glucosylation of Rap1­(A/B), Rap2­(A/B/C) and a partial glucosylation of Ral­(A/B) and (H/K/N)­Ras were detected. The glucosylation pattern of TcdA was compared to that of other CGT like TcdB, the variant TcdB from <i>C. difficile</i> strain VPI 1470 (TcdBF), and lethal toxin from <i>C. sordellii</i> (TcsL)

Vimentin is present at the cell surface of HT22 cells and J774A.1 macrophages.

<p>A) Intact HT22 cells were biotinylated for 1 h at 4°C. Whole cell lysates, cytosolic and particulate fractions were prepared. In addition, cell surface biotinylated proteins were enriched by precipitation with NeutrAvidin beads. The fractions and precipitation, respectively, were immunoblotted and probed with anti-vimentin. Biotinylation fraction represents the extracellular proteins exclusively. One representative Western blot experiment is shown (n = 3 independent experiments). Presence of vimentin at the cell surface of HT22 cells (B) and J774A.1 cells (C) was analyzed by FACS cytometry using anti-vimentin. Oregon green-488 conjugated goat anti-rabbit antibody alone served as negative control. Untreated cells were used as control.</p

Binding of C3 to HT22 cells after pronase treatment.

<p>A) Pronase pre-incubated HT22 cells were exposed to 100 or 500 nM of C3 for 1 h at 4°C. Subsequently, β-actin and bound C3 were detected by Western blot. NC = negative control without C3, PC = positive control lysate with 10 ng C3. One representative experiment is shown (n = 3 independent experiments). B) Pronase-treated HT22 cells were exposed to 500 nM of C3-E174Q-FITC for 1 h at 4°C and bound C3- E174Q-FITC was analyzed by FACS.</p

Vimentin Mediates Uptake of C3 Exoenzyme

<div><p><i>Clostridium botulinum</i> C3 exoenzyme (C3) selectively inactivates RhoA/B/C GTPases by ADP-ribosylation. Based on this substrate specificity C3 is a well-established tool in cell biology. C3 is taken up by eukaryotic cells although lacking an uptake and translocation domain. Based on different approaches vimentin was identified as membranous C3-interaction partner by mass spectrometry. Vimentin in fact was partly localized at the outer surface of hippocampal HT22 cells and J744A.1 macrophages. Domain analysis identified the rod domain as binding partner of C3. Vimentin was also involved in uptake of C3 as shown by knock down of vimentin in HT22 and J774A.1 cells. The involvement of vimentin in uptake of C3 was further supported by the findings that the vimentin disruptor acrylamide blocked uptake of C3. Vimentin is not only a major organizing element of the intermediate filament network but is also involved in both binding and uptake of C3 exoenzyme.</p></div

C3-overlay (binding of C3 to HT22 proteins).

<p>A) Whole cell lysate, cytosolic fraction or particulate fraction from HT22 cells were generated as described in material and methods followed by separation through SDS-PAGE and transfer onto nitrocellulose. Nitrocellulose was incubated with 10 µg/ml of C3 for 60 min at 4°C. After washing bound C3 was detected by anti-C3. Arrows indicate the protein of interest (55 kDa). B) The right panel shows the anti-C3 Western blot without C3-overlay. M = molecular mass marker, WCL = whole cell lysate, PF = particulate fraction, CF = cytosolic fraction, WCL +10 ng C3 = C3 was added to whole cells lysate prior to SDS-PAGE and blotting to generate a positive C3 signal.</p