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1.
Dr AR Holmes RD Cannon HF Jenkinson 《Journal of industrial microbiology & biotechnology》1995,15(3):208-213
The yeastCandida albicans coaggregates with a variety of streptococcal species, an interaction that may promote oral colonization by yeast cells.C. albicans andCandida tropicalis are the yeasts most frequently isolated from the human oral cavity and our data demonstrate that both these species bind toStreptococcus gordonii NCTC 7869 while two otherCandida species (Candida krusei andCandida kefyr) do not. Adherence ofC. albicans was greatest when the yeast had been grown at 30° C to mid-exponential growth phase. For 21 strains ofC. albicans there was a positive correlation between the ability to adhere toS. gordonii and adherence to experimental salivary pellicle. Whole saliva either stimulated or slightly inhibited adherence ofC. albicans toS. gordonii depending on the streptococcal growth conditions. The results suggest that the major salivary adhesins and coaggregation adhesins ofC. albicans are co-expressed. 相似文献
2.
Oomen CJ Hoogerhout P Bonvin AM Kuipers B Brugghe H Timmermans H Haseley SR van Alphen L Gros P 《Journal of molecular biology》2003,328(5):1083-1089
We present an in silico, structure-based approach for design and evaluation of conformationally restricted peptide-vaccines. In particular, we designed four cyclic peptides of ten or 11 residues mimicking the crystallographically observed beta-turn conformation of a predicted immunodominant loop of PorA from Neisseria meningitidis. Conformational correctness and stability of the peptide designs, as evaluated by molecular dynamics simulations, correctly predicted the immunogenicity of the peptides. We observed a peptide-induced functional antibody response that, remarkably, exceeded the response induced by the native protein in outer membrane vesicles, without losing specificity for related strains. The presented approach offers tools for a priori design and selection of peptide-vaccine candidates with full biological activity. This approach could be widely applicable: to outer membrane proteins of Gram-negative bacteria, and to other epitopes in a large range of pathogens. 相似文献
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Meiring HD Kuipers B van Gaans-van den Brink JA Poelen MC Timmermans H Baart G Brugghe H van Schie J Boog CJ de Jong AP van Els CA 《Journal of immunology (Baltimore, Md. : 1950)》2005,174(9):5636-5643
The meningococcal class I outer membrane protein porin A plays an important role in the development of T cell-dependent protective immunity against meningococcal serogroup B infection and is therefore a major component of candidate meningococcal vaccines. T cell epitopes from porin A are poorly characterized because of weak in vitro memory T cell responses against purified Ag and strain variation. We applied a novel strategy to identify relevant naturally processed and MHC class II-presented porin A epitopes, based on stable isotope labeling of Ag. Human immature HLA-DR1-positive dendritic cells were used for optimal uptake and MHC class II processing of (14)N- and (15)N-labeled isoforms of the neisserial porin A serosubtype P1.5-2,10 in bacterial outer membrane vesicles. HLA-DR1 bound peptides, obtained after 48 h of Ag processing, contained typical spectral doublets in mass spectrometry that could easily be assigned to four porin A regions, expressed at diverging densities ( approximately 30-4000 copies/per cell). Epitopes from two of these regions are recognized by HLA-DR1-restricted CD4(+) T cell lines and are conserved among different serosubtypes of meningococcal porin A. This mass tag-assisted approach provides a useful methodology for rapid identification of MHC class II presented bacterial CD4(+) T cell epitopes relevant for vaccine development. 相似文献
5.
Identification of formaldehyde-induced modifications in proteins: reactions with model peptides 总被引:1,自引:0,他引:1
Metz B Kersten GF Hoogerhout P Brugghe HF Timmermans HA de Jong A Meiring H ten Hove J Hennink WE Crommelin DJ Jiskoot W 《The Journal of biological chemistry》2004,279(8):6235-6243
Formaldehyde is a well known cross-linking agent that can inactivate, stabilize, or immobilize proteins. The purpose of this study was to map the chemical modifications occurring on each natural amino acid residue caused by formaldehyde. Therefore, model peptides were treated with excess formaldehyde, and the reaction products were analyzed by liquid chromatography-mass spectrometry. Formaldehyde was shown to react with the amino group of the N-terminal amino acid residue and the side-chains of arginine, cysteine, histidine, and lysine residues. Depending on the peptide sequence, methylol groups, Schiff-bases, and methylene bridges were formed. To study intermolecular cross-linking in more detail, cyanoborohydride or glycine was added to the reaction solution. The use of cyanoborohydride could easily distinguish between peptides containing a Schiff-base or a methylene bridge. Formaldehyde and glycine formed a Schiff-base adduct, which was rapidly attached to primary N-terminal amino groups, arginine and tyrosine residues, and, to a lesser degree, asparagine, glutamine, histidine, and tryptophan residues. Unexpected modifications were found in peptides containing a free N-terminal amino group or an arginine residue. Formaldehyde-glycine adducts reacted with the N terminus by means of two steps: the N terminus formed an imidazolidinone, and then the glycine was attached via a methylene bridge. Two covalent modifications occurred on an arginine-containing peptide: (i) the attachment of one glycine molecule to the arginine residue via two methylene bridges, and (ii) the coupling of two glycine molecules via four methylene bridges. Remarkably, formaldehyde did not generate intermolecular cross-links between two primary amino groups. In conclusion, the use of model peptides enabled us to determine the reactivity of each particular cross-link reaction as a function of the reaction conditions and to identify new reaction products after incubation with formaldehyde. 相似文献
6.
Dagmar Waberski Anke Döhring Florencia Ardón Nadine Ritter Holm Zerbe Hans-Joachim Schuberth Marion Hewicker-Trautwein Karl Fritz Weitze Ronald HF Hunter 《Acta veterinaria Scandinavica》2006,48(1):13-8
Whole boar semen or seminal plasma has been demonstrated to advance the time of ovulation in gilts. As a means of clarifying
this influence, the contribution of uterine lymphatics and their white cell populations has been examined. After duct visualisation
with Evan's blue, lymph was sampled from a mesometrial vessel in eight pre-ovulatory gilts whose uterine lumen was infused
simultaneously with whole semen in one ligated horn and saline in the contralateral ligated horn. Lymph was collected from
cannulated vessels for periods of up to four hours under general anaesthesia. Thereafter, mesometrial lymph nodes, utero-tubal
junction and uterine wall tissues were sampled. The proportion of nucleated cells in the sampled lymph increased towards the
end of the collection period, but erythrocytes were found in all instances preventing a meaningful differentiation and identification
of leukocytes. Prominent uterine lymph nodes were present in the mesometrium on both sides of the reproductive tract in 7
of 10 gilts. Differences in cellular contents were demonstrated between the side of the tract infused with semen and that
infused with saline control. Two of 4 gilts had lower values for CD4 (Cluster Differentiation) and 3 of 6 gilts higher values
for MHC II (Major Histocompatibility Complex) markers on the side challenged with semen. In contrast, values remained constant
for CD8 but ranged widely for CD18. Immunohistochemical analysis of uterine tissue samples for MHC II+ cells revealed significant
differences (P < 0.05) between the control and semen-treated ligated portions of the horns, as well as between the tissue
sample of uterine wall and that from the utero-tubal junction, but there were no significant differences for CD4+ cells. It
therefore remains plausible that semen-induced cytokines in the uterine lymph undergo counter-current transfer to the ipsilateral
ovary and accelerate the final maturation of pre-ovulatory Graafian follicles. 相似文献
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Solid-phase synthesis of DNA fragments containing the modified base 7-hydro-8-oxo-2'-deoxyguanosine. 总被引:2,自引:2,他引:0 下载免费PDF全文
H C Roelen C P Saris H F Brugghe H van den Elst J G Westra G A van der Marel J H van Boom 《Nucleic acids research》1991,19(16):4361-4369
The 5'-(4,4'-dimethoxytrityl) protected 3'-(2-cyanoethoxy)-N,N-diisopropylphosphoramidite of 7-hydro-8-oxo-2'-deoxy-guanosine, the exocyclic amino and lactam functions of which are protected with acetyl and diphenylcarbamoyl groups, respectively, has been prepared from the 8-bromo derivatives of deoxy- and riboguanosine. This synthon, in combination with standard d-nucleoside 3'-(2-cyanoethoxy)-N,N-diisopropylphosphoramidites, was applied successfully to a solid-phase synthesis. Well-defined oligodeoxyribonucleotides containing a 7-hydro-8-oxo-2'-deoxyguanosine residue at predetermined positions were obtained after deprotection with methanolic ammonia and purification by gel filtration. 相似文献
9.
Repair and replication of plasmids with site-specific 8-oxodG and 8-AAFdG residues in normal and repair-deficient human cells. 总被引:2,自引:0,他引:2 下载免费PDF全文
J C Klein M J Bleeker C P Saris H C Roelen H F Brugghe H van den Elst G A van der Marel J H van Boom J G Westra E Kriek 《Nucleic acids research》1992,20(17):4437-4443
The in vivo mutagenicity of 7-hydro-8-oxo-2'-deoxyguanosine (8-oxodG) and N-(guanin-8-yl)-N-acetyl-2-aminofluorene (8-AAFdG) in human cells was determined by transfecting various cell lines with plasmids that carried a single adduct at a defined site. 8-OxodG is one of the many DNA modifications formed by oxygen radicals, and was found to be highly miscoding during replication with purified DNA polymerases in vitro. Here we show that the frequency of mutations induced by 8-oxodG during replication in vivo is at most only 2% above background. The most predominant mutation found was a single G----T transversion. The frequency of this transversion was found to be 3 to 5-fold increased in excision repair deficient XP-A cells. Interestingly, also the replication of 8-oxodG containing plasmids was significantly impaired (approximately 4-fold) in the XP-A cells, but not in HeLa cells, normal fibroblasts or XP-A revertant cells. When 8-AAFdG containing plasmids were used, the mutation frequencies did not exceed background levels (less than 2%) with any of the cell lines tested. The presence of 8-AAFdG almost completely inhibited plasmid replication (more than 50-fold) in XP-A cells. Apparently, both 8-AAFdG and 8-oxodG are not or poorly repaired in these cells, causing a block of DNA replication. This suggests that both lesions are substrates for excision repair, although to a varying extent. 相似文献
10.
Marcia Berrêdo-Pinho Dario E Kalume Paloma R Correa Leonardo HF Gomes Melissa P Pereira Renata F da Silva Luiz RR Castello-Branco Wim M Degrave Leila Mendonça-Lima 《BMC microbiology》2011,11(1):80