Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.4 (trypsin)
 42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This study evaluates the accuracy of the PerioScan reagent card kit which uses BANA hydrolization to detect the presence of P. gingivalis, T. denticola, and B. forsythus in dental plaque during an experimental gingivitis in man. 32 healthy subjects underwent a phase of optimal oral hygiene before they abolished all oral hygiene practices for 21 days, but rinsed twice daily with a slurry of three different toothpastes. On days 0, 7, 14, and 21, full mouth Plaque and Gingival Index scores were assessed and, in addition, on days 0 and 21, sulcular plaque samples were obtained from the mesiobuccal aspects of the second premolars. The samples were placed on BANA reagent cards (PerioScan), and the result of the trypsin-like activity read after 15 minutes. Subsequently, the samples were processed for the detection of P. gingivalis, T. denticola and B. forsythus using ELISA. The Gingival Indices on day 21 indicated a development towards gingival inflammation. The frequencies of detection of the three periodontopathogens revealed by ELISA showed increased presence of P. gingivalis, B. forsythus and T. denticola on day 21. Changes in the composition of the microbiota were also indicated by the higher rate of positive BANA results at the end of the experimental gingivitis. Without considering further clinical diagnostic tests such as "bleeding on probing", this clinically simple test does not provide a prognostic indicator for the eventual onset of disease in cases with gingival inflammation. However, specificity was only 61% and the sensitivity was 41.7%.
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PMID:Application of BANA during experimental gingivitis. Application of N-alpha-benzoyl-DL-arginine 2 naphtilamide (BANA) hydrolysis to identify periodontopathic environments during experimental gingivitis in man. 147 Aug 87

Substrate impregnated paper discs were prepared using peptidyl derivatives of 7-amino-4-trifluoromethylcoumarin (AFC). After incubation with test solutions, the green, UV-induced fluorescence of AFC liberated by enzyme activity was distinguishable from the blue-violet fluorescence of the substrates. The AFC could then be coupled with p-dimethylaminocinnamaldehyde to form a colored Schiff base. Semi-quantiative assessments of disc fluorescence and color were made by comparison with AFC/substrate standards. Assays with discs impregnated with MeOSuc-Ala-Ala-Pro-Val-AFC, Z-Gly-Gly-Arg-AFC and Ala-Pro-AFC for elastase-, trypsin-, and dipeptidyl peptidase (DPP) IV-like activities respectively were evaluated using purified DPP IV and 100 eluates of crevicular fluid collected on filter paper strips from 10 gingivitis and periodontitis patients. The results showed that, within their working ranges, scores of disc fluorescence and color were reasonably accurate and reliable by comparison with enzyme activities measured in parallel quantitative fluorimetric assays with the same substrates. Using disc color, which was more sensitive than fluorescence, it was generally possible to measure all three enzyme activities in crevicular fluid samples from 5 periodontitis patients with varying degrees of gingival inflammation and pocketing. Disc color assays require no special apparatus and could be used for enzyme estimations in the clinical setting.
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PMID:A simple, combined fluorogenic and chromogenic method for the assay of proteases in gingival crevicular fluid. 214 76

The present investigation explored the hypothesis that elevated levels of certain enzymes in the gingival crevicular environment of individuals with poor oral hygiene and/or gingival inflammation may modify the surfaces of epithelial cells and thereby modulate the types of bacteria which attach and colonize. Buccal epithelial cells treated with neuraminidase and certain proteases were used as a model for study. Bacteria studied included Streptococcus sanguis and Streptococcus mitis which have been associated with gingival health, Actinomyces species which are increased in plaque associated with developing gingivitis, and Bacteroides gingivalis, Bacteroides intermedius, and Actinobacillus actinomycetemcomitans which are associated with destructive periodontal diseases. Treatment of epithelial cells with the enzymes studied produced selective effects on their receptivity for bacteria. Neuraminidase treatment of epithelial cells greatly reduced the attachment of all strains of S. sanguis and S. mitis studied. In contrast, the number of Actinomyces viscosus, A. naeslundii and A. israelii cells which attached was significantly increased. Neuraminidase treatment also appeared to enhance attachment of B. intermedius and B. gingivalis. Treatment of buccal cells with trypsin, chymotrypsin or papain also selectively affected bacterial attachment. Such protease treatment greatly reduced the numbers of streptococci and A. viscosus cells which attached, while the numbers of B. gingivalis and B. intermedius were significantly increased. Treatment of epithelial cells with preparations of lysosomal enzymes derived from human PMNs produced similar selective effects. The changes in bacterial adhesion observed by the enzyme treatments studied are consistent with the shifts in the composition of the gingival crevice flora which occur when oral hygiene is terminated and gingivitis develops.
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PMID:Selective modulation of bacterial attachment to oral epithelial cells by enzyme activities associated with poor oral hygiene. 214 77

Crevicular fluid was collected from gingivitis and periodontitis patients on filter paper strips and then eluted into buffer. The eluates hydrolyzed ZAlaArgArgAFC at alkaline pH and the effector response at pH 8.5 indicated serine proteinase activity. The results, particularly the substantial increase in activity produced by heparin, were suggestive of mast cell tryptase. They were also consistent with the properties of the tryptase-like enzyme identified in extracts of inflamed human gingiva (Cox and Eley, Arch Oral Biol, in press). Partial inhibition of crevicular fluid eluate activity by soybean trypsin inhibitor suggested the additional presence of a second trypsin-like enzyme which might be of host or bacterial origin.
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PMID:Tryptase-like activity in crevicular fluid from gingivitis and periodontitis patients. 252 68

Crevicular fluid samples were collected from 20 gingivitis and periodontitis patients using filter paper strips; these were then eluted into buffer. Portions of each sample were combined and the activities of this pooled eluate against different peptidyl derivatives of 7-amino-4-trifluoromethyl coumarin (AFC) were examined with respect to their pH profiles and effector responses. Ca-thepsin B- and L-like activity was detected with Bz-Val-Lys-Lys-Arg-AFC; elastase-like activity with MeOSuc-Ala-Ala-Pro-Val-AFC; tryptase-like activity with Z-Ala-Ala-Lys-AFC; trypsin-like activity with Z-Gly-Gly-Arg-AFC; and dipeptidyl peptidase (DPP) IV-like activity with Ala-Pro-AFC. The selectivity and sensitivity of these assays were improved by choice of appropriate conditions. The cathepsin B- and L-, elastase-, tryptase-, and trypsin-like activities all had properties consistent with those from host sources, whilst partial inactivation of the DPP IV-like activity by heat treatment (60 degrees C for 30 min) suggested that it may have represented a mixture of human and Bacteroides gingivalis enzymes. Individual patient eluates showed wide variations in enzyme concentrations, but generally elastase-like activity was by far the highest. The sensitivity of the assays with AFC-linked substrates was such that it should prove possible to measure all five different types of activity in crevicular fluid samples from local periodontal disease sites.
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PMID:Detection of cathepsin B- and L-, elastase-, tryptase-, trypsin-, and dipeptidyl peptidase IV-like activities in crevicular fluid from gingivitis and periodontitis patients with peptidyl derivatives of 7-amino-4-trifluoromethyl coumarin. 257 34

Inflamed gingiva contain a serine proteinase which could not previously be identified on the basis of its substrate specificity and inhibitor response. Using the substrate ZAlaArgArgAFC at alkaline pH, the enzyme was shown to be extracted more efficiently in high salt buffer. Inclusion of NaCl in assays, however, caused progressive reduction of activity. There was also inhibition by CaCl2, MgCl2 and 2 mM TosLysCH2Cl but not by 2 mM TosPheCH2Cl. Heparin produced significant activation. In gel filtrations with 1.0 M NaCl, activity appeared in fractions corresponding to a molecular weight of about 135,000. These properties are all consistent with tryptase from human mast cells. The enzyme may participate in both the connective tissue destruction and the inflammatory and immunological processes of gingivitis and periodontitis.
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PMID:Identification of a tryptase-like enzyme in extracts of inflamed human gingiva by effector and gel-filtration studies. 268 10

These activities were measured simultaneously, using synthetic fluorescent protease substrates, in gingival crevicular fluid collected at 6 pre-determined sites from 10 individuals with mild to moderate gingivitis. The three enzyme activities were detected in 85, 18 and 93% of the sites, respectively. The volume of fluid collected from discrete sites was significantly correlated with the total amount of substrate hydrolysed, but not with the specific rate of substrate hydrolysis. Log10 (total trypsin-like activity) was significantly correlated with the Gingival Index, Plaque Index and probing depth (r = 0.319, 0.423 and 0.336), while total glycylprolyl dipeptidase activity was significantly correlated with probing depth (r = 0.381). These findings add to knowledge of the biochemistry of gingival crevicular fluid, but the usefulness of such assays for diagnostic or monitoring purposes in periodontal diseases needs to be determined.
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PMID:Trypsin-like, chymotrypsin-like and glycylprolyl dipeptidase activities in gingival crevicular fluid from human periodontal sites with gingivitis. 269 44

Since they are found to be increased in lesions of acute necrotic ulcerative gingivitis or marginal periodontitis, agents for these diseases. In the present study, 38 pure cultured strains were obtained as a result of isolation and culture of samples collected from lesions of marginal periodontitis (periodontal pokets), and the biological and biochemical characteristics of these strains were investigated. 1) Light microscopy (including dark-field microscopy) and transmission electron microscopy (negative staining) were used for observation of the morphology and cellular structure of the strains. The cells had a spiral shape, and showed active movement. Based on the above findings the cultured strains were all confirmed to be spirochetes of small to medium size, being 0.08-0.24 micron in width. 2) Growth and motility of the strains were investigated on various types of culture medium. Intense growth and movement were noted in strains cultured in bovine liver exudate medium containing horse serum (pH 7.2) at 37 degrees C under anaerobic conditions produced by the evacuation-replacement method (95% N2, 5% CO2) for 3-7 days after inoculation. 3) Thirty-five strains were positive for indole production and decomposition of urea, mucin, hippuric acid and esculin. Production of hydrogen sulfied was observed in 31 strains. In decomposition tests for 17 carbohydrates, 17 strains were positive for galactose and 14 strains were positive for glucose, while 11 strains were positive for dextrin and 10 strains for fructose upon decomposition of soluble starch. Other carbohydrates were also decomposed by a few strains. 4) In an investigation of the production of alcohol and lower fatty acids, among the metabolic products detected by gas chromatography, a large amount of acetic acid and small amounts of ethanol, lactic acid, propionic acid, pyruvic acid were observed. 5) The results of enzyme activity tests using an API ZYM system indicated relatively high activities of esterase, esterase-lipase, alpha-glucosidase, alkaline phosphatase, trypsin and acid phosphatase.
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PMID:[Biological and biochemical characteristics of the oral spirochetes isolated from the focus of marginal periodontitis]. 276 48

The ability of Bacteroides gingivalis 381 to attach to hydroxyapatite (HA) beads, treated with either human type I or type IV collagen, or to particles of bovine bone collagen was studied. All preparations were blocked with human albumin prior to being incubated with 3H-thymidine-labeled B. gingivalis 381 cells. The presence of collagen on HA surfaces (C-HA) significantly promoted attachment of the organism. HA treated with Type IV collagen bound B. gingivalis cells more effectively than did HA treated with type I collagen. Attachment of two additional strains of B. gingivalis to HA was also promoted by collagen. Binding to type I or type IV C-HA occurred rapidly, and equilibrium was attained within 45 min. B. gingivalis 381 cells also bound to particles of bovine bone collagen, and this appeared to be biphasic. Heating the bacteria abolished their ability to bind to C-HA. Attachment of B. gingivalis 381 cells to HA treated with type I collagen was strongly inhibited by the presence of soluble type I or type IV collagen, or gelatin, but not by the presence of human albumin, salivary proline-rich protein 1, or saliva. Human serum, fibronectin, fibrinogen, certain protease inhibitors, and some peptides were also inhibitory. 3H-fibronectin bound to bovine bone collagen particles and blocked the attachment of 14C-B. gingivalis cells. Mild trypsin treatment of the fibronectin-collagen complex restored its ability to promote 14C-B. gingivalis attachment concomitant with the loss of 3H-fibronectin. We suggest that elevated levels of proteases in the gingival sulcus, such as are associated with poor oral hygiene and gingivitis, might remove fibronectin and expose collagen molecules in the basement membrane, thereby promoting the attachment of B. gingivalis cells and facilitating their invasion into gingival tissues.
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PMID:Attachment of Bacteroides gingivalis to collagenous substrata. 304 24

Leukotoxic activity in Actinobacillus (Haemophilus) actinomycetemcomitans isolated from patients with rapidly progressive periodontitis (RP), gingivitis (G), and juvenile periodontitis (JP), and several oral bacteria, was determined by observation of morphological changes in polymorphonuclear leukocytes (PMNs). Many A. actinomycetemcomitans isolates yielded both rough-surfaced and umbonate-shaped colonies (A-type), and smooth-surfaced and convex-shaped colonies (B-type), when stock cultures were streaked on agar medium. Both types of cells were identical in terms of Gram stain, cell morphology, sugar fermentation profile, nitrate reduction and cellular fatty acid composition. Sonic extracts were prepared from 32 A. actinomycetemcomitans strains isolated from patients and from 3 American Type Culture Collection (ATCC) strains. Sonic extracts from 8 isolates and 2 ATCC strains induced sphering of PMNs during a 45-50 min period of incubation at 37 C. Extracts from the other oral bacteria had no effects on PMN morphology. The sphered PMNs were found by their fluorochromatic-negative reactions to be damaged cells. The leukotoxic substance was heat-sensitive (56 C, 30 min), trypsin-sensitive and did not induce sphering of PMNs at 4 C. There was no clear correlation between colony type and leukotoxicity. Among 8 leukotoxic strains, 5 were isolates from an RP patient.
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PMID:Leukotoxic activity in Actinobacillus (Haemophilus) actinomycetemcomitans isolated from periodontal disease patients. 361 93


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