Isolation of human salivary polymorphonuclear leukocytes and their stimulation-coupled responses

Yamamoto, Masazumi; Saeki, Kiyomi; Utsumi, Kozo

doi:10.1016/0003-9861(91)90444-n

Cited by 79 publications

(51 citation statements)

References 32 publications

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“…Since the oral cavity is directly open to the external environment, P. gingivalis should occasionally be exposed to aerobic conditions. It has been reported that neutrophils in saliva, but not in peripheral blood, always produce active oxygen molecules (66). Active oxygen molecules produced by phagocytes in gingival epithelial cells (32) may have inhibitory effects on invading P. gingivalis.…”

Section: Vol 72 2006 P Gingivalis Virulence Factors In a Chemostatmentioning

confidence: 99%

Effects of Various Growth Conditions in a Chemostat on Expression of Virulence Factors inPorphyromonas gingivalis

Masuda

Murakami

Noguchi³

et al. 2006

Appl Environ Microbiol

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Porphyromonas gingivalis, one of the gram-negative organisms associated with periodontal disease, possesses potential virulence factors, including fimbriae, proteases, and major outer membrane proteins (OMPs). In this study, P. gingivalis ATCC 33277 was cultured in a chemostat under hemin excess and presumably peptidelimiting conditions to better understand the mechanisms of expression of the virulence factors upon environmental changes. At higher growth rates, the amounts of FimA and the 75-kDa protein, forming long and short fimbriae, respectively, increased significantly, whereas gingipains decreased in amount and activity. In a nutrient-limited medium, lesser amounts of the above two fimbrial proteins were observed, whereas clear differences were not found in the amounts of gingipains. In addition, two-dimensional electrophoresis revealed that proteins in cells were generally fewer in number during nutrient-limited growth. Under aeration, a considerable reduction in gingipain activity was found, whereas several proteins associated with intact cells significantly increased. However, the expression of major OMPs, such as RagA, RagB, and the OmpA-like proteins, was almost constant under all conditions tested. These results suggest that P. gingivalis may actively control expression of several virulence factors to survive in the widely fluctuating oral environment.It has been reported that more than 80% of U.S. adults have overt gingivitis and about half of them have periodontitis (1). In periodontal diseases, teeth may be lost after inflammation spreads to the supporting periodontal tissues. Not only local events such as inflammation and trauma but also systemic and environmental factors are involved in establishment of periodontal diseases (9). The formation and maturation of plaque, especially subgingival plaque in periodontal pockets, are also involved (61). Although there are more than 600 species of bacteria in the mouth (30), specific species of gram-negative anaerobic bacteria, spirochetes, and motile rods are known to participate in formation of subgingival plaque (20,60). Porphyromonas gingivalis, a gram-negative anaerobe, is considered to contribute to periodontitis, since it is detected frequently in patients with these diseases (56, 60).Asaccharolytic P. gingivalis mainly obtains nutrients by degradation of proteins. This bacterium is known to have many potential virulence factors, including fimbriae, proteases, hemagglutinins, lipopolysaccharide, capsule polysaccharides, and major outer membrane proteins (26, 31). P. gingivalis expresses two types of fimbriae, long fimbriae composed of FimA (fimbrilin) (67), and short fimbriae composed of a 75-kDa protein (21, 51, 68). Many studies have shown that FimA has various functions, which include attachment to and invasion of gingival epithelial cells (28, 64), coaggregation with other species of bacteria (2), attachment to salivary components (4), activation of fibroblasts and macrophages (24), induction of inflammatory cytokines, and bone resorption (23)....

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Section: Vol 72 2006 P Gingivalis Virulence Factors In a Chemostatmentioning

confidence: 99%

Effects of Various Growth Conditions in a Chemostat on Expression of Virulence Factors inPorphyromonas gingivalis

Masuda

Murakami

Noguchi³

et al. 2006

Appl Environ Microbiol

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“…Since leukocytes are normally present in the oral cavity (Al-Essa et al, 1994;Yamamoto et al, 1991), collected whole saliva usually contains not only salivary peroxidase but also myeloperoxidase from the leukocytes. Salivary peroxidase is a heme protein and can catalyze H 2 O 2 -dependent oxidation of SCN -and I -but not Cl - (Tenovuo, 1989), and these anions are salivary components.…”

mentioning

confidence: 99%

Oxidation of Quercetin by Salivary Components. I. Salivary Peroxidase-Dependent Oxidation of Quercetin and Characterization of the Oxidation Products.

Takahama

Hirota

Nishioka

et al. 2002

FSTR

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Quercetin glucosides ingested as components of a foodstuff can be transformed to quercetin, and the quercetin is oxidized by peroxidase in the oral cavity. One objective of this study was the characterization of the oxidation products. Although both salivary peroxidase and myeloperoxidase are contained in whole saliva, salivary peroxidase is mainly involved in the oxidation in the saliva. As the oxidation products, 2,3,5,7,3¢ ¢ ¢ ¢,4¢ ¢ ¢ ¢-hexahydroxyflavanone-like compound and two isomers of a compound, which was transformed to quercetin and the above flavanone by a HCl/heattreatment, were isolated. 3,4-Dihydroxybenzoic acid and 2,4,6-trihydroxyphenylglyoxylic acid were also detected in the oxidation products. When dialyzed saliva was used, the salivary peroxidase-dependent oxidation of quercetin and the formation of the oxidation products were partly inhibited by 1 mM thiocyanate which was a substrate for the peroxidase in the oral cavity. The mechanism and the significance of salivary peroxidase-dependent oxidation of quercetin in the oral cavity are discussed.

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“…It has been proposed that the exposure of A. actinomycetemcomitans to intermittent aerobic conditions (due to production of oxygen by neutrophils and phagocytes in the oral environment) might affect the expression of colonization determinants and, ultimately, the invasion of host cells (42,72). Interestingly, the present study found all three selected biofilm determinant genes to be transcribed at a higher level in anaerobically grown cells than in aerobically grown cells.…”

Section: Discussionmentioning

confidence: 39%

Transcriptional and Translational Analysis of Biofilm Determinants of Aggregatibacter actinomycetemcomitans in Response to Environmental Perturbation

2009

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Fimbriae, lipopolysaccharide (LPS), and extracellular polymeric substance (EPS) all contribute to biofilm formation by the periodontopathogen Aggregatibacter actinomycetemcomitans. To understand how individual biofilm determinants respond to changing environmental conditions, the transcription of genes responsible for fimbria, LPS, and EPS production, as well as the translation of these components, was determined in rough (Rv) and isogenic smooth (Sv) variants of A. actinomycetemcomitans cultured in half-strength and full-strength culture medium under anaerobic or aerobic conditions, and in iron-supplemented and iron-chelated medium. The transcription of tadV (fimbrial assembly), pgaC (extracellular polysaccharide synthesis), and orf8 or rmlB (lipopolysaccharide synthesis) was measured by real-time PCR. The amounts of fimbriae, LPS, and EPS were also estimated from stained sodium dodecyl sulfate-polyacrylamide gels and verified by Western blotting and enzyme-linked immunoadsorbent assay using specific antibodies. Each gene was significantly upregulated in the Rv compared to in the Sv. The transcription of fimbrial, LPS, and EPS genes in the Rv was increased approximately twofold in cells cultured in full-strength medium under anaerobic conditions compared to that in cells cultured under aerobic conditions. Under anaerobic conditions, the transcription of fimbrial and EPS enzymes was elevated in both Rv and Sv cells cultured in half-strength medium, compared to that in fullstrength medium. Iron chelation also increased the transcription and translation of all biofilm determinants compared to their expression with iron supplementation, yet the quantity of biofilm was not significantly changed by any environmental perturbation except iron limitation. Thus, anaerobic conditions, nutrient stress, and iron limitation each upregulate known biofilm determinants of A. actinomycetemcomitans to contribute to biofilm formation.

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Isolation of human salivary polymorphonuclear leukocytes and their stimulation-coupled responses

Cited by 79 publications

References 32 publications

Effects of Various Growth Conditions in a Chemostat on Expression of Virulence Factors inPorphyromonas gingivalis

Effects of Various Growth Conditions in a Chemostat on Expression of Virulence Factors inPorphyromonas gingivalis

Oxidation of Quercetin by Salivary Components. I. Salivary Peroxidase-Dependent Oxidation of Quercetin and Characterization of the Oxidation Products.

Transcriptional and Translational Analysis of Biofilm Determinants of Aggregatibacter actinomycetemcomitans in Response to Environmental Perturbation

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