lactoferrin and thiocyanate

We studied the effects of glycopyrrolate on oral mucous host defenses. Single IV doses of glycopyrrolate (4 microg/kg) or placebo were administered to 12 healthy volunteers in a randomized, double-blinded, cross-over study. Salivary flow rates and the concentrations/activities of total protein, amylase, and nonimmunologic (lysozyme, lactoferrin, myeloperoxidase, total salivary peroxidase, and thiocyanate) and immunologic (total immunoglobulin A, immunoglobulin G, and immunoglobulin M) mucous host defense factors were determined for paraffin-stimulated whole saliva before and 1, 3, 6, 12, 24, and 48 h after drug administration. Glycopyrrolate serum concentrations were determined before and 2, 4, 6, 10, 15, and 30 min and 1, 2, 3, 6, 12, and 24 h after IV drug injection. Salivary flow rates were decreased significantly for 12 h after glycopyrrolate injection, compared with saline injection. The concentrations of immunologic and nonimmunologic defense factors were increased in the glycopyrrolate group, and differences between the groups were found for all factors (P < 0.05-0.001) except lysozyme and total salivary peroxidase. In contrast, because of the reduced flow rate, the output of all defense factors into the saliva was decreased after glycopyrrolate injection, compared with saline injection. Glycopyrrolate thus decreases the output of salivary host defense factors into the oral cavity.. Glycopyrrolate induces long-lasting hyposalivation and decreases the secretion of salivary immunologic and nonimmunologic defense factors in healthy volunteers.

Topics: Adjuvants, Anesthesia; Adult; Amylases; Cross-Over Studies; Double-Blind Method; Glycopyrrolate; Humans; Immunoglobulins; Injections, Intravenous; Lactoferrin; Male; Mouth Mucosa; Muramidase; Peroxidase; Saliva; Salivary Proteins and Peptides; Thiocyanates

This study was designed to compare various salivary parameters between smokers and non-smokers and to determine the influence of a nicotine-containing chewing gum (used to aid in quit-smoking efforts) upon these same parameters. At the baseline examination, subjects were assigned to one of three groups: non-smokers who did not utilize any gum, smokers provided a nicotine-containing gum, and smokers provided a placebo gum. Saliva was collected from all subjects and analyzed for acidogenicity and buffer pH as well as for levels of thiocyanate, lactoperoxidase, lysozyme, lactoferrin, and secretory IgA. After 15 weeks of gum usage, saliva was again collected from each subject and the identical analyses performed. Significant differences were observed between smokers and non-smokers with regard to three parameters: The saliva of smokers contained greater concentrations of thiocyanate and lower concentrations of lactoferrin, at the baseline examination and after the 15-week test period. In addition, the CO content of alveolar air was higher in smokers at both examination periods. In contrast, the use of the nicotine gum per se had no effect on any of the test parameters.

Topics: Adult; Carbon Monoxide; Chewing Gum; Humans; Hydrogen-Ion Concentration; Immunoglobulin A, Secretory; Lactoferrin; Lactoperoxidase; Muramidase; Nicotine; Saliva; Smoking; Thiocyanates

Copper-containing plasma protein ceruloplasmin (Cp) forms a complex with lactoferrin (Lf), an iron-binding protein, and with the heme-containing myeloperoxidase (Mpo). In case of inflammation, Lf and Mpo are secreted from neutrophil granules. Among the plasma proteins, Cp seems to be the preferential partner of Lf and Mpo. After an intraperitoneal injection of Lf to rodents, the "Cp-Lf" complex has been shown to appear in their bloodstream. Cp prevents the interaction of Lf with protoplasts of Micrococcus luteus. Upon immunoprecipitation of Cp, the blood plasma becomes depleted of Lf and in a dose-dependent manner loses the capacity to inhibit the peroxidase activity of Mpo, but not the Mpo-catalyzed oxidation of thiocyanate in the (pseudo)halogenating cycle. Antimicrobial effect against E. coli displayed by a synergistic system that includes Lf and Mpo-H2O2-chloride, but not thiocyanate, as the substrate for Mpo is abrogated when Cp is added. Hence, Cp can be regarded as an anti-inflammatory factor that restrains the halogenating cycle and redirects the synergistic system Mpo-H2O2-chloride/thiocyanate to production of hypothiocyanate, which is relatively harmless for the human organism. Structure and functions of the "2Cp-2Lf-Mpo" complex and binary complexes Cp-Lf and 2Cp-Mpo in inflammation are discussed.

Topics: Animals; Antimicrobial Cationic Peptides; Ceruloplasmin; Chlorides; Escherichia coli; Humans; Hydrogen Peroxide; Injections, Intraperitoneal; Lactoferrin; Mice; Mice, Inbred C57BL; Micrococcus luteus; Models, Molecular; Peroxidase; Protein Interaction Domains and Motifs; Rats; Rats, Wistar; Thiocyanates

Caries as a multifactorial process is influenced by salivary defense. Cluster analyses should give additional information on the role of salivary variables in relation to caries increment.. Samples of unstimulated and stimulated whole saliva from 28 young adults (mean age 23.5+/-2.1 years) were analyzed for flow rate, pH and buffer variables, lysozyme, lactoferrin, peroxidase, thiocyanate, secretory immunoglobulin A, and total protein. The decayed, missing, and filled surfaces (DMFS) were recorded at baseline and after 4 years. Cluster analyses were executed on the basis of salivary data.. The mean caries increment (DeltaDMFS) over 4 years was 6.7+/-4.0 (range 1-16). In two-cluster processing, three out of four volunteers with low caries increments were grouped into one cluster. Only a few variables proved to be important for cluster characteristics.. The results suggest that over 4 years (1) the volunteers with very low caries increment (DeltaDMFS=1) are classified always together, (2) these volunteers do not form a separate cluster by themselves, (3) low caries increment was related to higher salivary flow rate and lower levels of lysozyme and lactoferrin for unstimulated saliva and (4) the partial pressure of CO(2) was of importance in stimulated saliva.

Topics: Adult; Buffers; Carbon Dioxide; Cluster Analysis; Dental Caries; DMF Index; Female; Humans; Hydrogen-Ion Concentration; Immunoglobulin A, Secretory; Incidence; Lactoferrin; Longitudinal Studies; Male; Muramidase; Partial Pressure; Saliva; Salivary Proteins and Peptides; Secretory Rate; Thiocyanates

Lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-peroxide-system are naturally occurring antimicrobial components of milk. The objective of this study was to examine, whether these components were responsible for negative results, when mastitis milk is cultured microbiologically. Quarter milk samples from 75 cows with clinical mastitis on a dairy farm in Brandenburg were submitted for microbiological culture and analysed for the content and the activities of the three components. Animals from all stages of lactation with clinical mastitis were included in the study. Animals were examined clinically and milk samples were collected prior to first treatment. Secretions from quarters with clinical mastitis were compared to those of neighbouring quarters without clinical mastitis. Secretions with positive cultural results were compared to those with negative results. The concentrations or activities of the three factors were significantly higher in the diseased quarters than in the quarters without clinical signs of mastitis. The concentration of lysozyme increased with severity of the clinical signs (local swelling and changes in secretion). The concentration of lactoferrin was significantly higher in quarters with slight alterations of glandular tissue than in quarters with medium or severe alterations (P < 0.05). LPS-activities did not correlate with the severity of clinical signs. No differences in the concentration of lactoferrin or LPS-activities were seen between mastitis with positive and negative culture results. The concentration of lysozyme was even higher in culturally positive samples than in negative samples (P < 0.05). Results from this study indicate that the three factors examined did not impair the results of microbiological culture of milk samples from quarters with clinical mastitis.

Topics: Animals; Cattle; Female; Fermentation; Hydrogen Peroxide; Lactoferrin; Lactoperoxidase; Mastitis, Bovine; Milk; Muramidase; Thiocyanates

Previous studies of the possible associations of salivary antimicrobial agents with dental caries have given controversial results, obviously mainly because almost all studies have been cross-sectional. Our aim was to find out, in a two-year longitudinal follow-up study, the associations among selected salivary non-immune and immune antimicrobial variables, cariogenic bacteria, and caries increment. The study population was comprised of 63 subjects, all of whom had their 13th birthday during the first study year. In addition to a comprehensive dental examination at baseline and after 2 yrs, paraffin-stimulated whole saliva samples were collected in a standardized way at six-month intervals. Saliva samples were analyzed for flow rate, buffer effect, lysozyme, lactoferrin, total peroxidase activity, hypothiocyanite, thiocyanate, agglutination rate, and total and specific anti-S. mutans IgA and IgG, as well as for numbers of total and mutans streptococci, lactobacilli, and total anaerobic bacteria. Cluster analysis and Spearman-Rank correlation coefficients were used to explore possible associations between and among the studied variables. During the two-year period, a statistically significant increase was observed in flow rate, thiocyanate, agglutination rate, anti-S. mutans IgA antibodies, lactobacilli, and total anaerobes, whereas lysozyme, lactoferrin, and total and anti-S. mutans IgG antibodies declined significantly. Based on various analyses, it can be concluded that, at baseline, total IgG and hypothiocyanite had an inverse relationship with subsequent two-year caries increment, anti-S. mutans IgG antibodies increased with caries development, and mutans streptococci and lactobacilli correlated positively with both baseline caries and caries increment. Total anaerobic microflora was consistently more abundant among caries-free individuals. In spite of the above associations, we conclude that none of the single antimicrobial agents as such has sufficiently strong power to have diagnostic significance in vivo with respect to future caries.

Topics: Adolescent; Agglutination; Anti-Bacterial Agents; Antibodies, Bacterial; Bacteria, Anaerobic; Buffers; Child; Cluster Analysis; Cohort Studies; Colony Count, Microbial; Cross-Sectional Studies; Dental Caries; DMF Index; Female; Finland; Follow-Up Studies; Humans; Immunoglobulin A, Secretory; Immunoglobulin G; Lactobacillus; Lactoferrin; Longitudinal Studies; Male; Muramidase; Peroxidases; Saliva; Secretory Rate; Streptococcus mutans; Thiocyanates

The number of decayed, missed and filled permanent teeth (DMFT), the degree of periodontal inflammation (Periodontal Status Index, PSI), stimulated salivary flow rate and the concentrations of total protein, lactoferrin, lysozyme, myeloperoxidase, salivary peroxidase, calcium, potassium, sodium and thiocyanate in whole saliva of 26 adult asthma patients were compared with those of 33 non-asthmatic controls. The saliva was also analysed for mutans streptococci, lactobacilli, total anaerobic flora and Candida spp. The mean PSI (p < 0.05; 95% confidence interval for the difference between means (95% CI) 2.47-25.30) was higher and the mean stimulated salivary flow rate (p < or = 0.05; 95% CI 0.57-0.55) was lower in the asthmatic group than in the control group. No differences were found between the groups in non-immune defense factors, except for myeloperoxidase. The myeloperoxidase concentrations were higher in asthmatics than in non-asthmatics (p < 0.05; 95% CI 4.4-134.0 ng/ml). No differences in microbial counts were found. It was concluded that stimulated salivary flow rates decrease while myeloperoxidase concentrations increase in adult asthmatic patients compared with non-asthmatic adults. The higher concentrations of myeloperoxidase are explained by a higher PSI in asthmatics.

Topics: Adult; Asthma; Bacteria, Anaerobic; Calcium; Candida; Colony Count, Microbial; Confidence Intervals; DMF Index; Female; Humans; Lactobacillus; Lactoferrin; Male; Middle Aged; Muramidase; Periodontal Index; Periodontitis; Peroxidase; Peroxidases; Potassium; Saliva; Salivary Proteins and Peptides; Secretory Rate; Sodium; Streptococcus mutans; Thiocyanates

The aim of the present study was to evaluate the salivary secretion rate and composition in a group of 16 children and young adults (6-27 years) with Papillon-Lefèvre Syndrome (PLS), and to compare the findings with a group (n = 16) of healthy controls. Unstimulated and stimulated whole saliva was collected at least 2 h after meals and the secretion rate determined. The stimulated saliva was assessed for buffer capacity, total protein, peroxidase and hexosamine, while the unstimulated samples were evaluated for total protein, lysozyme, thiocyanate, lactoferrin and salivary IgA. Both the unstimulated (p < 0.01) and stimulated (p < 0.05) saliva secretion rates were significantly lower among the PLS patients compared with the controls. Furthermore salivary buffer capacity was significantly (p < 0.01) lower in the PLS patients. The total protein content in saliva was comparatively high in the study group, while the concentrations of immunoglobulins and non-immunoglobulins were within normal ranges. When calculating the output of the assessed antimicrobial factors, the mean peroxidase level in stimulated whole saliva was found to be significantly (p < 0.01) lower in the PLS patients than in the healthy controls. In conclusion, the present study indicates an impaired water secretion and a somewhat altered saliva gland function in children and young adults with PLS.

Topics: Adolescent; Adult; Case-Control Studies; Child; Child, Preschool; Female; Hexosamines; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Muramidase; Papillon-Lefevre Disease; Periodontitis; Peroxidase; Saliva; Salivary Glands; Salivary Proteins and Peptides; Secretory Rate; Statistics, Nonparametric; Thiocyanates; Water-Electrolyte Balance

Resting and stimulated whole saliva was collected from 94 children aged 12-14 years and analyzed for thiocyanate, hypothiocyanite, 'free' and 'total' lysozyme, lactoferrin and secretory IgA. Clinical assessments of the amounts of plaque and gingival inflammation were made, and plaque was collected for determination of dry weight. An inverse relationship was observed between salivary thiocyanate concentrations in both resting and stimulated saliva and the amounts of plaque and gingival inflammation in these subjects (p < 0.05). Lactoferrin concentration in stimulated saliva was directly related to the amounts of plaque and gingivitis (p < 0.05). 'Total' lysozyme concentration in stimulated saliva was directly related to the amount of plaque (p < 0.05), and the 'free' lysozyme concentration in the same saliva was directly related to the amount of gingivitis (p < 0.05). The direct relationship observed between clinical measurements and both lysozyme and lactoferrin concentrations in saliva may have been due to contributions from gingival crevicular fluid. Cluster analysis identified three groups of subjects with different profiles in resting whole saliva, and in particular with different levels of secretory IgA. A statistically significant difference was observed in the quantity of plaque collected from subjects in two of these groups (p < 0.05). These results from cluster analysis using resting whole saliva from children confirmed the findings of a previous study with young adults.

Topics: Adolescent; Child; Cluster Analysis; Dental Plaque; Gingivitis; Humans; Immunoglobulin A, Secretory; Lactoferrin; Multivariate Analysis; Muramidase; Saliva; Salivary Proteins and Peptides; Secretory Rate; Thiocyanates

We have studied the possible relationship between indigenous salivary antimicrobial agents, indigenous mutans streptococci and the capability of added mutans streptococci to grow in saliva. Stimulated whole saliva was collected from 19 healthy donors. Saliva samples were sterilized, supplemented with glucose and inoculated with Streptococcus mutans or Streptococcus sobrinus. The mixtures were incubated for 20 h followed by counting of viable cells. Saliva samples were analysed, both before and after sterilization, for indigenous antimicrobial agents and the bacterial flora. The subjects could be divided into two groups: those (n = 9) whose saliva promoted and those (n = 10) whose saliva inhibited the growth of the inoculated streptococci. A statistically significant correlation (+0.82, p < 0.001) was found between the numbers of viable cells of S. mutans and S. sobrinus after incubation in saliva. The sterilization procedure reduced the content of all antimicrobial proteins. Salivary antimicrobial factors, or levels of indigenous mutans streptococci, did not differ between the two groups. We conclude that none of the individual salivary antimicrobial factors alone can explain the large individual differences in growth-promoting or growth-inhibiting patterns of saliva on S. mutans and S. sobrinus. Inter-individually, saliva either supports or inhibits the growth of mutans streptococci, indicating a similar response of these two species in relation to the properties of saliva.

Topics: Adult; Anti-Bacterial Agents; Antibodies, Bacterial; Female; Humans; Hydrogen-Ion Concentration; Immunoglobulin A, Secretory; Immunoglobulin G; Lactoferrin; Male; Muramidase; Peroxidase; Peroxidases; Saliva; Sterilization; Streptococcus mutans; Streptococcus sobrinus; Thiocyanates

To assess the effects of smokeless tobacco on the secretory immune system and dental caries, we examined users of smokeless tobacco and non-tobacco users. There were no significant differences in the prevalence of DMFS between users and non-users. There was significantly more salivary IgA, IgA2 and J-chain in users. Levels of salivary lysozyme and lactoferrin were significantly lower in users than controls. Because there was no difference in levels of secretory component in relation to the increased IgA levels of smokeless tobacco users, this suggests an effect of smokeless tobacco on secretory epithelial cells responsible for synthesis of secretory component, lysozyme and lactoferrin, and for the packaging of secretory component on IgA. There were only slight differences in salivary or serum antibody levels to Streptococcus mutans. These findings indicate that although smokeless tobacco has a significant influence on the synthesis of secretory IgA, the numbers of DMFS were similar between smokeless tobacco users and controls.

Topics: Adolescent; Adult; Amylases; Antibodies, Bacterial; Blood; Glucosyltransferases; Humans; Immunoglobulin J-Chains; Immunoglobulins; Lactoferrin; Male; Muramidase; Plants, Toxic; Saliva; Salivary Proteins and Peptides; Secretory Component; Streptococcus mutans; Thiocyanates; Tobacco, Smokeless; Virulence

Saliva antimicrobial proteins may interact in a common system to influence the oral ecology. Clinical studies of antimicrobial protein action thus may require a multiple-protein approach. Multivariate statistical methods have been used to describe possible patterns of interaction for lysozyme, lactoferrin, salivary peroxidase and secretory IgA in stimulated parotid saliva. However, oral microbes are most likely to encounter antimicrobial proteins in mixed resting saliva. Relationships among levels of lysozyme, lactoferrin, salivary peroxidase, and secretory IgA therefore were investigated in whole saliva from 216 subjects, and an attempt made to relate interperson variation in those proteins to differences in health and status, and dental plaque accumulation and composition. All proteins were significantly (alpha = 0.05) correlated with each other (r = 0.38-0.52, p less than 0.001). There was only one axis of common variation among proteins, and that axis was significantly correlated (p less than 0.001) with total protein (r = 0.84) and flow rate (r = -0.56). That pattern deviated from the previous finding that proteins of acinar origin tended to vary independently from proteins of ductal origin in stimulated parotid saliva. The difference between parotid and whole saliva may reflect constitutive secretion of all proteins at low levels of stimulation. Common variation of unstimulated saliva proteins suggests that antimicrobial actions can be compared in subjects at population extremes. There were no significant associations between antimicrobial proteins in whole saliva and measures of health status or plaque accumulation. However, the proportions of Streptococcus sanguis were significantly correlated with lysozyme (r = -0.26), lactoferrin (r = -0.34), peroxidase (r = -0.30), total protein (r = -0.37), flow rate (r = 0.24) and principal-components scores (r = -0.33) in a subset of subjects (n = 85) where commercial biochemical tests were used to supplement species identification by colony morphology. Those findings may indicate that saliva antimicrobial proteins can affect the composition of dental plaque.

Topics: Adolescent; Adult; Bacteria; Bacterial Physiological Phenomena; Dental Plaque; Dental Plaque Index; Female; Health Status; Humans; Immunoglobulin A, Secretory; Lactoferrin; Male; Muramidase; Oral Health; Peroxidases; Saliva; Salivary Proteins and Peptides; Secretory Rate; Streptococcus; Streptococcus mutans; Streptococcus sanguis; Thiocyanates

Lactoferrin (LF) is an iron-binding glycoprotein common to exocrine secretions and the specific granules of neutrophils. Each molecule is capable of high-affinity coordinate-binding of two ferric ions with two bicarbonate or carbonic anions. The initial aspect of the present study was directed at determining the nature of anion involvement in LF bactericidal activity. It was found that selective anions were capable of inhibiting the expression of bactericidal activity by LF on S. mutans 10449. The ability to block LF expression was directly related to the capacity of the anion to serve as a coordinate ion in iron-binding by the transferrin molecules. These data support the hypothesis that the LF target site on the bacterial surface is anionic. There has been controversy in the literature regarding LF involvement in hydroxy radical generation. The second phase of these studies indicated that treatment of S. mutans with LF under anaerobic conditions abrogated the bactericidal effect of this molecule. LF-killing could be enhanced by the presence of thiocyanate and inhibited by catalase and lactoperoxidase; however, bovine serum albumin was equally effective as an inhibitor. The apparent requirement for oxygen in LF bactericidal effect on S. mutans is not inconsistent with a hydroxy radical mechanism.

Topics: Anaerobiosis; Anions; Carbon Dioxide; Humans; Hydrogen Peroxide; Lactoferrin; Lactoglobulins; Oxygen; Streptococcus mutans; Thiocyanates; Time Factors