Generation of bovine immune colostrum against Streptococcus mutans and Streptococcus sobrinus and its effect on glucose uptake and extracellular polysaccharide formation by mutans streptococci

Due to potential side-effects of active immunization by cariogenic mutans streptococci, oral administration of passively-derived antibodies could be a more acceptable way to reduce colonization and virulence of these microorganisms in human dentition. The aim of this study was to produce antistreptococcal immunoglobulins into bovine colostrum and explore the possible antibacterial mechanisms of these immunoglobulins against mutans streptococci. Specific serum IgG antibodies to whole cell antigens of both Streptococcus mutans and Streptococcus sobrinus increased rapidly in cows during immunization and were high also in the final whey-product. Low concentration (0.5% w/v) of bovine immune preparation inhibited significantly the incorporation of [14C]glucose by both S. mutans and S. sobrinus. Higher concentration (> 1%) was needed to inhibit the glucosyltransferase or fructosyltransferase activities of these bacteria. No such inhibitory effects were observed with the control preparation from the non-immunized cows. Our results indicate that bovine immune colostrum has a significant inhibitory potential against mutans streptococci, apparently dependent on the presence of specific IgG antibodies against S. mutans and S. sobrinus.     

The role of the Streptococcus mutans glucosyltransferases in the sucrose-dependent attachment to smooth surfaces: essential role of the GtfC enzyme

Previous results have indicated that the glucosyltransferase activities of mutans streptococci are required for sucrose-dependent colonization of tooth surfaces. We have constructed mutants of Streptococcus mutans GS5 that are altered in varying combinations of the three gtf genes present in this organism. A quantitative in vitro sucrose-dependent attachment system was used to demonstrate that the inactivation of the gtfC gene drastically reduced adherence to smooth surfaces. By contrast, inactivation of the gtfB gene resulted in a smaller, but significant, reduction in attachment while the gtfD mutant was only marginally affected. Furthermore, production of only the glucosyltransferase C enzyme allowed for attachment although at reduced levels compared to the wild-type organism. The results from reintroduction of single copies of each of the gtf genes into a mutant of strain GS5 lacking glucosyltransferase activity also demonstrated the crucial role of the glucosyltransferase C enzyme in colonization. These results suggest a unique role for the glucosyltransferase C enzyme in the sucrose-dependent colonization of tooth surfaces by S. mutans strains.     

Effect of Cervitec varnish on the salivary Streptococcus mutans levels in the patients with fixed orthodontic appliances

The aim of the present study was to assess the efficiency of a 1% chlorhexidine-containing varnish (Cervitec, Vivadent, Liechtenstein) on the levels of Streptococcus mutans in saliva of patients with fixed orthodontic appliances using the Dentocult-SM (Vivadent, Liechtenstein) technique for the microbiological investigation. Eighty subjects participated in the study and, divided randomly into two equal groups in which one group was treated with the placebo varnish (Vivadent, Liechtenstein) for negative controls. Streptococcus mutans in saliva of the subject was sampled and enumerated by using the Dentocult-SM dip-slide technique for periods of one, two four and twelve weeks after a single varnish application. The results were evaluated statistically. After the chlorhexidine containing varnish treatment the levels of Streptococcus mutans in saliva were significantly reduced after one week (p < 0.01) and continued reduction for one month (p < 0.05). After twelve weeks Streptococcus mutans levels in saliva were given a relative increase. No significant suppression was found in the placebo group (p > 0.05). The results suggested that Cervitec varnish reduces salivary Streptococcus mutans levels and that the application should be repeated every 3 months to get antibacterial effect.     

Effect of an enzymatic rinse on salivary levels of Streptococcus mutans and lactobacilli in periodontally treated patients

Root surface caries is prevalent in patients with both treated and untreated periodontal disease. The major etiologic factor has been identified as microbial plaque. In periodontally treated patients, significantly higher root caries prevalence and incidence have been found in patients with high levels of Streptococcus mutans and Lactobacilli in saliva. Reducing the levels of S. mutans and Lactobacilli in saliva may lower the risk of root caries development. The purpose of this investigation was to determine the effect of an oral enzymatic rinse on the salivary counts of S. mutans and Lactobacilli in periodontally treated patients. Fifteen adult subjects participated in a double-blind, cross-over designed clinical trial. Each subject had previously undergone comprehensive periodontal therapy and had been maintained on a regular program of supportive periodontal therapy. Paraffin-stimulated whole saliva was collected from each participant. Each subject was then randomly given either the enzymatic rinse product or a control rinse and instructed to rinse with one tablespoonful twice a day for 2 weeks, after which saliva samples were taken. After a washout period, salivary samples were again taken, and the subjects received the alternate rinse product. Two weeks later, final salivary samples were taken. The salivary samples were serially diluted and incubated aerobically on selective culture media. S. mutans and Lactobacilli were counted on the basis of colonial morphology. Pretreatment and posttreatment salivary counts of S. mutans and Lactobacilli were analyzed using the Wilcoxon matched-pairs signed-rank test at the 5% level of significance. Analysis of data revealed that neither the test nor the control rinse significantly lowered salivary counts of either species in the sample population.     

Assessment of salivary Lactobacillus and Streptococcus mutans counts following sodium fluoride mouthrinsing in Egyptian children

The salivary Lactobacillus and Streptococcus mutans counts were assessed in 100 Egyptian children, initially before mouthrinsing with 0.05% sodium fluoride solution, 24 hours and 48 hours following rinsing. A statistically significant reduction in Lactobacillus and Streptococcus mutans counts of 49% and 44% respectively was obtained 24 hours following rinsing. This percentage decrease in both bacterial counts was reduced to 27% and 31% respectively, 48 hours following rinsing.     

Inverse correlation between the proportion of salivary bacteria inhibiting Streptococcus mutans and the percentage of untreated carious teeth

To evaluate the role of inhibitory substances produced by bacteria in the oral cavity, we estimated, by a deferred test on Todd-Hewitt agar enriched with hemin and vitamin K, the proportion of bacteria that inhibited or stimulated the growth of Streptococcus mutans and Porphyromonas gingivalis, from the saliva of 109 patients (54 males and 55 females) attending our dental clinics. The patients, aged from 8 to 75 years old (mean: 31 +/- 18 years), were randomly selected whatever the reason for their visit. The results, evaluated with the Spearman rank test, indicated that there was no statistically significant (P > 0.05) correlation between the proportion of salivary bacteria inhibiting or stimulating P. gingivalis with the Community Periodontal Index of Treatment Needs (CPITN), the number of carious, missing and filled teeth, or with the decayed, missing and filled teeth index (DMFT). Also, no statistically significant correlation was observed between the proportion of salivary bacteria stimulating the growth of S. mutans and the above mentioned health indexes. However, a statistically significant (P < 0.005) negative correlation was found between the percentage of cultivated bacteria that inhibit S. mutans and the percentage of untreated carious teeth as well as with the CPITN. The results thus indicate a possible role for inhibitory substances produced by bacteria in the maintenance of oral health.     

The effect of antibody to Streptococcus mutans in saliva on caries activity and S. mutans adhering

An indirect enzyme-linked immunosorbent assay (ELISA) was used for measuring IgA antibody to whole cell of Streptococcus mutans (serotype c) in saliva. 36 parotid salivary samples of human were collected from two groups: Caries free (CF) and caries sensitive (CS). The result shows that the IgA antibody to S. mutans in CF group was higher than those in CS group (P < 0.05). The saliva antibody was gained from the rabbits by injection with S. mutans (serotype c), and the adhesion of S. mutans--3H on the surface of hydroxylapatite beads treated by rabbit's saliva was measured. The results show that the saliva with immunity could inhibit the S. mutans to adhere on the HA beads (P < 0.05). It means saliva antibody may prevent caries through inhibition of S. mutans from adhesion.     

Influence of secretory immunoglobulin A and purified secretory component on dextran-sucrase activity of Streptococcus mutans

The net stimulation of dextransucrase EC 2.4.1.5) activity from Streptococcus mutans HS6 by dextran, secretory immunoglobulin A, or secretory component was investigated. Approximately equal stimulation resulted from treatment with these three components.     

Concentrated bovine colostral whey proteins from Streptococcus mutans/Strep. sobrinus immunized cows inhibit the adherence of Strep. mutans and promote the aggregation of mutans streptococci

The aim of this study was to examine the effect of bovine colostral whey proteins from cows immunized with Streptococcus mutans/Strep. sobrinus on the adherence and aggregation of caries-inducing bacteria, i.e., mutants streptococci. Both adherence and aggregation are important phenomena in the bacterial colonization of the human oral cavity. In all adherence experiments there was a significant difference between treatments by immune product (IP; from immunized cows) and a control product (CP; a similar product from non-immunized cows). The adherence of 35S-labelled Strep. mutans cells (serotype c) to parotid saliva-coated hydroxyapatite (SHA) was dose-dependently inhibited by both IP and CP if SHA was coated with either product before exposure to bacteria, but markedly lower concentrations of IP than CP were effective. When instead of SHA the bacterial cells were pretreated with IP or CP, only IP strongly and dose-dependently inhibited streptococcal adherence. When bacteria, IP or CP, and SHA were incubated simultaneously, a significant difference between IP and CP treatments was again found. Further, IP effectively aggregated both Strep. mutans and Strep. sobrinus cells, whereas hardly any effect was seen with CP. Both IP and CP aggregated the control bacterium Strep. sanguis, which affected the adherence of the pretreated bacteria.     

Preparations of antigens and immunoadsorbents corresponding to the Streptococcus group A cell-wall polysaccharide

The allyl glycosides of a tri-, penta- and hexasaccharide corresponding to the Streptococcus Group A cell-wall polysaccharide were coupled to solid or soluble supports to give immunoaffinity columns and neoglycoproteins, respectively. Cysteamine hydrochloride was added to the allyl glycosides and the resulting cysteamine adducts were used for subsequent coupling to linkers via the amine functionality. The tri- and penta-saccharide cysteamine adducts were coupled directly to the azalactone-derivatized 3M Emphase Biosupport Medium AB 1 to yield two affinity columns. The penta- and hexa- saccharides were coupled to bovine serum albumin or ovalbumin via the conjugate addition of the epsilon-amino groups of lysines on the proteins with the N-acryloylated sugars or the oligosaccharide-squarate adducts, derived in turn from the cysteamine adducts. The efficiency of the above methods is compared.     

Influence of glycoproteins B, C and D on the conversion of virus-to-cell attachment from heparin sensitivity to resistance

In two municipalities in Lower Saxony statistically valid clusters were observed, which attracted great public interest. Committees were set up to initiate a large variety of on-the-spot-investigations. Finally, it was decided to conduct a case control study throughout Lower Saxony to explore potential risk factors which might explain the observed clusters. A limited number of already established and currently discussed hypotheses was chosen for investigation. The study was based on patients registrated at the German Children's Cancer Registry (GCCR). For each child with leukemia diseased between 1988 and 1993 two population-based controls (local and state controls) and one tumour control were selected. Data were collected by means of a questionnaire self-administered by the parents and a telephone interview. In addition, measurements of electromagnetic fields and radon were performed and inspections of the child's birth record were made for the purpose of confirming a potential association between parenteral vitamin K prophylaxis and leukemia or tumours. This paper presents the concept and basic considerations of the study, its design and statistical evaluation. Response rates and a summary of results will be presented, too. The paper will serve as a reference for subsequent publications about more detailed analyses of specific potential risk factors. 425 parents of diseased children and 610 of non-diseased children were asked for participation. The rates of response were 82% for families with diseased and 71% for families with non-diseased children. In total, 781 parents participated in the study. The most important results are as follows: The Greaves' hypothesis (9, 10) was supported by the following results: In children diseased with leukemia, vaccinations were less frequent, virus-related infections occurred more rarely, these children were more frequently first-born children and more frequently breast-feed, and they possibly had fewer contacts with other children in infancy. Our data do not show a significant association between parenteral vitamin K prophylaxis and leukemia or tumours (17). Measurements of electromagnetic fields show a weak association between high-level exposure and an increased risk of developing leukemias. Children who were X-rayed more than four times or were born prematurely and had received intensive care treatment show a positive association with occurrence of leukemia, but the number of these patients was very small and these factors are not independent. The only potential job-related hazard was paternal exposure to plastic and resin fumes. The incidence of miscarriages and abortions was increased in mothers of children with leukemia. Our study did not confirm the hypotheses, that leukemia is associated with gestational age, with consumption of alcohol, nicotine, and medicaments during pregnancy. Nor was any association detected with exposure to wood preservatives and insecticides or with a high socio-economic status. The number of patients living in municipalities with increased incidence of leukemia was too small to show statistically valid results. However, it is noteworthy that some of the above-mentioned risk factors were observed also in these children. One of the purposes of a nation-wide case control study which is currently performed at the GCCR is to validate and complete these results.     

Effects of surfactants on glucosyltransferase production and in vitro sucrose-dependent colonization by Streptococcus mutans

OBJECTIVE: To study the distribution of myocardial structural elements in a univentricular experimental model (Halobatrachus didactylus) and the changes induced by acute exposure to a sublethal concentrations of cadmium. DESIGN: 15 specimens of H. didactylus (marine teleost) were divided in three groups: CTRL: the control group, the fish were injected with a saline solution; 24 H: 1 mg/kg of cadmium chloride was injected and fish were sacrificed after 1 day post injection; 7 D: fish where subjected to the same cadmium concentration and sacrificed after 7 days post injection. INTERVENTIONS: Histological sections (5 microns) of ventricle were coloured by PicroSirius method and used to determine the fraction area occupied by the several myocardium structural elements, using specific software Optimas Bioscan 5.2. MEASUREMENTS AND RESULTS: The different structural elements of H. didactylus myocardium displayed in different ways and its distribution along the ventricular wall remained unchanged on exposure to cadmium. The structural elements of H. didactylus myocardium showed a characteristic pattern: collagen type I revealed a heterogeneous distribution, with fractional area values greater in epicardial and endocardial layers; collagen type III, has a homogeneous distribution with the myocardial wall thickness with lesser fractional area values than collagen I; cardiac muscle tissue showed a tendency to occupy a greater fractional area in the epicardial-endocardial region. Cadmium toxicity resulted in an increase in the fractional area occupied by muscle tissue to the detriment of the area occupied by the intertrabecular spaces (lacunae), with no alterations in the other structural components. CONCLUSIONS: On our H. didactylus fish model, cadmium may have induced a myocardial edema which resulted in an increase in the fractional area occupied by muscle tissue, with no alterations in other structural components of the myocardium or in the relative ventricular mass.     

Isolation, characterization and sequence analysis of the scrK gene encoding fructokinase of Streptococcus mutans

A gene encoding an ATP-dependent fructokinase from Streptococcus mutans GS-5 was identified within a 2 kb DNA fragment immediately downstream from the scrA gene. The gene cloned in Escherichia coli also expressed mannokinase activity. Insertional inactivation of this gene in S. mutans markedly decreased both fructokinase and mannokinase activities. Nucleotide sequence analysis of the 2 kb fragment revealed an ORF starting 199 bp downstream from the scrA gene, preceded by potential ribosome-binding (Shine-Dalgarno) and promoter-like sequences. This ORF specified a putative protein of 293 amino acids with a calculated M(r) of 31,681. The deduced amino acid sequence of the fructokinase gene, scrK, from S. mutans exhibited no significant similarity to fructokinase genes from Klebsiella pneumoniae, E. coli plasmid pUR400 or Vibrio alginolyticus, but was similar to a comparable gene from Zymomonas mobilis.     

Effect of long-term therapies with penicillin and sulfadiazine on Streptococcus mutans and lactobaccilli in dental plaque

Plaque samples were obtained from 13 children receiving long-term therapy with benzathine penicillin for the prevention of rheumatic fever recurrences, 31 children receiving oral sulfadiazine for the same purpose, and 29 untreated siblings. The therapies were found to have no effect upon the proportions of Streptococcus mutans or lactobacilli in dental plaque, upon the percentage of children harboring the organisms, nor upon the susceptibility of the organisms to penicillin and sulfadiazine. Of the S. mutans strains tested, 97% had a minimal inhibitory concentration of penicillin G of less than 48 ng/ml and, of the lactobacillus strains tested, 96.8% had a minimal inhibitory concentration of less than 1,600 ng/ml. All strains of both organisms were profoundly resistant to sulfadiazine.     

Effect of humic acid on formation and several properties of the extracellular polysaccharide of Achromobacter delicatulus

The reaction of turning over in the ophiuroid A. kochii Lütken begins when the ambulacral legs are detached from the substrate. At the bottom, when aboral side of the animal touches the substrate, turning over begins with oral flexion of three arms. In water, turning over begins with aboral flexion of all the arms. The organism takes the shape of a ball, its center of gravity being shifted to the side of a disc which outer surface is formed by the oral part. As a result, when dropped into the water with its oral side upwards, the ophiuroid passively turns over due to the gravity force. Single arm isolated together with the adjacent part of the nervous ring is capable of turning over. After sectioning the nervous ring, coordinated activity of the arms during turning over is disturbed.     

Expression of plasminogen activator pla of Yersinia pestis enhances bacterial attachment to the mammalian extracellular matrix

The effect of the plasminogen activator Pla of Yersinia pestis on the adhesiveness of bacteria to the mammalian extracellular matrix was determined. Y. pestis KIM D27 harbors the 9.5-kb plasmid pPCP1, encoding Pla and pesticin; the strain efficiently adhered to the reconstituted basement membrane preparation Matrigel, to the extracellular matrix prepared from human lung NCI-H292 epithelial cells, as well as to immobilized laminin. The isogenic strain Y. pestis KIM D34 lacking pPCP1 exhibited lower adhesiveness to both matrix preparations and to laminin. Both strains showed weak adherence to type I, IV, and V collagens as well as to human plasma and cellular fibronectin. The Pla-expressing recombinant Escherichia coli LE392(pC4006) exhibited specific adhesiveness to both extracellular matrix preparations as well as to laminin. The Pla-expressing strains showed a low-affinity adherence to another basement membrane component, heparan sulfate proteoglycan, but not to chondroitin sulfate proteoglycan. The degradation of radiolabeled laminin, heparan sulfate proteoglycan, or human lung extracellular matrix by the Pla-expressing recombinant E. coli required the presence of plasminogen, and degradation was inhibited by the plasmin inhibitors aprotinin and alpha2-antiplasmin. Our results indicate a function of Pla in enhancing bacterial adhesion to extracellular matrices. Y. pestis also exhibits a low level of Pla-independent adhesiveness to extracellular matrices.     

Auger electron spectroscopy and its use for the characterization of titanium and hydroxyapatite surfaces

This review paper provides the basic background and underlying theory behind Auger electron spectroscopy (AES). Among the many surface analytical tools, AES has been shown to be very effective for surface composition analysis. These analyses are critically needed to better understand the interactions between the host and implant. The use of AES for titanium (Ti) and hydroxyapatite (HA) biomaterials characterization is demonstrated in this paper. The relative peak heights of TiL(2,3)M(2,3)V can be used as 'fingerprints' for TiO2 surfaces which have undergone different degrees of reduction. Similarly, for HA coatings, a shift in the phosphorus Auger peaks to a higher kinetic energy indicates the presence of a phosphate group, with strong P-O bonds. Depth compositional profiling and thin-film analysis can be performed using AES. In our studies, oxide thicknesses on Ti surfaces range from 36.8 +/- 7.4 A to 436 +/- 49 A depending on the surface treatment. Depth profiling can also be used to determine the subsurface composition of biomaterials. For HA coatings, a phosphorus concentration at the oxide/metal interface has been observed to be higher than at the outermost oxide surface. The HA coatings have also been observed to coexist within the titanium oxide, suggesting the occurrence of chemical bonding between the coatings and the metallic substrates. However, like other analytical tools, AES has its limitations. The electron beam damage can severely limit useful analysis of organic and biological materials and occasionally ceramic materials. Carbide buildup during long beam exposure times has been shown to affect the relative peak-to-peak intensities of the oxygen and metal Auger signals. The determination of film thickness requires a standard of known thickness and depth profiling of overlapping peaks can be very problematic. Even with these limitation, AES can be a powerful analytical tool for the characterization of biomaterial surfaces.