Effects of Lactobacillus salivarius WB21 combined with green tea catechins on dental caries, periodontitis, and oral malodor.

OBJECTIVE: To evaluate the combined use of Lactobacillus salivarius WB21 and (-)-epigallocatechin gallate (EGCg) for oral health maintenance. DESIGN: The effects of L. salivarius WB21 on growth of Streptococcus mutans, the insoluble glucan produced by S. mutans, and on growth of Porphyromonas gingivalis were evaluated in vitro. In addition, the susceptibility of five oral pathogenic bacteria and L. salivarius WB21 to EGCg, the inhibiting effect of EGCg on methyl mercaptan, and the effects of L. salivarius WB21 and EGCg in combination on growth of P. gingivalis were examined. RESULTS: Lactobacillus salivarius WB21 showed concentration-dependent inhibition of the growth of S. mutans. Addition of L. salivarius WB21 inhibited production of the insoluble glucan by S. mutans (p < 0.001). A filtrate of L. salivarius WB21 culture solution inhibited growth of P. gingivalis (p < 0.001 vs. control), and this effect was enhanced when it was used in combination with EGCg (p < 0.001 vs. the addition of L. salivarius WB21). In addition, EGCg directly inhibited methyl mercaptan in a concentration-dependent manner (p < 0.001). Concerning bacterial susceptibility to EGCg, growth of P. gingivalis, Prevotella intermedia, and Fusobacterium nucleatum was inhibited at 2.5 mg/mL of EGCg, while that of L. salivarius WB21 was inhibited at 25 mg/mL EGCg. CONCLUSIONS: Our results imply that L. salivarius WB21 may be useful for controlling dental caries, periodontitis, and oral malodor. In addition, the effects of L. salivarius WB21 on periodontitis and oral malodor may be synergistically enhanced by use in combination with EGCg.

Modelling error distribution in the ground reaction force during an induced-acceleration analysis of running in rear-foot strikers.

The objective of this study was to develop and evaluate a methodology for quantifying the contributions of modelling error terms, as well as individual joint torque, gravitational force and motion-dependent terms, to the generation of ground reaction force (GRF), whose true value can be measured with high accuracy using a force platform. Dynamic contributions to the GRF were derived from the combination of (1) the equations of motion for the individual segments, (2) the equations for constraint conditions arising from the connection of adjacent segments at joints, and (3) the equations for anatomical constraint axes at certain joints. The contribution of the error term was divided into four components caused by fluctuation of segment lengths, geometric variation in the constraint joint axes, and residual joint force and moment errors. The proposed methodology was applied to the running motion of thirteen rear-foot strikers at a constant speed of 3.3 m/s. Modelling errors arose primarily from fluctuations in support leg segment lengths and rapid movement of the virtual joint between the foot and ground during the first 20% of stance phase. The magnitudes of these error contributions to the vertical and anterior/posterior components of the GRF are presented alongside the non-error contributions, of which the joint torque term was the largest.

Probiotic effects of orally administered Lactobacillus salivarius WB21-containing tablets on periodontopathic bacteria: a double-blinded, placebo-controlled, randomized clinical trial.

AIM: This study was designed to evaluate whether the oral administration of lactobacilli could change the bacterial population in supra/subgingival plaque. MATERIAL AND METHODS: Sixty-six healthy volunteers without severe periodontitis were randomized into two groups to receive lactobacilli or placebo for 8 weeks (8W): the test group (n=34) received 2.01 x 10(9) CFU/day of Lactobacillus salivarius WB21 and xylitol in tablets; the control group (n=32) received placebo with xylitol. Supra/subgingival plaque samples were collected at the baseline and after 4 weeks (4W) and 8W. The bacterial amounts in plaque samples were analysed by quantitative real-time polymerase chain reaction. RESULTS: The numerical sum of five selected periodontopathic bacteria in the test group was decreased significantly in subgingival plaque at 4W [odds ratio (OR)=3.13, 95% confidence intervals (CI)=1.28-7.65, p=0.012]. Multivariate analysis showed that significantly higher odds were obtained for the reduction of Tannerella forsythia in subgingival plaque of the test group at both 4W (OR=6.69, 95% CI=2.51-17.9, p<0.001) and 8W (OR=3.67, 95% CI=1.45-9.26, p=0.006). CONCLUSION: Oral administration of probiotic lactobacilli reduced the numerical sum of five selected periodontopathic bacteria and could contribute to the beneficial effects on periodontal conditions.

Improvement of periodontal condition by probiotics with Lactobacillus salivarius WB21: a randomized, double-blind, placebo-controlled study.

AIM: This randomized clinical study was designed to evaluate the effect of probiotic intervention using lactobacilli on the periodontal condition of volunteers without severe periodontitis. MATERIAL AND METHODS: Freeze-dried Lactobacillus salivarius WB21 (WB21)-containing tablets or a placebo were given to volunteers in a double-blind randomized study. A total of 66 volunteers were finally enrolled and randomly assigned to receive tablets containing WB21 (6.7 x 10(8) CFU) with xylitol or xylitol alone (placebo) three times a day for 8 weeks. Periodontal clinical parameters and whole saliva samples were obtained at baseline (BL), 4 weeks, and the end of the interventional period (8 weeks). Salivary lactoferrin (Lf) levels were measured by enzyme-linked immunosorbent assay. Lactobacilli in saliva and plaque samples was detected by semi-quantitative RT-PCR using 16S rRNA primers. RESULTS: Periodontal clinical parameters were improved in both groups after an 8-week intervention. Current smokers in the test group showed a significantly greater improvement of plaque index and probing pocket depth from BL when compared with those in the placebo group. Salivary Lf level was also significantly decreased in the test group smokers. CONCLUSION: Our results indicate that probiotics could be useful in the improvement/maintenance of oral health in subjects at a high risk of periodontal disease.

Antibiotic effects of WP-0405, a thermo-setting ofloxacin gel, on methicillin-resistant Staphylococcus aureus keratitis in rabbits.

PURPOSE: The chemotherapeutic effects and pharmacokinetics properties of WP-0405 (a thermo-setting in situ 0.3% ofloxacin-containing ophthalmic gel) and ofloxacin (a conventional 0.3% ofloxacin solution) on methicillin-resistant Staphylococcus aureus (MRSA) keratitis were compared in a rabbit model. METHOD: The single-instillation pharmacokinetics of WP-0405 and ofloxacin in the cornea, aqueous humor, conjunctiva, and iris-ciliary body were determined in normal rabbit eyes. To compare the duration of antimicrobial action, WP-0405 or ofloxacin was instilled oncedaily in an early-treatment model of keratitis, and corneas were either removed immediately or 4 or 8 h postinstillation. In another experiment, WP-0405 was instilled two or three times daily to compare its antibiotic efficacy with three-times daily instillation of ofloxacin in the same early-treatment model of keratitis; corneas were then removed after determining the extent of the abscess area. In another experiment, WP-0405 was instilled four or eight times daily to compare its effects with eight-times daily instillation of ofloxacin in a late-treatment model of keratitis, and corneas were removed. The number of viable bacteria in the corneas was determined in all experiments. RESULTS: Cmax and AUC0- in tissues treated with WP-0405 were 1.5-3.4-fold and 1.8-2.9-fold greater than those treated with ofloxacin, respectively. WP-0405 significantly reduced the number of viable bacteria for up to 8 h after a single instillation. WP-0405 not only significantly reduced the number of viable bacteria, but also the size of the abscess area at the same frequency of instillation. When compared to ofloxacin, WP-0405 exhibited an approximately equivalent antibiotic effect, with fewer administrations. CONCLUSIONS: As a result of its pharmacokinetics, WP-0405 had a more potent, longer-acting antibiotic effect than did ofloxacin. Furthermore, because of its lower required instillation frequency, which would improve patient compliance, WP-0405 has great potential therapeutic benefits.

Catalase, a specific antigen in the feces of human subjects infected with Helicobacter pylori.

Recently, we reported the production of three new monoclonal antibodies with high specificity for a Helicobacter pylori antigen suitable for diagnosis of H. pylori infection. The aim of the present study was to identify the antigen recognized by these monoclonal antibodies concerning both H. pylori and the feces of human subjects infected with H. pylori. The cellular antigen was purified from an H. pylori cell extract by immunoaffinity column chromatography with the monoclonal antibody as a ligand. The amino-terminal amino acid sequences (eight residues) of the purified antigen and H. pylori catalase were the same. The molecular weights of native and subunit, specific catalase activity, and UV and visible spectra of the purified antigen were in good agreement with those of H. pylori catalase. The human fecal antigens were purified from two fecal samples of two H. pylori-positive subjects by ammonium sulfate precipitation, CM-Sephadex C(50) chromatography, and the same immunoaffinity chromatography used for the H. pylori cellular antigen. The fecal antigens had catalase activity. The amino-terminal amino acid sequences (five residues) of the human fecal antigen and H. pylori catalase were the same. The monoclonal antibodies reacted with the native cellular antigen, but did not react with the denatured antigen, human catalase, and bovine catalase. The results show that the target antigen of the monoclonal antibodies is native H. pylori catalase and that the monoclonal antibodies are able to specifically detect the antigen, which exists in an intact form, retaining the catalase activity in human feces.

Production and application of new monoclonal antibodies specific for a fecal Helicobacter pylori antigen.

The aim of the present study was to establish monoclonal antibodies that could be used to produce a diagnostic test composed of one kind of monoclonal antibody recognizing a fecal Helicobacter pylori antigen. The need to develop such a test arose from disadvantages of the diagnostic test that uses a polyclonal antibody or plural kinds of monoclonal antibodies, such as the lower specificity for H. pylori antigen and the difficulty of reproduction with consistent quality. Mice were immunized with sonicated cells of the coccoid form of H. pylori, and fecal samples from H. pylori-positive subjects were screened by a direct sandwich enzyme immunoassay (EIA) for antibody production from 32 hybridoma clones. The three stable clones produced antibodies (21G2, 41A5, and 82B9) that reacted with the same soluble antigen. Gel filtration chromatography showed that the molecular masses of the cellular antigen and the fecal antigen were the same, 260 kDa. The antigen was labile in response to sodium dodecyl sulfate and heat treatments. A single-step direct sandwich EIA using a single monoclonal antibody, 21G2, was developed. The EIA could detect the antigen in 41 H. pylori clinical isolates and in fecal samples from seven H. pylori-positive subjects. Several kinds of Helicobacter species (Helicobacter felis, Helicobacter hepaticus, Helicobacter mustelae, and Helicobacter cinaedi) except H. pylori, major bacteria in feces (Campylobacter jejuni, Bacteroides vulgatus, Bifidobacterium breve, Bifidobacterium infantis, and Escherichia coli), and fecal samples from six H. pylori-negative subjects showed negative results. These results indicate that the new monoclonal antibodies and the new specific EIA would be useful as a noninvasive method of diagnosis of H. pylori infection.