Changes in oral microflora following 0.3% cetylpyridinium chloride-containing mouth spray intervention in adult volunteers after professional oral care: Randomized clinical study.

OBJECTIVES: This study explored the changes in bacterial flora composition and total bacterial count in the saliva and tongue coating, along with the change in the tongue coating index (TCI) following an intervention with 0.3% cetylpyridinium chloride (CPC) mouth spray after professional oral care. MATERIALS AND METHODS: Fifty-two adult volunteers aged 30-60 years were equally divided into CPC spray (n = 26) and control (n = 26) groups. All subjects underwent scaling and polishing. The CPC spray group was administered four puffs of CPC spray to the tongue dorsum four times a day for 3 weeks. The control group performed only routine daily oral care (brushing) and did not use any other spray. Bacteriological evaluation of saliva and tongue coating was performed using 16S ribosomal RNA gene sequencing and quantitative polymerase chain reaction. The tongue coating was evaluated to calculate the TCI. A per-protocol analysis was conducted for 44 subjects (CPC spray group, n = 23; control group, n = 21). RESULTS: At 1 and 3 weeks after CPC spray use, the flora of the saliva and tongue coating changed; the genus Haemophilus was dominant in the CPC spray group, whereas the genus Saccharibacteria was dominant in the control group. The sampling time differed among individual participants, which may have affected the bacterial counts. There was no significant intragroup change in TCI in either group. CONCLUSIONS: CPC spray affected the bacterial flora in the saliva and tongue coating, particularly with respect to an increase in the abundance of Haemophilus. However, CPC spray did not change the TCI. These results suggest that it may be optimal to combine CPC spray with a physical cleaning method such as using a tongue brush or scraper. Clinical Trial Registration: University Hospital Medical Information Network UMIN000041140.

A Novel Antihypertensive Peptide Identified in Thermolysin-Digested Rice Bran.

SCOPE: Hypertension is a risk factor for arteriosclerosis. In this study, we investigate the antihypertensive effect of protease-digested rice bran in a spontaneously hypertension rat (SHR) model. We also purify a novel antihypertensive peptide from the digest. METHODS AND RESULTS: Thermolysin-digested rice bran (TRB) is administered to SHRs for 4 weeks, and systolic blood pressure (SBP) was measured weekly using the tail-cuff method. TRB shows an antihypertensive effect in a dose-dependent manner. TRB also reduces angiotensin I-converting enzyme (ACE) activity in lung tissue and serum troponin I levels. TRB is fractionated by HPLC and ACE-inhibitory activity in the HPLC fractions is measured. Peptides LRA and YY are identified from the two fractions with the strongest ACE-inhibitory activity. Amino acid sequence of these peptides are found in a vicilin-like seed storage protein, and identified in rice bran protein using the peptide mass fingerprint method. We confirm that LRA and YY are cleaved by thermolysin digestion of a model synthetic peptide. Orally administered LRA (0.25 mg kg-1 ) or YY (0.5 mg kg-1 ) lowers the SBP of SHRs at 4 h after administration. CONCLUSION: We identify a novel, orally active antihypertensive peptide, LRA from the digest of rice bran protein.

Supplementation of broccoli or Bifidobacterium longum-fermented broccoli suppresses serum lipid peroxidation and osteoclast differentiation on alveolar bone surface in rats fed a high-cholesterol diet.

High-cholesterol diet enhances osteoclastic activity on alveolar bone by increasing serum lipid peroxidation. We hypothesized that supplementation with dietary antioxidants, such as found in broccoli and its fermented products, might suppress increases in serum lipid peroxidation, contributing to the inhibition of osteoclastic activity after high-cholesterol diet intake. The purpose of the present study was to investigate the effects of broccoli and fermented broccoli consumption on serum lipid peroxidation and osteoclast differentiation in alveolar bone of rats fed a high-cholesterol diet. In this 12-week study, rats were divided into 4 groups (n = 6/group): a control group (fed regular diet) and 3 experimental groups (fed a high-cholesterol [1% wt/wt] diet, or a high-cholesterol diet supplemented with either broccoli powder [5% wt/wt] or Bifidobacterium longum-fermented broccoli powder [5% wt/wt]). Serum hexanoyl-lysine (HEL) levels were measured as a parameter of lipid peroxidation. The number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in alveolar bone was enumerated to evaluate osteoclast differentiation. When compared with regular diet, the high-cholesterol diet increased serum HEL levels and resulted in a higher number of TRAP-positive osteoclasts at 12 weeks. The high-cholesterol diet supplemented with broccoli or B. longum-fermented broccoli showed lower levels of serum HEL and fewer TRAP-positive osteoclasts than the high-cholesterol diet at 12 weeks. In conclusion, consumption of broccoli, or its fermented product, inhibited the effects of a high-cholesterol diet on osteoclast differentiation in rat alveolar bone by suppressing serum lipid peroxidation.

Human trial of liposomal lactoferrin supplementation for periodontal disease.

New approaches to periodontal health have been in strong demand in addition to conventional local plaque control. In this study, liposomal bovine lactoferrin (L-bLF) was orally administered to subjects with periodontal disease to investigate whether it could be a useful treatment. L-bLF composed of soy phosphatidylcholine was given as a supplement for four weeks in tablet form (180 mg bLF/d) to twelve subjects with multiple sites of more than 3 mm probing depth (PD). PD, bleeding on probing (BOP), gingival crevicular fluid (GCF) volume and the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and monocyte chemoattractant protein-1 (MCP-1) in GCF were evaluated for 51 sites with more than 4 mm PD in five subjects. Blood samples of all subjects were collected 0, 2 and 4 weeks after supplementation. Isolated peripheral blood mononuclear cells (PBMCs) were incubated for 24 h with or without lipopolysaccharide (LPS) (100 ng/ml) from Porphyromonas gingivalis, and TNF-α, IL-1ß, IL-6 and MCP-1 in the culture media were measured. Toll-like receptor 2 (TLR2) and TLR4 mRNA expressions of isolated PBMCs were also quantitatively analyzed using real-time reverse transcription-polymerase chain reaction (RT-PCR). The PD was significantly reduced by L-bLF supplementation, but the BOP and GCF volume were not significantly changed. The MCP-1 level in GCF was significantly reduced, while levels of other cytokines were not changed. Four-week L-bLF supplementation also showed significant decreases of LPS-induced cytokine production from PBMCs. Relative gene expressions of TLR2 and TLR4 did not change. These results suggest that L-bLF supplementation can be effective in the treatment of periodontal disease, although prospective controlled large-scale studies are required.

Morphological study on dental caries induced in WBN/KobSlc rats (Rattus norvegicus) fed a standard laboratory diet.

In our previous studies, WBN/KobSlc was characterized as a rat strain in which only males began to develop pancreatitis, and then presented with diabetic symptoms. In the course of studying their pancreatic inflammation, we detected molar caries in prediabetic males feeding on a standard diet (CRF-1) widely used for experimental animals. The purpose of this study is to confirm whether the WBN/KobSlc strain is caries-susceptible to the diet reported to be non-cariogenic, and to examine the effect of a prediabetic condition on their dental caries. For a morphological study, 25 male WBN/KobSlc rats aged 3.2-7.8 months and 24 females of the same strain aged 3.3-6.6 months were used, along with 10 males and 10 females of 8.2-month-old F344 rats. Marked dental caries were detected in the mandibular molars of male and female WBN/KobSlc rats regardless of pancreatitis, although no similar changes were observed in any teeth of the F344 strain fed the same diet. Soft X-ray examination revealed that the caries began in the crown and progressed horizontally and vertically, and that a severe radiolucent lesion extensively expanded to the entire crown, corresponding to a macroscopically deleted molar. The caries had gradually developed mainly in the second mandibular molar from more than 3.5 months of age, while none were seen in any rats before that time. The WBN/KobSlc rats were caries-susceptible even to the standard laboratory diet, and pancreatitis was not directly associated with the onset of dental caries in this strain.

Bifidobacterium longum-fermented broccoli supernatant inhibited the growth of Candida albicans and some pathogenic bacteria in vitro.

The aim of this study is to develop a growth inhibitory material against some pathogenic microorganisms, using beneficial bacteria such as Bifidobacterium species and certain types of vegetables which can be good substrates for the growth of the beneficial bacteria. At first, various vegetable juices were screened for the growth promotion of Bifidobacterium longum etc. Among the vegetables tested, broccoli (Brassica oleracea L. var. botrytis L.) and cabbage (Brassica oleracea L. var. capitata L.) showed excellent growth promoting activities for B. longum. Secondly, the B. longum-fermented broccoli (BFB) and Lactobacillus pentosus-fermented broccoli (LFB) supernatants were prepared and the growth inhibitory activities against Candida albicans were determined. Both of them showed dose-dependent, growth inhibitory effects, and the effect of BFB was superior to LFB. It was thought that the superior effect of BFB could be mainly attributed to the acids, especially acetic acid, produced by B. longum. BFB also inhibited some pathogenic bacteria such as Streptococcus mutans and Porphylomonas gingivalis. In conclusion, broccoli was found to be a good growth-promoting substance for B. longum. The fermented product, BFB, appears to be a usable material that inhibits the growth of C. albicans and some pathogenic bacteria.

[LDL-cholesterol-lowering effect of a mixed green vegetable and fruit beverage containing broccoli and cabbage in hypercholesterolemic subjects].

The serum LDL-cholesterol (LDL-C)-lowering effects of two types of canned beverage containing mixed green vegetables and fruits, with or without broccoli and cabbage (B&C), were examined in a randomized double-blind study design. Seventy-seven adults subjects with mild to moderate hypercholesterolemia participated in this study after giving their informed consent. The subjects were randomly divided into two groups. One group(test group) was allocated a test sample, containing B&C as the main materials. Another group(control group) was allocated a placebo sample made from the same materials but without B&C. The subjects were administered 2 cans of the assigned sample (160 g contents/can) per day for 12 weeks. Forty-nine out of 77 subjects, whose LDL-C levels were greater than or equal to 140 mg/dl and less than 180 mg/dl, were analyzed for the effectiveness. Serum LDL-C levels in the test group were significantly(p < 0.05) reduced at 3, 6 and 9 weeks after administration in comparison to the baseline levels (155.7 mg/dl in average). The average LDL-C value at 9 weeks was 142.5 mg/dl and the reduction rate was 8.5%. But serum LDL-C levels in the control group were not significantly reduced. Significant differences(p < 0.05) between the groups were observed in the LDL-C levels at 6 and 9 weeks and also in the total cholesterol levels at 9 week. Thus daily intake of the beverage tested containing B&C are useful for lowering serum LDL-C levels in hypercholesterolemic subjects.

A mixed green vegetable and fruit beverage decreased the serum level of low-density lipoprotein cholesterol in hypercholesterolemic patients.

The effects of a canned mixed green vegetable and fruit beverage, containing broccoli (Brassica oleracea L. var. botrytis L.) and cabbage (Brassica oleracea L. var. capitata L.) as main materials, on serum lipid levels in hypercholesterolemic patients were investigated. Thirty-one adult subjects were administered two cans of the beverage (160 g/can) per day for 3 weeks. Their serum total cholesterol (TC) levels significantly decreased from 6.7 +/- 0.8 to 6.1 +/- 0.6 mmol/L, and, more strikingly, the level of low-density lipoprotein cholesterol (LDL-C) significantly decreased from 4.4 +/- 0.8 to 3.8 +/- 0.7 mmol/L. At 9 weeks after the cessation of the administration, these levels had returned to the preadministration levels. Furthermore, 14 other subjects who were administered one can of the sample for 12 weeks also showed a significant reduction in the levels of serum TC and LDL-C. Thus, daily consumption of this mixed green vegetable and fruit beverage may be useful in lowering serum TC and LDL-C levels in hypercholesterolemic patients.