VOC Profiles of Saliva in Assessment of Halitosis and Submandibular Abscesses Using HS-SPME-GC/MS Technique.

Halitosis and submandibular abscesses are examples of mouth-related diseases with the possible bacterial origin. Salivary volatile organic compounds (VOCs) are potential biomarkers of them, once they can be addressed as metabolites of bacterial activity. Healthy patients (n = 15), subjects with submandibular abscesses located in fascial deep space (n = 10), and subjects with halitosis (n = 5) were enrolled in the study. Saliva samples were subjected to headspace solid-phase microextraction (HS-SPME) and gas chromatography coupled to mass spectrometry (GC/MS) analysis. A total number of 164 VOCs was detected by the developed methodology, 23 specific for halitosis and 41 for abscess. Halitosis' profiles were characterized by a larger number of sulfur compounds, while for abscess they had a higher variety of alcohols, aldehydes, and hydrocarbons-biomarkers of inflammatory processes. Principal components analysis allowed visualization of clusters formed according to the evaluated conditions. Kruskal-Wallis test indicated that 39 VOCs presented differentiated responses between the studied groups, with statistical relevance (p < 0.05). Random forest was applied, and a prediction model based on eight VOCs (2-butanone, methyl thioacetate, 2-methylbutanoic acid, S-methyl pentanethioate, dimethyl tetrasulfide, indolizine, pentadecane, and octadecanal) provided 100% of sensitivity, 82% of specificity, and 91% of balanced accuracy, indicating the specific presence of submandibular abscess.

Microbiological results of improvement in periodontal condition by administration of oral probiotics.

Oral bacteria that degrade sulphur-containing amino acids (cysteine, cystine, and methionine) produce volatile sulphur compounds (VSCs = hydrogen sulphide, methyl mercaptan, and dimethyl sulphide) highly correlated with halitosis. When these bacteria are given the right environment, i.e. periodontal disease, cariogenic biofilm or food source they can grow in number very quickly and will start to convert proteins to VSC that, together with volatile fatty acids are largely responsible for oral malodor. Recently, the prevention of dental caries and periodontal diseases using various probiotics has been attempted. The purpose of this study was to investigate the effects of probiotics based on in vitro analysis, such as antibacterial activity, and to evaluate the neutralizing effect of probiotics on halitosis, the levels of VSCs were measured by gas chromatography. .

Immediate results of photodynamic therapy for the treatment of halitosis in adolescents: a randomized, controlled, clinical trial.

UNLABELLED: Light with or without chemical agents has been used to induce therapeutic and antimicrobial effects. With photodynamic therapy, the antimicrobial effect is confined to areas covered by a photosensitive dye and irradiated with light. The aim of the present study was to evaluate the effect of photodynamic therapy for the treatment of halitosis in adolescents through the analysis of volatile sulfur compounds, especially sulfide. A controlled, clinical trial was conducted with 45 adolescents randomly allocated to three groups: group 1, photodynamic therapy administered to the dorsum of the tongue; group 2, treatment with a tongue scraper; and group 3, treatment with a tongue scraper combined with photodynamic therapy. The diagnosis of halitosis was performed using gas chromatography before and after treatment. Comparisons were made using the Kruskal-Wallis test followed by the Student-Newman-Keuls test, with the level of significance set at 5 % (p < 0.05). After treatment, a statistically significant reduction in halitosis was found in all groups (p < 0.001). The greatest reduction in total sulfides (median = 0) occurred with the combination of tongue scraper and photodynamic therapy. The present study describes a novel option for the treatment of halitosis in adolescents with an immediate effect that does not involve the mechanical aggression of the lingual papillae that occurs with conventional treatment. TRIAL REGISTRATION: Photodynamic Therapy in Adolescents Halitosis ( https://clinicaltrials.gov/ct2/show/NCT02007993?term=NCT02007993&rank=1 )Number: NCT02007993FUNDING:FAPESPNumber: 2013/13032-8.

Short-term effect of strontium- and zinc-containing toothpastes and mouthrinses on volatile sulphur compounds in morning breath: a randomized, double-blind, cross-over clinical study.

Zinc (Zn) reduces the formation of volatile sulphur compounds (VSCs) associated with oral malodour. Although strontium (Sr) is included in some products for reducing dental hypersensitivity, it may also have anti-halitosis properties. This randomized, double-blind, cross-over clinical study compared the anti-VSC effect of brushing with commercial toothpastes and rinses containing Zn and Sr. The volunteers (n = 30) either brushed/rinsed with/without tongue brushing using Zn-containing toothpaste/rinse, Sr-containing toothpaste/rinse, or placebo (control). Volatile sulphur compounds [hydrogen sulphide (H2 S) and methyl mercaptan (CH3 SH)] were measured, in morning breath, using gas chromatography. The anti-VSC effects of the test toothpastes and test rinses were significantly better than the anti-VSC effects of the respective controls. Toothbrushing with test toothpastes gave median reductions, compared with the control, of 70% for H2 S and 55-57% for CH3 SH. Rinsing with the Sr- and Zn-containing solutions had the same anti-VSC effect as toothbrushing and tooth- and tongue brushing with the Sr- and Zn-containing toothpastes. Zinc-containing rinse resulted in a significantly higher median salivary level of Zn compared with brushing with Zn-containing toothpaste, although this effect did not correlate with the anti-VSC effect. It can be concluded that the Sr- and Zn-containing toothpastes and the Zn- and Sr-containing rinses, when used in the evening, are equally effective in reducing morning-breath VSCs the following day.

Association Between Oral Hygiene Status and Halitosis Among Smokers and Nonsmokers.

PURPOSE: To study the association of smoking with poor oral hygiene status and halitosis in a comparative cross-sectional study. MATERIALS AND METHODS: 100 smokers and 100 nonsmokers ages 18-50 years were recruited for this study in Kota Bharu, Malaysia. Oral hygiene (good/fair vs poor) was determined using the Simplified Oral Hygiene Index, and the halitosis level was measured using a Halimeter. Subjects were instructed to refrain from consuming foods containing garlic, onions, strong spices, alcohol and using mouthwashes 48 h prior to the examination. The halitosis levels were quantified by recording volatile sulphur compounds (VSCs) three times at 3-min intervals, resulting in a mean halitosis score. Various statistical analyses were performed, ranging from simple frequency analysis to multivariable modelling. RESULTS: The proportions of subjects with poor oral hygiene and high halitosis were 24.0% and 41.5%, respectively. According to bivariate analyses, both problems were significantly less frequent among younger adults (halitosis), females, subjects with higher education, those with adequate habits to maintain good oral hygiene, those who had recent dental visits and those self-reporting fewer health problems. The percentages of poor oral hygiene and high halitosis were significantly higher in smokers (p < 0.001). However, almost all these variables failed to show significance in the multivariate analyses, with the exceptions of smoking for both poor oral hygiene and halitosis, education for poor oral hygiene, and age, self-reported health problems and time since the previous dental visit for halitosis. CONCLUSION: These findings demonstrate a significantly higher level of halitosis and poorer oral hygiene in smokers than nonsmokers.

The Negative Effects of Volatile Sulphur Compounds.

Oral malodor has been studied extensively in humans but not necessarily to the same degree in our veterinary patients where malodor constitutes a significant problem. Breath malodor may originate from the mouth, or from an extra oral source, originating from other organ systems such as gastrointestinal, respiratory, or even systemic disease. Oral malodor is a result of microbial metabolism of exogenous and endogenous proteinaceous substrates leading to the production of compounds such as indole, skatole, tyramine, cadaverine, puterescine, mercaptans, and sulphides. Volatile sulphur compounds have been shown to be the main cause of oral malodor. Although most clients perceive oral malodor to be primarily a cosmetic problem, there is an increasing volume of evidence in human dental literature demonstrating that volatile sulphur compounds produced by bacteria, even at low concentrations, are toxic to tissues and play a role in the pathogenesis of periodontitis. This article reviews the current available literature in human dentistry looking at these negative effects. No veterinary studies have been conducted looking at the negative effects of volatile sulphur compounds specifically, but as this article highlights, we should be aware of the potential negative effects of volatile sulphur compounds and consider this an area of future research.

Volatile sulphur compound levels and related factors in patients with chronic renal failure.

AIMS: To analyse specific volatile sulphur compound(VSC) levels in a group of chronic renal failure (CRF) patients and determine the relationship between these VSC levels and organoleptic measurements, blood urea nitrogen (BUN) levels, dental and periodontal conditions, salivary flow rate, and tongue coating scores. MATERIALS AND METHODS: One examiner performed organoleptic and VSC measurements on fifty patients with CRF before and after haemodialysis (HD) and controls. DMFT and CPITN indexes, tongue coating scores, salivary flow rates were measured. Comparisons were performed using the Mann-Whitney U, Wilcoxon signed-ranks, and chi-square tests. Spearman correlation coefficient was used to analyse correlations. RESULTS: Before HD, the mean dimethyl sulphide level was 1.04 ± 1.20 in the CRF patients and 0.51 ± 0.65 in controls, with a significant difference. The mean hydrogen sulphide, methyl mercaptan and dimethyl sulphide levels in CRF patients were 1.47 ± 3.04, 1.03 ± 1.85, and 1.04 ± 1.20, respectively, before HD; and 0.53 ± 1.65, 0.48 ± 1.27, and 0.56 ± 0.85, respectively, after HD; with the differences being significant. Methyl mercaptan levels increased with an increase in HD duration. Tongue coating and organoleptic measurements were significantly correlated with methyl mercaptan. CONCLUSIONS: Dimethyl sulphide is the main VSC in extraoral blood-borne halitosis; but methyl mercaptan may also contribute to this type of halitosis. A decreased salivary flow rate and an increased pH of the biofilm matrix may be a significant parameter for VSC levels in CRF patients.

Tongue-Coating Microbial and Metabolic Characteristics in Halitosis.

Halitosis is a common oral condition, which leads to social embarrassment and affects quality of life. Cumulative evidence has suggested the association of tongue-coating microbiome with the development of intraoral halitosis. The dynamic variations of tongue-coating microbiota and metabolites in halitosis have not been fully elucidated. Therefore, the present study aimed to determine the tongue-coating microbial and metabolic characteristics in halitosis subjects without other oral diseases using metagenomics and metabolomics analysis. The participants underwent oral examination, halitosis assessment, and tongue-coating sample collection for the microbiome and metabolome analysis. It was found that the microbiota richness and diversity were significantly elevated in the halitosis group. Furthermore, species from Actinomyces, Prevotella, Veillonella, and Solobacterium were significantly more abundant in the halitosis group. However, the Rothia and Streptococcus species exhibited opposite tendencies. Eleven Kyoto Encyclopedia of Genes and Genomes pathways were significantly enriched in the halitosis tongue coatings, including cysteine and methionine metabolism. Functional genes related to sulfur, indole, skatole, and cadaverine metabolic processes (such as serA, metH, metK and dsrAB) were identified to be more abundant in the halitosis samples. The metabolome analysis revealed that indole-3-acetic, ornithine, and L-tryptophan were significantly elevated in the halitosis samples. Furthermore, it was observed that the values of volatile sulfur compounds and indole-3-acetic abundances were positively correlated. The multiomics analysis identified the metagenomic and metabolomic characteristics to differentiate halitosis from healthy individuals using the least absolute shrinkage and selection operator logistic regression and random forest classifier. A total of 19 species and 39 metabolites were identified as features in halitosis patients, which included indole-3-acetic acid, Bacillus altitudinis, Candidatus Saccharibacteria, and Actinomyces species. In conclusion, an evident shift in microbiome and metabolome characteristics was observed in the halitosis tongue coating, which may have a potential etiological significance and provide novel insights into the mechanism for halitosis.

Efficacy of different mouthrinse formulations in reducing oral malodour: a randomized clinical trial.

AIMS: The aim of this study was to assess the efficacy of mouthrinses formulations in oral malodour. MATERIAL & METHODS: This single-centre, double-blind, randomized, parallel group clinical trial compared the efficacy of Halita™ and meridol(®) with and without zinc lactate versus negative and positive control. Volunteers with confirmed oral malodour (18/group) rinsed with one mouthrinse during 7 days (15 ml, 2x/day for 1 min.). 15 min. after a first rinse (masking effect), and after 7 days (therapeutic effect) the change in organoleptic scores and level of sulphur compounds was recorded. RESULTS: All rinses showed a masking effect (OLS 1 to 2 values reduced), only the rinses with antimicrobial ingredients showed a therapeutic effect (OLS 1 to 1.5 value less). The addition of zinc resulted in a more pronounced masking effect. Halita™ and meridol(®) with zinc showed the best therapeutic effect. CONCLUSION: Although the masking effect of the rinses can be attributed partially to a dilution and the effect of aromas, the therapeutic effect should be linked to the anti-microbial action of active ingredients and counter action of zinc ions on VSC. A complete resolution of the unpleasant breath by additional mechanical intervention remains to be proven.

Influence of gender and stress on the volatile sulfur compounds and stress biomarkers production.

BACKGROUND: Stress and menstrual cycle have been described as factors influencing bad breath, as they can alter oral homeostasis and contribute to the production of volatile sulfur compounds (VSC). OBJECTIVE: Considering that the experimenter's and volunteer's gender may influence the volunteer's responses to stress, the aim of this work was to evaluate the influence of stress and gender on the production of VSC and salivary biomarkers. METHODS: The experimental acute stress was induced by the Video-Recorded Stroop Color-Word Test (VRSCWT). The VSC, salivary proteins, and cardiovascular parameters were measured before and after VRSCWT. RESULTS: The VRSCWT induced significant increase in total VSC, hydrogen sulfide, and blood pressure values in men and women. Women presented higher values of both these compounds than men. The increase in systolic blood pressure was more pronounced when subjects were evaluated by an experimenter of the opposite gender. When women were evaluated by a member of the opposite gender, they showed significant increases in salivary alpha-amylase and cortisol compared with baseline values. CONCLUSION: Thus, the results showed that VRSCWT induced acute stress, which increased VSC production, and these effects were shown to be influenced by the gender.

Chewing gum containing allyl isothiocyanate from mustard seed extract is effective in reducing volatile sulfur compounds responsible for oral malodor.

PURPOSE: To evaluate the in vivo effect of chewing gum containing allyl isothiocyanate alone, and in combination with zinc salts on reduction of the level of volatile sulfur compounds responsible for oral malodor. METHODS: 15 healthy volunteers between the ages of 20-50 chewed either an experimental gum or a placebo gum for 12 minutes. Their mouth air was analyzed for volatile sulfur compounds by a gas chromatograph at baseline, immediately after chewing, and at 60, 120 and 180 minutes after treatment. RESULTS: The study revealed that allyl isothiocyanate, a constituent of mustard seed extract, can effectively reduce the concentration of volatile sulfur compounds in mouth air. Chewing gum containing 0.1% zinc lactate and 0.01% of allyl isothiocyanate eliminated 89%, 55.5%, 48% and 24% of the total VSC concentration immediately after chewing and at 1, 2, and 3 hours after chewing, respectively.

Short term clinical efficacy of new meridol HALITOSIS tooth & tongue gel in combination with a tongue cleaner to reduce oral malodor.

OBJECTIVE: This study evaluated the short term efficacy of tongue cleaning with meridol HALITOSIS tooth & tongue gel in comparison to mechanical tongue cleaning alone and untreated after five and 60 minutes in patients with an oral cause of bad breath. METHODS: Fifty-four male and female subjects with an intra-oral cause of halitosis (organoleptic ratings > or = 2 and volatile sulphur compounds > or = 50 ppb) participated in this crossover study and were assigned to six different treatment sequences (ABC, ACB, BAC, BCA, CAB, CBA) with no treatment (A), mechanical tongue cleaning alone (B), and tongue cleaning with tooth & tongue gel applied to the tongue cleaner (C). Efficacy was assessed by organoleptic ratings and volatile sulphur compound measurements five and 60 minutes after treatment RESULTS: Cleaning the tongue with tooth & tongue gel applied onto the tongue cleaner resulted in significantly reduced organoleptic ratings (p < 0.001 for the five-minute assessment; p = 0.001 for the 60-minute assessment) and volatile sulphur compounds (H2S + CH3SH: p = 0.005 for the five-minute assessment; p = 0.003 for the 60-minute assessment) compared to no treatment at the five- and 60-minute assessment time points, while mechanical tongue cleaning alone was less effective in reducing organoleptic ratings (p = 0.008 for the five-minute assessment; p = 0.144 for the 60-minute assessment) and volatile sulphur compounds (H2S + CH3SH: p = 0.261 for the five-minute assessment; p = 0.365 for the 60-minute assessment). CONCLUSIONS: Single tongue cleaning with meridol HALITOSIS tooth & tongue gel had a positive effect on halitosis five and 60 minutes after treatment. Tongue cleaning with tooth & tongue gel in combination with other oral hygiene procedures is a promising approach to control halitosis.

Oral malodorous compound causes caspase-8 and -9 mediated programmed cell death in osteoblasts.

BACKGROUND AND OBJECTIVE: Hydrogen sulfide (H(2) S) is one of two volatile sulfur compounds that are known to be the main cause of oral malodor; the other is methyl mercaptan. Other known volatiles existing in mouth air do not contribute significantly to oral malodor originating in the oral cavity. Hydrogen sulfide is also known to be an etiological factor in periodontal disease. However, the effects of H(2) S on alveolar bone remain unclear. The objectives of this study were to determine the apoptotic effects of H(2) S on osteoblasts and to verify the apoptotic molecular pathways. MATERIAL AND METHODS: A clonal murine calvaria cell line was incubated with 50 ng/mL of H(2) S. To detect apoptosis, the cells were analysed by flow cytometry and ELISA. Mitochondrial membrane depolarization was assessed using flow cytometry as well. ELISA was used to evaluate the release of cytochrome c into the cytosol and to assess Fas ligand, p53, tumor necrosis factor α, interleukin IL1-α IL-ß, IL-2, IL-4, IL-10, interferon-γ, granulocyte-colony stimulating factor and granulocyte-macrophage colony stimulating factor. Caspase-3, -8 and -9 activities were estimated. Expression of BAX and Bcl-2 was assessed by real-time quantitative RT-PCR. DNA fragmentation was detected by single-cell gel electrophoresis. Fas receptors were evaluated by western blotting. RESULTS: After H(2) S incubation, apoptotic levels increased significantly in a time-dependent manner. Mitochondrial membrane depolarization, the release of cytochrome c, p53 and caspase-3, -8 and -9 and DNA fragmentation were all significantly greater. BAX gene activity was upregulated, whereas Bcl-2 remained low. Fas ligand/Fas receptor, tumor necrosis factor α and other cytokines were not increased to a significant degree. CONCLUSION: At less-than-pathological concentrations in gingival crevicular fluid, H(2) S induces apoptosis in osteoblasts. The molecular mechanisms underlying the apoptotic process include p53, a mitochondrial pathway and caspase-8 activation.

Clinical utility of a novel colorimetric chair side test for oral malodour.

AIM: The aim of this study was to evaluate the utility of a simple colorimetric chair side test detecting amines in saliva as an adjunct test in the oral malodour diagnosis. MATERIAL AND METHODS: Non-stimulated saliva samples were collected from 100 volunteers with different degrees of oral malodour. The amount of amines detected by the test was estimated clinically (colorimetric test), confirmed semi quantitatively in the laboratory (standard addition method) and consequently compared with (i) the organoleptic score (OLS); (ii) the volatile sulphur compounds (VSCs) levels (OralChroma(™)) and (iii) the amount of amines detected by means of gas chromatography - mass spectroscopy. RESULTS: The chair side test correlated well (Spearman correlation coefficient: 0.46-0.77), with the OLS, the level of VSCs, and the amines determined by using gas chromatography - mass spectroscopy. The results of the new test for patients with and without oral malodour were significantly different (Mann-Whitney U-test, p < 0.0001). The sensitivity, specificity, and positive and negative predictive value of this chair side test were similar to those of the VSCs evaluations. CONCLUSIONS: These results support the "fit for purpose" of the new chair side test as adjunctive diagnostic tool for oral malodour.

Daily variation of oral malodour and related factors in community-dwelling elderly Thai.

OBJECTIVES: The purposes of this study were (i) to estimate the prevalence of oral malodour, (ii) to evaluate the daily variation of oral malodour and (iii) to assess associations of volatile sulphur compound (VSC) concentrations with socio-demographics, health behaviours and oral health status in community-dwelling elderly Thai. METHODS: The subjects were 428 dentate elderly people (67.6 ± 5.6 years) living in Phitsaulok, Thailand. Information on their socio-demographics, general health and health behaviours was obtained by a questionnaire. Their dental condition, periodontal status and tongue coating were clinically examined. Their flow rates and the pH of unstimulated saliva were also assessed. Oral malodour was measured at four different times of day using an Oral Chroma™. RESULTS: The proportions of subjects diagnosed with oral malodour using the thresholds of H(2)S, CH(3)SH and (CH(3))(2) S were 60.5%, 62.9% and 80.7%, respectively. Concentrations of H(2)S showed significant daily variation. Linear regression analysis demonstrated the following significant associations: (i) oral malodour from H(2)S and thickness of the tongue coating, (ii) oral malodour from CH(3)SH and periodontal pocket depth of 5 mm or more and the presence of gingival bleeding and (iii) oral malodour from (CH(3))(2)S and systemic disease, medications and thickness of the tongue coating. DISCUSSION: Oral malodour was shown to be prevalent among the elderly. Daily variation was observed in the concentration of H(2)S. Tongue coating, periodontal disease, systemic diseases and medications were related to oral malodour. Therefore, these factors should be taken into consideration in oral malodour treatment and prevention programmes for the elderly.

The tongue biofilm metatranscriptome identifies metabolic pathways associated with the presence or absence of halitosis.

Intra-oral halitosis usually results from the production of volatile sulfur compounds, such as methyl mercaptan and hydrogen sulfide, by the tongue microbiota. There are currently no reports on the microbial gene-expression profiles of the tongue microbiota in halitosis. In this study, we performed RNAseq of tongue coating samples from individuals with and without halitosis. The activity of Streptococcus (including S. parasanguinis), Veillonella (including V. dispar) and Rothia (including R. mucilaginosa) was associated with halitosis-free individuals while Prevotella (including P. shahi), Fusobacterium (including F. nucleatum) and Leptotrichia were associated with halitosis. Interestingly, the metatranscriptome of patients that only had halitosis levels of methyl mercaptan was similar to that of halitosis-free individuals. Finally, gene expression profiles showed a significant over-expression of genes involved in L-cysteine and L-homocysteine synthesis, as well as nitrate reduction genes, in halitosis-free individuals and an over-expression of genes responsible for cysteine degradation into hydrogen sulfide in halitosis patients.

Effect of Underwater Exercise on Salivary Metabolites of Older Persons With Disability.

BACKGROUND/AIM: Underwater exercise is aimed at preventing aging, maintaining, and improving motor function, and improving physical function. However, its rehabilitation effects have not been well evaluated. In order to gain insight into the molecular basis of its rehabilitation effects, possible changes in the salivary metabolites of four older persons with disability (mean age: 72.5 years) during underwater exercise were investigated. MATERIALS AND METHODS: Halitosis was measured by Breathtron; salivary bacterial number by bacterial counter; amino acids by amino acid analyzer; 8-oxoguanine by ELISA; and intracellular metabolites by capillary electrophoresis, time-of-flight mass spectrometry, liquid chromatography, and triode quadrupole mass spectrometry. RESULTS: Underwater exercise induced apparent declines in two major salivary amino acids (glycine and proline) and bacterial numbers in the cheek mucosa and salivary, without apparent changes in the halitosis and urine 8-oxoguanine concentration. Older subjects showed higher concentrations of most of 166 metabolites compared to young volunteers (mean age: 38.8 years old). Fifteen compounds were significantly reduced with the progression of underwater exercise. CONCLUSION: Improvement of upright balance function with underwater exercise is correlated with several salivary components.

Effects of a composition containing lactoferrin and lactoperoxidase on oral malodor and salivary bacteria: a randomized, double-blind, crossover, placebo-controlled clinical trial.

We report a clinical trial of the effects of test tablets containing bovine lactoferrin and lactoperoxidase on oral malodor and salivary bacteria. Fifteen subjects with volatile sulfur compounds (VSCs) in mouth air above the olfactory threshold (H(2)S >1.5 or CH(3)SH >0.5 ng/10 ml) as detected by gas chromatography were enrolled in the trial. Either a test or a placebo tablet was ingested twice at 1-h intervals in two crossover phases. Mouth air was monitored for VSC levels at the baseline before ingestion of a tablet, 10 min after the first ingestion, 1 h (just before the second ingestion), and 2 h after the first ingestion. Whole saliva was analyzed at the baseline and at 2 h for bacterial numbers. At 10 min, the level of CH(3)SH was significantly lower in the test group (median [interquartile range] = 0.28 [0.00-0.68] ng/10 ml) compared to that in the placebo group (0.73 [0.47-1.00] ng/10 ml; P = 0.011). The median concentration of CH(3)SH in the test group was below the olfactory threshold after 10 min until 2 h, whereas the level in the placebo group was above the threshold during the experimental period. No difference in the numbers of salivary bacteria was detected by culturing or quantitative PCR, but terminal restriction fragment length polymorphism detected one fragment with a significantly lower copy number at 2 h in the test group (mean ± standard error, 4.89 ± 0.11 log(10) copies/10 µl) compared to that in the placebo group (5.38 ± 0.15 log(10) copies/10 µl; P = 0.033). These results indicate a suppressive effect of the test composition on oral malodor and suggest an influence on oral bacteria.

Relationship between oral malodor and the menstrual cycle.

BACKGROUND AND OBJECTIVE: Sex hormones have been suggested to be important modifying factors that may influence the pathogenesis of periodontal disease. This study examined changes in volatile sulfur compounds (VSC) levels, clinical parameters and bacterial levels during the menstrual cycle. MATERIAL AND METHODS: The study group consisted of 10 female subjects with periodontitis and 12 periodontally healthy female subjects. Clinical and bacterial measurements were performed for all subjects during the ovulation and follicular phases of the menstrual cycle. RESULTS: Bleeding on probing (BOP) was significantly increased in the ovulation phase in periodontitis subjects but not in healthy subjects. The VSC levels in subjects with periodontitis increased 2.2-fold in the ovulation phase compared with the follicular phase. In the ovulation phase, VSC levels and BOP were significantly higher in subjects with periodontitis than in healthy subjects. The number, and salivary levels, of Prevotella intermedia in subjects with periodontitis were significantly higher in the ovulation phase than in the follicular phase. CONCLUSION: The present study indicated changes in VSC, BOP and P. intermedia during the menstrual cycles of women with periodontitis.

Oral malodorous compound triggers mitochondrial-dependent apoptosis and causes genomic DNA damage in human gingival epithelial cells.

BACKGROUND AND OBJECTIVE: Volatile sulfur compounds are the main compounds causing halitosis. One of these compounds, hydrogen sulfide (H(2)S), which is responsible for physiological halitosis, is reported also to have periodontal pathogenic activities. Hydrogen sulfide has been shown to activate the apoptotic process in different tissues. Apoptosis plays an important role in the development of periodontitis. The aim of this study was to determine whether H(2)S causes apoptosis in human gingival epithelial cells and to examine the cellular signaling pathway initiating the process. MATERIAL AND METHODS: Human gingival epithelial cells were incubated with 50 ng/mL H(2)S in air contining 5% CO(2) for 24, 48 or 72 h. To detect apoptosis, the cells were stained with annexin V and 7-amino actinomycin D, and analyzed using flow cytometry. Reactive oxygen species, mitochondrial membrane depolarization and release of cytochrome C into the cytosol were assessed using flow cytometry and enzyme-linked immunosorbent assay. Activity levels for the key apoptotic enzymes caspase-9, -8 and -3 were also determined. Genomic DNA damage was detected using single-cell gel electrophoresis. RESULTS: Apoptosis was significantly increased to 24.5 +/- 5.7 at 24 h and 41.5 +/- 8.9% at 48 h (p < 0.01). Reactive oxygen species were enhanced and mitochondrial membrane depolarization was collapsed. Cytochrome C release was dramatically increased (0.12 +/- 0.02 vs. 0.02 +/- 0.01 at 24 h and 0.21 +/- 0.02 vs. 0.02 +/- 0.01 ng/mL at 48 h; p < 0.05). Caspase-9 and -3 were strongly activated, while caspase-8 activity remained low. The percentage of DNA strand breaks increased, especially at 48 h. CONCLUSION: Hydrogen sulfide induces apoptosis in human gingival epithelial cells by activating the mitochondrial pathway.