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One of the most severe complications of lung resection is postoperative pneumonia, and its prevention and prediction are critical. Exhaled acetone and isoprene are thought to be related to metabolism; however, little is known on their relationship with bacteria living in the oral cavity or their meaning in the acute phase in perioperative lung cancer patients. We measured acetone and isoprene in exhaled breath of 13 Japanese patients with lung cancer (3 women and 10 men, age range 62-82 years, mean 72.4 years) before breakfast during hospitalization, and compared with two acute-phase proteins, C-reactive protein (CRP) and albumin in blood serum, as well as the total number of bacteria in saliva and their activity to produce acetone and isoprene. Before operation, intensive oral care was carried out for each patient to prevent postoperative pneumonia, and swallowing and cough reflexes were measured for 12 of 13 patients to assess risk of postoperative pneumonia. Breath and saliva were sampled before intensive oral care (T1), after oral care but before operation (T2), and after operation (T3) during hospitalization. The total number of oral bacteria in saliva decreased significantly from T1 to T2 among 13 patients. No acetone or isoprene was detected from saliva after in vitro incubation under anaerobic or aerobic conditions, but both acetone and isoprene were detected in breath. After operation, breath acetone correlated significantly with CRP (Spearman's ρ = 0.559, P = 0.03), but not with albumin. Breath isoprene correlated significantly with albumin (Spearman's ρ = 0.659, P = 0.008), but not with CRP after operation. Although the number of subjects was small, our results support the hypothesis that breath acetone and isoprene may be related with these acute-phase proteins, which reflect inflammatory reactions and subsequent changes in metabolism in the early postoperative phase of lung resection.

Acetaldehyde is a potential carcinogen for esophageal cancer, and some oral microorganisms produce acetaldehyde from ethanol or glucose. In this prospective study, we examined the influence of professional oral care on acetaldehyde levels in mouth air of esophageal cancer patients. Acetaldehyde concentrations in mouth air and breath were measured by a portable gas chromatograph, and acetaldehyde production from oral microbiota was also evaluated. Samples were taken from 21 esophageal cancer patients (median age 68 years) and 20 age-matched healthy volunteers (control group) before and after oral care. Post-operative samples were also taken from 17 patients who had undergone surgery. All samples (mouth air, breath, and saliva) were collected 2 to 3 hours after lunch. Oral microbial samples were prepared from saliva. Genotype analysis of alcohol dehydrogenase 1B (ADH1B) and aldehyde dehydrogenase-2 (ALDH2) genes revealed no significant differences in the genotypes between the two groups. In the control group, acetaldehyde levels in mouth air showed no significant changes after oral care, while the amount of microbial acetaldehyde production from ethanol was significantly decreased. By contrast, among the patients, acetaldehyde levels in mouth air were significantly decreased after oral care and after operation, while the amount of microbial acetaldehyde production from ethanol showed no significant changes. Moreover, microbial acetaldehyde production from glucose was significantly decreased after operation. Overall, oral health was poorer in the patient group. In conclusion, professional oral care for esophageal cancer patients is effective for reducing acetaldehyde levels in mouth air due to the reduction of microbial count.

Acetaldehyde is known to be carcinogenic and produced by oral bacteria. Thus, bacterial acetaldehyde production might contribute to oral cancer. Therefore, we examined bacterial acetaldehyde production from ethanol and glucose under various conditions mimicking the oral cavity and clarified the metabolic pathways responsible for bacterial acetaldehyde production. Streptococcus mitis, S. salivarius, S. mutans, Neisseria mucosa and N. sicca were used. The bacterial metabolism was conducted at pH 5.0–8.0 under aerobic and anaerobic conditions. The production of acetaldehyde and organic acids was measured with gas chromatography and HPLC, respectively. Bacterial enzymes were also assessed. All of the bacteria except for S. mutans exhibited their greatest acetaldehyde production from ethanol at neutral to alkaline pH under aerobic conditions. S. mutans demonstrated the greatest acetaldehyde from glucose under anaerobic conditions, although the level was much lower than that from ethanol. Alcohol dehydrogenase and NADH oxidase were detected in all of the bacteria. This study revealed that oral indigenous bacteria, Streptococcus and Neisseria can produce acetaldehyde, and that such acetaldehyde production is affected by environmental conditions. It was suggested that alcohol dehydrogenase and NADH oxidase are involved in ethanol-derived acetaldehyde production and that the branched-pathway from pyruvate is involved in glucose-derived acetaldehyde production.

One of the most severe complications of lung resection is postoperative pneumonia, and its prevention and prediction are critical. Exhaled acetone and isoprene are thought to be related to metabolism; however, little is known on their relationship with bacteria living in the oral cavity or their meaning in the acute phase in perioperative lung cancer patients. We measured acetone and isoprene in exhaled breath of 13 Japanese patients with lung cancer (3 women and 10 men, age range 62-82 years, mean 72.4 years) before breakfast during hospitalization, and compared with two acute-phase proteins, C-reactive protein (CRP) and albumin in blood serum, as well as the total number of bacteria in saliva and their activity to produce acetone and isoprene. Before operation, intensive oral care was carried out for each patient to prevent postoperative pneumonia, and swallowing and cough reflexes were measured for 12 of 13 patients to assess risk of postoperative pneumonia. Breath and saliva were sampled before intensive oral care (T1), after oral care but before operation (T2), and after operation (T3) during hospitalization. The total number of oral bacteria in saliva decreased significantly from T1 to T2 among 13 patients. No acetone or isoprene was detected from saliva after in vitro incubation under anaerobic or aerobic conditions, but both acetone and isoprene were detected in breath. After operation, breath acetone correlated significantly with CRP (Spearman's ρ = 0.559, P = 0.03), but not with albumin. Breath isoprene correlated significantly with albumin (Spearman's ρ = 0.659, P = 0.008), but not with CRP after operation. Although the number of subjects was small, our results support the hypothesis that breath acetone and isoprene may be related with these acute-phase proteins, which reflect inflammatory reactions and subsequent changes in metabolism in the early postoperative phase of lung resection.

Background Since most oral malodor originates from microbial activities in the mouth, the role of microorganisms in producing malodorous gases has been clarified by numerous studies, accompanied by the development of analytical techniques for treatment of halitosis. Highlight Oral microorganisms should be controlled to prevent aspiration pneumonia, especially for elderly perioperative patients. Malodorous gases from the mouth can be an indicator of oral or systemic conditions among patients in intensive care units. Recently, malodorous gases originating from oral microorganisms have been reported as a causal factor in carcinogenesis. Conclusion Further analysis of oral malodor might be useful in accessing the risk of aspiration pneumonia and oral cancer.

Objectives The source of the bacteria involved in silent aspiration remains to be completely defined. This study aimed to obtain reliable evidence on silent aspiration of oral bacteria in elderly patients. Methods After obtaining informed consent, the cough and swallowing reflexes of patients were assessed. Bronchial fluids from patients undergoing lung resections were collected with a micro-sampling probe, and α-amylase activity of bronchial fluids was measured to estimate the degree of silent aspiration. The bronchial fluids were cultured aerobically and anaerobically on blood agar plates, and colonies were identified by 16S rRNA gene sequencing. Additionally, whole saliva bacterial amounts and composition were analyzed. Results Six patients (72.2±5.8 years) exhibited an impaired swallowing reflex and 5 (75.4±7.9 years) had a normal swallowing reflex, while all patients had a normal cough reflex. α-Amylase activity was detected in bronchial fluids of both the impaired and normal reflex groups. The amount of anaerobic bacteria in bronchial fluids in the impaired reflex group [(3.0±3.5)×10 4 ] was higher than in the normal reflex group [(2.5±5.3)×10 4 ], although the difference was not significant. Actinomyces , Gemella , Streptococcus , Rothia , Mogibacterium, and Campylobacter were the predominant bacterial species in bronchial fluids of the impaired reflex group, while Streptococcus , Lactobacillus , Veillonella , and Actinomyces were predominant in the normal reflex group. Conclusions Our results suggest that bacteria in bronchial fluids associated with silent aspiration are derived from saliva, and that the bronchial fluids of elderly patients with an impaired swallowing reflex may have a characteristic microbiota.

Postoperative pneumonia may occur when upper respiratory tract protective reflexes such as cough and/or swallowing reflexes are impaired; thus, silent aspiration of oral bacteria may be a causative factor in postoperative pneumonia. This study aimed to quantify and identify bacteria in intraoperative bronchial fluids and to evaluate the relationship between impairment of cough/swallowing reflexes and silent aspiration of oral bacteria in elderly patients. After obtaining informed consent, cough and swallowing reflexes were assessed using an ultrasonic nebulizer and a nasal catheter, respectively. Using a micro-sampling probe, intraoperative bronchial fluids were collected from nine subjects with pulmonary carcinoma and cultured anaerobically on blood agar plates. After 7 days, CFUs were counted and isolated bacteria were identified by 16S rRNA gene sequencing. Four subjects (aged 71.0 ± 8.4 years) had impaired swallowing reflexes with normal cough reflexes, whereas five subjects (73.6 ± 6.5 years) had normal cough and swallowing reflexes. The bacterial counts (mean CFU ± SD) tended to be higher in intraoperative bronchial fluids of subjects with impaired swallowing reflexes ([5.1 ± 7.7] × 10(5)) than in those of subjects with normal reflexes ([1.2 ± 1.9] × 10(5)); however, this difference was not statistically significant. Predominant isolates from intraoperative bronchial fluids were Streptococcus (41.8%), Veillonella (11.4%), Gemella (8.9%), Porphyromonas (7.6%), Olsenella (6.3%) and Eikenella (6.3%). These findings indicate that intraoperative bronchial fluids contain bacteria, probably derived from the oral microbiota, and suggest that silent aspiration of oral bacteria occurs in elderly patients irrespective of impairment of swallowing reflex.

Pneumonia in elderly people is mainly caused by silent aspiration due to an age-related impairment of cough and swallowing reflexes. Because most of the patients with lung cancer are elderly people, we hypothesized that the age-related impairment of these protective reflexes might exist or occur in patients undergoing lung surgery, and cause postoperative pneumonia. We revealed that many elderly patients showed depressed swallowing reflex even before surgery and transient attenuation of cough reflex after surgery, and that postoperative pneumonia occurred only in the patients whose cough and/or swallowing reflex was abnormal postoperatively. Then, we prospectively showed that 30 elderly patients who received perioperative intensive oral care, including professional assessment of oral status, dental cleaning, and patient education for self-oral care by dentists, followed by intensive oral care by intensive care unit nurses, and encouragement of self-oral care by floor nurses, did not develop pneumonia after lung resection. In this study, we retrospectively reviewed the execution status of professional oral care by dentists and the occurrence of postoperative pneumonia in 159 consecutive patients aged 65 or older undergoing lung resection from 2013 to 2014. Thoracic surgeons in our institute asked dentists to provide professional oral care before lung resection only in 30.3% of the subjects in 2013, and 45.8% in 2014. Postoperative pneumonia occurred in 3 out of 76 subjects(3.9%)in 2013, and 1 out of 83(1.2%) in 2014. In 2013, 1 patient who did not receive preoperative professional oral care developed aspiration pneumonia postoperatively followed by acute exacerbation of idiopathic pulmonary fibrosis and in-hospital death. We need to make an effective system to provide preoperative professional oral care by dentists especially for elderly patients and high-risk patients before lung resection.

Based on the hypothesis that the complex pathological and immunological manifestations of rheumatoid arthritis (RA) and the related diseases are under the control of multiple gene loci with allelic polymorphism, a recombinant congenic mouse strain was prepared between an MRL/Mp-lpr/lpr (MRL/lpr) strain, which develops arthritis resembling RA, and a non-arthritic strain C3H/HeJ-lpr/lpr (C3H/lpr). In MRL/lpr x (MRL/lpr x C3H/lpr) F1 mice, the mice developing severe arthritis were selected based on joint swelling to further continue intercrosses, and then an McH-lpr/lpr-RA1 (McH/lpr-RA1) strain was established and its histopathological phenotypes of joints and autoimmune traits were analyzed. Arthritis in McH/lpr-RA1 mice developed at a higher incidence by 20 weeks of age, compared with that in the MRL/lpr mice, who had severe synovitis (ankle, 60.3%; knee, 65.1%), and also fibrous and fibrocartilaginous lesions of articular ligamenta resembling enthesopathy (ankle, 79.4%; knee, 38.1%), resulting in ankylosis. The lymphoproliferative disorder was less, and serum levels of IgG and IgG autoantibodies including anti-dsDNA and rheumatoid factor were lower than those of both MRL/lpr and C3H/lpr strains. McH/lpr-RA1 mice may provide a new insight into the study of RA regarding the common genomic spectrum of seronegative RA and enthesopathy.

Background: The spontaneous onset of macroscopic arthropathy in the ankle of the particular F 1 mice descended from two Fas -deficient strains of mice; a mutant substrain of MRL/Mp. Fas lpr (MRL/rpl) and C3H/He. Fas lpr (C3H/lpr) was recently observed. Aim: To histopathologically characterise and genetically interpret the unique inheritance mode of disease in this arthropathy model. Methods: MRL/rpl, C3H/lpr, (MRL/rpl × C3H/lpr; MC) F 1 , (C3H/lpr × MRL/rpl; CM) F 1 and MCF 2 mice were bred under specific pathogen-free conditions. Histopathological grade of arthropathy was determined at 6 months by examination under a light microscope. To search for a linkage locus to the arthropathy, the whole genome of selected 48 male MCF 2 mice with 71 polymorphic microsatellite markers was scanned, followed by quantitative trait locus analysis. Results: The incidence of microscopically defined arthropathy in the male and female MCF 1 groups was 100% and 19.4%, respectively. No incidence was observed in the parental strains, MRL/rpl and C3H/lpr, and in CMF 1 mice. In the MCF 1 mice, the arthropathy mainly affected the ankle joints and was histopathologically characterised by marked entheseal proliferation with chondrocytic differentiation and ossification in the ankle joints, the manifestations similar to ankylosing enthesitis reported previously. An MRL/rpl-derived autosomal dominant susceptibility locus was mapped in the distal of D7Mit68 (60 cM) to the ankylosis onset. Conclusion: The MCF 1 mice stably develop spontaneous ankylosing disorders in the ankle, with a male predominance. The unique inheritance mode of ankylosis is possibly interpreted by the genetic interaction between the autosomal dominant locus and a Y-linked locus.

Although the pathogenesis of leukoplakia has been unclear, carcinogenic transformation is postulated to result from alterations of apoptotic signal transduction proteins in epithelial cells. The pathogenesis of oral lichen planus (OLP) has also been unclear, but apoptotic changes of the epithelial cells in OLP have been reported. In the present study, we used a histochemical approach to describe human keratinocyte-expression of several apoptotic signaling proteins in leukoplakia, in OLP, and in normal oral mucosa as a control. Mucosal biopsies from patients with leukoplakia (n=13), OLP (n=10), and normal oral mucosa (n=9) were frozen, sectioned and immunostained with monoclonal antibodies to wild-type (wt) tumor suppressive protein p53, cyclin-dependent kinase inhibitor p21WAF1/CIP1 and the oncoproteins MDM2, and Bcl-2. Apoptosis was assessed in all cases by the TUNEL method. MDM2 and Bcl-2 expression in keratinocytes were quantitatively greater in leukoplakia than in OLP. Wt-p53 and p21WAF1/CIP1 expression was quantitatively greater in keratinocytes in OLP than in leukoplakia. Keratinocyte maturation appeared histologically normal in OLP, even though wt-p53 and p21WAF1/CIP1 were expressed in these cells. Altered keratinocyte maturation was seen in leukoplakia lesions expressing MDM2 and Bcl-2. No significant difference for the number of apoptotic epithelial cells was observed between leukoplakia and OLP, in spite of the divergent outcomes of the apoptotic signaling proteins.

Synthesis, storage, and secretion of the proinflammatory cytokine interleukin-1β (IL-1β) and the anti-inflammatory cytokine IL-6 have not been established in normal exocrine gland secretory cells. Parotid glands and isolated acinar cells prepared from BALB/c mice were homogenized for RNA isolation and reverse transcription-polymerase chain reaction (RT-PCR). IL-1β and IL-6 enzyme-linked immunosorbent assays (ELISAs) were done on supernatants prepared from mouse parotid acinar cell (MPAC) preparations unstimulated or stimulated between 0 and 10 min with 10−5M norepinephrine at 37°C. MPACs were fixed in paraformaldehyde, frozen sectioned for light and electron microscopy, and labeled with antibodies to IL-1β and IL-6. Mouse specific riboprobes to IL-1 and IL-6 were used for in situ hybridization. RT-PCR yielded the expected IL-1 (336-bp) and IL-6 (614-bp) mRNA products. By ELISA, stimulated MPACs showed a significant increase in IL-1β ( P < 0.03) and IL-6 ( P < 0.01) release into supernatants by 10 min that paralleled the time course of amylase release. In situ hybridization showed the presence of transcripts for IL-1 and IL-6 in glandular epithelial cells. Gold-labeled IL-1β and IL-6 were significantly higher ( P < 0.01) in granules than in the nucleus and cytoplasm. This study shows that MPACs synthesize IL-1β and IL-6 and release these cytokines from their granules after α- and β-adrenergic stimulation.

SUMMARY The subcellular events responsible for release of mediators by mast cells may help to clarify roles for mast cells in health and disease. In this study we show that the granule-associated protease chymase is also within cytoplasmic vesicles in appropriately stimulated rat peritoneal mast cells. Rat peritoneal mast cells were recovered before or 1‐10 sec after exposure to the secretogogue compound 48/80 (10 m g/ml) and then were examined by radioimmunoassay to quantify histamine release or were processed, using routine methods for postembedding immunoelectron microscopy, to identify the subcellular localization of chymase. In comparison to unstimulated cells, compound 48/80 stimulated cells in two independent experiments showed an increase (15%, 28%) in the surface area of the cell and a decrease (12%, 6%) in the surface area of the total granule compartment before degranulation channel formation. These global cellular changes occurred in a background of transient but significant ( p , 0.01) increases in the area and number of chymase-immunoreactive vesicles per m m 2 cytoplasm. These changes were detectable at 5 or 7 sec after stimulation with compound 48/80 but returned to near prestimulation levels by 9 or 10 sec after addition of compound 48/80 (total cumulative histamine release was 28% by 8 sec and 47% by 14 sec). These observations suggest that vesicles participate in the early stages of regulated secretion of chymase from rat peritoneal mast cells. (J Histochem Cytochem 45:1379‐1391, 1997)

Reduction of flow rate of saliva secretion causes xerostomia, which hardly compromises the quality of life of the patients. Thus, clinical diagnosis of xerostomia should be accurate and practicable to prevent and to strengthen preemptive oral health care in the early stage of xerostomia. There are two type of saliva, resting saliva and stimulated saliva, according to physical status. Because the functions of these two saliva are different, it is needed to measure the flow rates of both saliva for the diagnosis of xerostomia. The flow rate of stimulated saliva was significantly correlated with age, gender, number of sound tooth and body height. Among the healthy subjects, the multiple linear regression analysis indicated that the flow rate of stimulated saliva was correlated with body weight and gender. This indicated that the new criteria of flow rate of saliva would be needed for the therapeutic diagnosis of xerostomia.

Difference of breath acetone between type 1 and type 2 diabetes mellitus (DM) is examined under informed consent. We measure acetone in mouth air and in deep breath by Gas Analyzer® XG-100V (New Cosmos Electric Co., Ltd., Osaka, Japan) and volatile sulfide compound in mouth air by Breathtron® (New Cosmos Electric Co., Ltd., Osaka, Japan), after meal. The acetone ratio of deep exhaled breath to mouth air and concentration of volatile sulfide compound (VSC) are compared between type 1 and type 2 DM patients. The acetone ratio of deep exhaled breath to mouth air is significantly higher in type 1 DM than type 2 DM patients. The concentration of VSC shows no significant difference. Though more cases should be studied, there is a possibility that the acetone ratio might reflect metabolic disorder of type 1 DM patients.

Recently, -G1cNAcβ1-6Man-(β1-6) branched N-glycosidic oligosaccharides have been reported to play a role in tumor malignancy. In this study, to examine the expression of β1-6 branched oligosaccharides on oral squamous cell carcinoma cells, lectin histochemical staining with phaseolus vulgaris leucoagglutinin (L-PHA), which has a high affinity for β1-6 branched oligosaccharides, was performed on frozen tissue sections prepared from 34 patients with oral squamous cell carcinomas.The β1-6 branched oligosaccharides were localized on the surface of the tumor cells as follows: 1. All of the tumor cells were positive for staining (positive); 2. A population of the tumor cells were positive but the remainder were negative (weakly positive), 3 All of the tumor cells were negative (negative). Statistical analysis revealed that the incidence of metastasis to regional lymph nodes in the cases positive for L-PHA staining on the tumor cells was higher than that in cases negative for the L-PHA staining. These results suggested that the increased expression of β1-6 branched oligosaccharides of oral squamous cell carcinoma may enhance the metastatic potential to regional lymph nodes. On the other hand, previous studies have shown that β1-6 branched oligosaccharides may contribute to the ability of tumor cells to escape recognition by macrophages. Therefore, the number of CD14 positive cells, regarded to be a population of macrophages, which invaded into the stromal tissue adjacent to the tumor was also examined. As a result, smaller numbers of the CD14 positive cells were observed in the stroma adjacent to tumors positive for L-PHA staining compared with the stroma adjacent to tumors which were weakly positive or negative for L-PHA staining. These results suggested a role of β1-6 branched oligosaccharides in the recognition of oral squamous cell carcinoma cells by macrophages.

Oral health examinations were held in summer continuously for 2 years with general health checks in a rural town. We included oral health check, practice of oral cleaning, and measurement of oral malodor in the examination. Aim of this study was to examine the difference of impression in the oral health examination between younger and older participants by questionnaires. No difference was found between male and female ratio in each generation. Also, no difference was seen as for ratio of joiners of oral cleaning and ratio of their satisfaction after the practice. Significant difference between two generations was seen as for strong satisfaction ratio with explanation about the result of oral malodor, and effective motivation ratio for oral hygiene caused by measurement of oral malodor. The oral health examination held with general checks with measurement of oral malodor seemed more impressive to older generation.

Swallowing, which is important for eating and drinking for survival, is one of the basic functions of the mouth. The purpose of this study was to develop a safe and easily-handleable measuring device to evaluate swallowing processes in younger generations. An intra-oral pressure-measuring device estimated the intra-oral positive and negative pressures produced by sucking or blowing. The results indicated that the maximal positive pressure or minimal negative pressure increased or decreased depending upon age. This suggests that the function to produce intra-oral negative pressure, which was made with lip and faucial isthmus closure for sucking, and positive pressure, which was made with lip closure for blowing, was developed with age or physical strength. This simple measuring system of oral pressure was useful in fieldworks for screening oral dysfunction among all generations, not depending upon the condition of tooth or dentition.

Halitosis is a common oral concern and patients often seek help from dentists. The purpose of this study is to evaluate psychological characteristics of halitosis patients and their changes with treatment. The subjects consisted of 150 halitosis patients and 64 non-halitosis patients. They were requested to complete Egogram and the halitosis scale questionnaires, before and after treatment. The results indicated that halitosis patients improved in the objective recognition of their malodor level, their obsession with halitosis, and interpersonal communication after treatment. Therefore, the ideal dentist attitude for halitosis treatment is to help patients overcome their obsession by examining their breath level repeatedly and encourage them to develop social skills. However, these subjects had a tendency for social phobia, and its removal was not easy. Therefore, it would be essential that dentists continue to accept their complaints and relapse of their obsession with bad breath sincerely.

We carried out residents-attractive dental health examinations by increasing the learning contents with systemic health examination of a rural town in Japan. Town staffs or some residents who had finished training programs participated in the dental examinations as staffs. Oral malodor was measured to motivate oral hygiene of examinees. We prepared questionnaires and asked the impression of examinees about “the dental examination with learning” 7 months after the examination. Questionnaires showed that 85% of dental examinees recognized it as satisfactory. This fact indicates the possibility that the new dental examination can be a model for oral health promotion in the community.

Objective For measuring oral malodor in daily clinical practice and in field study, we developed and evaluated a highly sensitive portable monitor system. Methods We examined sensitivity and specificity of the sensor for volatile sulfur compounds (VSC) and obstructive gases, such as ethanol, acetone, and acetaldehyde. Each mouth air provided by 46 people was measured by this monitor, gas chromatography (GC), and olfactory panel and compared with each other. Based on the result, we used the monitor for mass health examination of a rural town with standardized measuring. Results The sensor detected hydrogen sulfide, methyl mercaptan, and dimethyl sulfide with 10–1000 times higher sensitivity than the other gases. The monitor's specificity was significantly improved by a VSC-selective filter. There were significant correlations between VSC concentration by the sulfide monitor and by GC, and by organoleptic score. Thirty-six percent of 969 examinees had oral malodor in a rural town. Seventy-eight percent of 969 examinees were motivated to take care of their oral condition by oral malodor measuring with the monitor. Conclusions The portable sulfide monitor was useful to promote oral health care not only in clinics, but also in field study. The simple and quick operation system and the standardized measuring make it one of parameters of oral condition.

Ketone bodies including acetone are disease biomarkers for diabetes that sometimes causes severe ketoacidosis. The present study was undertaken to clarify the significance of exhaled acetone and plasma ketone bodies at bedside in a clinical setting. The oral glucose tolerance test (OGTT) was performed in 10 healthy Japanese volunteers (five females and five males). Exhaled breath acetone and volatile sulfide compounds (VSCs) in mouth air were measured simultaneously with blood sampling during the OGTT using a portable gas chromatograph equipped with an In2O3 thick-film type gas sensor and a VSC monitor. Acetone, β-hydroxybutyrate (β-OHB) and acetoacetate (AcAc) in blood plasma as well as glucose and insulin were examined. Oral conditions were examined based on the Community Periodontal Index (CPI) by one dentist. In addition, the same type of analysis was applied to two uncontrolled type 2 diabetes mellitus patients hospitalized at Tohoku University Hospital. Exhaled acetone was measured at the same time as blood withdrawal in the morning before breakfast and at night before bed at the beginning, the middle, and the end of hospitalization. All volunteers showed normal OGTT patterns with no ketonuria and periodontitis; however, there were significant correlations between breath acetone and plasma β-ΟΗΒ and between breath acetone and plasma AcAc under fasting conditions. Breath acetone of the type 2 diabetes mellitus patients showed positive correlations with plasma glucose when the level of plasma glucose tended to decrease during hospitalization. In spite of a very limited number of cases, our results support the idea that exhaled breath acetone may be related to plasma β-OHB and AcAc, which reflect glucose metabolism in the body.

The deficiencies of nucleotide excision repair (NER) factors are involved in rare genetic diseases such as xeroderma pigmentosum (XP) with increased risk of developing cancer on sun-exposed areas of the skin. However, the abnormality of NER factors in human sporadic carcinoma remains unclear. Loss of heterozygosity (LOH) analysis, using the microdissected tissues, for the XPA, XPB, XPC, XPD, XPE, XPF, XPG and the transcription-coupled repair factor, Cockayne syndrome B (CSB) revealed that NER factors were abnormal in 30.0% (3/10 cases) of oral squamous cell carcinomas. Furthermore, 10.0% of oral carcinomas exhibited LOH for NER factors without LOH for tumor suppressor genes such as p53, FHIT, APC, BRCA1, BRCA2 and DCC. These observations raise the possibility that alterations of NER factors may be involved in carcinogenesis in human oral squamous cell carcinoma.

Hyperplastic gingival tissues show the histopathological characteristics of thick parakeratinized squamous epithelia with acanthosis and rete pegs elongated into the lamina propria. However, the pathogenic factors that contribute to the epithelial morphogenesis of this disease are obscure and remain to be studied.We immunohistochemically examined the expression of both c-Myc and bcl-2 oncoprotein, which can exert influence on the epithelial morphogenesis and homeostasis, in 12 hyperplastic gingival tissues induced by nifedipine and phenytoin as well as 5 control tissues using avidin-biotin-peroxidase complex methods.Four specimens out of 5 nifedipine-induced and 5 out of 7 phenytoin-induced hyperplastic gingival tissues revealed the expression of c-Myc oncoprotein, whereas no significant immunostaining of c-Myc oncoprotein was found in 5 control tissues. The c-Myc oncoprotein-positive cells were observed to be localized in the basal and suprabasal cell layers of the hyperplastic gingival epithelia. Although all of the 12 hyperplastic gingival epithelia showed the distribution of bcl-2 oncoprotein in the basal and suprabasal layer cells, in 5 control epithelia the bcl-2 oncoprotein expression was slight and confined to the basal layer cells.The results of our present study indicate that the synergistic overexpression of c-Myc and bcl-2 oncoprotein may be related to the pathogenesis of gingival hyperplasia induced by nifedipine and phenytoin, especially to the morphogenesis of hyperplastic epithelia.

Although it has been thought that drug-induced gingival hyperplasia is not related to tumorigenesis, recent case reports have shown that squamous cell carcinomas may arise in gingival hyperplasia induced by cyclosporin and phenytoin. The possible implications between the pathogenesis of this disease and tumorigenesis have not been elucidated and remain to be studied.We immunohistochemically examined the expression of tumor-related markers such as p53 protein and Ki-67 antigen in 11 hyperplastic gingival tissues induced by nifedipine and phenytoin, as well as 5 control tissues using an avidin-biotin-peroxidase complex method.Two specimens out of 4 nifedipine-induced and 4 out of 7 phenytoin-induced hyperplastic gingival tissues revealed the expression of p53 protein in the nuclei of epithelial cells, while no expression of p53 protein was observed in the epithelia of the 5 non-hyperplastic control tissues. The immunoreactions against p53 protein showed sporadic distribution in the suprabasal layers of hyperplastic epithelia. The mean percentage of epithelial cells expressing Ki-67 antigen in the hyperplastic gingival tissues was more than 10% higher than that in the controls. The expression of Ki-67 antigen was suppressed in the typical rete pegs deeply elongated into lamina propria of hyperplastic gingival tissues. Intense immunostaining of Ki-67 antigen was found in fibroblasts of hyperplastic gingival tissues, while that of the control tissues was negligible.The expression of p53 protein in gingival hyperplasia suggests that the pathogenesis of this disease is involved with impaired DNA, while the growth arrest observed in the hyperplastic epithelia with typically elongated rete pegs as expressed with Ki-67 antigen may prevent the invasive expansion of epithelial cells undergoing DNA damage within gingival tissues and may consequently suppress tumorigenic progression.

Relationship between bacterial flora and H2S levels in breath air was investigated in this study. Oral malodor levels in 10 subjects (age: 21–56 years) were assessed with gas chromatography, Breathtron® and organoleptic score. Based on these assessments, subjects were divided into 2 groups; oral malodorous and nonodorous groups. Tongue coating were sampled and spread onto Fastidious Anaerobe Agar plates containing 0.05% cysteine, 0.12% glutathione and 0.02% lead acetate, and were then incubated anaerobically at 37 °C for 2 weeks. Bacteria forming black or gray colonies were designated as H2S-producing phenotypes, and were identified by 16S rRNA gene sequencing. The numbers of total bacteria (p

The aim of this study was to examine the relation between levels of salivary mutans streptococci and HLA gene polymorphisms in dental caries-resistant and caries-susceptible human subjects. We analyzed HLA alleles at the HLA loci (DRB1, DQA1, and DQB1) in 89 subjects. Thirty-two subjects had high caries susceptibility, and 57 subjects were caries free. Salivary levels of mutans streptococci were measured with a strip mutans test. Comparisons of salivary mutans streptococci levels between certain allele positive and negative and between caries-resistant and caries-susceptible subjects were tested by means of Mann–Whitney U -test. In caries-resistant or caries-susceptible subjects there were significant differences of salivary mutans streptococci levels between positive allele and negative allele in HLA-DQB1*05 ( p = 0.01), DRB1*01 ( p = 0.02), DRB1*14 ( p = 0.04), DQA1*01 ( p = 0.007), and DQA1*04 cluster ( p = 0.01), respectively. Moreover, we found significant difference of salivary mutans streptococci levels in HLA-DRB1*04, *08, *09, and *12 alleles between two dental caries statuses, respectively. On the other hand, we did not find such difference in HLA-DRB1*01, *13, *14, and *15 alleles.

Immunohistochemical localization of TGFβ, bFGF and their receptors, c-myc and bcl-2 oncoproteins, and Ki-67 antigen was examined in 9 hyperplastic gingival tissues induced by nifedipine and phenytoin, and 5 non-hyperplastic control tissues to elucidate the pathogenesis of drug-induced gingival hyperplasia. The results of our present study indicate that the overexpression of TGFβ, bFGF is related to the morphogenesis of hyperplastic lamina propria, whereas the intensive immunolocalization of c-myc and bcl-2 oncoprotein contributes to that of hyperplastic gingival epithelia. The tissue growth features of hyperplastic gingivae which were shown by Ki-67 antigen expression seem to reflect the involvement of these ligands in the pathogenesis of drug-induced gingival hyperplasia.

Approximately 90% of oral malodour is believed to originate from foul-smelling gases such as volatile sulphur compounds (H 2 S, CH 3 SH, CH 3 SCH 3 ), produced by oral bacteria. To detect the volatile sulphur compounds (VSC) objectively and easily is necessary for better diagnosis of halitosis especially for private clinics or field study. We have developed a bad-breath monitor, Breathtron™, equipped with a semiconductor gas sensor and a special filter for removing obstructive gases. 16 gases were used to reveal sensing characteristic of this sensor. The sensitivity of the sensor was high enough to detect 0.01 ppm VSC. The mouth air of 42 volunteers was analyzed. Adequate correlation was found between Breathtron™ vs. gas chromatography ( r = 0.817). The correlation between Breathtron™ and olfactory panel was also confirmed ( r = 0.644). Furthermore, we examined 869 adults (386 males and 483 females) in a Japanese rural town under informed consent. Halitosis was seen in about 40% of the examinees. The amount of tongue coating and the number of present teeth were found to be related to halitosis. Breathtron™ can be used widely at private clinics or even in field study of halitosis for its portable size, simple operation, and short measuring time.

We developed a portable device (Breathtron™) to measure volatile sulfur compounds (VSC). The aim of this study was to elucidate the circadian changes of VSC using this device. Five male students of Tohoku University Graduate School of Dentistry participated in the experiment; the VSC of the participants was measured for 2 days. VSC levels showed drastic changes according to meals and brushings. VSC level was the highest in the early morning, significantly reduced after meals and gradually increased until next meals. The results of the present study show that circadian changes exist in VSC level.

The expression of -GlcNAc beta 1-6Man-(beta 1-6) branched oligosaccharides in carcinoma cells has been considered to influence their metastatic potentials. In the present paper, the lectin histochemistry of oral squamous cell carcinomas obtained in biopsy from 34 patients with Phaseolus vulgaris leukoagglutinin (L-PHA), which potentially binds to N-glycosidic carbohydrates with beta 1-6 linked lactosamin antennae, was studied in order to analyze the relationship between their staining patterns and metastases. The L-PHA-binding oligosaccharides of the carcinomas were expressed on the cell surface in the following patterns: (i) all cells were positive for the staining ('positive'); (ii) some cells were positive but the rest of the carcinoma cells were negative ('weakly positive'); and (iii) all were negative ('negative'). Statistical analysis revealed that the incidence of the metastasis to regional lymph nodes in the 'positive' cases was significantly higher than that in the 'negative' cases. Moreover, the number of the CD14 positive cells including macrophages in the stroma adjacent to the carcinomas in the 'positive' cases was less than that in the 'negative' or 'weakly positive' cases. The expression of L-PHA-binding oligosaccharides in oral squamous cell carcinoma may be responsible for their metastatic potential to regional lymph nodes, possibly including their ability to escape macrophage recognition.

A case of sialolithiasis in a Blandin's gland duct is reported. Although minor salivary glands calculi are not uncommon, Blandin's glands are rare sites of occurrence. In this case, a calculus without decalcification is reported. Not only clinico-pathological features but also electron probe micro analyzer plus energy dispersive X-ray spectroscopy studies are presented. These studies reveal that phosphorus is contained in only the basophilic part of the calculus and sulphur in both the basophilic and the eosinophilic parts.