Aging-associated stem/progenitor cell dysfunction in the salivary glands of mice.

Although stem cell aging leads to a decline in tissue homeostasis and regenerative capacity, it remains unclear whether salivary gland stem cell function changes during this process. However, the salivary glands are gradually replaced by connective tissue during aging. Here, we show a decline in the stem cell ability of CD133-positive stem/progenitor cells in the salivary glands of aged mice. The CD133-positive cells were isolated from young, adult, and aged mice. The number of CD133-positive cells was significantly decreased in aged mice. They also showed a lower sphere formation capacity compared to young and adult mice. RNA sequencing revealed that CD133-positive cells in aged mice exhibited lower gene expression of several aging-related genes, including FoxO3a, than those in young and adult mice. Salivary gland cells infected with a recombinant lentivirus encoding the FoxO3a gene showed a reduction in oxidative stress induced by hydrogen peroxide compared with those infected with a control virus. Thus, FoxO3a may inhibit stem cell aging via oxidative stress.

Second-Degree Atrioventricular Block Occurring After Tooth Extraction.

Although cardiac arrhythmias are occasionally associated with dental extractions and dental anesthesia, atrioventricular block is rarely seen during dental procedures. We report a rare case of type I second-degree atrioventricular block (Wenckebach phenomenon) occurring after bilateral extraction of impacted mandibular third molars under general anesthesia in a 16-year-old Japanese girl. Under consultation with a cardiovascular physician, we carefully monitored the patient's vital signs postoperatively, including blood pressure, oxygen saturation, and electrocardiogram, using a bedside monitor. Her postoperative course was uneventful. A 12-lead electrocardiogram the following day revealed no abnormality. In this case, we hypothesize that extubation of the nasotracheal tube or oral/pharyngeal suction might have triggered a vagal reflex that caused type I second-degree atrioventricular block. Our experience indicates that standard cardiovascular monitoring should be used for patients undergoing dental treatment under general anesthesia, even for young, healthy patients, to prevent and detect cardiovascular emergencies.

Long-Term Follow-Up of SAPHO Syndrome for 15 Years Led to a Diagnosis of Temporomandibular Joint Pain and Trismus.

Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome is a systemic disease with symptoms of pustular skin disease and sterile osteoarticular lesions. This disease rarely involves the temporomandibular joint (TMJ). Although it is a disease with a good long-term prognosis, its treatment remains challenging. We describe a case with long-term follow-up of SAPHO syndrome for 15 years in which TMJ pain and trismus led to the diagnosis. A 30-year-old woman with TMJ pain and trismus was referred to our department. Her medical history included palmoplantar pustulosis. Sterile inflammation in the left TMJ and diffuse sclerosing osteomyelitis of the mandible were observed. Thus, she was diagnosed with SAPHO syndrome. The symptoms of severe TMJ pain, trismus, and left cheek swelling presented three times in the 15 years. Symptomatic treatment with nonsteroidal anti-inflammatory drugs, antibiotics, corticosteroids, and bisphosphonates was administered several times. There has been no relapse of symptoms over the past nine years. The patient must be continuously kept under observation to look for the relapse of symptoms.

Interleukin-1 (IL-1) Immunohistochemistry Assay in Oral Squamous Cell Carcinoma.

This section describes an immunohistochemistry method to analyze interleukin-1 (IL-1) in oral squamous cell carcinoma. The described protocol has been optimized for IL-1 detection in formalin-fixed, paraffin-embedded oral tissue sections by light microscopy. A few common pitfalls and problems associated with immunohistochemical staining are discussed.

CAD/CAM splint and surgical navigation allows accurate maxillary segment positioning in Le Fort I osteotomy.

PURPOSE: To evaluate the accuracy of the maxillary segment positioning method using a splint fabricated by computer-aided design/computer-aided manufacturing (CAD/CAM) and surgical navigation in patients who required two-jaw surgery. METHODS: Subjects were 35 patients requiring two-jaw surgery. A 3-dimensional (3D) skull model was prepared using cone-beam computed tomography (CBCT) data and dentition model scan data. Two-jaw surgery was simulated using this model, and a splint for maxillary positioning was fabricated by CAD/CAM. Using coordinate transformation software, the coordinate axis of surgical simulation data was merged with the navigation system, and data were imported to the navigation system. The maxillary segment was placed using the CAD/CAM splint, and consistency of the maxillary segment position with that planned by simulation was confirmed using the navigation system. CBCT taken at 4 weeks postoperatively and the prediction image fabricated using surgical simulation were superimposed. Predicted movement distances (PMD) at 6 arbitrary measurement points and actual movement distance (AMD) in surgery were measured. Differences of 3D measurements between the surgical simulation and postoperative results were evaluated. RESULTS: No significant differences were seen between PMD and AMD at most measurement points on the X and Y axes. Although significant differences between PMD and AMD were seen on the Z axis, no difference was evident between linear distance on the estimated image and postoperative CBCT image at most measurement points in 3D space. Mean error at measurement points between the PMD and AMD ranged from 0.57 mm to 0.78 mm on the X axis, 0.64 mm-1.03 mm on the Y axis, and 0.84 mm-0.90 mm in the Z axis. CONCLUSION: Position of the maxillary segment moved by the CAD/CAM splint in Le Fort I osteotomy was almost consistent with the position established by simulation using the navigation system, confirming clinical accuracy.

Comparison of codon usage and tRNAs in mitochondrial genomes of Candida species.

To gain insight into the nature of the mitochondrial genomes (mtDNA) of different Candida species, the synonymous codon usage bias of mitochondrial protein coding genes and the tRNAs in C. albicans, C. parapsilosis, C. stellata, C. glabrata and the closely related yeast Saccharomyces cerevisiae were analyzed. Common features of the mtDNA in Candida species are a strong A+T pressure on protein coding genes, and insufficient mitochondrial tRNA species are encoded to perform protein synthesis. The wobble site of the anticodon is always U for the NNR (NNA and NNG) codon families, which are dominated by A-ending codons, and always G for the NNY (NNC and NNU) codon families, which is dominated by U-ending codons, and always U for the NNN (NNA, NNU, NNC and NNG) codon families, which are dominated by A-ending codons and U-ending codons. Patterns of synonymous codon usage of Candida species can be classified into three groups: (1) optimal codon-anticodon usage, Glu, Lys, Leu (translated by anti-codon UAA), Gln, Arg (translated by anti-codon UCU) and Trp are containing NNR codons. NNA, whose corresponding tRNA is encoded in the mtDNA, is used preferentially. (2) Non-optimal codon-anticodon usage, Cys, Asp, Phe, His, Asn, Ser (translated by anti-codon GCU) and Tyr are containing NNY codons. The NNU codon, whose corresponding tRNA is not encoded in the mtDNA, is used preferentially. (3) Combined codon-anticodon usage, Ala, Gly, Leu (translated by anti-codon UAG), Pro, Ser (translated by anti-codon UGA), Thr and Val are containing NNN codons. NNA (tRNA encoded in the mtDNA) and NNU (tRNA not encoded in the mtDNA) are used preferentially. In conclusion, we propose that in Candida species, codons containing A or U at third position are used preferentially, regardless of whether corresponding tRNAs are encoded in the mtDNA. These results might be useful in understanding the common features of the mtDNA in Candida species and patterns of synonymous codon usage.

Orthodontic Treatment and Maxillary Anterior Segmental Distraction Osteogenesis of a Subject with Williams-Beuren Syndrome and Isolated Cleft Palate: A Long-Term Follow-Up from the Age of 5 to 24 Years.

Williams-Beuren syndrome (WBS) is a rare multisystem disorder caused by a hemizygous deletion of the elastin gene on chromosome 7q11.23. WBS patients have characteristic skeletal features and dental anomalies accompanied by mental retardation, a friendly outgoing personality, and mild to moderate intellectual disability or learning problems. In this case report, we present the combined orthodontic and surgical treatment of a WBS patient with an isolated cleft palate through a long-term follow-up from the age of 5 to 24 years. During the period of active treatment, comprehensive orthodontic treatment combined with maxillary anterior segmental distraction osteogenesis and prosthetic treatment using dental implants were effective in dramatically improving the patient's malocclusion. The patient's mental abilities and the cooperation shown by the patient and her family were crucial for the success of this complex and long-term treatment course.

Concomitant chemo-radio-immunotherapy has a lethal therapeutic effect on tongue carcinomas independent of the clinical stage and histological characteristics of the tumor.

We examined the effect of concomitant chemo-radio-immunotherapy on 80 patients with tongue carcinoma. Disappearance of the tumor without recurrence was observed in 21 patients (38.9%) in intravenous infusion chemotherapy group (A) and in 20 patients (76.9%) in intra-arterial infusion chemotherapy group (B) (P<0.005). A total of 41 patients (51.3%) were free from the tumor after the combined therapy. Along with the good therapeutic effect, oral function was preserved with minimal impairment of speech and mastication. Tumor stage, the mode of tumor cell invasion and tumor cell differentiation were not correlated with the therapeutic effect. In addition, the expression of p53, p21(Cip1/WAF1) and proliferating cell nuclear antigen did not differ between the patients with lethal and non-lethal effects. The 5-year-survival rate was 56.8% in Group A, 76.9% in Group B and 59.6% overall. Thus, combined chemo-radio-immunotherapy, especially intra-arterial infusion, may bring a universal therapeutic effect in tongue carcinoma regardless of the tumor stage and the expression of cell phase-regulating proteins.

Clinicopathological implications of vascular endothelial growth factor 165b expression in oral squamous cell carcinoma stroma.

Vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. VEGF165b was recently isolated as the anti-angiogenic VEGF splice variant. In the present study, we examined the association between VEGF165b expression and clinicopathological characteristics in order to determine how VEGF165b produced from oral squamous cell carcinoma (OSCC) affects the stromal cell biological activity. We examined the relationships between the expressions of both VEGF isoforms in normal human dermal fibroblasts (NHDFs) and OSCC cell lines (HSC2, 3, 4 and SAS). Our analyses indicated that both the mRNA and protein expression levels of VEGF165b in the HSC2 and SAS cells were higher than those in the NHDFs. VEGF165b did not promote cell growth or invasive capabilities, but it induced the cell adhesive capabilities to ECM. Although strong expression of the VEGF165 isoforms in tumor cells of OSCC tissues was observed, there was no significant difference in the VEGF165b expression level among the various degrees of malignancy. OSCC cells secrete VEGF165b into the stroma, and this factor may contribute to the process of anti-angiogenesis by inhibiting gelatinase-expressing cells and activating cell adhesive capabilities to ECM, such as that of fibroblasts surrounding tumor cells.

Protective effects of EGCG on salivary gland cells treated with gamma-radiation or cis-platinum(II)diammine dichloride.

Dysfunction of salivary glands is often associated with aging and cancer therapy. Green tea polyphenols were previously found to protect normal epithelial cells from reactive oxygen species, and to induce apoptosis in tumor cells. The current study investigated whether -(-) epigallocatechin-3-gallate (EGCG), the major green tea polyphenol, protects normal salivary gland cells from the effects of gamma-irradiation and the chemotherapy drug cis-platinum(II)diammine dichloride (CDDP). Human immortalized salivary acinar and ductal cells, and oral squamous cell carcinoma cells were irradiated with gamma-rays or treated with CDDP, with or without pretreatment with EGCG, followed by MTT and BrdU incorporation assays. The results demonstrated that EGCG protected the normal salivary gland cells from chemical or irradiation-induced damage. However, protection of oral cancer cells by EGCG was also observed if EGCG was at physiologically achievable salivary concentrations but not at higher concentrations, suggesting that the combination of green tea consumption with cancer therapy requires further evaluation.

Interleukin-1 (IL-1) immunohistochemistry assay in oral squamous cell carcinoma.

This chapter describes an immunohistochemistry method to analyze interleukin-1 (IL-1) in oral squamous cell carcinoma. The described protocol has been optimized for IL-1 detection in formalin-fixed paraffin-embedded oral tissue sections by light microscopy. A few common pitfalls and problems associated with immunohistochemical staining are discussed.

Intramuscular hemangioma with phleboliths of the tongue.

Intramuscular hemangioma (IMH) is relatively rare benign tumor of vascular origin. Phleboliths are calcified thrombi found in the presence of hemangioma. The main treatment of the hemangioma is a surgical extirpation based on location, accessibility, and cosmetic considerations. We herein report a rare case of IMH with phleboliths of the tongue with clinical, imaging, and histopathological findings.

Effectiveness of piezoelectric surgery in reducing surgical complications after bilateral sagittal split osteotomy.

Our aim was to investigate the effectiveness of piezoelectric surgery, where the osteotomy is made using ultrasonic vibration, in reducing surgical complications after bilateral sagittal split osteotomy (BSSO). Fifty-nine patients with skeletal mandibular prognathism who had mandibular setback with BSSO between January 2009 and April 2011 were included in the study. Piezosurgery was used in 29 cases, and the bone was split using a separator. In the remaining 30 cases, a Lindeman bur was used for the osteotomy and a chisel was used to split the bone. The amount of intraoperative bleeding and the Semmes Weinstein test scores were used as objective variables to evaluate the degree of neurosensory disturbance, and sex, age, use of piezosurgery, degree of setback, operating time, and method of fixation were used as explanatory variables. We used analysis of covariance (ANCOVA) to assess the significance of differences. Intraoperative bleeding was significantly less with age (p=0.003), and longer when operating time was prolonged (p=0.017), and was not influenced by the use of piezosurgery. The Semmes Weinstein test score significantly increased with age (p=0.01), and was significantly greater when piezoelectric surgery was used (p=0.008), and at 3 months, there were signs of more neurosensory disturbance in older patients and those who had had piezoelectric surgery. In this retrospective non-random study piezoelectric surgery reduced neither blood loss nor the incidence of neurosensory disturbance in BSSO.

The potential anticancer activity of extracts derived from the roots of Scutellaria baicalensis on human oral squamous cell carcinoma cells.

Various herb products derived from plants have potent biological effects including anticancer activity. In the present study, the antitumor activity of a herbal product derived from the Scutellaria baicalensis (S. baicalensis) was examined, using in vitro assays in a human oral squamous cell carcinoma (OSCC) cell line. Results showed that S. baicalensis root extract at the concentration of 100 µg/ml inhibited monolayer- and anchorage-independent growth in human OSCC cell lines, while not affecting the adhering abilities of cells. This suggested that it did not alter the expression of any of the adhesion receptors that mediate cell-extracellular matrix (ECM) interactions. The S. baicalensis root extract demonstrated potent cytostatic and apoptotic effects due to the downregulation of the cyclin-dependent kinase 4 expression and its partner cyclin D1, resulting in G1 arrest and poly (ADP-ribose) polymerase (PARP) cleavage. Additionally, the S. baicalensis root extract was found to have blocked vascular endothelial growth factor (VEGF)-induced migration and tube formation in human endothelial cells. Taken together, these results demonstrate that as a herbal product, the S. baicalensis root extract is a potential inhibitor of tumori- and angiogenesis and may be valuable in the development of pharmaceutical medications for the treatment of oral squamous cell carcinoma.

The root bark of Paeonia moutan is a potential anticancer agent in human oral squamous cell carcinoma cells.

Currently there is growing use of complementary and alternative anticancer medicines worldwide, and considerable interest in finding anticancer drugs among Chinese medicinal herbs. The aim of this study was to determine the antitumor activity of the root bark of Paeonia moutan (RBPM) in human squamous cell carcinoma (OSCC) cells. Cell lines derived from human oral squamous cell carcinoma (HSC2, 3, 4, SAS) were tested with different concentrations of RBPM (1-100 µg/ml) using a series of in vitro assay systems. RBPM at a concentration of 100 µg/ml inhibited monolayer and anchorage-independent growth, and interrupted coordinated migration. RBPM activated the phosphorylation of extracellular signal-regulated kinase (ERK) and serine/threonine kinase AKT in 30 min; then, at a later stage (after 6 hours) exhibited potent cytostatic, pro-apoptotic effects through the down-regulation of the expression of cyclin-dependent kinase 4 and its partner cyclin D1, and poly(ADP-ribose) polymerase cleavage. We found direct evidence that RBPM induces apoptotic cell death via DNA fragmentation. Taken together, the antitumor activity of RBPM was demonstrated through antiproliferative and apoptotic effects.

Evaluation of anticancer activities of benzo[c]phenanthridine alkaloid sanguinarine in oral squamous cell carcinoma cell line.

While the effects of benzo[c]phenanthridine alkaloids (QBA), known mainly as sanguinarine and chelerythrine, on the inhibition of some kinds of cancer cell proliferation have been established, the effect on oral squamous cell is not known. Here, the antitumor activity of sanguinarine was demonstrated using in vitro assay systems in SAS, a human oral squamous cell carcinoma (OSCC) cell line. The anti-proliferative and -invasive effects were confirmed with IC50 values in the concentration range of 0.75-1.0 µM by MTT assay and invasive assay, respectively. Sanguinarine was also able to suppress cell anchorage-independent growth, whereas it did not affect the cells' adhering capabilities. Finally, sanguinarine induced apoptotic cell death by activating caspase and altering the Bcl-2/Bax ratio. Taken together, these results indicate that sanguinarine is a potential inhibitor of tumorigenesis and suggest that it may be valuable in the development of new anticancer drugs for the treatment of OSCC.

Isolated metastasis from hepatocellular carcinoma to the mandibular condyle with no evidence of any other metastases: a case report.

Regional metastasis of hepatocellular carcinoma (HCC), the most common primary tumour of the liver to the mandible is rare. We present a case of a HCC metastasis to the mandibular condyle with no other extrahepatic metastases sites in a 59-year-old man. Incidental extrahepatic lesions in more uncommon sites should perhaps be investigated as potential areas for metastases, even if metastatic disease has not been found in the more common areas.

Polymorphic mutations of the Mn-SOD gene in intact human lymphocytes and oral squamous cell carcinoma cell lines.

Mutations of the superoxide dismutase (SOD) genes are associated with neoplastic and non-neoplastic diseases. However, the existence of polymorphic mutations of manganese SOD (Mn-SOD) has not been explored in squamous cell carcinoma (SCC) cells or in normal cells. In the present study, we examined mutations in the 5' flanking region of the Mn-SOD gene and Mn-SOD mRNA using 10 human oral SCC (OSC) cell lines and intact lymphocytes obtained from 10 healthy donors and one patient with OSC. The polymerase chain reaction products of DNA obtained from lymphocytes revealed insertions at many sites (-1833, -1575, -1093, -1056, -325, -318, and -310) in 10 of the 11 donors. Transitions and (or) transversions were also observed at -1638 and -216 in lymphocytes from six donors and one donor, respectively. In DNA obtained from OSC cells, insertions and transitions and (or) transversions were more frequent than those in DNA from lymphocytes. In addition, deletions at -1341 and -1288 were observed in all lines except for one line. In these mutations, the transcription factor binding sites were not involved except for the AP-2 binding site (-102) in three cell lines. In Mn-SOD mRNA, Val at -9 position was varied to Ala in lymphocytes from two donors and three OSC cell lines, respectively. In the remaining cell lines, Mn-SOD mRNA from lymphocytes and OSC cell lines revealed heterozygosity (Ala/Val) and homozygosity (Val/Nal), respectively. The Mn-SOD activities in lymphocytes were 3.8-5.8 x 10(-4) U/10(6) cells and the activities in OSC cell lines were 1.8-8.3 x 10(-4) U/10(6) cells. These Mn-SOD activities were not correlated with the mutations of DNA and mRNA. From these results, it is indicated that polymorphic mutations of Mn-SOD exist in human normal cells and that the deletions might be obtained in the course of malignant transformation of OSC although decrease in Mn-SOD activity is not involved in the transformation.

Tyrosine-nitration of caspase 3 and cytochrome c does not suppress apoptosis induction in squamous cell carcinoma cells.

The influence of tyrosine nitration of cytochrome c and caspase 3 on apoptosis induction was investigated in an established squamous carcinoma cell line, OSC-4. The intracellular NO and O2(-) levels were increased up to about 110-120% and 140-180% of the control levels, respectively, after the treatment of OSC-4 cells with 5-FU (100 microg/ml), PLM (10 microg/ml), CDDP (10 microg/ml), or gamma-rays (20 Gy). The treatment of OSC-4 cells with ONOO(-) (1 mM) and the above anticancer agents induced tyrosine nitration of 14, 32 kDa protein among others and nitration of tyrosine residues of cytochrome c and caspase 3 was identified by the Western blotting of immunoprecipitates obtained by antibodies to these proapoptotic proteins. When cytochrome c and procaspase 3 were treated with ONOO(-), tyrosine nitration was increased in a ONOO(-)-dose dependent manner. Tyrosine nitration of cleaved (17 kDa) caspase 3, however, was not induced by ONOO(-). Procaspase 3 in the cytosol of HeLa cells was activated by the addition of ONOO(-)-treated as well as ONOO(-)-untreated cytochrome c. In addition, cleavage of ICAD and PARP were not suppressed in OSC-4 cells by pretreatment with ONOO(-). Activity of cleaved caspase 3 was not suppressed at low concentrations or by treatment with ONOO(-) or NO donors, SIN-1 and SNP. Furthermore, apoptosis of OSC-4 cells by the anticancer agents was not suppressed by ONOO(-). In conclusion, these results suggest that nitration of tyrosine residues of cytochrome c and procaspase 3 is induced by chemoradiotherapy but their nitration does not suppress cancer cell apoptosis.