Shotgun metagenomic analysis of saliva microbiome suggests Mogibacterium as a factor associated with chronic bacterial osteomyelitis.

Osteomyelitis of the jaw is a severe inflammatory disorder that affects bones, and it is categorized into two main types: chronic bacterial and nonbacterial osteomyelitis. Although previous studies have investigated the association between these diseases and the oral microbiome, the specific taxa associated with each disease remain unknown. In this study, we conducted shotgun metagenome sequencing (≥10 Gb from ≥66,395,670 reads per sample) of bulk DNA extracted from saliva obtained from patients with chronic bacterial osteomyelitis (N = 5) and chronic nonbacterial osteomyelitis (N = 10). We then compared the taxonomic composition of the metagenome in terms of both taxonomic and sequence abundances with that of healthy controls (N = 5). Taxonomic profiling revealed a statistically significant increase in both the taxonomic and sequence abundance of Mogibacterium in cases of chronic bacterial osteomyelitis; however, such enrichment was not observed in chronic nonbacterial osteomyelitis. We also compared a previously reported core saliva microbiome (59 genera) with our data and found that out of the 74 genera detected in this study, 47 (including Mogibacterium) were not included in the previous meta-analysis. Additionally, we analyzed a core-genome tree of Mogibacterium from chronic bacterial osteomyelitis and healthy control samples along with a reference complete genome and found that Mogibacterium from both groups was indistinguishable at the core-genome and pan-genome levels. Although limited by the small sample size, our study provides novel evidence of a significant increase in Mogibacterium abundance in the chronic bacterial osteomyelitis group. Moreover, our study presents a comparative analysis of the taxonomic and sequence abundances of all genera detected using deep salivary shotgun metagenome data. The distinct enrichment of Mogibacterium suggests its potential as a marker to distinguish between patients with chronic nonbacterial osteomyelitis and chronic bacterial osteomyelitis, particularly at the early stages when differences are unclear.

Usefulness of quantitative bone SPECT/CT for evaluating medication-related osteonecrosis of the jaw treatment response.

OBJECTIVE: For assessment of therapeutic response in medication-related osteonecrosis of the jaw (MRONJ) cases, the clinical usefulness of quantitative bone single-photon computed tomography-computed tomography (SPECT/CT) results was investigated. MATERIALS AND METHODS: Sixteen patients (18 lesions) with a clinical diagnosis of MRONJ underwent bone SPECT/CT scanning before and during/after anti-inflammatory therapy given for 3 or more months. The GI-BONE software package was used to determine standard uptake values (SUVs), including maximum (SUVmax), peak (SUVpeak), and mean (SUVmean), and metabolic bone volume (MBV) and also total bone uptake (TBU). In both responders (downstage) and non-responders (upstage or no change), differences in quantitative values between the first and second SPECT/CT examinations were analyzed using a Wilcoxon test. RESULTS: Following therapy, significant reductions in SUVmax, SUVpeak, SUVmean, MBV and TBU values for 11 lesions were noted in the responders after therapy (p = 0.003, p = 0.006, p = 0.004, p = 0.003, and p = 0.002, respectively). On the other hand, those for the seven lesions in the non-responder group were not significantly different (p = 0.17, p = 0.16, p = 0.26, p = 0.96, and p = 0.12, respectively). Results for SUVmax change showed sensitivity and specificity values of 45.5% and 85.7%, respectively, for differentiating responders from non-responders, with - 37.3% the optimal cutoff value. Those for MBV change were 72.7 and 85.7%, respectively, with - 29.4% the optimal cutoff value. Those for TBU change were 81.8% and 85.7%, respectively, with - 36.3% the optimal cutoff value. CONCLUSION: The present findings showed that therapeutic response in MRONJ cases could be determined by use of quantitative SUV, MBV, and TBU values based on bone SPECT/CT findings.

Whole blood transcriptome profiling identifies gene expression subnetworks and a key gene characteristic of the rare type of osteomyelitis.

Chronic non-bacterial osteomyelitis (CNO) is a rare and severe inflammatory bone disorder that can occur in the jaw. It is often associated with systemic conditions including autoimmune deficiency. Medical management of patients and establishment of a correct diagnosis are difficult as the etiology of the disease remains unknown. Therefore, little is known about the disease characteristics at the gene expression level. Here, we explored aspects of CNO based on whole blood RNA sequencing (>6 Gb per sample) of 11 patients and 9 healthy controls in Japan and on a recently developed method that is applicable to small datasets, can estimate a directed gene network, and extract a subnetwork of genes underlying patient characteristics. We identified nine subnetworks, comprising 26 differentially regulated edges and 36 genes, with the gene encoding glycophorin C (GYPC) presenting the highest discrimination ability. The expression of the gene was mostly lower in patients with CNO than in the healthy controls, suggesting an abnormal status of red cells in patients with CNO. This study enhances our understanding of CNO at the transcriptome level and further provides a framework for whole blood RNA sequencing and analysis of data obtained for a better diagnosis of the disease.

Usefulness of quantitative bone SPECT/CT for medication-related osteonecrosis of the jaw in clinical settings.

OBJECTIVE: This study was conducted to investigate the clinical utility of quantitative bone single-photon computed tomography-computed tomography (SPECT/CT) for detection and classification for medication-related osteonecrosis of the jaw (MRONJ). MATERIALS AND METHODS: Fifty-nine patients (69 lesions) clinically diagnosed as MRONJ by four specialists of Japanese Society of Oral Surgery according to the AAOMS diagnostic criteria and who underwent bone SPECT/CT were enrolled. One reader determined standard uptake values (SUVs), including maximum (SUVmax), peak (SUVpeak), and mean (SUVmean), as well as metabolic bone volume (MBV), representing total volume above threshold, and total bone uptake (TBU), calculated as MBV × SUVmean, using the GI-BONE software package. One-way repeated-measures analysis of variance and subsequent post hoc analysis were employed to compare quantitative values between clinical stages. To check reproducibility of values, another reader calculated these quantitative values. RESULTS: Mean SUVmax values for stage 0 (n = 21), 1 (n = 13), 2 (n = 25), and 3 (n = 10) were 5.82 ± 3.20, 5.46 ± 3.79, 8.16 ± 3.93, and 10.57 ± 8.43, respectively, while values for MBV were 9.52 ± 6.33, 11.36 ± 7.32, 12.4 ± 8.21, and 17.84 ± 16.94, respectively, and for TBU were 40.60 ± 46.97, 53.70 ± 77.26, 62.37 ± 42.91, and 102.01 ± 74.52, respectively. There were significant differences for SUVmax, SUVpeak, and SUVmean between clinical stages (p = 0.024, p = 0.027, p = 0.039, respectively). Subsequent post hoc analysis showed that SUVmax and SUVpeak of stage 3 were significantly higher than those of stage 0 (p = 0.046, 0.045, respectively). MBV and TBU showed a tendency to increase with increased stage, though differences between stages were not significant (p = 0.15, p = 0.053, respectively). Little differences of mean SUVmax, SUVpeak, SUVmean, MBV, and TBU between two readers were observed (- 3.10%, - 0.26%, - 4.24%%, 0.69%, and - 3.42%, respectively). The intraclass correlation coefficients (ICCs) of SUVmax, SUVpeak, SUVmean, MBV, and TBU were 0.985, 0.990, 0.980, 0.994, and 0.994, respectively (almost perfect for all values). CONCLUSION: As objective and reliable indicators, SUVmax and SUVpeak derived from quantitative bone SPECT/CT results are useful for detection of early status disease, as well as staging in MRONJ patients.

Usefulness of Quantitative Bone SPECT/CT for Evaluating Treatment Response in a Patient with Mandibular Osteomyelitis.

We report here a case of mandibular osteomyelitis in a 63-year-old female in which quantitative values determined using bone SPECT/CT were useful to evaluate response to antibiotic therapy, hyperbaric oxygen therapy, and sequestomy. After finishing therapy, the chief complaints were well relieved, and posttreatment Tc-99m HMDP bone SPECT/CT examination showed decreased uptake. The maximum standardized uptake value (SUV), peak SUV, mean SUV, metabolic bone volume, and total bone uptake of the untreated lesion were 6.26, 5.16, 3.97, and 11.86 mL and 42.21, respectively, which were decreased to 4.65, 3.90, 2.77, and 9.67 mL and 26.80, respectively, following hyperbaric oxygen therapy and antibiotic administration, and were moreover decreased to 4.28, 3.67, 2.75, and 6.24 mL and 17.19, respectively, after sequestomy. In comparison with pretreatment situation, those parameters were decreased by -25.7, -24.4, -30.2, -18.5, and -36.5%, respectively, following hyperbaric oxygen therapy and antibiotic administration, and moreover by -31.6, -28.9, -30.7, -47.4, and -59.3, respectively, after sequestomy, likely reflecting treatment response. Quantitative bone SPECT/CT may be useful to evaluate bone inflammatory activity and treatment response in a patient with mandibular osteomyelitis.

Utility of intraoral stents in external beam radiotherapy for head and neck cancer.

AIM: This study aimed to assess the utility and stability of intraoral stent during intensity-modulated radiation therapy (IMRT). BACKGROUND: The benefits of intraoral stents in radiotherapy are unclear. MATERIALS AND METHODS: We analyzed 386 setup errors in 12 patients who received IMRT for head and neck cancers without intraoral stents (intraoral stent [-]) and 183 setup errors in 6 patients who received IMRT with intraoral stents (intraoral stent [+]). All patients were matched according to the immobilization method (masks and boards). Setup errors were measured as the distance from the initial setup based on the marking on the skin and mask to the corrected position based on bone matching on cone beam computed tomography. RESULTS: The mean interfractional setup errors in the right-left, craniocaudal, anterior-posterior (AP), and three-dimensional (3D) directions were -0.33, 0.08, -0.25, and 2.75 mm in the intraoral stent (-) group and -0.37, 0.24, -0.63, and 2.42 mm in the intraoral stent (+) group, respectively (P = 0.50, 0.65, 0.01, and 0.02, respectively). The systematic errors for the same directions were 0.89, 1.46, 1.15, and 0.88 mm in the intraoral stent (-) group and 0.62, 1.69, 0.68, and 0.56 mm in the intraoral stents (+) group, respectively. The random errors were 1.43, 1.43, 1.44, and 1.22 mm in the intraoral stent (-) group and 1.06, 1.11, 1.05, and 0.92 mm in the intraoral stents (+) group, respectively. CONCLUSION: Setup errors can be significantly reduced in the AP and 3D-directions by using intraoral stents.

Change in tongue pressure in patients with head and neck cancer after surgical resection.

Tongue pressure is reportedly associated with dysphagia. This study investigated relationships among characteristics of head and neck cancer, tongue pressure and dysphagia screening tests performed in patients with head and neck cancer during the acute phase after surgical resection. Fifty-seven patients (36 men, 21 women; age range 26-95 years) underwent surgical resection and dysphagia screening tests (Repetitive Saliva Swallowing Test, Water Swallowing Test, Modified Water Swallowing Test and Food Test) and pre- and postoperative measurement of tongue pressure at 5 time points (preoperatively, and 1-2 weeks and 1, 2, and 3 months postoperatively). Progression of cancer (stage), tracheotomy, surgical reconstruction, chemotherapy, radiotherapy and neck dissection were factors associated with postoperative tongue pressure. Data were analyzed by linear mixed-effect model, Spearman correlation coefficient and receiver operating characteristic (ROC) curve. Tongue pressure was significantly reduced 1-2 weeks after surgery, and recovered over time. Changes in tongue pressure were significantly associated with stage, radiotherapy and reconstruction. All screening tests showed a significant relationship with tongue pressure. Analysis of ROC and area under the effect curve suggested that a tongue pressure of 15 kPa can be used as a cut-off value to detect dysphagia after surgery for head and neck cancer. Our results suggest that tongue pressure evaluation might offer a safe, useful and objective tool to assess dysphagia immediately postoperatively in patients with head and neck cancer.

Osteonecrosis of the jaw in an AIDS patient: a case report.

We report a rare case of osteonecrosis of the jaw following necrotizing gingivitis in a Japanese AIDS patient. Intraoral examination showed exposed necrotic bone in the left mandible and spontaneous loss of teeth. This patient was successfully treated with highly active anti-retroviral therapy combined with minimally invasive surgical procedures to remove the osteonecrosis of the jaw.

Unicystic ameloblastoma metastasizing to multiple cervical lymph nodes.

Ameloblastoma is the most common odontogenic tumor, but the incidence of its metastasis is extremely low. We report a case of unicystic ameloblastoma metastasizing to the cervical lymph nodes. This patient pointed out a radiolucent cystic lesion with impacted wisdom tooth in the left mandibular region, and recieved enucleation of the cystic lesion and removal of the wisdom tooth. Histopathogical diagnosis was unicystic ameloblastoma. Three years later, this patient complained of a swelling in the left submandibular region. A CT scan showed a bilobed cystic mass measuring 30 mm in diameter compressing the submandibular gland, and we performed extirpation of the mass with the submandibular gland and associated lymph nodes. Histologically, the lesion was cystic and lymph follicles were seen in the cyst-like wall. The laminated epithelium of cyst wall was ameloblastomatous epithelium, and two lymph nodes associated with cystic lesion also included ameloblastomatous epithelium. This is the first report of metastasizing unicystic ameloblastoma.

Diffuse sclerosing osteomyelitis of the mandible successfully treated with pamidronate: a long-term follow-up report.

Diffuse sclerosing osteomyelitis of the mandible (DSOM) is an uncommon chronic inflammatory disease of bone that is refractory to conventional treatments, such as antibiotics, nonsteroidal anti-inflammatory drugs, and decortication. We report a case of chronic DSOM of 15 years' duration in a 61-year-old woman that was successfully treated with a single infusion of pamidronate. Persistent, intractable pain resolved 3 days after infusion. Intense accumulation on (99m)Tc scintigraphy decreased 2 months after infusion, and almost disappeared after 3 years. Panoramic radiography demonstrated a clear decrease in pathologic changes, close to that of normal bone architecture, which has not been reported in DSOM treated with bisphosphonates.

Up-regulation of neutrophil gelatinase-associated lipocalin in oral squamous cell carcinoma: relation to cell differentiation.

Neutrophil gelatinase-associated lipocalin (NGAL, also known as lipocalin2, LCN2) is a secreted glycoprotein with increased expression in solid tumors. The expression and functions of NGAL in oral cancer, however, remain unclear. We investigated the expression of NGAL in oral cancer tissues and oral cancer cell lines. By immunohistochemical examinations, NGAL expression was strongly up-regulated in well-differentiated OSCC tissues and moderately to weakly up-regulated in moderately to poorly differentiated OSCC tissues. In contrast, NGAL expression was weak or very weak in normal mucosa and leukoplakia. By western blot analysis, NGAL expression levels positively correlated with cell morphology patterns and loss of E-cadherin. In addition, the enzymatic activity of the NGAL/MMP-9 complex significantly correlated with the results obtained by zymographic analysis. In conclusion, NGAL expression is high in well-differentiated cancer, suggesting that NGAL may be a useful diagnostic marker of tumor-cell differentiation.

Novel PTCH1 mutations in Japanese Nevoid basal cell carcinoma syndrome patients: two familial and three sporadic cases including the first Japanese patient with medulloblastoma.

Nevoid basal cell carcinoma syndrome (NBCCS), also known as Gorlin syndrome, is inherited in an autosomal dominant mode, and is characterized by a combination of developmental abnormalities and predisposition to form a variety of tumors. The hedgehog receptor Patched1 (PTCH1) has been identified as the gene mutated in NBCCS. We analyzed PTCH1 in two familial and three sporadic Japanese NBCCS cases, and identified five germline mutations in PTCH1. Two cases have a nonsense mutation (c.3058C>T and c.2760C>A), one a splice site mutation (c.584+2T>G), one a 1 bp insertion (c.2712_2713insA) and one a 1 bp deletion (c.980Gdel). All mutations induce truncation of the PTCH1 protein or could induce nonsense-mediated mRNA decay. The 11-year-old male patient with splice-site mutation (c.584+2T>G) had medulloblastoma (MB) at the age of 1 year. This is the first NBCCS patient with molecularly defined MB in Japan.

[Severe advanced lower gingival carcinoma effectively reduced by chemoradiation with S-1--report of a case].

Severe advanced head and neck carcinoma which can not be removed via surgical procedure combined with a large lymph node metastasis has a poor prognosis. We administered concurrent chemoradiotherapy with S-1 for a lower gingival carcinoma. As a direct result, we discovered that the treatment greatly reduced the size of tumor, and we consider that this treatment prolonged the patient.s life. The treatment results suggest that the so-called dormancy state of the tumor was continued. In this case study, radiotherapy with S-1 showed a highly effective response from the viewpoint of QOL improvement.

Changes of histological and biological features by serial passages in a human adenoid cystic carcinoma line transplantable in nude mice.

The basic histologic patterns of adenoid cystic carcinoma (ACC) are classified into three types (tubular, cribriform and solid), but clinical significance of the histological type is unclear. We have successfully established a human tumor line derived from ACC that is serially transplantable in nude mice. This tumor showed an increased growth rate as the passage levels proceeded, and the histological type was changed from a cribriform pattern in the initial stage to a solid one. In this study, we investigated the relationship between histological type and biological characteristics by analyzing the serially transplantable ACC tumor model. As a result, the tumor growth rate at the 15th passage level was increased approximately 5-fold compared with that at the initial passage level. In the histological type, approximately 30% of the cribriform pattern in the initial level was changed to a solid one at the 15th passage level, and the PCNA labeling index was elevated 4-fold. Concomitant with this, expression of Ki-67, p53 and bcl-2 proteins was increased, and apoptotic cells were decreased as demonstrated by the TUNEL method. From these findings, it was suggested that cell proliferation and histological change of this ACC tumor are related to the inhibition of apoptosis. This tumor line would provide a useful model for investigating the biological behavior of ACC.

Establishment of a new cell line with neuronal differentiation derived from small cell neuroendocrine carcinoma of the maxillary sinus.

OBJECTIVE: It is well known that small cell neuroendocrine carcinoma (SNEC) arising at extrapulmonary sites has a poor prognosis and an interesting biological characterization. To understand biological characterization and elucidation of the origin of the histogenesis of SNEC, we report the establishment of a new SNEC cell line and characteristics of neuroendocrine properties including neuronal differentiation by treatment with dibutyryl cyclic AMP (db-cAMP). METHODS: We established a new cell line (SNEC-MI) derived from SNEC of the maxillary sinus by a modified spill-out method, and verified neuroendocrine properties including neuronal differentiation by immunocytochemical and immunoblotting methods. RESULTS: The established cell line showed spherical or spindle shape in monolayer culture and was positive for neuron-specific enolase (NSE), neuronal cell adhesion protein (N-CAM, CD56) and gastrin-releasing peptide. NSE was also demonstrated in the cultured medium and dense-core neuroendocrine granules were detected ultrastructurally in the cytoplasm. Treatment of cells with db-cAMP markedly induced the development and elongation of neuronal processes, which formed a netlike arrangement. Characterization of these elongated neuronal processes revealed them immunoreacting intensely with high molecular-weight neurofilament, and a time-dependent increase of microtubule-associated protein-2 in cell lysates. CONCLUSIONS: These findings indicated that this cell line possesses the capability to differentiate into neuronal cells, and supported the hypothesis that extrapulmonary SNEC might be derived from a pluripotent stem cell.