Flavinated SDHA underlies the change in intrinsic optical properties of oral cancers.

The molecular basis of reduced autofluorescence in oral squamous cell carcinoma (OSCC) cells relative to normal cells has been speculated to be due to lower levels of free flavin adenine dinucleotide (FAD). This speculation, along with differences in the intrinsic optical properties of extracellular collagen, lies at the foundation of the design of currently-used clinical optical detection devices. Here, we report that free FAD levels may not account for differences in autofluorescence of OSCC cells, but that the differences relate to FAD as a co-factor for flavination. Autofluorescence from a 70 kDa flavoprotein, succinate dehydrogenase A (SDHA), was found to be responsible for changes in optical properties within the FAD spectral region, with lower levels of flavinated SDHA in OSCC cells. Since flavinated SDHA is required for functional complexation with succinate dehydrogenase B (SDHB), decreased SDHB levels were observed in human OSCC tissue relative to normal tissues. Accordingly, the metabolism of OSCC cells was found to be significantly altered relative to normal cells, revealing vulnerabilities for both diagnosis and targeted therapy. Optimizing non-invasive tools based on optical and metabolic signatures of cancers will enable more precise and early diagnosis leading to improved outcomes in patients.

Flavinated SDHA Underlies the Change in Intrinsic Optical Properties of Oral Cancers.

The molecular basis of reduced autofluorescence in oral squamous cell carcinoma (OSCC) cells relative to normal cells has been speculated to be due to lower levels of free flavin adenine dinucleotide (FAD). This speculation, along with differences in the intrinsic optical properties of extracellular collagen, lie at the foundation of the design of currently-used clinical optical detection devices. Here, we report that free FAD levels may not account for differences in autofluorescence of OSCC cells, but that the differences relate to FAD as a co-factor for flavination. Autofluorescence from a 70 kDa flavoprotein, succinate dehydrogenase A (SDHA), was found to be responsible for changes in optical properties within the FAD spectral region with lower levels of flavinated SDHA in OSCC cells. Since flavinated SDHA is required for functional complexation with succinate dehydrogenase B (SDHB), decreased SDHB levels were observed in human OSCC tissue relative to normal tissues. Accordingly, the metabolism of OSCC cells was found to be significantly altered relative to normal cells, revealing vulnerabilities for both diagnosis and targeted therapy. Optimizing non-invasive tools based on optical and metabolic signatures of cancers will enable more precise and early diagnosis leading to improved outcomes in patients.

Three-dimensional Culture Using Atelocollagen Sponge and Self-assembling Peptide Hydrogel.

This study aimed to assess the combined application of two biomaterials, a selfassembling peptide hydrogel (SPH) and an atelocollagen sponge (ACS). The ACS was combined with SPH (PuraMatrixⓇ or PanaceaGelⓇ) and its osteogenic effects on mouse osteoblastic cell line MC3T3 then evaluated. Each type of SPH was successfully incorporated into the ACS. The MC3T3 cells showed uniform distribution within the scaffold. No necrotic cells were observed throughout the experimental procedures. When the SPH was combined with the ACS, the MC3T3 cells differentiated toward the osteo-lineage, expressing Alp, Runx2, Osx, Bsp, and Oc. PanaceaGelⓇ exhibited a stronger osteogenic effect on the cells than PuraMatrixⓇ.

Second Primary Metachronous Malignancies Occurring in Oral Cavity of Young Adult-A Case Report.

The incidence of oral cancer in Japan is increasing. Interestingly, the number of young patients with oral cancer is also rising. A 19-year-old man with no history of smoking or drinking alcohol presented with a 20×15-mm elastic, hard, protruding mass with a white surface on the right-hand margin of the tongue. A biopsy resulted in a diagnosis of a well-differentiated squamous cell carcinoma of the tongue, for which a partial resection was subsequently performed. During regular follow-up, the patient demonstrated no clinical or imaging abnormalities until 4 years and 9 months later, when erosion was observed at the right palatoglossal arch. A malignant tumor of the right palatoglossal arch was diagnosed based on cytology and imaging findings, and total resection of the lesion performed. Histopathological examination of the resected lesion revealed a moderately differentiated squamous cell carcinoma. Epithelial dysplasia on the right-hand margin of the tongue was diagnosed 4 years and 9 months after the second surgery and was subsequently resected. The patient's condition has been favorable for 7 years since the diagnosis of the second cancer, with no noted recurrence. This case emphasizes the importance of follow-up after initial treatment, as even young people, who are likely to have to endure long-lasting consequences from treatment, can develop metachronous cancer in the oral cavity.

Early Detection of Leukoplakic Oral Squamous Cell Carcinoma Using 4NQO-induced Rat Tongue Cancer Model: Study Utilizing Fluorescence Intensity and Histopathological Evaluation.

Early identification of leukoplakic oral squamous cell carcinoma (OSCC) is difficult. The purpose of this study was to determine whether it was possible to detect change from normal epithelium to leukoplakic OSCC using a fluorescence visualization (FV) device in a 4-nitroquinoline 1-oxide (4NQO) -induced rat tongue cancer model. If successful, this would facilitate early detection of OSCC. The rats (3 groups of 5) were administered 50 ppm 4NQO in their drinking water over a period of 10, 15, or 20 weeks. Five non-treated rats were used as a control group. Images of their tongues obtained by FV were analyzed for change in fluorescence intensity (FI) using image analysis software. Immunoreaction for anti-CK13, anti-CK17, and anti-E-cadherin antibodies was also histopathologically evaluated. Receiver operating characteristic (ROC) analysis was used to calculate the cut-off values, sensitivity, specificity, and area under the curve. The most marked change in FI was found between the control and 10-week groups, with an increase observed in its average value and range in the latter. These findings differed from those characteristic of leukoplakia. No significant difference was observed in the positive cell rate for immunoreaction for anti-CK13 or anti-CK17 antibodies between the control and 10-week groups. A significant decrease was observed in the positive pixel ratio of immunoreaction for anti-E-cadherin antibody in the 10-week group in comparison with in the control group (p <0.05). These results showed that disruption of intercellular adhesion could be observed at 10 weeks. In the ROC analysis, the FI cut-off value in the 10-week and control groups was 51.9, sensitivity 95.5%, and specificity 96.9%. This indicated that normal epithelium could be accurately distinguished from low-grade dysplasia with high probability. These results demonstrate that analysis of change in FI as measured by FV could facilitate early detection of leukoplakic OSCC.

RIOK2 Contributes to Cell Growth and Protein Synthesis in Human Oral Squamous Cell Carcinoma.

Ribosomes are responsible for the protein synthesis that maintains cellular homeostasis and is required for the rapid cellular division of cancer cells. However, the role of ribosome biogenesis mediators in the malignant behavior of tongue squamous cell carcinoma (TSCC) is unknown. In this study, we found that the expression of RIOK2, a key enzyme involved in the maturation steps of the pre-40S ribosomal complex, was significantly associated with poorer overall survival in patients with TSCC. Further, multivariate analysis revealed that RIOK2 is an independent prognostic factor (hazard ratio, 3.53; 95% confidence interval, 1.19-10.91). Inhibition of RIOK2 expression by siRNA decreased cell growth and S6 ribosomal protein expression in oral squamous cell carcinoma cell lines. RIOK2 knockdown also led to a significant decrease in the protein synthesis in cancer cells. RIOK2 has potential application as a novel therapeutic target for TSCC treatment.

Three-Dimensional Evaluation of Pharyngeal Morphology in Bimaxillary Surgery with and without Horseshoe Osteotomy in Skeletal Class III Cases.

Repositioning of the jaw in orthognathic treatment generates changes in the soft tissues of the maxillofacial region, with consequent changes in the airway. The purpose of this study was to determine how type of orthognathic surgical procedure affected the 3-dimensional morphology of the upper airway. Forty patients were divided into the following 2 groups according to the type of surgical procedure used: a horseshoe osteotomy (HS) group (20 patients, comprising 11 men and 9 women; average age 24.3±4.5 years) who underwent bimaxillary surgery; and a LeFort I osteotomy (LF) group (20 patients, comprising 8 men and 12 women; average age 22.5±4.6 years) who also underwent bimaxillary surgery. Cephalometric measurements were taken and 3-dimensional pharyngeal morphology evaluated in each group. The amounts of maxilla rotation, posterior maxilla impaction, and mandibular setback all revealed a significantly larger value in the HS group. Evaluation of pharyngeal volume revealed a significant decrease in the upper pharyngeal segment in the LF group. A significant decrease in the lower pharyngeal segment was observed in both groups. Differences were noted in postoperative pharyngeal morphology between the two groups. The results of this study suggest that HS has less effect on the upper pharyngeal segment, regardless of the amount of posterior maxilla impaction.

Three-Dimensional Analysis of Soft and Hard Tissue Changes following Orthognathic Surgery.

Change in soft tissue in relation to that in hard tissue following orthognathic surgery was evaluated. Twenty-five patients were enrolled in the study. The diagnosis in all was jaw deformity (maxillary retrusion and mandibular protrusion) and all underwent a Le Fort I osteotomy and bilateral sagittal splitting ramus osteotomy. Three-dimensional (3D) computer-aided design (CAD) models (polygon models) of the hard and soft tissue of the maxilla and mandible were constructed and superimposed. Reference points were established on the pre- and postoperative hard and soft tissues. Specific elements of each reference point were divided into X, Y, and Z components, respectively, and the distances in each direction and 3D distance (normal distance) measured. The Wilcoxon signed-rank test was used to determine differences in the mean values for the distance moved of each element as the error between pre- and postoperatively. The results revealed statistically significant differences in the Y-direction in the maxilla and the X- and Z-directions in the mandible. A significant difference was also observed in the 3D distances of the maxilla and mandible. Little evidence was found of linearity between the amount of hard and soft tissue movement in the X- and Z-directions in the maxilla. This means that 3D movement in the maxilla was masked more by changes in the morphology of the soft tissue than in the mandible, making it less evident. The results of this study suggest that the 3D analysis method used enables changes in hard and soft tissues to be understood qualitatively, and that it can be used in diagnosis and treatment in orthognathic surgery. It may also be useful in simulation of morphological change in soft tissue.

Possible Therapeutic Strategy Involving the Purine Synthesis Pathway Regulated by ITK in Tongue Squamous Cell Carcinoma.

The epidermal growth factor receptor is the only available tyrosine kinase molecular target for treating oral cancer. To improve the prognosis of tongue squamous cell carcinoma (TSCC) patients, a novel molecular target for tyrosine kinases is thus needed. We examined the expression of interleukin-2-inducible T-cell kinase (ITK) using immunohistochemistry, and the biological function of ITK was investigated using biochemical, phosphoproteomic, and metabolomic analyses. We found that ITK is overexpressed in TSCC patients with poor outcomes. The proliferation of oral cancer cell lines expressing ITK via transfection exhibited significant increases in three-dimensional culture assays and murine inoculation models with athymic male nude mice as compared with mock control cells. Suppressing the kinase activity using chemical inhibitors significantly reduced the increase in cell growth induced by ITK expression. Phosphoproteomic analyses revealed that ITK expression triggered phosphorylation of a novel tyrosine residue in trifunctional purine biosynthetic protein adenosine-3, an enzyme in the purine biosynthesis pathway. A significant increase in de novo biosynthesis of purines was observed in cells expressing ITK, which was abolished by the ITK inhibitor. ITK thus represents a potentially useful target for treating TSCC through modulation of purine biosynthesis.

Over-expression of PDE5 in Oral Squamous Cell Carcinoma - Effect of Sildenafil Citrate.

BACKGROUND/AIM: Phosphodiesterase 5 (PDE5) holds clinical relevance in several pathological states, including lung, breast, and prostate cancer. In this study, we examined PDE5 expression in oral squamous cell carcinoma (OSCC)-derived cell lines and tissues, and the anti-tumour effect of PDE5 inhibitor, sildenafil citrate (SC). MATERIALS AND METHODS: Cell proliferation, cell invasion, and gap closure assays were performed in six OSCC-derived cell lines upon treatment with varying concentrations of SC. PDE5 expression was determined in primary OSCC tissues by western blotting and immunohistochemistry. RESULTS: Elevated PDE5 expression was observed in all cell lines. A concentration-dependent decrease in cell viability, invasion rate, and migration was observed after SC treatment. A significant correlation (p=0.05) was observed between elevated PDE5 expression and lymphatic infiltration in OSCC tissues. CONCLUSION: PDE5 plays an important role in carcinogenesis of OSCC, and the specific inhibition of PDE5 may be an effective chemotherapeutic strategy.

Persistent bone resorption lacunae on necrotic bone distinguish bisphosphonate-related osteonecrosis of jaw from denosumab-related osteonecrosis.

BACKGROUND: Bisphosphonate and denosumab are widely used for the treatment of osteoporosis and bone metastasis of cancer to prevent excessive bone resorption. Osteonecrosis of the jaw is a serious adverse effect of bisphosphonate or denosumab referred to as bisphosphonate-related osteonecrosis of the jaw (BRONJ) or denosumab-related osteonecrosis of the jaw (DRONJ), respectively. Since bisphosphonate and denosumab inhibit bone resorption by different mechanism, we evaluated whether these drug types result in different histopathological characteristics related to bone resorption. MATERIALS AND METHODS: We histopathologically investigated 10 cases of BRONJ, DRONJ, and suppurative osteomyelitis. Paraffin sections prepared from decalcified dissected jaw bones were used for histopathological observation, second harmonic generation imaging, and bone histomorphometry. The samples were also observed by a scanning electron microscope. RESULTS: Numerous bone resorption lacunae were observed on the necrotic bone surface in almost all cases of BRONJ; however, such resorption lacunae were limited in DRONJ and suppurative osteomyelitis. Prominent bone resorption lacunae were also confirmed by second harmonic generation imaging and scanning electron microscopy in BRONJ, but not in DRONJ or suppurative osteomyelitis. As determined by bone histomorphometry, the number of bone resorption lacunae and the length of the erosion surface of resorption lacunae were significantly higher in BRONJ group than in the DRONJ and suppurative osteomyelitis groups. These parameters were correlated between the necrotic bones and the vital bones in BRONJ. CONCLUSIONS: Persistent bone resorption lacunae on the necrotic bone surface are unique to BRONJ, providing a basis for distinguishing BRONJ from DRONJ and OM in histopathological diagnosis.

Verification of the junctional dose for irradiation of the chest wall and supraclavicular regions under the circumstances of advanced technologies.

A single-isocenter half-beam technique is commonly used when irradiating the chest wall and supraclavicular regions in patients with high-risk breast cancer. However, several studies have reported that underdosage can occur at the junction of the chest wall and supraclavicular regions due to a "tongue-and-groove" effect. This study verified the efficacy of an open leaf technique (OL-tech) that involves placing a multileaf collimator 5 mm outside from the beam central axis to remove the effect of the multileaf collimator in a single-isocenter half-beam technique. We compared the junction doses of the OL-tech with those of a conventional technique (C-tech) in square and clinical plans, using 4 and 10 MV x-rays in the Clinac iX and 6 and 10 MV x-rays in the Trilogy accelerators (Varian Medical Systems, Palo Alto, CA). EBT3 radiochromic films were used for measurements. Measurements were performed at a depth of 3 cm when verifying field matching. The EBT3 films in the square plan indicated junction doses for the C-tech of 78.3% with the Clinac iX accelerator and 73.6% with the Trilogy accelerator. By contrast, the corresponding doses for the OL-tech were 107.2% and 99.8%, respectively. In the clinical plan, the junction doses for the C-tech were 76.5% with the Clinac iX accelerator and 72.6% with the Trilogy accelerator; the corresponding doses for the OL-tech were 108.3% and 101.7%, respectively. As with the square plan, variations in the junction dose were much smaller using the OL-tech than using the C-tech. Our results suggest that the OL-tech can be useful for improving dose homogeneity at the junction of the chest wall and supraclavicular regions.

Heat shock protein 90 as a molecular target for therapy in oral squamous cell carcinoma: Inhibitory effects of 17­DMAG and ganetespib on tumor cells.

Heat shock protein 90 (HSP90) expression is upregulated in numerous types of cancer. However, its role as a candidate for molecular targeted therapy in oral squamous cell carcinoma (OSCC) cells is poorly understood. In the present study, a common upstream search was performed using molecular network analysis software for proteins with expression abnormalities that were found in a proteomic analysis of six OSCC cell lines. HSP90 was identified as a target protein. In clinical samples, high frequencies of HSP90­high expression were detected via immunohistochemistry (26/58; 45%). Furthermore, the HSP90 expression status was associated with cervical lymph node metastasis (P=0.015). Furthermore, the potential of HSP90 as a candidate for molecular targeted therapy in OSCC cells was investigated using the HSP90 inhibitors 17­dimethylaminoethylamino­17­demethoxygeldanamycin (17­DMAG) and ganetespib. KON cells, which strongly express HSP90, were treated with the HSP90 inhibitors. The numbers of living cells in the 17­DMAG and ganetespib­treated groups were lower than those in the non­treated group. The cells treated with the inhibitors demonstrated reduced cell viability and migration, and this was associated with markedly decreased levels of the HSP90 target proteins EGFR, phospho­EGFR, phospho­MEK and phospho­MAPK in the treated groups compared with the non­treated group. To the best of our knowledge, this was the first study to investigate the effects of 17­DMAG and ganetespib on OSCC cells. The present results indicated the potential of HSP90 as a useful candidate for molecular targeted therapy in OSCC. However, additional studies with larger sample sizes are required to confirm these findings.

Sialo-cutaneous fistula with ectopic submandibular gland sialolith, revealing a hidden ipsilateral enlarged and elongated styloid process: a consideration based on CT findings.

Sialolithiasis is a common salivary pathology, and an uncommon complication of sialadenitis and sialolithiasis is the formation of fistulous tracts to other compartments. Submandibular gland sialo-oral fistulae are not particularly remarkable, given the location of the gland and Wharton's duct, but submandibular sialolith-associated fistulae to other cervico-facial compartments (transcervical sialo-cutaneous and sialo-pharyngeal fistulae) are much less common. We report herein an unusual case of a 49-year-old obese man with sialo-cutaneous fistula containing a large, ectopic sialolith in subcutaneous tissue that was expected to undergo spontaneous elimination, but revealed hidden Eagle syndrome featuring an ipsilateral enlarged, elongated styloid process. Furthermore, we offer a thorough review of the literature regarding sialo-fistulae and highlight the relationship between an abnormal styloid process and submandibular sialadenitis with sialolithiasis and new tract formation based on computed tomography.

Countermeasure and opportunistic screening systems for oral cancer.

OBJECTIVES: Tokyo Dental College started oral cancer screening in cooperation with a local dental association in 1992. Reveal the usefulness of Countermeasure and Opportunistic Screening Systems for Oral Cancer. The actual results of countermeasure and opportunistic oral cancer screening systems are reported. MATERIALS AND METHODS: Countermeasure screening for the public was performed in each region, and opportunistic screening was performed in a general dental clinic of a cooperating physician. RESULTS: In countermeasure screening, 19,721 persons were checked from 1992 to 2018; the gender ratio was 1:3. The close examination rate was 4.45%. The detection rates of oral cancer and oral potentially malignant disorders were 0.13% and 1.85%, respectively. In opportunistic screening, 29,912 persons were checked from 2006 to 2018; the gender ratio was 2:3. The close examination rate was 2.33%. The detection rates of oral cancer and oral potentially malignant disorders were 0.08% and 2.15%, respectively. The close examination rate was significantly lower in opportunistic screening than in countermeasure screening. The oral cancer detection rates and the positive predictive value for cancer were equivalent. In addition, the detection rate of oral potentially malignant disorders was significantly higher in opportunistic screening than in countermeasure screening. CONCLUSION: Oral cancer detection rates were equivalent between countermeasure and opportunistic screenings, and opportunistic screening were more effective on number of participants and the close examination rate, and the detection rate of oral potentially malignant disorders.

A Case of a Stafne Bone Defect Associated with Sublingual Glands in the Lingual Side of the Mandible.

A Stafne bone defect from the mandibular anterior to the premolar region is an extremely rare case. A case of a Stafne bone defect extending from the mandibular anterior to the premolar region was presented. Computed tomography (CT) and magnetic resonance imaging (MRI) suggested that salivary gland tissue connected to the sublingual glands was involved in the formation of the cavity. The patient was a 68-year-old man who was examined at our hospital's emergency outpatient department after a traffic accident. He was referred to our department for the treatment of contusions of the lips and oral cavity. A bone defect in the lingual side of the mandible from the right anterior to the right premolar region was incidentally detected on CT. CT showed a rounded cavity in the lingual side of the mandible that had a lingual opening, was monocystic, and had a cortical margin. The margin of the cavity was relatively dull and regular. MRI showed that the tissue filling the cavity in the lingual side of the mandible had similar signal intensity as the sublingual glands and was contiguous with the normal sublingual glands. Based on these findings, the bone defect was diagnosed as a Stafne bone defect filled with salivary gland tissue connected to the sublingual gland tissue.

A novel GNAS-mutated human induced pluripotent stem cell model for understanding GNAS-mutated tumors.

A missense mutation of the guanine nucleotide binding protein alpha stimulating activity polypeptide 1 (GNAS) gene, typically Arg201Cys or Arg201His (R201H/R201C), leads to constitutive activation of the Gsα-cyclic AMP (cAMP) signaling pathway that causes several diseases. However, no germline mutations of GNAS have been identified to date, likely due to their lethality, and no robust human cell models have been generated. Therefore, the aim of this study was to generate GNAS-mutated disease-specific induced pluripotent stem cells as a model for these diseases. We then analyzed the functionality of this induced pluripotent stem cell model and differentiated epithelial cells. We generated disease-specific induced pluripotent stem cells by introducing a mutation in GNAS with the clustered regularly interspaced short palindromic repeats (CRISPR) nickase method, which has lower off-target effects than the conventional CRISPR/Cas9 method. We designed the target vector to contain the R201H mutation in GNAS, which was transfected into human control induced pluripotent stem cells (Nips-B2) by electroporation. We confirmed the establishment of GNASR201H-mutated (GNASR201H/+) induced pluripotent stem cells that exhibited a pluripotent stem cell phenotype. We analyzed the effect of the mutation on cAMP production, and further generated teratomas for immunohistochemical analysis of the luminal epithelial structure. GNAS-mutated induced pluripotent stem cells showed significantly higher levels of intracellular cAMP, which remained elevated state for a long time upon hormonal stimulation with parathyroid hormone or adrenocorticotropic hormone. Immunohistochemical analysis revealed that several mucins, including MUC1, 2, and MUC5AC, are expressed in cytokeratin 18 (CK18)-positive epithelial cells. However, we found few CK18-positive cells in mutated induced pluripotent stem cell-derived teratoma tissues, and reduced MUCINs expression in mutated epithelial cells. There was no difference in CDX2 expression; however, mutated epithelial cells were positive for CEA and CA19-9 expression. GNASR201H-mutated induced pluripotent stem cells and GNASR201H-mutated epithelial cells have distinct phenotypic and differentiation characteristics. We successfully established GNASR201H-mutated human induced pluripotent stem cells with increased cAMP production. Considering the differentiation potential of induced pluripotent stem cells, these cells will be useful as a model for elucidating the pathological mechanisms of GNAS-mutated diseases.

Non-Invasive Early Detection of Oral Cancers Using Fluorescence Visualization with Optical Instruments.

BACKGROUND: Oral cancer screening is important for early detection and early treatment, which help improve survival rates. Biopsy is the gold standard for a definitive diagnosis but is invasive and painful, while fluorescence visualization is non-invasive, convenient, and real-time, and examinations can be repeated using optical instruments. The purpose of this study was to clarify the usefulness of fluorescence visualization in oral cancer screening. METHODS: A total of 502 patients, who were examined using fluorescence visualization with optical instruments in our hospitals between 2014 and 2019, were enrolled in this study. The final diagnosis was performed by pathological examination. Fluorescence visualization was analyzed using subjective and objective evaluations. RESULTS: Subjective evaluations for detecting oral cancer offered 96.8% sensitivity and 48.4% specificity. Regarding the objective evaluations, sensitivity and specificity were 43.7% and 84.6% for mean green value, 55.2% and 67.0% for median green value, 82.0% and 44.2% for coefficient of variation of value, 59.6% and 45.3% for skewness, and 85.1% and 75.8% for value ratio. For the sub-analysis of oral cancer, all factors on objective and subjective evaluation showed no significant difference. CONCLUSIONS: Fluorescence visualization with subjective and objective evaluation is useful for oral cancer screening.

O-Glycan-Altered Extracellular Vesicles: A Specific Serum Marker Elevated in Pancreatic Cancer.

Pancreatic cancer (PC) is among the most lethal malignancies due to an often delayed and difficult initial diagnosis. Therefore, the development of a novel, early stage, diagnostic PC marker in liquid biopsies is of great significance. In this study, we analyzed the differential glycomic profiling of extracellular vesicles (EVs) derived from serum (two cohorts including 117 PC patients and 98 normal controls) using lectin microarray. The glyco-candidates of PC-specific EVs were quantified using a high-sensitive exosome-counting system, ExoCounter. An absolute quantification system for altered glycan-containing EVs elevated in PC serum was established. EVs recognized by O-glycan-binding lectins ABA or ACA were identified as candidate markers by lectin microarray. Quantitative analyses using ExoCounter revealed that the ABA- or ACA-positive EVs were significantly increased in the culture of PC cell lines or in the serum of PC patients including carbohydrate antigen 19-9 negative patients with high area under curve values. The elevated numbers of EVs in PC serum returned to normal levels after pancreatectomy. Histological examination confirmed that the tumors stained with ABA/ACA. These specific EVs with O-glycans recognized by ABA/ACA are elevated in PC sera and can act as potential biomarkers in a liquid biopsy for PC patients screening.

Utilization of desktop 3D printer-fabricated "Cost-Effective" 3D models in orthognathic surgery.

BACKGROUND: In daily practice, three-dimensional patient-specific jawbone models (3D models) are a useful tool in surgical planning and simulation, resident training, patient education, and communication between the physicians in charge. The progressive improvements of the hardware and software have made it easy to obtain 3D models. Recently, in the field of oral and maxillofacial surgery, there are many reports on the benefits of 3D models. We introduced a desktop 3D printer in our department, and after a prolonged struggle, we successfully constructed an environment for the "in-house" fabrication of the previously outsourced 3D models that were initially outsourced. Through various efforts, it is now possible to supply inexpensive 3D models stably, and thus ensure safety and precision in surgeries. We report the cases in which inexpensive 3D models were used for orthodontic surgical simulation and discuss the surgical outcomes. REVIEW: We explained the specific CT scanning considerations for 3D printing, 3D printing failures, and how to deal with them. We also used 3D models fabricated in our system to determine the contribution to the surgery. Based on the surgical outcomes of the two operators, we compared the operating time and the amount of bleeding for 25 patients who underwent surgery using a 3D model in preoperative simulations and 20 patients without using a 3D model. There was a statistically significant difference in the operating time between the two groups. CONCLUSIONS: In this article, we present, with surgical examples, our in-house practice of 3D simulation at low costs, the reality of 3D model fabrication, problems to be resolved, and some future prospects.