Enhanced production of IL-2 from anti-CD3 antibody-stimulated mouse spleen cells by caffeic acid phenethyl ester, a major component of Chinese propolis.

OBJECTIVES: We evaluated the immune-modulatory effects of Chinese propolis (CP) and its major constituent, caffeic acid phenethyl ester (CAPE), on the cytokine production of anti-CD3 antibody-stimulated mouse spleen cells. METHODS: Mouse spleen cells stimulated by anti-CD3 monoclonal antibody were co-cultured with CP, CAPE, and HC030031, a specific antagonist of the TRPA1 Ca2+-permeable cation channel. Cytokine production was assayed by enzyme-linked immunosorbent assay. Interleukin (IL)-2 mRNA expression was examined by reverse transcription-quantitative polymerase chain reaction. RESULTS: In stimulated spleen cells treated with 1/16,000 CP diluent, IL-2 production was markedly enhanced, while IL-4 and IL-10 productions were not significantly affected. In contrast, interferon (IFN)-γ, IL-6, and IL-17 productions were markedly reduced. These effects of CP were reproduced by the CAPE treatment. A time-course observation demonstrated that, compared to control cells, IL-2 mRNA expression and production were significantly enhanced in the spleen cells stimulated by CAPE; however, IL-2 production was markedly delayed compared to that in the untreated control cells. The enhancement of IL-2 production by CAPE was scarcely alleviated by the addition of HC030031. These effects of CAPE upon IL-2 mRNA production were abolished in spleen cells without anti-CD3 antibody stimulation. CONCLUSIONS: CAPE is an important regulator of CP for cytokine regulation in anti-CD3 antibody-stimulated spleen cells. The agent specifically reduced IFN-γ, IL-6, and IL-17 and slightly enhanced Th2 cytokine production while significantly enhancing IL-2 production at the transcriptional level.

Synthesis of ß-tricalcium phosphate by modifying the heating process of a dental casting mold.

This study investigated a novel method for artificial synthesis of ß-tricalcium phosphate (ß-TCP). The binder of the phosphate-bonded investment was replaced with calcium oxide instead of magnesium oxide and sintered in an electric furnace. The water/powder mixing ratio for hardening was determined using preliminary experiments. Thermal analysis was performed to check the thermal behavior of the sample tested. In addition, the fired sample was analyzed using an X-ray diffraction apparatus to identify the compounds after sintering. The hardened sample exhibited multiple compounds, including unreacted components, post which, new compounds were generated by heating. Peaks of calcium pyrophosphate and ß-TCP were confirmed at 800ºC and 1,300ºC, respectively. ß-TCP could be easily synthesized within the limited study by sintering at 1,300ºC both monoammonium phosphate and calcium oxide. Experimental results suggest that ß-TCP can be easily synthesized by simulating the conventional dental casting process.

Augmented secretion of IL-1α from mouse oral squamous cell carcinoma (OSCC) vcells caused by serum deprivation and hypoxia promotes immune suppressive activity of mesenchymal stromal cells.

OBJECTIVES: We have previously reported that mouse oral squamous carcinoma (OSCC), Sq-1979-1, produces interleukin (IL)-1α, which specifically enhances the immunosuppressive activity of co-cultured mesenchymal stromal 10T1/2 cells. This study assessed the conditions promoting the production of IL-1α in Sq-1979-1 cells, which could further enhance the immunosuppressive function of 10T1/2 cells, and evaluated its expression in OSCC tissues. METHODS: The expression of IL-1α was examined by RT-PCR, western blotting, and enzyme-linked immune sorbent assay (ELISA). The interferon (IFN)- γ-producing capability of anti-CD3 antibody-stimulated mouse spleen cells co-cultured with 10T1/2 cells and conditioned medium (CM) from Sq-1979-1 cells was examined by ELISA. The function of IL-1α was examined using an anti-IL1α antibody. Immunohistochemical analysis of the OSCC tissues was performed. RESULTS: The production of IL-1α from Sq-1979-1 cells was synergistically enhanced in lower serum (0.5% or 1.0% FBS) at the transcriptional level, and under hypoxia (1.0% oxygen) at the release level compared to that in the control medium supplemented with 10% FBS under normoxia. The IFN-γ-producing capability of stimulated spleen cells co-cultured with 10T1/2 cells was significantly reduced in the CMs prepared with the lower serum or under hypoxia. These functions of CMs were completely abolished by the anti-IL-1α antibody. The expression of IL-1α in OSCC tissues was prominent in the midst of a carcinomatous cellular lesion or a nearby necrotic lesion, where a supply deficiency could occur. CONCLUSION: s: IL-1α production by Sq-1979-1 cells was synergistically augmented under low serum and hypoxic conditions, which could promote the immunosuppressive activity of mesenchymal cells.

Usefulness of dual-energy computed tomography for oral cancer image.

OBJECTIVES: We aimed to compare dual-energy computed tomography (DECT) virtual monochromatic imaging (VMI) and iodine density imaging (IDI) of oral cancers in terms of visual scoring and tumour volume estimation. MATERIALS AND METHODS: Nine patients diagnosed with oral cancer who underwent DECT VMI and IDI were enrolled. One radiation oncologist, one head and neck surgeon and nine oral surgeons evaluated image clarity and quality in each patient in terms of metal artefacts due to dental prosthesis, internal tumour structure, tumour-organ boundary and total quality of images for diagnosis. Tumour volume was estimated using VMI, IDI and magnetic resonance imaging (MRI). RESULTS: The mean score for image artefact was significantly higher for IDI than for VMI in three observers, the mean score for internal structure was significantly higher for IDI than for VMI in five, the mean score for tumour-organ boundary was significantly higher for IDI than for VMI in two and the mean score for total quality of images for diagnosis was significantly higher for IDI than for VMI in five. Standard deviation of estimated tumour volume was not significantly different between VMI and IDI, but that of MRI was significantly lowest in three images. CONCLUSIONS: In DECT for oral cancer, IDI has a visual image superior to VMI; thus, we recommend the use of IDI. TRIAL REGISTRATION: Clinical trial number: UMIN000038994.

Relationship between Oral Assessment Guide score and hypoalbuminemia in newly hospitalized patients.

This cross-sectional study investigated the relationship between Oral Assessment Guide (OAG) scores and malnutrition in newly hospitalized patients. A total of 880 hospitalized adults were enrolled. Hypoalbuminemia was defined as serum albumin less than 3.5 g/dL. Patients with hypoalbuminemia were older (P < 0.001), had a higher prevalence of respiratory diseases (P < 0.01), a higher prevalence of digestive diseases (P < 0.01), a lower prevalence of oral feeding (P < 0.001), a lower body mass index (P < 0.001), and higher OAG scores (P < 0.001) than those without hypoalbuminemia. Multivariate logistic regression analyses showed that the prevalence of hypoalbuminemia was significantly related to age (odds ratio [OR] = 1.05, P < 0.001), absence of oral feeding (OR = 2.72, P < 0.001), presence of respiratory diseases (OR = 2.53, P < 0.01), presence of digestive diseases (OR = 1.64, P < 0.01), and OAG scores (OR = 1.14, P < 0.01). Regarding OAG scores, the OR of hypoalbuminemia was greater in patients with disorders (scores 2 or 3) of swallowing (vs. score 1, OR = 1.83, P < 0.05) and saliva (vs. score 1, OR = 1.51, P < 0.05). There appears to be a positive association between OAG scores and hypoalbuminemia in hospitalized patients.

Immunomodulatory aspects in the progression and treatment of oral malignancy.

Inflammation substantially affects the risk of oral malignancy. Pro-inflammatory cytokine, interferon (IFN)-γ, confers anti-tumor activity using several different mechanisms. Conversely, higher expression of interleukin (IL)-17 is associated with worse prognosis. Monocyte chemotactic protein (MCP)-1 correlates positively with poor long-term survival of head and neck squamous cell carcinoma (HNSCC) patients. IL-1α affects cancer associated fibroblasts and macrophages, and promote several malignant phenotypes including immune suppression. Some anti-inflammatory cytokines, including IL-10 and transforming growth factor (TGF)-ß, relate to pro-tumoral activities. Among immune checkpoint modulators, programmed death (PD-)1 and PD-ligand (L)1 facilitate oral squamous cell carcinoma (OSCC) cell evasion from immune surveillance, and the expression status of these has a prognostic value. OSCCs contain tumor associated macrophages (TAMs) as major stromal cells of their tumor microenvironment. Among the two distinctive states, M2 macrophages support tumor invasion, metastasis and immune suppression. Crosstalk between TAMs and OSCC or cancer-associated fibroblasts (CAF) plays an important role in the progression of OSCC. Clinical trials with blocking antibodies against IL-1α or melanoma-associated antigens have been reported as therapeutic approaches against OSCCs. The most promising approach activating antitumor immunity is the blockade of PD-1/PD-L1 axis. Manipulating the polarization of pro-tumorigenic macrophages has been reported as a novel therapeutic approach.

Muraminomicins, new lipo-nucleoside antibiotics from Streptosporangium sp. SANK 60501-structure elucidations of muraminomicins and supply of the core component for derivatization.

We screened for bacterial phospho-N-acetylmuramyl-pentapeptide-translocase (MraY: EC 2.7.8.13) inhibitors with the aim of discovering novel antibiotics and observed inhibitory activity in the culture broth of an actinomycete, SANK 60501. The active compounds, muraminomicins A, B, C, D, E1, E2, F, G, H, and I exhibited strong inhibitory activity against MraY with IC50 values of 0.0105, 0.0068, 0.0104, 0.0099, 0.0115, 0.0109, 0.0089, 0.0134, 0.0186, and 0.0094 µg ml-1, respectively. Although muraminomicin F exhibited favorable antibacterial activity against drug-resistant Gram-positive bacteria, this activity was reduced with the addition of serum. To efficiently supply the core component for chemical modification studies, production was carried out in a controlled trial by adding myristic acid to the medium, and a purification method suitable for large-scale production was successfully developed.

Perturbation-Based Proteomic Correlation Profiling as a Target Deconvolution Methodology.

Molecular target identification of small molecules, so-called target deconvolution, is a major obstacle to phenotype-based drug discovery. Here, we developed an approach called perturbation-based proteomic correlation profiling (PPCP) utilizing the correlation between protein quantity and binding activity of compounds under cellular perturbation by gene silencing and successfully identified lanosterol synthase as a molecular target of TGF-ß pathway inhibitor. This PPCP concept was extended to the use of a cell line panel and provides a new option for target deconvolution.

Gene expression analyses associated with malignant phenotypes of metastatic sub-clones derived from a mouse oral squamous cell carcinoma Sq-1979 cell line.

To elucidate the genetic events that occur during the development of OSCC, the present study established a model of oral malignancy using a mouse oral squamous cell carcinoma (OSCC) Sq-1979 cell line. Sq-1979 cells were implanted into syngeneic C3H mice. Subsequently, 233 cells and metastatic sub-clones (L cells) from primary OSCC, as well as the metastasized lymph node tissues of Sq-1979-implanted mice were established. Compared with parental Sq-1979 and 233 cells, the majority of L cells exhibited a higher proliferation rate and transplantability, and conferred a lower survival rate on the implanted mice. To investigate the genetic background of L cells, preferentially expressed genes in L cells were identified by cDNA microarray and reverse transcription-polymerase chain reaction analyses. The expression of FYN-binding protein (Fyb), solute carrier family 16 member 13 (Slc16a13), keratin 7, transmembrane portion 173 and Slc44a3 mRNAs was significantly elevated in L cells compared with that in Sq1979 and 233 cells. The mRNA expression was also evaluated in human OSCC and leukoplakia (LP) tissues. Among the 5 aforementioned mRNAs, the expression of FYB and SLC16A13 was significantly higher in OSCC than in LP tissues. Furthermore, the expression of SLC16A13 mRNA was significantly elevated in highly invasive OSCCs, which were classified as grades 3 and 4 by the Yamamoto-Kohama (YK) classification of invasion, compared with those in lower grades (YK-1 and -2). The model proposed in the present study could thus describe essential marker genes for the diagnosis of oral malignancies.

Orofacial Pain Associated with Vasospastic Angina: A Case Report.

The primary symptom of ischemic heart disease is typically chest pain, but in some cases, this pain may radiate to the maxillofacial region. This article describes the case of a 44-year-old man with orofacial pain of cardiac origin. The patient was suspected to be suffering from cardiac disease by the oral and maxillofacial surgeon and was referred to a cardiologist, where he received a heart examination. The patient was diagnosed by means of cardiac catheterization as having coronary spastic angina. During catheterization, intracoronary ergonovine maleate induced orofacial pain that was almost the same in character and intensity as the patient's first episode. The orofacial pain was considered to be telalgia from coronary spastic angina. The patient started medication on the same day as the diagnosis. There was no recurrence of any symptoms. These findings indicate that in such cases, the dentist may contribute to identifying ischemic heart disease and should refer the patient to a cardiologist.

Symmetrical lipomatosis of the tongue: Case report and literature review.

Multiple symmetric lipomatosis is rare and characterized by diffuse growth and nonencapsulated lipomas. It is usually found in the posterior neck and upper trunk, and the entity is known as "benign symmetric lipomatosis," "Madelung disease," and "Launois-Bensaude syndrome." Symmetric lipomatosis of the tongue was first described by Desmond and is an extremely rare condition. A 74-year-old man complained of painless tongue swelling and difficulty speaking. Clinical findings revealed no tumor masses on the trunk, limbs, or head and neck region. Intraoral findings included soft yellowish masses with a smooth surface without erosions on the side of the tongue bilaterally. They were 30 mm in diameter. An incisional biopsy was taken from the mass, and the lipoma was diagnosed. The bilateral tongue lesions were resected under general anesthesia. Intraoperative findings revealed adipose tissues interspersed with lingual muscles and no capsulation. The lesion was finally diagnosed as symmetric lipomatosis of the tongue based on clinical findings and radiological and histologic examination.

Producing Capabilities of Interferon-gamma and Interleukin-10 in Peripheral Blood from Oral Squamous Cell Carcinoma Patients.

In order to evaluate the Th1 and Th2 responses of Oral Squamous Cell Carcinoma (OSCC) patients, we investigated the cytokine producing capability of peripheral blood (PB), and compared it with clinicopathological appearances of OSCC patients. The production of a Th1-type cytokine, interferon (IFN)-γ, from lipopolysaccharide (LPS)-stimulated PB correlated positively with the frequency of lymph node metastasis. We also investigated the production of a Th2-type cytokine, IL-10, however, no significant correlation was observed with the clinicopathological appearances. Our results suggested that the IFN-γ producing capability was specifically regulated and dependent on the regional metastatic potencies of OSCCs.

Increase of peripheral blood CD57+ T-cells in patients with oral squamous cell carcinoma.

BACKGROUND/AIM: The subset of T-cells positive for expression of cluster of differentiation (CD) 57 has been associated with various cancer phenotypes. However, the presence of CD57(+) T-cells in patients with oral squamous cell carcinoma (OSCC) has yet to be confirmed. In the present study, we examined the diagnostic significance of the presence of CD57(+) T-cells in peripheral blood (PB) from patients with OSCC. MATERIALS AND METHODS: The subset of CD57(+) T-cells in PB was analyzed in 43 patients with OSCC by fluorescence-activated cell sorting (FACS) analysis. RESULTS: The proportion of CD57(+) T-cells, including both CD8(+) and CD4(+) subsets, significantly increased with clinical stage, especially in parallel with tumor size. CONCLUSION: Our results suggest that an increase in the population of CD57(+) T-cells is a potent prognostic marker and may also influence the systemic immunity of patients with OSCC.

Primary intraosseous carcinoma arising from an odontogenic cyst: A case report.

Cyst-like lesions in the mandible rarely develop into malignancies, and the reported incidence is between 0.3 and 2%. The present study describes a rare case of primary intraosseous squamous cell carcinoma of the mandible arising from an odontogenic cyst. A 59-year-old female was referred to Asahi University Murakami Memorial Hospital (Gifu, Japan), with acute pain in the right molars. An initial examination revealed buccal swelling and paresthesia of the mental nerve. Following an intraoral examination, the oral mucosa was confirmed to be normal, however, percussion pain was experienced between the lower right first premolar and second molar. Panoramic radiography revealed a retained lower right wisdom tooth and an irregular radiolucent area between the lower right molar and a mandibular angle with unclear margins. Computed tomography revealed diffuse bone resorption and an extensive loss of cortical bone on the buccal and lingual sides. A biopsy was performed and the pathological diagnosis was of a squamous cell carcinoma arising from the epithelial lining of the odontogenic cyst. Radical dissection was subsequently performed, however, histopathological examination of the resected specimen revealed neither invasion into the surrounding tissues penetrating the periosteum nor lymph node metastasis at the right submandibular lesion. Following the pathological diagnosis of primary intraosseous carcinoma (PIOC), the patient received 6,000 Gy radiation as post-operative radiotherapy and chemotherapy with oral administration of tegafur, gimeracil and oteracil potassium. The patient is currently undergoing follow-up examinations. Although PIOC arising from an odontogenic cyst is rare, it should be considered as a differential diagnosis for radiolucency of the jaw bone, particularly in older patients exhibiting a history of cystic lesions.

Barium-induced appendicitis mimicking accidental ingestion of a dental metal crown in radiological findings.

Barium appendicitis is a rare complication that has only been reported in a small number of case reports in the medical literature. A 57-year-old male presented to the emergency room with a sudden onset of sharp right lower quadrant abdominal pain. He had undergone contrast barium examination of his stomach 2 months previously as part of a periodic examination for gastric cancer. The radiological findings showed that the shape and radiopaque levels were similar to those of a dental metal crown silhouette. The patient was strongly suspected to have a localized intra-abdominal abscess due to ileocaecal perforation with a foreign body such as a dental metal crown. Emergency surgery revealed acute phlegmonous appendicitis. The resected specimen demonstrated a phlegmonous appendix which contained solid coproma. Pathological diagnosis and composition analysis confirmed the onset of appendicitis to be a result of the patient's ingestion of barium sulfate.

Anti-Pseudomonas aeruginosa compound, 1,2,3,4-tetrahydro-1,3,5-triazine derivative, exerts its action by primarily targeting MreB.

In order to find new anti-Pseudomonas agents, we carried out whole-cell based P. aeruginosa growth assay, and identified 1,2,3,4-tetrahydro-1,3,5-triazine (Compound A). This compound showed anti-Pseudomonas activity against wild as well as pumpless strain equally at a same concentration. Also, this compound was structurally very similar to A22, which is known to inhibit the bacterial actin-like protein MreB. By the analysis of resistant strains, the primary target of this compound in P. aeruginosa was definitely confirmed to be MreB. In addition, these compounds showed a bacteriostatic effect, and induced the morphology changes in P. aeruginosa from rod shape to sphere shape, which leads to be clinically favorable in terms of susceptibility to phagocytosis and release of endotoxin. These results display that Compound A is a very attractive compound which shows anti-P. aeruginosa activity based on inhibition of MreB without being affected by efflux pumps, and could provide a new step toward development of new promising anti-Pseudomonas agents, MreB inhibitors.