The Vitamin D3 analogue calcipotriol suppresses CpG-activated TLR9-MyD88 signalling in murine plasmacytoid dendritic cells.

BACKGROUND: Plasmacytoid dendritic cells (pDCs) are involved in the pathogenesis of psoriasis by secreting interferon-α. Vitamin D3 analogues are widely used to treat psoriasis, and the representative analogue calcipotriol (CAL) uniquely downregulates the cytokine production and chemotactic activity of pDCs. However, the molecular mechanism of action of CAL is not well understood. AIM: To investigate effects of CAL on the Toll-like receptor 9-myeloid differentiation primary response gene 88 (TLR9-MyD88) signalling pathway, which induces cytokine production, in murine pDCs. METHODS: pDCs were isolated from mouse spleen cells by negative selection or were generated from mouse bone-marrow cells, and were stimulated with CpG-oligodeoxynucleotide (ODN) with or without CAL for 24 h. mRNA expression of TLR9 and MyD88 was assessed by real-time PCR, and the amount of TLR9 was measured by western blotting. RESULTS: CAL suppressed the CpG-ODN-induced increased expression of MyD88 and TLR9 in pDCs. CONCLUSIONS: CAL may downregulate pDCs by inhibiting TLR9-MyD88 signalling.

Expression of toll-like receptor 2, NOD2 and dectin-1 and stimulatory effects of their ligands and histamine in normal human keratinocytes.

BACKGROUND: Epidermal keratinocytes are involved in the skin innate immunity and express toll-like receptors (TLRs) and other innate immune proteins. The epidermis is continuously exposed to pathogenic gram-positive bacteria or fungi. However, few studies have examined the function and expression of innate immune proteins in keratinocytes. Histamine, which is well known for itch and allergy, is closely associated with innate immunity, but its influence on epidermal innate immunity is still unclear. OBJECTIVES: To clarify the expression of innate immune proteins in keratinocytes stimulated by ligand pathogen-associated molecules, and the function of histamine in this process. METHODS: We investigated the effects of lipopeptide (MALP-2, 1-100 ng mL(-1); ligand for TLR2), peptidoglycan (PGN, 0.02-2 microg mL(-1); ligand for NOD2) and beta-glucan (1-100 microg mL(-1); ligand for dectin-1) in the presence or absence of histamine on mRNA expression of TLR2, NOD2 and dectin-1 as well as human beta-defensin 2 by quantitative real-time polymerase chain reaction in cultured normal human epidermal keratinocytes. TLR2 expression was also examined at the cell surface and intracellularly, as determined by flow cytometry and confocal microscopy. The quantities of interleukin (IL)-1alpha and IL-8 produced by keratinocytes were measured using enzyme-linked immunosorbent assay. RESULTS: At the mRNA level, TLR2 was enhanced by PGN but not by its ligand MALP-2 or by beta-glucan; NOD2 was easily induced by all three ligands; and dectin-1 was enhanced by its ligand beta-glucan. These enhanced expressions were further augmented by histamine at 1 microg mL(-1). While the surface expression of TLR2 was barely detectable by flow cytometry even after stimulation, the intracellular expression of TLR2 was apparently elevated by PGN and further promoted by histamine. A confocal microscopic analysis also revealed the enhanced expression of TLR2 in the cytoplasm. The expression of TLR2, NOD2 and dectin-1 was functional, as these pathogen-associated molecules induced the production of IL-1alpha, IL-8 and defensin, and again, histamine greatly enhanced this production. CONCLUSIONS: Our study demonstrated that the expression of functional innate immune receptors is augmented by the pathogen-associated molecules in a ligand-feed forward or nonrelated manner in keratinocytes, and histamine promotes their expression and the resultant production of cytokines and defensins.

Relationship between patient characteristics, mandibular head morphology and thickness of the roof of the glenoid fossa in symptomatic temporomandibular joints.

OBJECTIVE: The purpose of this clinical study was to investigate the minimum thickness of the roof of the glenoid fossa (RGF) of grossly normal temporomandibular joints (TMJ) and to correlate this with patient gender, age and the morphological classification of the mandibular head. METHODS: The study was performed on 191 TMJs from 109 patients (25 male and 84 female, age range 3-79 years, mean age 28.1 years) who visited Nihon University Dental Hospital, Japan with suspected TMJ disorders. The patients underwent cone beam computed tomography (3DX CT) to enable observation of the morphological features of the mandibular head. The minimum thickness of the RGF was measured using frontal section images acquired by CT. The morphology of the mandibular heads was classified according to the method of Yale and colleagues. Mean linear measurements were used for statistical analyses of patient gender, age and mandibular head morphology. RESULTS: The average minimum thickness of the RGF was 0.79 mm. No significant difference in thickness was found between male and female patients. In addition, no differences were recorded as a result of variation in age or mandibular head morphology. CONCLUSIONS: These results indicate that RGF thickness is not significantly correlated with gender, age, or mandibular head morphology, at least in this cohort of patients.

Reproducibility and accuracy of measuring unerupted teeth using limited cone beam X-ray CT.

OBJECTIVE: The purpose of this investigation was to determine the reproducibility among observers and accuracy of the measurement of the tooth crown width of unerupted teeth using limited area cone beam X-ray CT. METHODS: 3DX multi-image micro-CT (3DX, Morita Co., Kyoto, Japan) images of impacted supernumerary teeth in the median maxillary region taken prior to extraction were used for the samples. The width of the tooth on the 3DX image was measured five times by five individual observers. Significant differences in values among the observers in the measurement were determined by one-way analysis of variance for examining reproducibility. The measurement results of the ten samples on 3DX images were compared with the laboratory measurements using a three-dimensional co-ordinate measuring apparatus, using the Wilcoxon signed-rank sum test. RESULTS: There was no significant difference among the observers in the measurement (P>0.05). The measurement results shown on 3DX images were significantly larger than those of the laboratory measurements (P<0.05). The mean difference was +0.088 mm. CONCLUSIONS: 3DX has high reproducibility for measuring the tooth crown width of unerupted teeth. While 3DX measurement values were larger than the laboratory measurements, the difference is clinically insignificant.

Relationship between functional disc position and mandibular displacement in adolescent females: posteroanterior cephalograms and magnetic resonance imaging retrospective study.

The purpose of this study was to investigate the relationship between the disc positions of temporomandibular joints (TMJ), the vertical and lateral mandibular displacement (VMD and LMD, respectively) and age in female adolescents with signs and symptoms of the temporomandibular disorders (TMD). The VMD and LMD were assessed, using posteroanterior (PA) cephalograms. The disc positions were assessed by magnetic resonance imaging (MRI) and categorized as follows: normal disc position, functional disc displacement and functional disc dislocation. Excluding patients with osteoarthritis, the total number of subjects was 54 female adolescents who were grouped into three: the bilateral normal disc position group, the unilateral or bilateral functional disc displacement group, and the unilateral or bilateral functional disc dislocation group. We compared the extent of VMD and LMD between the three groups, and investigated their correlation with age. Results indicate that functional disc displacement and dislocation are related to mandibular displacement, and VMD did not correlate with age but LMD did correlate with age. This study suggests that the onset of disc displacement is related to the mandibular displacement and disturbs normal growth of the mandible three-dimensionally.

[Squamous cell carcinoma of the bladder--a case report].

A 63-year-old male was referred to us for investigation of gross hematuria and miction pain. DIP showed a filling defect on the left side of the bladder. This patient underwent through clinical and endoscopic staging which included chest X-ray and abdominal and pelvic CT scan. Biopsy of the tumor resulted in a diagnosis of squamous cell carcinoma of the bladder, grade 2. Total cystectomy and lymphadenectomy were carried out. Unfortunately this patient died 6 months after surgery because of local recurrence. We believe that future research should be directed toward adjunctive studies with radiation and chemotherapy.