Microfold (M) cells residing in the follicle-associated epithelium of mucosa-associated lymphoid tissues are specialized for sampling luminal antigens to initiate mucosal immune responses. In the past decade, glycoprotein 2 (GP2) and Tnfaip2 were identified as reliable markers for M cells in the Peyer's patches of the intestine. Furthermore, RANKL–RANK signaling, as well as the canonical and non-canonical NFκB pathways downstream, is essential for M-cell differentiation from the intestinal stem cells. However, the molecular characterization and differentiation mechanisms of M cells in the lower respiratory tract, where organized lymphoid tissues exist rarely, remain to be fully elucidated. Therefore, this study aimed to explore M cells in the lower respiratory tract in terms of their specific molecular markers, differentiation mechanism, and functions. Immunofluorescence analysis revealed a small number of M cells expressing GP2, Tnfaip2, and RANK is present in the lower respiratory tract of healthy mice. The intraperitoneal administration of RANKL in mice effectively induced M cells, which have a high capacity to take up luminal substrates, in the lower respiratory epithelium. The airway M cells associated with lymphoid follicles were frequently detected in the pathologically induced bronchus-associated lymphoid tissue (iBALT) in the murine models of autoimmune disease as well as pulmonary emphysema. These findings demonstrate that RANKL is a common inducer of M cells in the airway and digestive tracts and that M cells are associated with the respiratory disease. We also established a two-dimensional culture method for airway M cells from the tracheal epithelium in the presence of RANKL successfully. This model may be useful for functional studies of M cells in the sampling of antigens at airway mucosal surfaces.

Objective: To evaluate and compare the effect of each congenital and postnatal treatment factors in treatment outcome based on dental arch relationship (DAR) of four different populations at a time using GOSLON yardstick.  Material and Methods: 432 unilateral cleft lip and palate subjects (Japanese, Malay ethnic group, Bangladeshi and Pakistani populations) age 5 to 12 years were taken before orthodontic treatment and alveolar bone grafting. The DAR was assessed by GOSLON Yardstick. Independent t-test was performed to compare the GOSLON score for each factors. A multiple comparison (Anova) was also conducted between the GOSLON score of four different populations. The significance level was set at p<0.05.  Results: Statistically no significant different was found among the congenital factors. However, the family history of Class III malocclusion showed most likely to associate with, though there was no significant difference (p=0.069). Significant difference revealed between two techniques of cheiloplasty in both Malay ethnic group and Pakistani population (p=0.038 and p=0.000, respectively). Gender and Techniques of palatoplasty also showed significant difference in Pakistani population (p=0.026 and p=0.000, respectively). Japanese and Bangladeshi population showed no significant differences. Also no significant differences found between the GOSLON score and different countries.  Conclusion: The treatment outcome based on DAR significantly varies in individual population [Malay ethnic group (cheiloplasty); Pakistan (gender, cheiloplasty and palatoplasty)].

Objective: To evaluate the craniofacial morphology of Japanese patients with unilateral cleft lip and palate and to evaluate the multiple congenital factors that affects craniofacial morphology in unilateral cleft lip and palate patients. Material and Methods: Lateral cephalograms of 140 subjects with unilateral cleft lip and palate were taken before any orthodontic treatment and alveolar bone graft. Subjects mean age was 6.85 +/- 1.56 years. Primary surgeries performed by surgeons from Hokkaido University Hospital. The craniofacial morphology of the 140 subjects was assessed by angular and linear cephalometric measurements. Gender, side of cleft, complete/incomplete type of cleft, presence/absence of lateral incisor in the affected side, family history of cleft and family history of skeletal Class III was chosen as congenital factors. To compare the assessments using congenital factors affecting craniofacial morphology in the unilateral cleft lip and palate subjects, angular and linear cephalometric measured values from each individual subject (control group) were converted into Z scores in relation to the means and standard deviation of the two parameters. Results: Twelve out of 13 craniofacial morphology outcomes were insignificant in 5 out of 6 dependent variables. Only 1 dependent variable showed 6/15 significant differences. Conclusion: Current study revealed the evidence that there were almost no significant differences in the craniofacial morphology outcome among various congenital factors. This will provide base line information and help determine the effectiveness of such factors.

Mandibular prognathism is a phenotype of facial deformity seen in populations around the world, but with higher incidence among East Asian populations. Five genome-wide nonparametric linkage analyses and a genome-wide association study to identify susceptibility loci of the phenotype have shown inconsistent results. To explore variants related to mandibular prognathism, we undertook whole-exome sequencing in a Japanese pedigree. The pedigree was ascertained as mandibular prognathism. The pedigree comprised 15 individuals from 4 generations. Four affected individuals across 2 generations and 5 unaffected individuals were chosen for whole-exome sequencing. Five non-synonymous single-nucleotide variants (SNVs) of UBASH3B, OR6M1, OR8D4, OR8B4, and BEST3 genes were detected in all 4 affected individuals, but in none of the 5 unaffected individuals. A non-synonymous SNV of the BEST3 gene, Chr12(GRCh37):g.70048878G > T, NM_032735.2:c.1816C > A, p.(L606I), was identified as rare missense variant. BEST3 is located on chromosome 12q15 and encodes bestrophin 3 from the bestrophin family of anion channels. The 4 other non-synonymous SNVs of UBASH3B, OR6M1, OR8D4, and OR8B4 were not considered plausible candidates for mandibular prognathism. Our whole-exome sequencing implicates a rare non-synonymous SNV of BEST3 as a candidate for mandibular prognathism in the Japanese pedigree.

This study investigates the significance of the expression and dynamics of podoplanin in mechanostress and mineralization in cultured murine osteoblasts. Podoplanin increased in osteoblasts subjected to straining in non-mineralization medium, suggesting that the mechanostress alone is a podoplanin induction factor. In osteoblasts subjected to vertical elongation straining in the mineralization medium, the mRNA amounts of podoplanin, osteopontin, and osteocalcin were significantly larger than those in cells not subjected to straining, suggesting that mechanostress is the cause of a synergistic effect in the expression of these proteins. In osteoblasts in the mineralization medium, significant increases in osteocalcin mRNA occurred earlier in cells subjected to straining than in the cells not subjected to straining, suggesting that the mechanostress is a critical factor to enhance the expression of osteocalcin. Western blot and ELISA analysis showed increased podoplanin production in osteoblasts with longer durations of straining. There was significantly less mineralization product in osteoblasts with antibodies for podoplanin, osteopontin, and osteocalcin. There was also less osteopontin and osteocalcin produced in osteoblasts with anti-podoplanin. These findings suggest that mechanostress induces the production of podoplanin in osteoblasts and that podoplanin may play a role in mineralization in cooperation with bone-associated proteins.

In a previous genome-wide association study, plexin A2 (PLXNA2) was suggested as one of the candidate genes for mandibular prognathism. PLXNA2 encodes plexin A2, a member of the plexin-A family of semaphorin co-receptors. Semaphorin 3A (sema3A) exerts an osteoprotective effect. However, to the best of our knowledge, there have been no previous studies examining the role of sema3A or plexin A2 on human chondrocytes. The objectives of the present study were to examine the function of sema3A and its receptor, plexin A2, in human chondrocytes. Normal human chondrocytes were cultured in media with either a high (100 ng/ml) or a low (1 ng/ml) concentration of sema3A, or without sema3A as a control. Cells and extracellular matrices were assayed for concentrations of protein and parathyroid hormone-related peptide receptor 1 (PTH-R1) using a bicinchoninic acid assay and an enzyme immunoassay, respectively. At culture day 7, the high and low concentrations of exogenous sema3A significantly increased the protein content compared with the control (P=0.0008 and 0.00002, respectively). At culture day 14, a high concentration of exogenous sema3A significantly increased the protein content and decreased the concentration of PTH-R1 compared with the control (P=0.002). The present study revealed novel results that exogenous sema3A suppresses the expression of PTH-R1 in human proliferative chondrocytes and suggested that sema3A may affect human chondrocytes via its receptor, plexin A2.

INTRODUCTION:The aims of this study were to determine the difference in electromyography (EMG) of the orbicularis oris muscles between subjects with lip incompetence and lip competence and to elucidate the effectiveness of hypoxic lip training with EMG. METHODS:Twenty-five subjects (14 men, 11 women; mean age, 24.5 ± 2.6 years) were divided into 2 groups by sealed lip ratio: 13 subjects with lip incompetence (LI) and 12 subjects with lip competence (LC). Integral values of EMG for the orbicularis oris muscle during lip closing were estimated (baseline, T1). Then the subjects in the LI group performed the hypoxic lip training every day for 4 weeks. The EMG recordings were repeated during the training (T2), immediately after the training (T3), 4 weeks after the training (T4), and 8 weeks after the training (T5). RESULTS:In the LI group, the EMG values during lip closing were significantly higher than those of the LC group, and the values of EMG during lip closing at T3 were significantly lower than those at T1. In the posttraining period, the values of EMG at T4 and T5 were significantly lower than those at T1. The results suggested that subjects with lip incompetence need greater orbicularis oris muscle activity to keep their lips closed. CONCLUSIONS:The suggested standardized lip training could be used as a training method for patients with lip incompetence.

In this study, we investigated on suppression of metal ions elution on dental orthodontic wire. To suppress metal ion elution from stainless wire, two types of surface chemical modifications were carried out. The obtained wire were immersed into several aqueous solution then the eluted ions (Cr and Ni, which are well known to cause a metal allergy) were determined using coupled plasma mass spectrometry (ICP-MS). By both electropolymerization and self-assembly monolayer formation, the ion elution was effectively reduced. These results suggested that these surface modifications succeeded to suppress the metal ions.

We investigated in controlled dye-release behavior of nanosized silica particles containing nanocavities (Nanoporous silica, NPS). To determine this, NPS were mixed with glass ionomer cement (GIC), which is a medical material used as a matrix. The dye-release behavior was observed using a UV-visible spectrometer. After cationic dye was charged into GIC pellet containing NPS, the pellet could gradually release cationic dye for up to two weeks. To understand the dependence of electric charge on the dye-release behavior, three types of dyes with different charge were also investigated. Dyes having a neutral or negative electric charge were quickly released from the pellet within a couple of days. These results suggest that the nanocavities present in NPS can selectively bind cationic dyes and allow for their gradual release. This result reveals the excellent sustained dye-release property of NPS.

Lip incompetence, which is a state in which the upper and lower lips are constantly apart, is thought to induce mouth breathing that may result in the development of gingivitis and periodontal disease. In the orthodontic field, lip incompetence has major problems for diagnosis, when determining the treatment goal and at the time of retention. We have been performing studies on lip incompetence and have obtained the following results : (1) it is possible to predict to some extent the state of lip closure during sleep by examination only in an awake state, (2) anterior teeth are positioned forward in individuals with lip incompetence, and (3) a training method called Button Pull is effective as a myofunctional therapy for improving lip incompetence, but there remains a need to clarify the background.

Oligosilane-functionalized C60 fullerenes [namely, C60–(SiH2)n–H, n = 1–4] have been investigated by the density functional theory (DFT) method to elucidate the structures and electronic states of oligosilane-radical added fullerene. The DFT calculation showed that oligosilane radicals bind to the carbon atom of C60 in the on-top site, and a strong Si–C heterojunction is formed. The binding energies of oligosilane radicals to C60 were calculated to be 24.6–28.2 kcal/mol at the CAM-B3LYP/6-311G(d,p) level. The electronic states of oligosilane-functionalized fullerenes C60–(SiH2)n–H are discussed on the basis of theoretical results.

Background:Since it is difficult to precisely control the magnitude of force in orthodontic practice, controlling the duration of force is expected to prevent tissue damage. The use of an actuator as an appliance for intermittent force application is a possible solution for controlling the duration. However, effectiveness of an actuator for controlling the duration is still not clear.Aim:To determine whether a short loading duration of intermittent force has the potential to maintain a sound condition of the capillaries and induce dilation of capillaries, which is a requisite phenomenon for orthodontic tooth movement.Materials and methods:Six-week-old male hamsters with a dorsal skinfold chamber attached to an intermittent loading device were used. In three experimental groups, application of intermittent pressure that involved repetition of loading and unloading was performed. The durations of loading/unloading were 1 second/9 seconds (group T10), 1 second/19 seconds (group T20), and 1 second/29 seconds (group T30). Vessels were examined using a fluorescence microscope and a stereomicroscope for 5 days.Results:Widths of capillaries in groups T10 and T20 increased significantly (P < 0.01). In contrast, widths of capillaries in group T30 showed no significant difference from those in the control group. Only group T10 showed bleeding, obvious destruction of vessels, and a significant increase in the rate of disappearance of vessels.Conclusion:In the epidermis, although conditions are limited, a short loading duration of intermittent force maintains a sound condition of the capillaries at the tissue level and induces dilation of capillaries.

To elucidate which of elevated serum concentration of inorganic phosphate (Pi) or disrupted signaling linked to alpha klotho/fibroblast growth factor 23 (FGF23) is a predominant regulator for senescence-related degeneration seen in alpha Klotho-deficient mice, we have examined histological alteration of the periodontal tissues in the mandibular interalveolar septum of alpha Klotho-deficient mice fed with Pi-insufficient diet. We prepared six groups of mice: wild-type, kl/kl, and alpha Klotho(-/-) mice with normal diet or low-Pi diet. As a consequence, kl/kl(norPi) and alpha Klotho(-/-norPi) mice showed the same abnormalities in periodontal tissues: intensely stained areas with hematoxylin in the interalveolar septum, dispersed localization of alkaline phosphatase-positive osteoblasts and tartrate-resistant acid phosphatase-reactive osteoclasts, and accumulation of dentin matrix protein 1 in the osteocytic lacunae. Although kl/kl(lowPi) mice improved these histological abnormalities, alpha Klotho(-/-lowPi) mice failed to normalize those. Gene expression of alpha Klotho was shown to be increased in kl/kl(lowPi) specimens. It seems likely that histological abnormalities of kl/kl mice have been improved by the rescued expression of alpha Klotho, rather than low concentration of serum Pi. Thus, the histological malformation in periodontal tissues in alpha Klotho-deficient mice appears to be due to not only increased concentration of Pi but also disrupted alpha klotho/FGF23 signaling.