Tadashi Noda

Sour taste stimulation facilitates reflex swallowing from the pharynx and larynx in the rat

Chemical stimulation of the pharynx and larynx is effective in eliciting reflex swallowing. A sour taste bolus facilitates the onset of swallowing in patients with neurogenic dysphagia, but the mechanism of the facilitation has not been clarified. We investigated the effect of sour solutions on the elicitation of reflex swallowing in anesthetized rats. The main ducts of salivary glands were ligated to avoid the effect of saliva. A small amount of water, sour solutions, and other taste solutions were applied to the mucosa of the pharyngolaryngeal region. Acetic acid and citric acid, which provide a sour taste, had a stronger effect on evoking reflex swallowing as compared with other taste solutions. The effectiveness of these acids increased with increasing concentrations. We also examined the contribution of the superior laryngeal nerve (SLN) and the pharyngeal branch of the glossopharyngeal nerve (GPNph) to reflex swallowing. Acetic acid was greatly effective in evoking swallowing in both the region innervated by the SLN and the GPNph. On the other hand, water was effective in the SLN region but only slightly effective in the GPNph region. The results indicate that stimulation of the pharyngolaryngeal region with sour solutions facilitates reflex swallowing, suggesting that the facilitation may be due to increases of sensory inputs via the SLN and GPNph.

Localization of H[+]-ATPase and carbonic anhydrase II in ameloblasts at maturation

The localization of vacuolar-type H+-ATPase and carbonic anhydrase II (CA II) in rat incisor enamel organs at maturation was examined by light and electron microscopy. The immunoreactivity for both vacuolar-type H+-ATPase and CA II was intense on the ruffled border of ruffle-ended ameloblasts (RA), but moderate at the distal end of smooth-ended ameloblasts (SA). Immuno-gold particles indicated that CA II was not confined to the ruffled border of RA alone, but also distributed in the cytoplasm of RA and SA. These findings suggest that RA may secrete protons produced by CA II via the ruffled border into enamel by active transport of vacuolar-type H+-ATPase. Secreted protons may activate hydrolytic enzymes to degrade the organic components of enamel matrix. Vacuolar-type H+-ATPase on vesicles of SA suggests that a specific configuration of ruffled borders in RA may be formed by the fusion of vesicle membranes in the distal end of cytoplasm of SA.

Responses of immunocompetent cells in the dental pulp to replantation during the regeneration process in rat molars

Abstract. Responses of immunocompetent cells to tooth replantation during the regeneration process of the dental pulp in rat molars were investigated by immunocytochemistry using antibodies to class II major histocompatibility complex (MHC) molecules (OX6 antibody), monocyte/macrophage lineage cells (ED1 antibody) and protein gene product 9.5 (PGP 9.5), as well as by histochemical reaction for periodic acid-Schiff (PAS). Tooth replantation caused an increase in both the number of OX6- and ED1-positive cells and their immunointensity in the replanted pulp, but almost all PGP 9.5-immunoreactive nerves diminished in the initial stages. By postoperative day 3, many OX6- and ED1-immunopositive cells had accumulated along the pulp-dentin border to extend their cytoplasmic processes into the dentinal tubules in successful cases. Once reparative dentin formation had begun after postoperative day 7, OX6- and ED1-immmunopositive cells became scattered in the odontoblast layer, while reinnervation was found in the coronal pulp. The temporal appearance of these immunocompetent cells at the pulp-dentin border suggests their participation in odontoblast differentiation as well as in initial defense reactions during the pulpal regeneration process. On postoperative day 14, the replanted pulp showed three regeneration patterns: (1) reparative dentin, (2) bone-like tissue formation, and (3) an intermediate form between these. In all cases, PAS-reactive cells such as polymorphonuclear leukocytes (PML) and mesenchymal cells occurred in the pulp space. However, the prolonged stagnation of inflammatory cells was also discernible in the latter two cases. Thus, the findings on PAS reaction suggest that the migration of the dental follicle-derived cells into the pulp space and the subsequent total death of the proper pulpal cells are decisive factors for eliciting bone-like tissue formation in the replanted pulp.

Molecular basis of perinatal hypophosphatasia with tissue-nonspecific alkaline phosphatase bearing a conservative replacement of valine by alanine at position 406. Structural importance of the crown domain.

Hypophosphatasia, a congenital metabolic disease related to the tissue‐nonspecific alkaline phosphatase gene (TNSALP), is characterized by reduced serum alkaline phosphatase levels and defective mineralization of hard tissues. A replacement of valine with alanine at position 406, located in the crown domain of TNSALP, was reported in a perinatal form of hypophosphatasia. To understand the molecular defect of the TNSALP (V406A) molecule, we examined this missense mutant protein in transiently transfected COS‐1 cells and in stable CHO‐K1 Tet‐On cells. Compared with the wild‐type enzyme, the mutant protein showed a markedly reduced alkaline phosphatase activity. This was not the result of defective transport and resultant degradation of TNSALP (V406A) in the endoplasmic reticulum, as the majority of newly synthesized TNSALP (V406A) was conveyed to the Golgi apparatus and incorporated into a cold detergent insoluble fraction (raft) at a rate similar to that of the wild‐type TNSALP. TNSALP (V406A) consisted of a dimer, as judged by sucrose gradient centrifugation, suggestive of its proper folding and correct assembly, although this mutant showed increased susceptibility to digestion by trypsin or proteinase K. When purified as a glycosylphosphatidylinositol‐anchorless soluble form, the mutant protein exhibited a remarkably lower Kcat/Km value compared with that of the wild‐type TNSALP. Interestingly, leucine and isoleucine, but not phenylalanine, were able to substitute for valine, pointing to the indispensable role of residues with a longer aliphatic side chain at position 406 of TNSALP. Taken together, this particular mutation highlights the structural importance of the crown domain with respect to the catalytic function of TNSALP.

Role of the pharyngeal branch of the vagus nerve in laryngeal elevation and UES pressure during swallowing in rabbits.

Elevation of the larynx during swallowing plays an important role in protecting the laryngeal inlet and in the opening of the upper esophageal sphincter (UES). The thyrohyoid (TH) muscle is the most important muscle for laryngeal elevation, and it is thought to be innervated by the thyrohyoid branch. However, in preliminary studies we found that laryngeal elevation was severely disturbed after sectioning of the pharyngeal branch of the vagus nerve (X-ph). In the present study, we examined the role of the X-ph in laryngeal elevation and the contribution of this nerve to UES pressure. Ten male rabbits under anesthesia were used. Sectioning of the X-ph not only abolished the electromyographic activities of the TH and cricopharyngeus (CP) muscles, it also greatly reduced the maximal value of laryngeal elevation during swallowing. On the other hand, sectioning of the hypoglossal nerve, which contains the thyrohyoid branch, produced no appreciable change in the electromyographic activity of either muscle and it reduced the maximal value of the elevation only slightly. These results indicate that the X-ph innervates the TH and CP muscles and suggest that the X-ph plays an important role in elevating the larynx and in regulating the UES pressure in rabbits.

The development of terminal Schwann cells associated with periodontal Ruffini endings in the rat incisor ligament.

The postnatal development of the terminal Schwann cell, an analogue of the lamellar cell in cutaneous sensory receptors, was examined by histochemistry for non-specific cholinesterase and immunohistochemistry for S-100 protein in the periodontal Ruffini endings of the rat incisor. Double immunohistochemistry for S-100 protein and protein gene product 9.5 (PGP 9.5) was also performed to examine the relationship between terminal Schwann cells and axons. Histochemistry for non-specific cholinesterase was able to demonstrate the age-related development of the terminal Schwann cells; the morphology and distribution of the developing terminal Schwann cells became almost identical to those in adults during postnatal days 15-18. Axons showing PGP 9.5-like immunoreactivity elongated and expanded after arrangement of terminal Schwann cells in the alveolus-related part. This suggests that the terminal Schwann cell is important in the development and maturation of the periodontal Ruffini endings.

Eruption disturbances in Japanese children and adolescents

Abstract The aims of this report were to determine the nature of eruption disturbances and to establish the pattern of management for these teeth in a group of Japanese children and adolescents. Data were collected from the clinical records of patients in the Pediatric Dental Clinic of Niigata University Medical and Dental Hospital. There were 700 patients (364 males and 336 females) and 748 teeth (26 primary teeth and 722 permanent teeth) who were treated for eruption disturbances between 1979 and 2002. During this period, 17,365 new patients visited the clinic, of whom approximately 4% had eruption disturbances. The most frequently affected permanent teeth were the maxillary central incisors (38.6%), followed by the maxillary canines (15.4%). Of the permanent teeth, 52.5% were in males and 47.5% in females. Although there were more cases of maxillary central incisors in males than in females, the number of cases involving maxillary canines was higher in females. Whereas delayed eruption was noted in 76.7% of maxillary central incisors, 18.6% were impacted and 4.7% demonstrated abnormal direction/position which required treatment. In contrast, a higher percentage (64.8%) of maxillary canines that showed abnormal direction/position was treated. Eruption disturbances were found in 40% of the patients during routine examinations. This suggests that regular routine examinations are important for pediatric dentists to detect and treat eruption disturbances early.

Comparison of SLN-evoked swallows during rest and chewing in the freely behaving rabbit.

Interactions between the swallowing central neural pathway and the chewing central neural pathway were examined in freely behaving, unanesthetized rabbits. Pharyngeal swallows were elicited by electrical stimulation of the superior laryngeal nerve (SLN) and defined by thyrohyoid muscle (TH) activity in the electromyogram (EMG). Recordings were obtained from rabbits at rest and during chewing. The number of swallows elicited by the SLN stimulation was significantly increased (P<0.001) during quiet oral function (at rest) and during chewing. The increased number of swallows from each baseline was similar, signifying that the effect of the SLN stimulation was similar in generating swallowing in both groups. The swallows induced with SLN stimulation were very similar to natural swallows as defined by the temporal pattern of the EMG duration and the timing of EMG activities. Our results suggest that: (1). the peripheral inputs to the swallowing pathway may rarely be modulated by the chewing pathway in the generation of swallows; (2). the swallowing pathway and the chewing pathway may interact at the level of the rhythm generators; (3). each animal has its own threshold for eliciting pharyngeal swallowing, and the threshold may be independent of the number of chews.

Changes in the mouthful weights of familiar foods with age of five years, eight years and adults

Abstract For effective mastication and swallowing, to take optimum mouthful food regularly is necessary. The purpose of this study is to compare the weights and the variance of a mouthful food between children and adults. We studied mouthful weight and the coefficient variation in 5-year-old children (n=10), 8-year-old children (n=10) and adults (n=10) while they were eating rice, bread, sausage and apple on two different days. The test foods were served in random order 2–4 hours after lunch. Each portion was weighed before and after each bite to measure the mouthful weight, and the weight of the last bite of each portion was eliminated from the data. The mean weight and coefficient variation in each subject were calculated. The results showed the mean weight was largest in the adults, intermediate in the 8-year-old children, and smallest in the 5-year-old children for all test foods (ANOVA). Moreover, the mean coefficient variation among the weights of the groups revealed that mouthful weight within an individual varied most widely in the 5-year-old children and most narrowly in the adults, and that 8-year-old children could show the same coefficient variation of mouthful weight as adults in rice and apple (Steel-Dwass test). Our results suggest that mouthful weight becomes larger and more regularly with age.

A diagnostic proposal to support early treatment of ectopically erupting maxillary canines

Although early diagnosis is essential for the correction of ectopically erupting canines, it is difficult to determine whether the condition of a canine is within acceptable normal limits, particularly in children aged 10 years or younger. The purpose of the present study was to ascertain the appropriate time for a preventive procedures, such as extraction of the predecessor, in order to avert impaction of maxillary canines. Orthopantomograms of 225 normal children without dental anomalies were examined to compare the eruption stages of mandibular and maxillary canines. Sixty-four patients with 68 unerupted and/or impacted maxillary canines were examined. In normal children, mandibular canines exhibited earlier movement toward oral emergence than maxillary canines on the same side. This tendency was particularly prominent after exfoliation of the mandibular primary canine. In contrast, nearly half of the patients with anomalies were treated before oral emergence of the mandibular canine through simple procedures such as extraction of the primary canine or exposure of the affected canine. The present findings may provide a useful supplemental rule for early diagnosis and aid in decisions whether to treat ectopically erupting maxillary canines, particularly when the anomaly is obscure.