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Biomedical subjects

T Inai

Publications and source records attributed to T Inai.

At least 19 recordsLinked to original sources

Effects of defect configuration, size, and remaining teeth on masticatory function in post-maxillectomy patients.

The aim of this study was to investigate the correlation between the masticatory function and the maxillary defect configuration, size, and remaining teeth in post-maxillectomy patients restored with an obturator prosthesis. Fifty consecutive post-maxillectomy patients (mean age 67.0) participated in this study. The subjects consisted of 26 dentate and 24 edentulous patients. Data were collected from the patients' clinical records, diagnostic casts, and a questionnaire on masticatory function to evaluate the defect configuration, size, and the masticatory function scores associated with wearing obturator prostheses. The defect size was evaluated by the ratio of defect area to the horizontal impression area. The defect configuration was assessed according to Aramany's classification and separated into unilateral and bilateral defects. The multiple regression analysis and the Mann-Whitney U-test (P<0.05) were used to assess statistical significance. The Spearman's correlation coefficient by rank test was also used to detect correlation. The following conclusions were made: (i) The presence of teeth in the maxillary dentition and the different type of the defect configuration had significant correlation with the masticatory function score (r=0.616). (ii) The masticatory function scores of the subjects differed significantly with the presence of teeth in the maxillary dentition and the different types of defect configurations (P=0.005, P=0.002, respectively). (iii) There was significant correlation between the masticatory function score and the size of the defect area in the edentulous group (r=-0.648, P=0.001). The presence of teeth, the size and configuration of the defect influenced the masticatory function of post-maxillectomy patients with obturator prostheses.

Adult↗

Differential expression of major gap junction proteins, connexins 26 and 32, in rat mammary glands during pregnancy and lactation.

We examined the expression pattern of two major gap junction proteins, connexin 26 (Cx26) and connexin 32 (Cx32), in rat mammary glands during pregnancy and lactation. Immunohistochemically the two different Cxs were coexpressed in acinar cells and were independently modulated according to the physiological cell activity. Western blot analysis demonstrated that Cx26 gradually increased from early pregnancy, while Cx32 rapidly and dramatically increased at 16 h after parturition, and that both Cxs reached a maximum early in lactation. Increased expression of both Cxs was confirmed by Northern blot analysis showing that their mRNA transcripts were significantly induced on the day of parturition. We also analyzed double-immunofluorescent staining for Cx26 and Cx32 on a confocal laser scanning microscope, in order to examine colocalization of these Cxs in situ. Cx26 immunoreactivity mostly overlapped with Cx32-positive sites in acinar cells of lactating mammary glands, indicating that both Cxs were colocalized together in the same gap junctional plaques in lactation. These results suggest that upregulation of Cx26 and Cx32 in acinar cells at lactating stages, with colocalization in the same gap junctional plaques, may be important for control of secretion by acinar cells in rat mammary glands.

Animals↗

Dental rehabilitation using endosseous implants and orthognathic surgery in patients with cleft lip and palate: report of two cases.

We describe the use of endosseous implants in the autogenous particulate cancellous bone and marrow grafted alveoli after orthognathic surgery for dental rehabilitation of patients with cleft lip and palate. This procedure has been applied to two patients and produced good results functionally and aesthetically. The results are encouraging and indicate that implant placement after orthognathic surgery is useful for patients both with congenital missing teeth and retrognathic maxillae.

Adult↗

Claudin-1 contributes to the epithelial barrier function in MDCK cells.

Tight junctions (TJs) create a paracellular permeability barrier and also act as a fence preventing intermixing of proteins and lipids between the apical and basolateral plasma membranes. Recently, claudin-1 has been identified as an integral membrane protein localizing at TJs, and introduced claudin-1 can form TJ-like networks in fibroblasts. To investigate the function of claudin-1, MDCK cells were transfected with a mammalian expression vector containing myc-tagged mouse claudin-1, and four stable clones were obtained. The myc-tagged claudin-1 precisely colocalized with both occludin and ZO-1 at cell-cell contact sites, indicating that exogenous claudin-1 was properly targeted to the TJs. Immunoblot analysis revealed that overexpression of claudin-1 increased expression of ZO-1 but not of occludin or ZO-2. The barrier functions of these cells were evaluated by transepithelial electrical resistance (TER) and paracellular flux. Claudin-1-expressing cells exhibited about four times higher TER than wild-type MDCK cells. Consistent with the increase of TER, the cells overexpressing claudin-1 showed reduced paracellular flux, estimated at 4 and 40 kD FITC-dextrans. These results suggest that claudin-1 is involved in the barrier function at TJs.

Animals↗

Possible involvement of histamine in muscular fatigue in temporomandibular disorders: animal and human studies.

As an approach to clarifying the molecular basis of pain and fatigue in muscles involved in temporomandibular disorders, we examined the activity of histidine decarboxylase (HDC), the enzyme which forms histamine, in the masseter muscles of mice. In the resting muscle, HDC activity was very low. Direct electrical stimulation of the muscle markedly elevated HDC activity. HDC activity rose within 3 hrs of the electrical stimulation, peaked at 6 to 8 hrs, and then gradually declined. Intraperitoneal injection of a small amount of interleukin-1 (IL-1) (from 1 to 10 microg/kg) produced a similar elevation of HDC activity in the masseter muscle. We also examined the effect of an antihistamine, chlorphenylamine (CP), on temporomandibular disorders in humans and compared it with that of an anti-inflammatory analgesic, flurbiprofen (FB). Two groups received one or the other of the drugs daily for 7 days, and they were asked about their signs and symptoms before and after the treatment. A positive evaluation of their treatment was made by 74% of the CP group, but by only 48% of the FB group. Although the effects of CP on the limitation of mouth-opening and on joint noise were negligible, about 50% of the CP group answered positively concerning the drug's effect on spontaneous pain or pain induced by chewing or mouth-opening. The positive evaluation for CP (50%) in relieving associated symptoms (headache or shoulder stiffness) was significantly greater than for FB (13%). FB showed effectiveness similar to but sometimes weaker than that of CP on several symptoms. On the basis of these and previous results and the known actions of histamine, we propose that the histamine newly formed following the induction of HDC activity, which is itself mediated by IL-1, may be involved in inducing pain and, possibly, stiffness in muscles in temporomandibular disorders.

Adult↗

Distribution of gap junction protein connexin 37 in smooth muscle cells of the rat trachea and pulmonary artery.

Connexin 37, one of the gap junction protein families, has been detected by Northern blotting in various organs and tissues, and found to be especially abundant in the lung. However, detailed information on the precise types of cells which express connexin 37 has not been previously published. We therefore prepared site-specific connexin 37 antibodies and examined the distribution of connexin 37 immunohistochemically. Connexin 37 was detected in endothelial cells, in the tunica media of both the pulmonary artery and the aorta, and in the smooth muscle layer of the trachea and bronchioles. In the tracheal smooth muscle layer, connexin 37 overlapped with desmin-positive areas, but was clearly segregated from vimentin- and von Willebrand factor-positive areas. These results suggest that connexin 37 is expressed in smooth muscle cells in the trachea, but not in fibroblastic cells or endothelial cells. Connexin 37 was partially colocalized with connexin 43 in tracheal smooth muscle cells, and showed a gradual increase in expression during postnatal development. To our knowledge, this is the first report to be published regarding the expression of connexin 37 in smooth muscle cells.

Age Factors↗

[Postoperative prophylactic intravesical instillation of tetrahydropyranyl-adriamycin (THP) for superficial bladder cancer].

Intravesical instillation of tetrahydropyranyl-adriamycin (THP) was performed on 51 patients with superficial bladder cancer after transurethral resection (TUR) for prophylaxis of recurrence. The instillation was carried out with 20 mg of THP dissolved in 40 ml of distilled normal saline. Instillation was performed once 24 hours postoperatively, 9 times every 2 weeks, and 8 times every 4 weeks. These drugs were instilled for 30 to 60 minutes. The recurrence-free survival at 1, 2 and 3 years was 74.5%, 64.6% and 58.0%, respectively. Side effects of THP instillation were observed in only 4 cases (7.8%) as slight urinary frequency or micturition pain. Cases involving 5 or more tumors, or tumors measuring 3 cm or larger, more frequently demonstrated recurrence. The cases that did not respond to preoperative intravesical instillation of THP demonstrated a high frequency of recurrence. Intravesical instillation of THP as a prophylaxis against recurrence of superficial bladder cancer was effective in selected patients.

Administration, Intravesical↗

Use of an osseointegrated implant for dental rehabilitation after cleft repair by periosteoplasty: a case report.

OBJECTIVE: We report here on a patient with unilateral cleft lip and alveolus who underwent dental rehabilitation of cleft alveolus using an osseointegrated implant after cleft repair by periosteoplasty. The patient, whose lateral incisor was congenitally missing, had periosteoplasty, followed by excellent bone formation at the cleft alveolus within 2 years. After the completion of orthodontic alignment of the maxillary dental arch, a Brånemark single-tooth implant was placed into the bone-formed alveolus. CONCLUSION: This treatment procedure offers an option of dental rehabilitation for the alveolar clefts of patients with cleft lip and/or palate.

Alveolar Process↗

Immunohistochemical localization of connexin43 in the enamel organ of the rat upper incisor during ameloblast development.

We examined the localization of connexin (Cx)43 in the enamel organ during ameloblast development. The specificity of monoclonal anti-Cx43 antibody was elucidated by immunoblot analysis and immunoelectron microscopy. Gold particles for Cx43 were detected by immunoelectron microscopy on gap junctions but not on other structures such as desmosomes. Punctate and intense immunofluorescence for Cx43 was detected in all cell types of the enamel organ. Cx43 expression in ameloblasts showed a transient decrease and then increase during ameloblast development. Double staining of Cx43 and amelogenin, one of the enamel proteins, revealed that immunofluorescence for Cx43 markedly decreased in some late presecretory ameloblasts just prior to enamel formation. Moreover, the localization of Cx43 changed during enamel formation. Cx43 was distributed randomly on the lateral plasma membranes of presecretory ameloblasts, but tended to gather on those corresponding to the supranuclear regions of secretory ameloblasts. Immunofluorescence for Cx43 in maturation ameloblasts appeared linear rather than punctate. These results suggest that Cx43 in the late presecretory ameloblasts is degraded just before enamel formation and then newly synthesized Cx43 is redistributed during the secretory stage. These changes in Cx43 expression may be related to the cellular differentiation of ameloblasts.

Ameloblasts↗

[Mutagenicity studies of (+/-)-4-diethylamino-1,1,-dimethylbut-2-yn-1-yl 2-cyclohexyl-2-hydroxy-2-phenylacetate monohydrochloride monohydrate (NS-21), a novel drug for urinary frequency and incontinence].

The mutagenicity of (+/-)-4-diethylamino-1,1-dimethylbut-2-yn-1-yl 2-cyclohexyl-2-hydroxy-2-phenylacetate monohydrochloride monohydrate (NS-21), a new drug for the treatment of urinary frequency and incontinence, was investigated by the reverse mutation test in bacteria, the chromosome aberration test in vitro, and the micronucleus test in mice. The reverse mutation test was performed at a dose from 31.3 to 4000 micrograms/plate, at which dose cell killing was observed, using Salmonella typhimurium TA100, TA1535, TA98, and TA1537, and Escherichia coli WP2uvrA. NS-21 did not increase revertant colonies significantly in any of the test strains with or without metabolic activation system (S9 mix). The chromosome aberration test was carried out at a dose from 3.75 to 140 micrograms/ml, at which dose more than 50% cell proliferation was inhibited, using cultured Chinese hamster lung cells (CHL/IU). No significant increases of the frequencies of cells with chromosome aberrations were observed with or without S9 mix. The micronucleus test was conducted in the bone marrow cells of Slc : ddY male mice. Mice were given NS-21 by a single oral administration at doses of 0, 43.8, 87.5, 175, and 350 mg/kg, the geometric mean dose between the maximum tolerated dose and the minimum lethal dose. There were no significant increases in the frequencies of micronucleated polychromatic erythrocytes at any dose levels. These results show that NS-21 has no mutagenic activity in vitro or in vivo.

Animals↗

Changes in the phosphorylation states of connexin43 in myoepithelial cells of lactating rat mammary glands.

Using specific antibodies and cDNA probe, we examined the expression pattern of a major gap junction protein, connexin43 (Cx43), in rat mammary glands during pregnancy and lactation. Double immuno-fluorescence revealed that the labeling of Cx43 was superimposed in the alpha-smooth muscle actin-positive cells, suggesting that myoepithelial cell were interconnected by gap junctions formed of Cx43. Just after delivery, the Cx43-labeled plaques were enlarged and increased in intensity. Northern and Western blot analyses confirmed the dramatic induction of Cx43 at both mRNA and protein levels on the day of parturition. Cx43 mRNA transcript immediately declined, while the increase of Cx43 protein continued for a few days. During pregnancy, immunoblots showed two bands of almost equal amounts at 43 and 45 kDa. Following delivery, the 45-kDa band gradually increased in intensity with a concomitant decrease of the 43-kDa band. From the sixth day of lactation, Cx43 was always detected as a single band at 45 kDa. Alkaline phosphatase treatment of immunoprecipitated Cx43 revealed that both bands represented phosphorylated forms, thus indicating that Cx43 was naturally phosphorylated and that it altered its phosphorylation states during lactation stages. These results suggest that the induction of Cx43 with the changes in the phosphorylation states plays an important role in the lactating function of myoepithelial cells in rat mammary glands. This is the first report on the changes of Cx43 phosphorylation states during physiological stages in vivo.

Amino Acid Sequence↗

Influence of fluoride on secretory pathway of the secretory ameloblast in rat incisor tooth germs exposed to sodium fluoride.

Fluoride, which is an environmental toxicant, is a potent inducer of mottled enamel in humans and rats. To define the influence of fluoride on the secretory pathway in enamel fluorosis, mottled enamel was induced in the incisor tooth germs of rats by subcutaneous injections of sodium fluoride for 4 days, and then morphological and cytochemical changes of the secretory ameloblast were examined in the tooth germs with HRP-labeled lectin (Con A, GS-I, SBA and PNA) and En3 antibody labeling amelogenins. The accumulation of small vesicles on the route of the secretory pathway between the rER and the Golgi apparatus, disorder of Golgi stacks, and formation of abnormal large granules in distal cytoplasm were seen in the secretory ameloblast. Lectin staining patterns of the secretory ameloblast indicated the disturbance of the vesicular transport between the rER and the Golgi apparatus, and disorganization of the Golgi stack. Immunolabeling of the cell showed disruption of the sorting and fusion process on the secretory pathway. These results suggest that the fluoride disturbs the intracellular transport in the synthesis-secretory pathway of the ameloblast, and that this effect of fluoride on the synthesis-secretory pathway participates in the formation of enamel fluorosis.

Ameloblasts↗

Colocalization of connexin 43 and connexin 45 but absence of connexin 40 in granulosa cell gap junctions of rat ovary.

The expression and localization of gap junction family proteins (connexins) were examined in nonstimulated and gonadotrophin-stimulated ovarian follicles of immature rats. Immunoblot and RNA blot analysis showed the presence of connexin (Cx) 43, Cx40 and Cx45 in ovarian tissue. Of these connexin proteins, Cx43 and Cx45 were identified by immunofluorescent microscopy between granulosa cells in characteristic expression patterns related to follicular developmental stages, while Cx40 was not expressed in granulosa cells but was detected in blood vessels in ovarian stroma. In some plaques of gap junction between granulosa cells, Cx45 was found to be colocalized with Cx43. In immunofluorescent microscopy, the expression of Cx43 was increased with follicular growth, but decreased after induction of ovulation by injection of human chorionic gonadotrophin. In contrast, the Cx45 protein was constantly expressed through follicular development; however, after ovulation, no staining of Cx45 was detected in the corpus luteum. Dual expression and the functional role of Cx43 and Cx45 in cell-to-cell communication in ovarian granulosa cells at various developmental stages were discussed.

Animals↗

[Efficacy of granisetron rescue therapy for nausea and vomiting induced by cancer chemotherapies in urogenital malignant tumor].

In this study, the usefulness of granisetron rescue therapy for nausea and vomiting induced by cancer chemotherapies, including CDDP, was examined. Granisetron was given to twenty patients with urogenital malignant tumor by iv infusion for thirty minutes after the onset of nausea or vomiting. Nausea disappeared in 15 out of 20 patients (75%), 8 of whom (40%) experienced its disappearance while the granisetron was being administered. Vomiting was perfectly controlled in 5 out of 20 patients (25%) for 24 hours after the granisetron administration. No adverse event seemingly due to granisetron was observed. The result of this study confirmed the speedy effect granisetron on nausea induced by cancer chemotherapy including CDDP, but it stopped short of demonstrating sufficient efficacy for vomiting. Prophylactic use, therefore, seems more desirable in view of the patient's QOL, when a highly emetogenic anti-tumor drug, such as CDDP, is used.

Adult↗

Annexin V is localized in association with Z-line of rat cardiac myocytes.

OBJECTIVE: The aim of this study was to characterize a 33-kDa protein (p33) isolated from bovine liver and to determine the subcellular localization of the protein in rat cardiocytes as well as in non-cardiac tissues. METHODS: Cycles of calcium-induced precipitation coupled with EGTA-resolubilization were used to isolate crude annexins from bovine and rat tissues. Column chromatography was performed to purify the p33 from the crude annexins. The protein was identified as annexin V by partial amino acid sequence determination. Specificity of anti-annexin V antibody was examined by using Western blotting after one- or two-dimensional electrophoresis. For characterization of the protein, immunofluorescence microscopy and actin-binding assays were carried out. RESULTS: Immunofluorescence microscopy showed that annexin V was stained as a striated pattern along myofibrils on frozen sections of both atria and ventricles of adult rats. The striations were seen more clearly in cultured rat atrial myocytes. Examination of doubly stained cardiocytes with anti-annexin V and anti-alpha-actinin by a confocal laser scanning microscope suggests that annexin V is localized in association with the Z-line of rat cardiac myocytes. We also found that annexin V was stained intensely in actin-rich regions of non-cardiac tissues such as bile canaliculi of rat liver and brush border-terminal web region in the epithelial cells of both small intestine and kidney proximal tubular cells. F-actin binding experiments revealed that annexin V failed to bind to F-actin directly in vitro. CONCLUSION: Our results suggest that annexin V is a new component of the Z-line in rat cardiocytes and might be involved in regulation of its organization.

Actins↗

Immunohistochemical detection of an enamel protein-related epitope in rat bone at an early stage of osteogenesis.

Monoclonal antibody MI315 was produced against hamster tooth germ homogenate by in vitro immunization. It was found that MI315 reacted with enamel matrix, ameloblasts, and bone matrix at an early stage of osteogenesis. Decalcified tissues of rat femurs and mandibles were examined with MI315 using indirect immunofluorescence. In endochondral ossification of femurs, immunoreactivity was found in bone extracellular matrix (ECM) deposited on the surface of the cartilage core of primary spongiosa, but not in the cartilage core itself. In intramembranous ossification of 0-day-old rat mandibles, intense immunofluorescence was detected in bone ECM and a few young osteocytes, but not in osteoblasts. Immunoreactivity in bone ECM of 2-day-old rats decreased and almost disappeared from bone ECM of 4-day-old rats. Although in nondecalcified sections of 0-day-old rats, negligible immunofluorescence was detected in bone ECM which showed positive staining in decalcified tissues, the immunostaining appeared after decalcification using ethylenediaminetetraacetic acid (EDTA). These results indicate that a substance(s), which had a common epitope with an enamel-derived protein(s), existed in immature bone ECM of both endochondral and intramembranous ossification, and that it might be masked by bone mineral. Monoclonal antibody MI315 is a useful tool to investigate the time- and position-specific changes in osteogenesis and amelogenesis.

Animals↗

Demonstration of amelogenin in the enamel-free cusps of rat molar tooth germs: immunofluorescent and immunoelectron microscopic studies.

The enamel-free cusps of 1-4 day-old rat mandibular first molars were investigated using the monoclonal antibody En3 against rat amelogenin at light and electron microscopic levels in order to clarify whether the enamel-free cusp is virtually devoid of enamel. At 1 day after birth, there were presecretory ameloblast-like cells (PALCs), which were short and were not polarized, at the cusp tips. They were close to the outer enamel epithelium. Hematoxylin positive enamel matrix was not distinctly observed in the enamel-free cusp by light microscopy, but almost continuous immunofluorescence for amelogenin was detected at the interface between PALCs and dentin. The penetration of immunopositive material toward the dental pulp was also observed in the enamel-free cusp. At 4 day after birth, both in the frontal section and in the horizontal section, almost continuous immunofluorescence was recognized at the interface between PALCs and dentin in the enamel-free cusp. The penetration of amelogenin toward the dental pulp was not seen in the enamel-free cusp. By immunoelectron microscopy, immunolabelling was recognized in the Golgi apparatus of PALCs, in a layer of amorphous material at the interface between PALCs and dentin, and in stippled material-like substance in the intercellular space between PALCs. Although no basement membrane was observed beneath PALCs, they did not have Tomes' processes. These investigations suggest that PALCs in the enamel-free cusp differentiate into the secretory cells and that they can synthesize and secrete the amorphous material containing amelogenin at the interface between PALCs and dentin. The penetration of amelogenin toward the dental pulp might play a role in the interaction between PALCs and odontoblasts in the enamel-free cusp and/or the initiation of mineralization of predentin.

Ameloblasts↗

Primary and secondary culture of rat ameloblasts in serum-free medium.

Enamel is the hardest tissue in vertebrates. Ameloblasts are derived from epithelial cells and are responsible for enamel formation. They secrete enamel matrix components in which amelogenins are the major proteins, the biochemical properties of which are well known. However, little is known about the characteristics of ameloblasts themselves or about the functions of amelogenins. In this study, we developed a novel primary and secondary culture system for ameloblasts using a monoclonal antibody which recognized amelogenin (En3). The cell layer on dentine removed from rat mandibular incisors was isolated and cultured in low calcium, serum-free medium. Primary culture was performed on collagen-coated culture plates and typically, two types of cells appeared. One major type changed morphology after the addition of a high concentration of calcium to the medium. Expression of amelogenin was shown as cytoplasmic particles in these cells using En3. In the secondary culture, expression of amelogenins was also observed. In this system, the cells grew and maintained the expression of amelogenin for about 3 weeks.

Ameloblasts↗