Biomedical subjects

M Shimono

Publications and source records attributed to M Shimono.

At least 19 recordsLinked to original sources

PubMed2007

Osteopontin as biomarker in early invasion by squamous cell carcinoma in tongue.

BACKGROUND: Osteopontin (OPN) expression in squamous cell carcinoma (SCC) of the tongue has not been clearly elucidated. METHODS: We selected 46 cases of tongue SCC and investigated the expression of OPN by immunohistochemical staining. The immunopositive reaction and score for each case were semiquantitatively evaluated. RESULTS: Scores were significantly higher in carcinoma nests than in neighboring normal epithelium or epithelial dysplasia. The OPN was expressed clearly in the cytoplasm of carcinoma cells. In cases of early invasive carcinoma, in particular, expression of OPN showed a remarkable increase at the invasion front compared with the non-invaded regions. However, there was no significant correlation between expression of OPN in the primary tumor nest and lymphatic metastasis, recurrence, or survival rate. CONCLUSION: This suggests that OPN is a useful biomarker of early invasion by SCC in tongue.

Adult↗

PubMed2006

Konjak mosaic virus: the complete nucleotide sequence of the genomic RNA and its comparison with other potyviruses.

Konjak mosaic virus (KoMV) belongs to the genus Potyvirus, family Potyviridae. The complete nucleotide sequence of KoMV F isolate (KoMV F) was determined. The genome is 9,544 nucleotides long excluding the 3' terminal poly A tail and encodes a typical potyviral 350-kDa polyprotein of 3,087 amino acids. Phylogenetic analysis using known potyvirus polyproteins shows that KoMV constitutes a branch with yam mosaic virus, close to another branch including Japanese yam mosaic virus, turnip mosaic virus, scallion mosaic virus and lettuce mosaic virus. The 3' terminal 1,842 nucleotides of a different isolate of KoMV, K-2, was also determined, covering the C-terminal 292 amino acids of the nuclear inclusion protein b (NIb), coat protein (CP), and the 3' untranslated region. The amino acid sequences of the KoMV F CP and the nucleotide sequences of the KoMV F 3' untranslated region showed 92.5 and 90.5% identity to the corresponding genes of K-2, 88.7-96.8 and 92.7-94.4% to those of Zantedeschia mosaic virus (ZaMV) isolates, 87.5-89.7% and 85.5-90.3% to those of Japanese hornwort mosaic virus (JHMV) isolates. These results showed that KoMV is a distinct potyvirus and that KoMV, ZaMV, and JHMV are members of the same potyvirus species. Considering that KoMV was the first of these to be described, ZaMV and JHMV may be considered isolates of KoMV.

Amorphophallus↗

PubMed2006

Morphology of Malassez's epithelial rest-like cells in the cementum: transmission electron microscopy, immunohistochemical, and TdT-mediated dUTP-biotin nick end labeling studies.

BACKGROUND AND OBJECTIVE: It is known that epithelial islands are embedded in the cementum during tooth root formation, but details of this process remain unknown. The purpose of this study was to investigate the dynamic characteristics of Malassez's epithelial rest cells in the cementum during tooth root formation in pigs in vivo. MATERIAL AND METHODS: The first molars of 6-mo-old pigs were used in this study. Specimens were decalcified before being embedded in paraffin. Paraffin sections were investigated using TdT-mediated dUTP-biotin nick end labeling (TUNEL), immunohistochemical, and ultrastructural techniques. RESULTS: Malassez's epithelial rest cells were located close to the root surface at the apical one-third of the periodontal ligament, and epithelial clusters surrounded by distinct lamina cementia were sometimes observed in the cementum. TUNEL-positive cells were detected only in the cementum. Malassez's epithelial rest cells in the periodontal ligament were completely surrounded by basement membranes, but epithelial clusters in the cementum were only intermittently surrounded by such membranes. Cytokeratin-positive cells in the superstratum of the cementum were directly connected by cementocytes and by desmosome-like structures. However, organelles were scarce in the cytokeratin-positive cells in the substratum of the cementum, and the matrix of the cementum was deposited in the cells. CONCLUSION: These results suggest that the majority of the fragmented Hertwig's root sheath remains in the periodontal ligament and that some cells, which are connected to cementoblasts, are embedded in the cementum and progress to apoptosis.

Animals↗

PubMed2006

Responses of rat pulp cells to heat stress in vitro.

Although heat stress can cause irritation in the dentin/pulp complex, little is known about the thermotolerance of pulp cells and their response to heat stress. We investigated cultured rat pulp cell responses to heat stress. Cells were subjected to a temperature of 42 degrees C for 30 minutes, and HSPs, alkaline phosphatase activity, and gap-junctional communication were determined at various time points. Although only low levels of HSP70 expression were detected before heat treatment, heat shock markedly induced HSP70 expression, with it gradually increasing at 1 hour after being heated. HSP25, however, showed no dramatic change. Gap junction protein connexin43 rapidly degraded after heat treatment, recovering to normal levels within the following 6 hours. Alkaline phosphatase activity decreased immediately after heat stress, recovering after 1 hour. These results indicate that dental pulp possesses protective factors, including HSPs, and that it can recover viability of intercellular communication and alkaline phosphatase activity after heat stress.

Alkaline Phosphatase↗

PubMed2005

Correlation between reduction of syndecan-1 expression and clinico-pathological parameters in squamous cell carcinoma of tongue.

We examined expression of syndecan-1 in squamous cell carcinoma (SCC) of tongue using immunohistochemistry. Forty-three cases of SCC arising in lateral border of tongue were investigated. From the immunohistochemical staining pattern, the cases were divided into two groups based on expression of syndecan-1 at the supra-peripheral cells of the tumor nest: Group A, completely or mainly positive; Group B, sporadically positive or negative. Most poorly differentiated SCC cases were classified into Group B (81.8%). The number of Group B cases in T1-2 was different from that in T3-4. The number of cases where syndecan-1 expression was reduced was much greater in T3-4, and represented the majority of Group B (86.7%). More than 80% of Grade 4D cases were in Group B (83.3%) based on the Yamamoto-Kohama criteria. These results indicate that reduction of syndecan-1 correlates to histological grade, tumor size and mode of invasion in tongue SCC.

Adult↗

PubMed2005

Involvement of aquaporin-5 water channel in osmoregulation in parotid secretory granules.

Aquaporins (AQPs) are a family of channel proteins that allow water or very small solutes to pass, functioning in tissues where the rapid and regulated transport of fluid is necessary, such as the kidney, lung, and salivary glands. Aquaporin-5 (AQP5) has been demonstrated to localize on the luminal surface of the acinar cells of the salivary glands. In this paper, we investigated the expression and function of AQP5 in the secretory granules of the rat parotid gland. AQP5 was detected in the secretory granule membranes by immunoblot analysis. The immunoelectron microscopy experiments confirmed that AQP5 was to be found in the secretory granule membrane. Anti-AQP5 antibody evoked lysis of the secretory granules but anti-aquaporin-1 antibody did not and AQP1 was not detected in the secretory granule membranes by immunoblot analysis. When chloride ions were removed from the solution prepared for suspending secretory granules, the granule lysis induced by anti-AQP5 antibody was inhibited. Furthermore, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, an anion channel blocker, blocked the anti-AQP5 antibody-induced secretory granule lysis. These results suggest that AQP5 is, expressed in the parotid gland secretory granule membrane and is involved in osmoregulation in the secretory granules.

Animals↗

PubMed2005

Reduction of osteocalcin expression in aged human dental pulp.

AIM: To investigate the expression of osteocalcin mRNA in young and in aged human dental pulp tissue to determine the characteristics of osteocalcin expression. METHODOLOGY: Human dental pulp tissues of the third molars were obtained from healthy young (17-23 years) and aged (>50 years) subjects, and total RNA was extracted. Osteocalcin mRNA expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between the value in young and aged pulp was statistically analysed. RESULTS: Osteocalcin mRNA was detected in all samples of human dental pulp tissue homogenates by RT-PCR analysis. Osteocalcin mRNA was expressed in young adult dental pulp but was decreased in aged human dental pulp. QRT-PCR analysis also showed a reduced expression of osteocalcin mRNA in aged human pulp. Expression of osteocalcin in young human pulp was significantly higher (about sixfold) than in aged pulp (P<0.01, Mann-Whitney U-test). CONCLUSION: Reduction of osteocalcin expression may be associated with the loss of viability in human dental pulp tissue, and may be a characteristic of aged human dental pulps.

Adolescent↗

PubMed2004

Effects of multigrooved surfaces on osteoblast-like cells in vitro: scanning electron microscopic observation and mRNA expression of osteopontin and osteocalcin.

This study evaluated the behavior of osteoblast-like cells on multigrooved surfaces consisting of a combination of microgrooves and macrogrooves. A polystyrene substrate was fabricated with multigrooves with 90-degree, V-shaped microgrooves with a 2-microm pitch cut on trapezoidal macrogrooves, which had a 50-microm ridge width, a 50-microm wall width, a 50-microm bottom width, and 25-microm depth. Smooth polystyrene substrates were also prepared as controls. Rat bone marrow cells were cultured as osteoblast-like cells on the substrates for morphological evaluation using a scanning electron microscope, and for biochemical evaluation using the quantitative reverse transcriptase-polymerase chain reaction technique for osteopontin and osteocalcin mRNA expression. After 8 days of incubation, the osteoblast-like cells were aligned parallel to the surface grooves on the multigrooved substrates. After 16 days of incubation, a dense mineralized extracellular matrix (ECM) was produced along the multigrooves. The ECM on the multigrooved surface appeared oriented more in the direction of the grooves than on the smooth surface, and trapezoid-shaped macrogrooves of the ECM were cast upside down. Although there were not significant differences, the osteopontin and osteocalcin mRNA expressions of the osteoblast-like cells on the multigrooved surfaces tended to be higher than on smooth surfaces. These results suggest that multigrooves could be used to control the orientation of mineralized ECM as well as of cells, and also to enhance the production of mineralized ECM.

Animals↗

PubMed2004

Cell proliferation and tumour suppressor gene expression in iodine unstained area surrounding oral squamous cell carcinoma.

The purpose of this study was to investigate the relationship between epithelial dysplasia unstained with iodine and the expression of proliferating cell nuclear antigen (PCNA) and/or tumour suppressor gene (p53) and the existence of glycogen. Thirty cases of squamous cell carcinomas arising from the buccal mucosa and floor of the mouth were examined. Iodine unstained areas were diagnosed histopathologically as mild, moderate or severe epithelial dysplasia. Normal oral mucosa stained with iodine was used as a control group. There was no histochemical difference in the distribution or ratio of PAS-positive cells between the control and the mild epithelial dysplasia groups, however PAS stained areas of the moderate and the severe dysplasia groups were significantly decreased. Ultrastructurally, glycogen granules were not recognized in the moderate or severe dysplastic epithelia. Immunoreactive ratios of PCNA and p53 in the moderate and severe dysplastic groups were significantly higher than those of the control and the mild dysplasia groups. The positive ratio of PCNA was higher than that of p53, although the immunostaining patterns of PCNA- and p53-positive cells were quite similar. These results suggest that mild dysplastic epithelia that are stained with iodine may be in the category of normal epithelia, whereas both moderate and severe dysplasia that are un-stained with iodine may be suspected of malignant lesions.

Carcinoma, Squamous Cell↗

PubMed2004

Reduction of connexin 43 expression in aged human dental pulp.

AIM: To investigate the expression of connexin 43 (CX43) mRNA in young and old human dental pulp tissues to determine the characteristics of CX43 expression. METHODOLOGY: Samples were obtained from human dental pulp of healthy young (17-23 years) and aged (>50 years) subjects. CX43 expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between value in the young pulp and that in the aged pulp was statistically analysed. RESULTS: RT-PCR analysis of human dental pulp tissue detected CX43 mRNA in all the samples. CX43 was abundantly expressed in young adult dental pulp, but expression of CX43 mRNA was dramatically decreased in aged human dental pulp. QRT-PCR analysis also showed the reduced expression of CX43 in aged pulp, and expression of CX43 in young pulp was significantly higher (about 10-fold, P < 0.01, Mann-Whitney U-test). CONCLUSION: Reduction of CX43 expression may be associated with the loss of viability in human dental pulp, and is considered as one characteristic of aged pulp.

Adolescent↗

PubMed2004

Proliferative activities of epithelial and connective tissue cells in the rat periodontal regeneration using argyrophilic nucleolar organizer regions staining.

BACKGROUND AND OBJECTIVE: It is still an open question why long junctional epithelium can proliferate and occupies the root surface following periodontal surgery or experimentally produced periodontitis, and why the epithelium repopulated once on the root surface is replaced by the connective tissue. The aim of this study is to investigate the proliferative activity of the newly formed regenerative connective tissue and long junctional epithelium during wound healing by staining argyrophilic proteins of the nucleolar organizer regions (AgNORs). METHODS: Regenerative connective tissue and long junctional epithelium were experimentally created by insertion of a rubber piece between maxillary molars of rats for 1 week. After removal of the rubber, AgNORs parameters including nuclear area (NA), AgNORs area (AA), AgNORs percentage nuclear area (APNA), AgNORs number (AN) and nuclear number (NN) in regenerative connective tissue and long junctional epithelium were measured and analyzed statistically. RESULTS: APNA in long junctional epithelium after 1 and 4 weeks was over two times greater than that in the regenerative connective tissue. AA in long junctional epithelium was significantly higher than in regenerative connective tissue at 1 and at 4 weeks post-treatment. AN was higher in the central portion than at the root surface except at 20 weeks. APNA and AA decreased remarkably in long junctional epithelium at 12 weeks post-treatment (approximately half at 4 weeks), whereas in regenerative connective tissue, they did not change distinctly. CONCLUSIONS: These results imply that long junctional epithelium cannot supply sufficient epithelial cells because of their significantly low rates of proliferation, consequently long junctional epithelium becomes shorter after 12 weeks, whereas the proliferative activity of regenerative connective tissue maintains the same level of proliferation, and ultimately long junctional epithelium is replaced by regenerative connective tissue.

Animals↗

PubMed2003

Effects of multigrooved surfaces on fibroblast behavior.

Microgrooves have been investigated as substrates for the control of cell alignment. However, they are relatively too narrow and shallow for controlling the orientation of extracellular matrices (ECM) such as collagen. Multigrooves, a combination of microgrooves and macrogrooves, are expected to be able to control the orientation of both cells and ECM. This study investigated a method for fabricating multigrooves and evaluated fibroblast behavior on these novel surfaces. Multigrooved patterns were fabricated on a gold-alloy metal die, in which 90-degree V-shaped microgrooves with a 2-microm pitch were cut on trapezoidal macrogrooves. The macrogrooves had a 50- microm ridge width, a 50-microm wall width, a 50-microm bottom width, and a 25-microm depth. The grooves were made by an ultraprecision micromachine using a single crystal diamond. This metal die served as a template for making surface replicas from polystyrene. Microgrooved and smooth polystyrene replicas also were prepared as comparative substrates. Mouse fibroblast L929 cells were cultured in each type of replica substrate for 7 to 21 days. After these periods, the cells were fixed with 2.5% glutaraldehyde, treated with conventional methods, and, finally, observed by SEM. Confocal laser scanning microscopy was performed to investigate ECM formation. The multigrooved metal die exhibited the desired sharp configuration without defects. The dimensional values of the multigrooves on the polystyrene replicas were almost the same as the designed values. The fibroblasts on the multigrooved and microgrooved substrates were aligned parallel to the surface grooves after 7 days of incubation. In contrast to the microgrooved and flat surfaces, a dense extracellular matrix was produced along the multigrooves after 21 days of incubation. These results suggest that multigrooves can control the orientation of ECM as well as cells and thus enhance the production of ECM.

Animals↗

PubMed2003

Oncocytic carcinoma arising in submandibular gland with immunohistochemical observations and review of the literature.

We report a case of oncocytic carcinoma arising in submandibular gland. The tumour occurred in the left submandibular gland of an 82-year-old Japanese man. Histologically, the tumour was mostly composed of large cells with eosinophilic granules in the cytoplasm and they were arranged in the solid sheets, islands with duct-like structure and cords. The tumour cells had aggressively invaded muscles and perineural tissues, and cervical lymphatic metastasis was frequently observed. Histochemically, the tumour cells were strongly positive for phosphotungstic acid-hematoxylin (PTAH) stain, and we diagnosed this malignant tumour as oncocytic carcinoma. Immunohistochemically, the tumour cells reacted positively for cytokeratin 7, 8, 19, epithelial membrane antigen (EMA), alpha-1-antichymotrypsin and carcinoembryonic antigen (CEA), but negative for cytokeratin 13, 14, smooth muscle actin (HHF35) and S-100 protein (S-100). Tumour was diagnosed as oncocytic carcinoma in submandibular gland. Its characteristics are discussed in term of its histopathological and immunohistochemical features.

Adenocarcinoma↗

PubMed2003

Oncocytic tumor in myoepithelioma arising from the grossopalatine gland.

Oncocytoma or oncocytic change in salivary glands normally occurs in old patients and mostly in the parotid gland, but those arising from the grossopalatine gland in young patients are extremely rare. The present case shows that oncocytic ductal structures were observed in myoepithelioma, consisting of spindle, plasmacytoid or epithelioid cells. The oncocytic tumor contained large amounts of eosinophilic granular cytoplasm and small nuclei.

Adenoma, Pleomorphic↗

PubMed2002

Bone response to calcium phosphate-coated and bisphosphonate-immobilized titanium implants.

Thin calcium phosphate (Ca-P) coatings have been introduced to overcome the shortcomings of plasma-sprayed Ca-P coatings. In our previous experiments, thin Ca-P coatings also enabled the immobilization of bisphosphonate, which is a drug used to treat osteoporosis. The present study was designed to evaluate the bone response to titanium implants treated with a thin Ca-P coating and bisphosphonate. Forty cylindrical commercially pure titanium implants with a length of 7 mm and a diameter of 3 mm were used as test implant fixtures. Three groups of surface-treated implants were prepared: (1) blasted with titanium powder and etched with a solution of 10% HF + 5% HNO3 (control); (2) modified with 0.5-microm thick Ca-P coatings and rapid heat-treating, and (3) immobilized with bisphosphonate by immersion in pamidronate disodium solution (10(-2) M) for 24 h at 37 degrees C. These surface-treated implants were inserted into edentulous areas in the mandibular molar region of five beagle dogs. After implantation periods of 4 and 12 weeks, the bone implant interface was evaluated histologically and histomorphometrically. All measurements were statistically evaluated using a one-way ANOVA and Fisher PLSD test for multiple comparisons among the means. Four weeks after the implantation, higher percentage of bone contact was found around the thin Ca-P-coated implants compared to that of the control group. The highest percentage of bone contact was found around the bisphosphonate-immobilized implants after 12 weeks of implantation. These data suggest that a thin coating of calcium phosphate followed by bisphosphonate-immobilization is effective in the promotion of osteogenesis on surfaces of dental implants.

Animals↗

PubMed2002

Cell proliferation and cell death in periodontal ligaments during orthodontic tooth movement.

The purpose of this study was to investigate cellular responses of periodontal ligaments during tooth movement. Twenty-eight male Sprague-Dawley rats, weighing 200-250 g each, were used. To create the orthodontic force, elastic rubber blocks (0.65 mm thick) were inserted between the maxillary first and second molars on both sides. On days 3, 7, 10, 14, 21 and 28 after rubber block insertion, histopathological changes in both the tension and the pressure sides were examined by immunohistochemistry using proliferating cell nuclear antigen (PCNA) and by the TUNEL method. The ratios of PCNA-positive cells on the tension side 3 and 7 days after rubber block insertion were higher than those on the pressure side. The ratios of PCNA-positive cells on the tension side were highest at day 3 after insertion and then decreased during the remainder of the experimental period. On the pressure side, the ratios of PCNA-positive cells increased up to day 10 post insertion, then decreased from 14 to 28 days. The ratios of TUNEL-positive cells on both the tension and the pressure sides increased throughout the entire experimental period. These results indicate that the periodontal ligaments on the tension side are able to respond more promptly to orthodontic forces than those on the pressure side. The data also suggest that the ratios of cell proliferation and of cell death are closely related to the regeneration and reconstruction of periodontal ligaments which reflect the orthodontic force.

Animals↗

PubMed2001

Expression and localization of connexin 43 in rat incisor odontoblasts.

We have examined the expression and localization of connexin 43 (CX43) in rat incisor odontoblasts using reverse transcriptase polymerase chain reaction, in situ hybridization and immunohistochemistry. The CX43 gene was expressed in odontoblasts, and levels of gene expression increased throughout the course of development. In contrast, CX43 was down-regulated at an incisal segment. In situ hybridization analysis showed no positive signal for CX43 RNA in the cytoplasm of differentiating dental papilla cells, but faint positive signals for CX43 RNA were observed in early pre-odontoblasts. Those signals were more intense in young and in old odontoblasts, but were less in short odontoblasts. CX43 could not be detected in differentiating dental papilla cells or in early pre-odontoblasts by immunohistochemical localization, but a positive reaction was found in the late pre-odontoblast stage where predentin had been produced. The positivity gradually increased during odontoblast maturation, and was highest in the layer of old odontoblasts. These results indicate that odontoblasts that secrete actively dentin matrix components are tightly in contact with each other by gap junctions as suggested by the intense CX43.

Animals↗

PubMed2001

Immunohistochemical localization of amelogenin in human odontogenic tumors, using a polyclonal antibody against bovine amelogenin.

In the present study, we investigated the localization of amelogenin in odontogenic tumors, using an anti-amelogenin polyclonal antibody. In order to make the antibody, antisera against an amelogenin fraction obtained from the enamel matrix of unerupted bovine tooth was raised in rabbits. By Western blot analysis, a main band of 25 kDa and six minor bands (6.8, 12, 18, 20, 23, and 27 kDa) were detected under nonreducing conditions. Immunoreactivity for the amelogenin was observed in ameloblasts and in the immature enamel matrix of 4-day-old rats. In odontogenic tumors, positive reactions for amelogenin were localized in limited areas in adenomatoid odontogenic tumor, calcifying odontogenic cyst, primary intraosseous carcinoma and odontoma. The strongest immunoreactions were shown in enamel matrices in odontomas. Small mineralized foci in epithelial nests showed positive reactions, and a few reactions were observed in epithelium adjacent to the mineralized foci. In calcifying odontogenic cysts, some ghost cells in the lining epithelium were strongly stained. The results indicate that the present antibody for amelogenin is useful for the determination of odontogenic tumors, especially in those in which small mineralized foci are present in the epithelial nests.

Amelogenin↗