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Biomedical subjects

K Motegi

Publications and source records attributed to K Motegi.

At least 37 records · Page 2Linked to original sources
PubMed2000

5-Fluorouracil induces apoptosis through the suppression of NF-kappaB activity in human salivary gland cancer cells.

Activation of the transcription factor NF-kappaB results in protection against apoptosis, and the chemotherapeutic agent 5-Fluorouracil (5-FU) exerts its cytotoxic effect through the induction of apoptosis. Thus, we examined whether 5-FU could induce apoptosis through the suppression of NF-kappaB activity. We found that upon treatment of a human salivary gland cancer cell line (cl-1) with 5-FU, the NF-kappaB activity was suppressed in a time-dependent manner. This inhibition was mediated by a prevention of the degradation of the inhibitory IkappaB-alpha protein. In addition, the expression of TRAF-2 and cIAP-1, which are transcriptionally regulated by NF-kappaB and function as anti-apoptotic molecules through the interruption of caspase pathway, was also inhibited by 5-FU. Finally, the activity of caspase-8 and caspase-3 showed a significant increase in response to 5-FU. By flow cytometric analysis, 5-FU did not affect the expression level of Fas on the cell surface. Thus, our results suggest that one of the molecular mechanisms involved in 5-FU-induced apoptosis in cl-1 cells may be due to the suppression of NF-kappaB activity, resulting in the activation of the pro-apoptotic pathway.

Antimetabolites, Antineoplastic↗
PubMed2000

Suppression of tumor necrosis factor alpha-induced matrix metalloproteinase 9 production by the introduction of a super-repressor form of inhibitor of nuclear factor kappaBalpha complementary DNA into immortalized human salivary gland acinar cells. Prevention of the destruction of the acinar structure in Sjögren's syndrome salivary glands.

OBJECTIVE: We have previously shown that specific enhancement in acinar cells of proteolytic activity induced by tumor necrosis factor alpha (TNFalpha) may be responsible for the destruction of the acinar structure in the salivary glands of patients with Sjögren's syndrome. Because matrix metalloproteinase 9 (MMP-9) is regulated by nuclear factor kappaB (NF-kappaB), we investigated the effect of a super-repressor form of inhibitor of nuclear factor kappaBalpha (srIkappaBalpha) on the suppression of TNFalpha-induced MMP-9 production in acinar cells. METHODS: Two srIkappaBalpha complementary DNA (cDNA)-transfected acinar cell clones (ACMT-6 and ACMT-7) and 1 empty vector-transfected cell clone (ACpRc-1) were established. After treatment of cell clones with TNFalpha, the expression of MMP-9 was examined. In addition, the effect of TNFalpha on cell growth and the morphogenetic behavior of cell clones cultured on type IV collagen-coated dishes were examined. RESULTS: TNFalpha induced the production of MMP-9 in the ACpRc-1 cell clone, but greatly suppressed MMP-9 production in ACMT-6 and ACMT-7 clones. No apparent cytotoxic effect of TNFalpha treatment was observed in these cell clones. When ACpRc-1 cells were seeded on type IV collagen-coated dishes in the presence of both TNFalpha and plasmin, type IV collagen interaction with the cells was lost and the cells entered apoptosis. However, even when ACMT-6 and ACMT-7 cells were cultured under the same culture conditions as those for ACpRc-1, these cell clones attached to the substrate and grew consistently without showing apoptosis. Conclusion. These observations indicate that suppression of TNFalpha-induced MMP-9 production by the introduction of srIkappaBalpha cDNA corrected the aberrant in vitro morphogenesis of acinar cells grown on type IV collagen.

Blotting, Western↗
PubMed2000

Confocal microscopy of Tomes' granular layer in dog premolar teeth.

Tomes' granular layer is the hypomineralized area of radicular dentin, but knowledge concerning it is limited. The present study was designed to investigate the structural characteristics of Tomes' granular layer in the dog's teeth by confocal microscopy. Permanent premolars of four beagles, two at 7 months and the other two at 14 months of age, were used for observation. During premolar root formation, the 7-month-old dogs were injected with calcein and alizarin red S for vital staining of dentin, and ground sections of the teeth were prepared. Both ground and decalcified-paraffin sections were made from the teeth of the 14-month-old dogs and stained with basic fuchsin or with hematoxylin and eosin. All sections were examined by fluorescence and confocal microscopy. In the ground sections, granules of Tomes' layer and dentinal tubules were stained with basic fuchsin and with calcein. The granules of Tomes' layer stained with calcein were seen only near the labeling lines by calcein. The granules of Tomes' layer appeared as bright spots in cross sections, and as lines in longitudinal sections. When the sections were cut tangentially through the surface of dentin, the granules of Tomes' layer showed a reticular structure. Most of the dentinal tubules were seen to pass between the granules and terminated in the dentin-cementum junction. Looped tubules were not found in this area. In the paraffin sections stained with hematoxylin and eosin, extracellular matrix of dentin showed fluorescence of various intensities and dentinal tubules appeared dark. At the surface of the radicular dentin, the granules of Tomes' layer appeared as fluorescent fibers running parallel to the surface of dentin in the longitudinal sections. The fibers appeared as bright spots in the cross sections and as a mesh in the tangential sections. In the periodontal ligament, collagen fibers showed intense fluorescence, whereas most cells were negative. From these results we conclude that Tomes' granular layer of dog's teeth may be the collagen fiber bundles that remained uncalcified or hypocalcified within the radicular dentin.

Aging↗
PubMed2000

Induction of the activity of the histamine-forming enzyme, histidine decarboxylase, in mice by IL-18 and by IL-18 plus IL-12.

OBJECTIVE AND DESIGN: IL-18 shares functional properties with IL-12, which induces an elevation of histidine decarboxylase (HDC) activity in mouse tissues. Therefore, we examined the effects of IL-18 and IL-18+IL-12 on HDC activity. METHODS: IL-18, IL-12 or IL-18+IL-12 was intraperitoneally injected into BALB/c and C3H/HeN mice and their HDC activities were measured. RESULTS: IL-18, at effective antitumour doses, induced HDC activity in lung, liver, spleen and bone. The IL-18-induced HDC elevation was more rapid than that induced by IL-12. As with IL-12, repeated injections of IL-18 produced more HDC activity than a single injection. Repeated injections of IL-18+IL-12 induced serious illness and produced marked HDC activity. However, an inhibitor of HDC did not prevent the illness. CONCLUSIONS: IL-18 may be involved in the stimulation of histamine synthesis during the course of immune responses, though an elevation of HDC activity is not itself the direct cause of the serious illness induced by IL-18+IL-12.

Animals↗
PubMed2000

Immunological activation of dermal Langerhans cells in contact with lymphocytes in a model of human inflamed skin.

Langerhans cells play an important role in the skin's immune system. Little is known, however, about the antigen-presenting capacity of Langerhans cells in the context of skin inflammation. By immunohistochemistry we investigated the phenotypic characteristics of epidermal and dermal Langerhans cells and their spatial relationship with infiltrating lymphocytes. We studied skin flaps autotransplanted to the oral cavity to fill a defect after maxillofacial cancer surgery. In 15 of 21 cases sampled for the present study, the skin flaps were severely inflamed by Candida albicans infection. In contrast to the normal skin, such inflamed skin showed a marked increase in CD1a(+) dermal Langerhans cells. Double immunohistochemistry revealed that dermal Langerhans cells abundantly expressed B7-2 (CD86), a representative costimulatory molecule, and CD83, a marker of mature dendritic cells. Furthermore, these dermal Langerhans cells were in close contact with CD4(+)/CD45RO(+) lymphocytes. This cell-to-cell contact was further visualized by immunoelectron microscopy. Langerhans cells were also observed within lymphatic vessels that were identified by the expression of vascular endothelial growth factor receptor-3. Ki-67 labeling indices were 4.2% in CD4(+) T cells and 0.8% in CD8(+) T cells within the dermis. Factor XIIIa(+) dermal dendrocytes were distributed outside the clusters of lymphocytes and were not in contact with them. Our observations indicate that dermal Langerhans cells in the inflamed skin are activated to express common phenotypes to mature dendritic cells so that they could stimulate neighboring memory CD4(+) T cells.

Antigens, CD↗
PubMed2000

Purification and properties of alanine racemase from crayfish Procambarus clarkii.

Fresh water crayfish Procambarus clarkii is known to accumulate D-alanine remarkably in muscle after seawater acclimation, accompanied by an increase in alanine racemase activity. We have purified alanine racemase from crayfish muscle to homogeneity. The enzyme is a monomeric protein with a molecular mass of 58 kDa. It is highly specific to alanine and does not racemize L-serine, L-aspartate, L-glutamate, L-valine and L-arginine. The enzyme shows the highest activity at pH 9.0 in the conversion of L- to D-alanine and at pH 8.5 in the reverse conversion. Properties such as amino acid sequence, quaternary structure, pyridoxal 5'-phosphate (PLP)-dependency, pH-dependency and kinetic parameters seem to be distinct from those of the microbial alanine racemases. Various salts including NaCl at concentrations around seawater level were potently inhibitory for the activity in both of L- to -D and D- to -L direction.

Alanine Racemase↗
PubMed2000

Involvement of interleukin-1 in the inflammatory actions of aminobisphosphonates in mice.

Aminobisphosphonates (aminoBPs) are potent inhibitors of bone resorption. However, they cause undesirable inflammatory reactions, including fever, in humans. Intraperitoneal injection of aminoBPs into mice also induces inflammatory reactions, including a prolonged elevation of the activity of the histamine-forming enzyme, histidine decarboxylase (HDC). Because interleukin-1 (IL-1) is a typical pyrogen and a strong inducer of HDC, we examined whether aminoBPs induce inflammatory reactions in mice deficient in genes for both IL-1alpha and IL-1beta (IL-1-KO mice). In control mice, aminoBPs induced an elevation of HDC activity and other inflammatory reactions (enlargement of the spleen, atrophy of the thymus, exudate in the thorax and increase in granulocytic cells in the peritoneal cavity). These responses were all weak or undetectable in IL-1-KO mice. We have previously shown that lipopolysaccharides (LPSs) from Escherichia coli and Prevotella intermedia (a prevalent gram-negative bacterium both in periodontitis and endodontal infections) are capable of inducing HDC activity in various tissues in mice. In control mice treated with an aminoBP, the LPS-induced elevations of serum IL-1 (alpha and beta) and tissue HDC activity were both markedly augmented. However, such an augmentation of HDC activity was small or undetectable in IL-1-KO mice. These results, taken together with our previous findings (i) suggest that IL-1 is involved in the aminoBP-induced inflammatory reactions and (ii) lead us to think that under some conditions, inflammatory reactions induced by gram-negative bacteria might be augmented in patients treated with an aminoBP. In this study, we also obtained a result suggesting that IL-1-deficiency might be compensated by a second, unidentified, mechanism serving to induce HDC in response to LPS when IL-1 is lacking.

Alendronate↗
PubMed2000

Kidney lipids in galactosylceramide synthase-deficient mice. Absence of galactosylsulfatide and compensatory increase in more polar sulfoglycolipids.

UDP-galactose:ceramide galactosyltransferase (CGT) catalyzes the final step in the synthesis of galactosylceramide (GalCer). It has previously been shown that CGT-deficient mice do not synthesize GalCer and its sulfated derivative GalCer I(3)-sulfate (galactosylsulfatide, SM4s) but form myelin containing glucosylceramide (GlcCer) and sphingomyelin with 2-hydroxy fatty acids. Because relatively high concentrations of GalCer and SM4s are present also in mammalian kidney, we analyzed the composition of lipids in the kidney of Cgt(-/-) and, as a control, Cgt(+/-) and wild-type mice. The homozygous mutant mice lacked GalCer, galabiaosylceramide (Ga(2)Cer), and SM4s. Yet, they did not show any major morphological or functional defects in the kidney. A slight increase in GlcCer containing 4-hydroxysphinganine was evident among neutral glycolipids. Intriguingly, more polar sulfoglycolipids, that is, lactosylceramide II(3)-sulfate (SM3) and gangliotetraosylceramide II(3),IV(3)-bis-sulfate (SB1a), were expressed at 2 to 3 times the normal levels in Cgt(-/-) mice, indicating upregulation of biosynthesis of SB1a from GlcCer via SM3. Given that SM4s is a major polar glycolipid constituting renal tubular membrane, the increase in SM3 and SB1a in the mice deficient in CGT and thus SM4s appears to be a compensatory process, which could partly restore kidney function in the knockout mice.

Animals↗
PubMed1999

TGF-beta1 inhibits NF-kappaB activity through induction of IkappaB-alpha expression in human salivary gland cells: a possible mechanism of growth suppression by TGF-beta1.

Transforming growth factor (TGF)-beta is the prototype of a large superfamily of signaling molecules involved in the inhibition of proliferation of multiple epithelial cell types. Although accumulated evidence indicates the mechanisms of the antimitogenic effect of TGF-beta in a variety of cell types, the signal transduction mechanism underlying the regulation of NF-kappaB transcription factor by TGF-beta is largely unknown. Because NF-kappaB is not only involved in inflammatory responses but also mediates cell growth, we have investigated the effect of TGF-beta1 on the activity of NF-kappaB and the role of the inhibitory IkappaB-alpha protein in the growth of the human salivary gland cell clones NS-SV-AC, HSGc, and cl-1. NF-kappaB, which is usually maintained in an inactive state by protein-protein interaction with IkappaB, was found to be constitutively active in salivary gland cell lines. Upon treatment of cell clones with TGF-beta1, the NF-kappaB activity in NS-SV-AC and HSGc, but not in cl-1, which lacks the expression of TGF-beta type II receptor, was suppressed. In NS-SV-AC and HSGc, this inhibition was mediated by the induction of IkappaB-alpha at the mRNA and protein levels. The blocking of NF-kappaB subunit with a specific antisense oligonucleotide reduced the growth rate of all of the cell clones, including cl-1. Introduction of a mutated form of IkappaB-alpha cDNA into NS-SV-AC suppressed the growth rate of this cell clone. These results indicate that TGF-beta1 downregulates NF-kappaB activity through the induction of IkappaB-alpha expression in human salivary gland cells and that inhibition of NF-kappaB activity suppresses the growth rate of these cells.

Apoptosis↗
PubMed1999

Ratio of primary and secondary bile acids in feces: possible marker for colorectal cancer?

PURPOSE: Increases in fecal bile acids may play a role in colorectal carcinogenesis. The authors tested the hypothesis that high concentrations of primary and secondary bile acids are more common in patients with colon cancer than in patients with other gastrointestinal diseases. METHODS: In this retrospective study the secondary bile acid deoxycholic acid and the primary bile acid cholic acid were measured in the feces by enzyme-linked immunoabsorbent assay in 63 patients with colorectal cancer, 24 patients with gastric cancer, 11 patients with biliary disorders, and 47 healthy volunteers. RESULTS: Preoperatively, the mean deoxycholic acid values tended to be higher and the cholic acid values were significantly lower in patients with colorectal cancer than in healthy subjects. Patients with other gastrointestinal diseases had lower deoxycholic acid and cholic acid values than healthy subjects. In healthy subjects the deoxycholic acid to cholic acid ratio ranged from 0.10 to 2.86 (mean, 0.88), but in almost two-thirds, the ratio did not exceed 1. In contrast, the mean preoperative ratio in patients with colorectal cancer was 2.26 (range, 0.06-7.17; P < 0.0001) and tended to be higher in patients with advanced cancer and in those with sigmoid and rectal tumors. If 1.1 is taken as the upper limit of normal for deoxycholic acid to cholic acid ratio, 67 percent of patients with colorectal cancer had an abnormal value preoperatively. CONCLUSION: A high deoxycholic acid concentration and deoxycholic acid to cholic acid ratio may be indicators of colorectal cancer. Further study is needed to improve sensitivity and specificity, perhaps by combining fecal bile acid measurements with other tests, and a large prospective trial may be warranted to determine whether these measurements have value in screening for this common cancer.

Adult↗
PubMed1999

Confocal microscopy of dentinal tubules in human tooth stained with alizarin red.

The present study was designed to analyze the structures of dentinal tubules by confocal microscopy. Undecalcified ground sections of human teeth were stained with alizarin red in 0.1% KOH aqueous solution, and examined by confocal microscopy. Alizarin red stained dentinal tubules, interglobular dentine, granular layer of Tomes, and the surface of dentine. Interglobular dentine was seen between the outer and middle layers of coronal dentine. At the outer layer of coronal dentine, the dentinal tubules were thin and showed numerous branches. At the middle layer of coronal dentine, dentinal tubules displayed two types. The type I tubules are the dentinal tubules that do not show any nodular structures and the type II tubules are the dentinal tubules that appear bamboo-like with many nodules. In the cross section through the type II tubules, the nodules appeared as fine circular tubules surrounding the dentinal tubules. The circular tubules of nodules adhered to one side of the dentinal tubules. When the fluorescence images were compared with the images taken by transmission light mode, the fluorescence of dentinal tubules was seen at the inner surface of dentinal tubules, and the fluorescence of nodules was seen at interface between peritubular and intertubular dentine. Most of the dentinal tubules were of the type II tubules in the teeth from older individuals, whereas the type II tubules were scarce in the teeth from younger individuals. At the inner layer of coronal dentine, the dentinal tubules have no nodules and branches were scarce. The dentinal tubules of radicular dentine were different from those of coronal dentine. Most of the dentinal tubules were the type I tubules. Numerous fine branches were seen at the outer and middle layers of radicular dentine. No interglobular dentine was seen in the root except at the cervical part, and the granular layer of Tomes was also positive with alizarin red. At the cervical part of the root, interglobular dentine was present and the dentinal tubules displayed types I and II.

Adult↗
PubMed1999

Long-term changes in clinical signs and symptoms and disc position and morphology in patients with nonreducing disc displacement in the temporomandibular joint.

PURPOSE: The purpose of this study was to examine disc position and configuration, as well as clinical findings, during the natural course of nonreducing disc displacement of the temporomandibular joint (TMJ). PATIENTS AND METHODS: The subjects were 21 patients (24 affected joints) for whom a diagnosis of anterior disc displacement without reduction in the TMJ had been made on arthrography, but who had received no treatment for more than 12 months. The disc position and configuration were examined at a mean follow-up of 27.1 months using magnetic resonance imaging. Clinical signs and symptoms were also examined. As a control, 30 joints in 15 persons who had no current or previous TMJ symptoms were studied. RESULTS: The patients' clinical signs and symptoms tended to be alleviated during the follow-up period. In all patients, the disc was displaced anteriorly on mouth closure, and it did not reduce into normal position during mouth opening at the follow-up, whereas normal disc position was found in all of the controls. In the patients, the disc deviated from the normal biconcave configuration found in all of the controls, frequently showing enlargement of the posterior band, even thickness, folding, and convex configurations in the follow-up magnetic resonance imaging. CONCLUSION: Disc displacement does not resolve, and continued disc deformity is common in the natural course of nonreducing disc displacement of the TMJ, although the clinical signs and symptoms tend to become alleviated.

Adolescent↗
PubMed1999

Unique inflammatory features noted in intraorally transferred skin flaps: correlation with Candida albicans infection.

OBJECTIVE: The purpose of this study was to evaluate how well intraorally transferred skin flaps endure their new surroundings. STUDY DESIGN: Biopsy specimens were taken from 20 patients who had undergone microsurgical reconstruction and as pretransferred skin from 5 of these patients at the time of surgery. The study used immunohistochemistry for immunocompetent cells, differentiation markers for the epidermis and desmosomal proteins, and immunoelectron microscopy for desmosomal protein, in addition to routine histologic examination, including Sudan IV, periodic acid Schiff, and Grocott stains. We also measured the thickness of the epidermis and stratum corneum. Oral swabs from the skin flaps were examined for the presence of yeasts, particularly Candida albicans, by means of a culture method. RESULTS: According to the results of periodic acid-Schiff and Grocott staining, 20 cases were divided into 2 groups: fungal element-positive cases (n = 15) and fungal element-negative cases (n = 5). All swabs from the former were positive for Candida albicans. In these fungus-positive cases, histopathologic evaluation revealed marked diminution of stratum corneum and pronounced epidermal hyperplasia. Immunohistochemistry demonstrated the dermal infiltration of numerous immunocompetent cells-CD4+, CD8+, CD20+, CD68+, neutrophil elastase+, and HLA-DR+ cells-and the scarce infiltration of IgA+ and IgG+ cells. There were scattered CD1a+, CD4+, CD8+, and HLA-DR+ cells and elastase+ neutrophils in the epidermis. Expression of cytokeratin subtypes (10, 14, 16, and 19), involucrin, and tenascin showed the characteristic features of epidermal proliferation. Enumeration of Ki-67+ keratinocytes showed an increase, indicating epidermal proliferation. Expression of desmoglein 1 and desmocollin 1 in the epidermal keratinocytes was decreased in comparison with that in the pretransferred skin. Immunoelectron microscopy for desmoglein 1 confirmed the reduced immunoreactive deposits along the desmosomal plaques. In the fungus-negative cases, all such changes were a great deal milder. CONCLUSIONS: Taken together, our results demonstrate that most intraorally transferred flaps are affected by an inflammatory process that is induced by the influence of the wet oral environment. They present psoriasiform tissue reactions characterized by epidermal hyperproliferation that are mostly due to Candida albicans infection.

Adult↗
PubMed1999

Occlusal contact area, occlusal pressure, bite force, and masticatory efficiency in patients with anterior disc displacement of the temporomandibular joint.

The occlusal contact area, occlusal pressure, bite force, and masticatory efficiency were measured in 48 patients with anterior disc displacement (ADD) of the temporomandibular joint (TMJ). The results were compared with those of 30 normal controls without TMJ dysfunction. The values of occlusal contact area, bite force, and masticatory efficiency measured in patients with ADD were significantly smaller than those measured in the controls, although there was no difference in occlusal pressure between the two groups. The results of the measurements of 22 patients with ADD with reduction were also compared with those of 26 patients with ADD without reduction. There was no difference in any measurement between these patients subgroups. The analysis of occlusal contact area, bite force, and masticatory efficiency appeared to be useful methods in documenting the fact that masticatory function was impaired in patients with ADD of the TMJ.

Adenosine Triphosphate↗
PubMed1999

Implantation of octacalcium phosphate (OCP) in rat skull defects enhances bone repair.

Synthetic octacalcium phosphate (OCP) enhances bone formation if implanted into the subperiosteal region of murine bone. Such implanted OCP may be resorbed and replaced by bone with time. We hypothesized that OCP could be used as an effective bone substitute. To test this hypothesis, we designed the present study to investigate if bone repair in a rat skull defect is enhanced by the implantation of OCP. Rats were divided into two groups: OCP-treated animals and untreated controls. Six rats from each group were fixed at 4, 12, and 24 weeks after implantation. A full-thickness standardized trephine defect was made in the parietal bone, and synthetic OCP was implanted into the defect. After being examined radiographically, the specimens were decalcified and processed for histology. OCP implantation significantly promoted bone repair compared with the controls. A statistical analysis showed an increase in the area of radiopacity within the skull defect between week 4 and week 12. Histologically, bone was formed on the implanted OCP and along the defect margin at week 4. At week 12, the implanted OCP was surrounded by newly formed bone. At week 24, the defect was almost completely filled with bone. In the control, bone formation was observed only along the defect margin. The present results demonstrate that OCP could be used as an effective bone substitute.

Animals↗
PubMed1999

Disc position and morphology in patients with nonreducing disc displacement treated by injection of sodium hyaluronate.

Disc position and morphology were examined in 21 patients (22 joints) with nonreducing disc displacement of the temporomandibular joint (TMJ) at more than 12 months after undergoing injection of sodium hyaluronate into the superior joint space. The patients' clinical signs and symptoms improved during the follow-up period. In all patients, the disc was displaced anteriorly on mouth closure and did not reduce into the normal position during mouth opening at follow-up, whereas a normal disc position was found in all the controls. In the patients, the disc deviated from the normal biconcave configuration found in all of the controls on the follow-up magnetic resonance imaging (MRI). Disc displacement apparently is persistent and continued disc deformity common in patients, although the clinical signs and symptoms improved.

Adolescent↗