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Biomedical subjects

T Deguchi

Publications and source records attributed to T Deguchi.

At least 19 recordsLinked to original sources
PubMed1996

Calbindin-D28k-immunoreactivity in the trigeminal ganglion neurons and molar tooth pulp of the rat.

The cell body size and coexpression of carbonic anhydrase (CA), calretinin (CR) and calcitonin gene-related peptide (CGRP) of primary neurons with calbindin-D28k (CB) was examined in the trigeminal ganglion (TG) of the rat. CB-immunoreactive (-ir) cells were mostly large and preferentially distributed in the maxillary and mandibular divisions of the TG. 48% of CB-ir TG cells exhibited enzyme CA activity. 10% of CB-ir TG cells contained CR-ir. Most TG cells coexpressing CB- and CR-irs were localized to the maxillary and mandibular divisions and exhibited CA activity. 6.5% of CB-ir TG cells coexisted with CGRP-ir. 46% of TG cells coexpressing CB and CGRP exhibited CA activity. The innervation of the molar tooth pulp by CB-ir TG primary neurons was also examined. CB-ir thick and smooth nerve fibers projected from the root pulp to the pulp horn and the roof of the pulp chamber, where they became thinner and rarely entered the subodontoblastic layer. However, they could not be traced to the odontoblastic layer, predentin or dentine. The distribution pattern of CB-ir pulpal fibers was different from that of CR-ir ones. The trigeminal neurons cells retrogradely labeled with fast blue (FB) from the maxillary molar tooth pulp contained CB- and CR-irs. 23% and 1% of the labeled cells were immunoreactive for CB and CR, respectively. The coexpression of CB- and CR-immunoreactivities (-irs) in FB-labeled cells was negligible. An immunoelectron microscopic method revealed that 21% of pulpal nerve fibers were immunoreactive for CB, and that all CB-ir nerve fibers in the root pulp were myelinated. The present study indicated that the tooth pulp primary neurons contained CB-ir but did not coexpress CB- and CR-irs and that these neurons projected their myelinated axons to the pulp.

Animals↗
PubMed1996

Cytogenetic analyses of hepatocellular carcinoma by in situ hybridization with a chromosome-specific DNA probe.

BACKGROUND: Numerical chromosome analysis has been established in solid tumors by using in situ hybridization (ISH) with a chromosome-specific probe. We analyzed human hepatocellular carcinoma (HCC) by ISH for chromosome 17 and investigated the correlation of its copy number with histologic malignancy, proliferative activity, p53 mutation, and DNA ploidy. METHODS: Chromosome 17 was hybridized with a pericentromere-specific DNA probe directly on the tumor cells isolated from paraffin blocks of 25 surgically resected HCCs. Proliferative activity was measured by Ki-67 immunohistochemistry, p53 mutation was analyzed by p53 immunohistochemistry, and DNA ploidy was estimated by cytofluorometry. RESULTS: Forty-four percent of the 25 HCCs showed numerical abnormality of chromosome 17. Many disomic cases had a less malignant histology, whereas many polysomic cases had a more malignant histology. The Ki-67 positive index of polysomic cases was higher than that of disomic cases. In 22 cases (88.0%), the copy number of chromosome 17 was well matched with DNA ploidy. However, the numerical abnormality of chromosome 17 did not show a significant correlation with p53 mutation. Two of four HCCs that showed histologic heterogeneity were also heterogenous on ploidy pattern and the copy number of chromosome 17. Conversely, there was one case in which only ISH could demonstrate heterogeneity, although the other features exhibited homogeneity. CONCLUSIONS: Numerical chromosome abnormalities correlated with the increase of histologic malignancy proliferative activity, and DNA ploidy. Moreover, ISH analysis was useful in assessing the intratumoral heterogeneity in HCC, especially when current methods failed to detect it. Thus, ISH provides information on important biologic features, such as malignant potential and intratumoral heterogeneity, in HCC.

Adult↗
PubMed1996

Cytodifferentiation of the odontoclast prior to the shedding of human deciduous teeth: an ultrastructural and cytochemical study.

BACKGROUND: In human deciduous teeth, odontoclastic resorption takes place at the pulpal surface of the coronal dentine prior to shedding, and this resorption shows clear time-related histological changes (Sahara et al., 1992). METHODS: Using this phenomenon as an observation system, we examined the cytodifferentiation of human odontoclasts by light and electron microscopy. For a histochemical marker of odontoclast differentiation and function, tartrate-resistant acid phosphatase (TRAP) activity was determined by light and electron microscopic enzyme histochemistry. RESULTS: As root resorption neared completion, TRAP-positive mononuclear cells were initially detected in the pulp chamber. They had abundant mitochondria, small lysosomes, and moderately developed rough endoplasmic reticulum throughout their cytoplasm. In these mononuclear cells, TRAP activity was localized in compartments of the biosynthetic pathway, i.e., in cisternae of the endoplasmic reticulum and Golgi lamellae, as well as small lysosomes. The TRAP-positive mononuclear cells first made contact with the predentine surface by their elongated cellular processes. After attachment, they spread out along the predentine surface and developed specialized membrane structures, clear zones, and ruffled borders. Next, they fused with each other on the predentine surface and formed typical multinucleate odontoclasts. After termination of their resorption function, the odontoclasts lost their ruffled borders and became detached from the resorbed surface. Most of the detached odontoclasts had numerous large pale vacuoles and secondary lysosomes and appeared to be in the process of degeneration. CONCLUSIONS: The present study demonstrates that: (1) odontoclasts differentiated from TRAP-positive mononuclear cells, which presumably originate from circulating progenitor cells, (2) membrane specialization of odontoclasts, i.e., development of a clear zone and ruffled border, is induced following their contact with the resorption surface, (3) multinucleation of odontoclasts takes place only after their attachment to the resorption surface, (4) mature multinucleate odontoclasts can resorb predentine as well as dentine in the same way as osteoclasts resorb bone, and (5) at the end of the resorption, odontoclasts gradually lose their ruffled borders and become detached from the resorbed surface.

Acid Phosphatase↗
PubMed1996

Failure to detect Mycoplasma fermentans, Mycoplasma penetrans, or Mycoplasma pirum in the urethra of patients with acute nongonococcal urethritis.

Urethral swab specimens collected from 108 male Japanese patients with acute nongonococcal urethritis (NGU) and from 50 Japanese men without NGU were examined for the presence of Mycoplasma fermentans, Mycoplasma penetrans, and Mycoplasma pirum by means of polymerase chain reaction-based assays. These mycoplasmas were not detected in any of the specimens, which suggests that they are unlikely to have a pathogenic role in acute NGU.

Acute Disease↗
PubMed1996

Morphologic adaptation of temporomandibular joint after chincup therapy.

The purpose of this study was to clarify the morphologic changes of the temporomandibular joint (TMJ) after chincup application, with longitudinal TMJ cephalometric laminagraphs. The subjects were 19 prepubertal patients with true, mild skeletal Class III malocclusions with anterior crossbite. All underwent chincup therapy from the beginning of treatment. The control subjects were 16 patients with functional anterior crossbite, with normal jaw relationships. Cephalometric laminagraphs and lateral cephalograms were obtained before and after treatment. Statistical analysis of the data revealed the results as follows: (1) The chincup therapy changed the direction of growth of the mandible, especially, the ramus swing-back. (2) The chincup group showed a relatively more slender mandibular neck compared with the control group. (3) The condylar heads were bent forward after the chincup application, and the glenoid fossa was deepened and widened. The clearance between condyles and fossae was decreased by the orthopedic force of the chincup appliance.

Adaptation, Physiological↗
PubMed1996

Stability of changes associated with chin cup treatment.

Twenty-four Japanese girls with anterior crossbite (Class III malocclusion) were selected for this study of the stability of changes associated with chin cup therapy. Pretreatment cephalometric measurements of the study sample were compared with those in a normal group. In addition, angular and linear measurements were also compared to assess the effectiveness of chin cup therapy in improving Class III skeletal components during the postretention period. The subjects showed characteristic values found in Class III malocclusion for SNB, ANB, and NPg to FH, and these values were significantly different from those in the normal group. The subjects who were past puberty showed more severe Class III skeletal patterns for ANB compared with the prepubertal subjects, and their initial Class III skeletal components showed more satisfactory improvement, including 2.0 degrees increase of SNA and 1.8 degrees increase of ANB during the postretention period.

Cephalometry↗
PubMed1996

Ciliary neurotrophic factor (CNTF) genotypes and CNTF contents in human sciatic nerves as measured by a sensitive enzyme-linked immunoassay.

To study the level of ciliary neurotrophic factor (CNTF) in human nervous tissues, we developed a sensitive enzyme-linked immunoassay using a specific antibody against human CNTF. This method allowed us to detect as little as 0.3 ng/ml of human CNTF with good linearity and accuracy. Using this method, CNTF levels were determined in human sciatic nerves obtained at autopsy from 21 amyotrophic lateral sclerosis (ALS) patients and 48 subjects who had died of other neurological diseases. CNTF genotypes were also determined. The results indicated that CNTF levels were high in the normal homozygotes and approximately halved in the heterozygote subjects. There was, however, no significant difference in CNTF levels in the sciatic nerves between ALS and other neurological disease patients, indicating that the CNTF level was mainly determined by its genotypes and that the level in the sciatic nerves was not reduced in ALS patients.

Amyotrophic Lateral Sclerosis↗
PubMed1996

Detection of low-level expression of P-glycoprotein in ACHN renal adenocarcinoma cells.

A highly sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay and a flow cytometric assay were used to examine ACHN cells for the expression of P-glycoprotein. The expression of P-glycoprotein was detected at the RNA and protein levels in ACHN cells by RT-PCR and flow cytometry, respectively. However, it was below the limit of detection by immunoblotting. The intracellular accumulation of adriamycin in ACHN cells was enhanced by verapamil, cyclosporin A and medroxyprogesterone acetate. Therefore, this study has demonstrated that low-level expression of P-glycoprotein detectable only by RT-PCR and flow cytometry plays a significant role in reducing the intracellular concentration of antitumor agents and thus contributes to the multidrug-resistant phenotype of ACHN cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

High-dose chemotherapy with peripheral blood stem-cell transplantation for hormone-refractory advanced carcinoma of the prostate: experience of two cases.

Two patients with hormone-refractory advanced prostate cancer were treated with high-dose chemotherapy and peripheral blood stem-cell transplantation. A satisfactory number of stem cells were collected by appropriately timed leukapheresis in these patients. The stem-cell samples collected from the first patient were positive for prostate-specific antigen messenger ribonucleic acid expression; such expression was not detected in the samples from the second patient. The first patient has remained in complete remission as of December 1995. The second patient died of the disease.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1996

Histological suture changes following retraction of the maxillary anterior bone segment after corticotomy.

By cephalometric and dental cast analyses, the effects of corticotomy in combination with 45-50 days retraction of the maxillary anterior segment were evaluated in five female Japanese monkeys (Macaca fuscata). In addition, microscopic changes of the related sutures were examined. According to the analyses of the dental casts, the retraction of the maxillary anterior segment in the experimental group was increased when compared with that of the control group. There was no change in occlusal molar relationship. The cephalometric analysis revealed that the "Point A" and "Metal Implant" on the mid-sagittal site between the upper central incisors in the experimental group showed more pronounced retraction than that in two untreated control animals. These results were histologically evaluated by changes of the suture structure: irregularity of functional arrangement of component cells and fibers, an increase in number of cells, resorption and formation of bone, and an enlargement of the suture width. Resorption of tooth roots and alveolar bone, and irregularity of the periodontal ligament were observed. In general, these histological changes appeared more extensive in the control specimens compared with the experimental specimens. The osseous histological changes were mainly observed in the compact bone area, while in the spongy bone area of both control and experimental specimens only minimal changes occurred. These results demonstrated the efficacy of combining a corticotomy procedure with retraction of the maxillary anterior bone segment by an orthodontic force.

Animals↗
PubMed1996

Quinolone-resistant Neisseria gonorrhoeae: correlation of alterations in the GyrA subunit of DNA gyrase and the ParC subunit of topoisomerase IV with antimicrobial susceptibility profiles.

Fifty-five clinical strains of Neisseria gonorrhoeae were examined for mutations in the gyrA and parC genes and for antimicrobial susceptibility profiles. The MICs of quinolones for 31 strains with alterations in GyrA were significantly higher than the MICs for 24 strains without such alterations. Eleven strains with alterations in both GyrA and ParC were significantly more resistant to fluoroquinolones than those with alterations in GyrA alone. The MICs of cephalosporins for these strains were also significantly higher than those for other strains.

Amino Acid Sequence↗
PubMed1996

Characterization of manganese peroxidases from the hyperlignolytic fungus IZU-154.

Four isozymes of manganese peroxidase (MnP) were identified in the culture fluid of the hyperlignolytic fungus IZU-154 under nitrogen starvation conditions. One of them was purified and characterized kinetically. The specific activity and Kcat/K(m) value of the MnP from IZU-154 were 1.6 times higher than those of the MnP from a typical lignin-degrading fungus, Phanerochaete chrysosporium. Two cDNAs encoding MnP isozymes from IZU-154 were isolated. The coding sequence of the two cDNAs, IZ-MnP1 cDNA and IZ-MnP2 cDNA, were 1,152 (384 amino acids) and 1,155 (385 amino acids) bp in length, respectively. They exhibit 96.2% identity at the nucleotide level and 95.1% identity at the amino acid level. Southern blot analysis indicated that two MnP isozyme genes exist in IZU-154 genomic DNA. The primary structures of two MnPs from IZU-154 were similar to those of MnPs from P. chrysosporium. The amino acid sequences including the important residues identified in MnPs from P. chrysosporium, such as the manganese-binding residues, the calcium-binding residues, the disulfide bonds, and the N-glycosylation site, were conserved in the two deduced IZ-MnPs. However, several discrepancies were found in the context around the distal histidine residue between MnP from IZU-154 and MnP from P. chrysosporium, which likely led to the difference in the kinetic parameters for MnP function.

Amino Acid Sequence↗
PubMed1996

Comparison among performances of a ligase chain reaction-based assay and two enzyme immunoassays in detecting Chlamydia trachomatis in urine specimens from men with nongonococcal urethritis.

We evaluated the performances of a ligase chain reaction (LCR)-based assay and two enzyme immunoassays (Chlamydiazyme and IDEIA) in the detection of Chlamydia trachomatis in urine specimens. We compared the results of testing urine specimens by these assays with those of urethral swab culture by examining samples from 131 men with nongonococcal urethritis. Discrepant results were analyzed by testing urethral swab specimens for C. trachomatis by a PCR-based assay. After the resolution of discrepant results, the sensitivity of urethral swab culture was 85.3%, whereas those of the LCR assay, Chlamydiazyme, and IDEIA with urine specimens were 94.1, 82.4, and 94.1%, respectively. The LCR assay and IDEIA were more sensitive than was urethral swab culture. In addition, the LCR assay, with a sensitivity equal to that of IDEIA, was more specific. Overall, the LCR assay proved to be superior to the enzyme immunoassays in detecting C. trachomatis in urine specimens. Testing urine specimens by LCR assay should be a helpful alternative method for diagnosing C. trachomatis urethral infection in men with nongonococcal urethritis.

Chlamydia Infections↗
PubMed1996

Rapid detection of point mutations of the Neisseria gonorrhoeae gyrA gene associated with decreased susceptibilities to quinolones.

Mutations in the gyrA gene resulting in amino acid changes at Ser-91 and Asp-95 are significantly associated with decreased susceptibilities to quinolones in Neisseria gonorrhoeae. To detect these mutations, we developed a rapid and simple assay based on amplification of the region of the gyrA gene containing the mutation sites by PCR and digestion of the PCR product with a restriction enzyme. A naturally occurring HinfI restriction site was present in the region containing the Ser-91 codon, and an artificial HinfI restriction site was created in the region containing the Asp-95 codon by the method of primer-specified restriction site modification. The mutations generating alterations at Ser-91 and Asp-95 were detected as restriction fragment length polymorphisms of the PCR products digested with HinfI. Fifty-five clinical strains of N. gonorrhoeae were examined for mutations in the gyrA gene by this method. Mutations at Ser-91 and/or Asp-95 were detected in all the 31 strains in which the mutations had been confirmed by DNA sequencing. Our method allows simultaneous testing of a large number of strains and provides results within 8 h. This rapid and simple assay could be a useful screening device for genetic alterations associated with decreased susceptibilities to quinolones in N. gonorrhoeae and could facilitate epidemiological studies on clinical isolates of N. gonorrhoeae with decreased susceptibilities to quinolones.

Base Sequence↗
PubMed1996

Interleukin (IL)-1 beta, IL-6, tumor necrosis factor-alpha, epidermal growth factor, and beta 2-microglobulin levels are elevated in gingival crevicular fluid during human orthodontic tooth movement.

Bone remodeling is a complex process regulated by several mediators. Recent work has revealed that cytokines and growth factors have significant effects on bone cell metabolism. However, little information is available concerning the production of cytokines during orthodontic tooth movement in human subjects, and there is no non-invasive model for determining the production of cytokines. Therefore, the purpose of this study was to identify and quantify the various cytokines in human gingival crevicular fluid (GCF), and to investigate the changes in their levels during orthodontic tooth movement. Twelve patients (mean age, 14.4 years) were used as subjects. An upper canine of each patient having one treatment for distal movement served as the experimental tooth, whereas the contralateral and antagonistic canines were used as controls. The GCF around the experimental and the two control teeth was taken from each subject immediately before activation, and at 1, 24, and 168 hr after the initiation of tooth movement. Cytokine levels were determined by ELISAs. The concentrations of interleukin (IL)-1 beta, IL-6, tumor necrosis factor-alpha, epidermal growth factor, and beta 2-microglobulin were significantly higher in the experimental group than in the controls at 24 hr after the experiment was initiated. All the cytokines remained at baseline levels throughout the experiment for the two control groups. In contrast to cytokine alteration, the amount of total protein in the GCF exhibited a gradual increase, but no significant difference was observed between the control and experimental groups. Since all cytokines in GCF play an important role in the bone remodeling processes in vitro, the present results indicate that the changes in cytokines in GCF are associated with orthodontic tooth movement.

Adolescent↗
PubMed1996

Trial production of titanium orthodontic brackets fabricated by metal injection molding (MIM) with sintering.

Safety and esthetics are two indispensable factors to consider when fabricating orthodontic brackets. However, these factors are not easily achieved when conventional techniques (including forging and casting) are used in the mass production of titanium brackets, albeit the brackets exhibit excellent biocompatibility. In the present study, orthodontic brackets were manufactured by metal powder injection molding with sintering. Brackets with three different base designs were made and subjected to compression shear tests for evaluation of their bonding strength to enamel substrate. The shapes given to the dimple of the base were spherical, oval, and grooved. The maximum shear forces for each type were 11.1 kgf, 7.6 kgf, and 18.5 kgf, respectively. The bonding strengths of the titanium bracket were equivalent to those obtained with conventional stainless steel brackets. Moreover, uniform distribution of Vickers hardness values (average, 240 +/- 40 Hv) measured at three locations indicated that the titanium bracket was uniformly sintered. Accordingly, titanium brackets thus fabricated exhibit a potential for clinical application.

Analysis of Variance↗