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Biomedical subjects

K Yamazaki

Publications and source records attributed to K Yamazaki.

At least 37 records · Page 2Linked to original sources

Gene expression analysis of the CD4+ T-cell clones derived from gingival tissues of periodontitis patients.

The function of T cells infiltrating periodontitis lesions is complex and has not been fully elucidated. Here, we established T-cell clones from the gingival tissues of periodontitis patients and examined their gene expression. A total of 57 and 101 T-cell clones were established by means of immobilized anti-CD3 antibody and IL-2 from gingival tissues and peripheral blood, respectively. The gingival T-cell clones were derived from three patients, and the peripheral blood T-cell clones from two of these patients and a further patient whose gingival T-cell clones were not established. Gingival tissues were also obtained from a further 19 periodontitis patients. The expression of cytokines and molecules related to both regulatory function and tissue destruction were examined by means of reverse-transcription polymerase chain reaction. All the gingival T-cell clones expressed mRNA for TGF-beta1, CTLA-4, and CD25, and all the T-cell clones from peripheral blood expressed IFN-gamma and TGF-beta1 mRNAs. Most but not all the T-cell clones from gingival tissues and peripheral blood expressed mRNA for IFN-gamma and, CD25 and CTLA-4, respectively. The frequency of T-cell clones and gingival tissues expressing FOXP3, a possible master gene for mouse CD4(+)CD25(+) regulatory T cells, was very high (97%, 93%, and 100% for gingival T-cell clones, peripheral blood T-cell clones, and gingival tissues, respectively). Whereas the frequency of IL-4-expressing T-cell clones was lower for gingival T-cell clones (70% vs. 87%), the frequency of the gingival T-cell clones expressing IL-10 and IL-17 was higher than peripheral blood T-cell clones (75% vs. 62% for IL-10, 51% vs. 11% for IL-17). A similar expression profile was observed for gingival T-cell clones compared with gingival tissue samples with the exception of IL-4 expression, where the frequency of positive samples was lower in the gingival tissues (70% vs. 11%). These results suggest that the individual T cells infiltrating gingival lesions can express mRNA for both Th1 and Th2 cytokines as well as regulatory cytokines simultaneously.

Antigens, CD↗

Post-challenge hyperinsulinaemia rather than hyperglycaemia is associated with the severity of coronary artery disease in patients without a previous diagnosis of diabetes mellitus.

OBJECTIVE: To ascertain the prevalence of abnormal glucose metabolism in patients with coronary artery disease (CAD) but no previous diagnosis of diabetes mellitus (DM) and to examine the relation between the severity of CAD and responses of glucose and insulin to the glucose tolerance test. METHODS AND RESULTS: Abnormalities of glucose metabolism and insulin response were analysed in 144 patients with CAD without a previous diagnosis of DM who underwent both coronary arteriography and 75 g oral glucose tolerance test. The proportions of impaired and diabetic glucose tolerance were very high (39% for impaired and 21% for diabetic glucose tolerance); only 40% had normal glucose tolerance. The parameters of glucose metabolism were not associated with the number of diseased coronary arteries or the presence of previous myocardial infarction (MI). However, the insulin concentration at 60 minutes or 120 minutes after glucose challenge, insulin area, and the ratio of insulin to glucose area were significantly higher in patients with significant coronary stenosis and with previous MI. Fasting glucose concentration and most conventional risk factors did not predict post-challenge hyperinsulinaemia. CONCLUSION: Patients with CAD without a previous diagnosis of DM had a high prevalence of abnormal glucose tolerance. Post-challenge hyperinsulinaemia was associated with the number of diseased coronary arteries and the presence of previous MI. The insulin response to the glucose challenge test requires further investigation as a potential risk factor for CAD and a potential target for intervention.

Blood Glucose↗

Increased E2F-1 expression via tumour cell proliferation and decreased apoptosis are correlated with adverse prognosis in patients with squamous cell carcinoma of the oesophagus.

BACKGROUND: The retinoblastoma (Rb) pathway, which governs cell cycle progression, is frequently genetically altered in cancer, causing deregulated expression of the E2F-1 transcription factor, which promotes DNA synthesis and cell cycle progression. Recent studies show that E2F-1 also participates in apoptosis induction in a p53 dependent or independent manner. Despite its crucial role and paradoxical effects on cell turnover, the function of E2F-1 in human cancer is unclear. AIMS: To evaluate E2F-1 expression using immunohistochemistry in 43 surgically resected oesophageal squamous cell carcinoma (OSCC) specimens. METHODS: This study analysed the association of E2F-1 with tumour cell proliferation and apoptosis and the upstream regulators modulating these processes, and its impact on patient outcome. Tumour cell proliferation and apoptosis were assessed as percentage of MIB-1 positive or apoptotic cells (MIB-1 labelling index (MI) and apoptotic index (AI)), respectively. RESULTS: Entire specimens showed abnormal expression of one or more upstream regulators of pRb/E2F-1. Although E2F-1 positivity was not associated with the expression of upstream regulators, it showed a linear and positive correlation with MI but not AI. Patients with high MI, low AI, or high E2F-1 positivity had significantly shorter recurrence free survival. By multivariate analysis, high MI and low AI were independently associated with recurrence free survival, but E2F-1 was not. CONCLUSIONS: Increased cell proliferation and decreased apoptosis are associated with adverse prognosis in patients with OSCC. Although E2F-1 remains a controversial prognostic factor, its expression was closely associated with tumour cell proliferation and might influence clinical outcome, mainly via cell cycle progression.

Adult↗

Regulatory T-cells infiltrate periodontal disease tissues.

CD4+CD25+ regulatory T (Tr) cells are critical in regulating the immune response and thereby play an important role in the defense against infection and control of autoimmune diseases. Our previous studies demonstrated the involvement of autoimmune responses in periodontitis. The aim of this study was to identify CD4+CD25+ Tr cells in periodontitis tissues and compare them with those in gingivitis tissues. Immunohistological analysis of CD4, CD25, and CTLA-4 and the gene expression analysis of FOXP3, TGF-beta1, and IL-10 on gingival biopsies revealed the presence of CD4+CD25+ Tr cells in all tissues. In periodontitis, the percentage of CD4+CD25+ Tr cells increased with increasing proportions of B-cells relative to T-cells. FOXP3, a characteristic marker for CD4+CD25+ Tr cells, TGF-beta1 and IL-10 were expressed more highly in periodontitis compared with gingivitis. These findings suggest that CD4+CD25+ Tr cells and possibly other regulatory T-cell populations do exist and may play regulatory roles in periodontal diseases.

Aged↗

Foci of amorphous/granulofilamentous matrix in the extracellular domain of tumours. 2. Immunohistochemical and immunogold characterization of a fibronectin-rich matrix component.

The term FEAM (foci of extracellular amorphous matrix) has been used for discretely outlined areas of moderately dense material having a filamentous/granular substructure located in the extracellular matrix of tumours. In spite of being widespread in mesenchymal tumours especially, and often abundant, they have received little attention in terms of structure, composition and origin. Mostly, they have been regarded as a variant or a product of lamina ('basement membrane material'). However, they also appear in tumours whose cells should and do lack a lamina, such as giant-cell fibroblastoma and solitary fibrous tumour. This paper describes their fine structure in a variety of predominantly mesenchymal tumours, and documents their composition using light microscope immunostaining and immunogold labelling. Small amounts of type IV collagen and laminin were found focally and inconsistently among the five tumours by light microscope immunostaining, but fibronectin was strongly and consistently identified. Strong fibronectin staining was also identified by immuno-electronmicroscopy. These data suggest that FEAM represent a fibronectin-rich matrix constituent, which might be a common final product of either lamina or the external component of the subplasmalemmal linear density (focal adhesion). There is little support light microscopically for a relationship to immune-complexes or cryoglobulins.

Biomarkers, Tumor↗

MCG measurement in the environment of active magnetic shield.

MCG (Magnetocardiography) measurement by a SQUID gradiometer was attempted with only active magnetic shielding (active shielding). A three-axis-canceling-coil active shielding system, where three 16-10-16 turns-coil sets were put in the orthogonal directions, produces a homogeneous magnetic field in a considerable volume surrounding the center. Fluxgate sensors were used as the reference sensors of the system. The system can reduce environmental magnetic noise at low frequencies of less than a few Hz, at 50 Hz and at 150 Hz. Reducing such disturbances stabilizes biomagnetic measurement conditions for SQUIDs in the absence of magnetically shielded rooms (MSR). After filtering and averaging the measured MCG data by a first-order SQUID gradiometer with only the active shielding during the daytime, the QRS complex and T wave was clearly presented.

Electricity↗

Magnetic noise rejection in the MCG using independent component analysis.

In magnetocardiogram (MCG) measurements, the magnetic noise from any magnetic wire, used to suture the sternum after heart surgery, becomes a problem. As one method of noise rejection, independent component analysis is seen here to be effective. In this study, MCG measurements were carried out under conditions where a wire was attached to a normal subject, also where no wire was attached. Signal processing by independent component analysis was carried out in order to reduce the effect of magnetic noise from the wire. From the comparison of the waveforms after this signal processing and the waveforms where the wire was not attached, it was apparent that the magnetic noise from the wire was reduced. Thus, we have shown that independent component analysis is effective for the reduction of magnetic noise from a magnetic wire, in MCG measurements.

Electricity↗

Active magnetic compensation composed of shielding panels.

Magnetically shielded rooms (MSRs) with materials of high permeability and active shield systems have been used to shield magnetic noise for biomagnetic measurements up to now. However, these techniques have various disadvantages. Therefore, we have developed a new shielding system composed of shielding panels using an active compensation technique. In this study, we evaluated the shielding performance of several unit panels attached together. Numerical and experimental approaches indicated that the shielding factor of a cubic model composed of 24 panels was 17 for uniform fields, and 7 for disturbances due to car movement. Furthermore, the compensation space is larger than that of an ordinary active system using large coils rather than panels. Moreover, the new active compensation system has the important advantage that panels of any shape can be assembled for occasional use because the unit panels are small and light.

Electromagnetic Fields↗

Gastric and intestinal phenotypic marker expression in gastric carcinomas and recurrence pattern after surgery-immunohistochemical analysis of 213 lesions.

Both gastric and intestinal phenotypic markers are known to be expressed in gastric carcinomas, irrespective of their histologic type. In the present study, the relation between gastric and intestinal phenotypic marker expression in gastric carcinomas and the recurrence pattern after surgery was examined. The phenotypic marker expression of the tumour was determined by examining the expression of human gastric mucin (HGM), MUC6, MUC2 and CD10 in 213 advanced gastric carcinomas in 213 patients who had undergone a curative resection (97 died from recurrent gastric carcinoma and 116 were alive without recurrence at the end of the follow-up period). Tumours were classified into gastric (G), gastric and intestinal mixed (GI), intestinal (I) or unclassified (UC) phenotypes according to the immunopositivity of HGM, MUC6, MUC2 and CD10 stainings. The incidence of HGM-positive tumours and MUC2-negative tumours was significantly higher in tumours with peritoneal recurrence than in tumours without recurrence (73.3%, 44 out of 60 cases vs 54.3%, 63 out of 116 (P=0.022); and 70.0%, 42 out of 60 vs 38.8%, 45 out of 116 (P=0.0002), respectively). The incidence of G-phenotype tumours was also significantly higher in tumours with peritoneal recurrence than in tumours without recurrence (58.3%, 35 out of 60 cases vs 28.4%, 33 out of 116 (P=0.0002)). The incidence of MUC2-negative tumours and CD10-positive tumours was significantly higher in tumours with haematogenous recurrence than in tumours without recurrence (62.5%, 20 out of 32 cases vs 38.8%, 45 out of 116 (P=0.028); and 43.8%, 14 out of 32 vs 23.3%, 27 out of 116 (P=0.039); respectively). Our present findings show that the gastric and intestinal phenotypic marker expression of the tumour, determined by immunohistochemical staining for HGM, MUC6, MUC2 and CD10, can be used to predict the pattern of gastric carcinoma recurrence after curative resection.

Aged↗

Hypoxia-induced nitric oxide protects chondrocytes from damage by hydrogen peroxide.

OBJECTIVE: Because articular cartilage has no vascular supply, chondrocytes are hypoxic under normal physiological conditions. Nitric oxide (NO) plays an important role in chondrocyte damage, such as apoptosis. Although oxygen stress with hydrogen peroxide was found to cause chondrocyte damage, these data were obtained under normoxic (21% O2) conditions. We investigated the effects of hypoxia on hydrogen peroxide-induced chondrocyte damage METHODS: Bovine articular chondrocytes were used in this study. Proteoglycan (PG) synthesis and the induction of apoptosis were analyzed with [(35)S]-sulfate incorporation and annexin V staining, respectively. The induction of NO was examined using a fluorescent probe and RT-PCR. RESULTS: Cells maintained at 5% O2 had the maximum PG synthesis. Under normoxic conditions, hydrogen peroxide inhibited PG synthesis and induced annexin V positive cells in a dose-dependent fashion. However, in those cells cultured under hypoxic (5%) conditions, the hydrogen peroxide-induced annexin V expression was attenuated. Chondrocytes exposed to hypoxia showed induction of NO. When the hypoxia-induced NO was inhibited, the hypoxia-enhanced PG synthesis was abolished and hydrogen peroxide clearly induced cell damage. CONCLUSIONS: Endogenous NO induced by hypoxia protects chondrocytes from apoptosis induced by an oxidative stress.

Animals↗

[3H]alpha,beta-methylene ATP binding to P2X purinoceptor is unaffected by volatile anaesthetics.

BACKGROUND AND OBJECTIVE: The P2X receptor is responsible for fast excitatory neurotransmission in the central nervous system. This receptor is suggested as one of the targets of volatile anaesthetics. The study was undertaken to examine the site of action of volatile anaesthetics, especially for P2X purinoceptor-mediated neurotransmission. METHODS: The effect of sevoflurane and isoflurane on [3H]alpha,beta-methylene ATP binding was investigated in rat crude synaptic membranes. The crude synaptic membranes were prepared from Sprague-Dawley rat brains by centrifugation and then incubated with volatile anaesthetics or P2 receptor antagonists. RESULTS: [3H]alpha,beta-methylene ATP binding was unchanged by either sevoflurane or isoflurane at clinically relevant concentrations. Suramin, a P2 antagonist, significantly (P < 0.05) decreased the binding of [3H]alpha,beta-methylene ATP in a dose-dependent manner (68.7 +/- 14.7% at 10 micromol, 49.5 +/- 6.4% at 50 micromol, 24.3 +/- 5.7% at 100 micromol, n = 10, mean +/- SD), whereas pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), another P2 antagonist, did not affect the binding. CONCLUSIONS: The suppressive effect of volatile anaesthetics on ATP-mediated excitatory synaptic transmission could be one site of action. However, the blockade of ATP binding to P2X receptors is not a mechanism of action of volatile anaesthetics.

Adenosine Triphosphate↗

T-cell clonality to Porphyromonas gingivalis and human heat shock protein 60s in patients with atherosclerosis and periodontitis.

Individuals with periodontitis have been reported to have a significantly increased risk of developing coronary heart disease. Several studies have demonstrated that the immune response to heat shock protein 60 (HSP60) may be involved in the pathogenesis of both atherosclerosis and chronic periodontitis. To investigate this possible link between these diseases, cellular and humoral immune responses to HSP60 in atherosclerosis patients were compared with those in periodontitis patients and healthy subjects using human and Porphyromonas gingivalis HSP60 (GroEL) as antigens. Antibody levels to both human and P. gingivalis HSP60s were the highest in atherosclerosis patients, followed by periodontitis patients and healthy subjects. Clonal analysis of the T cells clearly demonstrated the presence of not only human HSP60- but also P. gingivalis GroEL-reactive T-cell populations in the peripheral circulation of atherosclerosis patients. Furthermore, these HSP60-reactive T cells seemed to be present in atherosclerotic lesions in some patients. These results suggest that T-cell clones with the same specificity may be involved in the pathogenesis of the different diseases.

Adult↗

Effect of periodontal treatment on the serum antibody levels to heat shock proteins.

We have shown previously that both humoral and cellular immune responses to heat shock protein 60 (HSP60) are elevated in chronic periodontitis patients compared with non-diseased subjects. The aim of the present study was to determine whether periodontal treatment could influence the level of serum antibodies to human HSP60 and Porphyromonas gingivalis GroEL, a bacterial homologue of human HSP60. Sera were obtained from 21 patients with moderate to advanced chronic periodontitis at the baseline examination and again after completion of treatment. Antibody levels were determined using an enzyme-linked immunosorbent assay. The mean anti-P. gingivalis GroEL antibody levels were down-regulated significantly by periodontal treatment when recombinant P. gingivalis GroEL was used as an antigen, whereas antibody levels to P. gingivalis GroEL-specific peptide were significantly elevated following successful periodontal therapy. The mean level of anti-human HSP60 antibody remained unchanged although individual levels of antibody either increased or decreased after periodontal treatment, suggesting that synthesis of these antibodies might be regulated independently during the course of periodontal infection. Although their regulatory mechanisms in chronic infection are not understood, further study would provide insight not only into the role of these antibodies in the pathogenesis of periodontitis but also into the possible link between periodontitis and systemic diseases such as coronary heart disease.

Adult↗

Decontamination effect of electrolysed NaCl solutions on carp.

AIMS: To evaluate the efficacy of electrolysed NaCl solutions (EW) for disinfecting bacterial isolates from carp, and the potential application of EW to reducing the bacterial load in whole carp and carp fillets. METHODS AND RESULTS: EW was produced by using a two-compartment batch-type electrolysed apparatus. Pure cultures (in vitro), whole carp (skin surface) and carp fillets were treated with EW to detect its antimicrobial effects. The anodic solution [EW (+)] completely inhibited growth of the isolates. Furthermore, dipping the fish samples in EW (+) reduced the mean total count of aerobic bacteria on the skin of whole carp and in fillets by 2.8 and 2.0 log(10), respectively. The cathodic solution [EW (-)] also reduced growth of the isolates from carp by ca 1.0 log(10). Moreover, the total counts of aerobic bacteria in whole carp (on the skin) and fillets were reduced by 1.28 and 0.82 log(10), respectively. CONCLUSIONS: EW (+) has a strong bactericidal effect on bacteria isolated from carp. SIGNIFICANCE AND IMPACT OF THE STUDY: Treatment with EW (+) could extend the shelf life of these fish.

Animals↗

Endobronchial ultrasonography with guide-sheath for peripheral pulmonary lesions.

The usefulness of endobronchial ultrasonography (EBUS) with guide-sheath (GS) as a guide for transbronchial biopsy (TBB) for diagnosing peripheral pulmonary lesions (PPL)s and for improving diagnostic accuracy was evaluated in this study. EBUS-GS-guided TBB was performed in 24 patients with 24 PPLs of < or =30 mm in diameter (average diameter=18.4 mm). A 20-MHz radial-type ultrasound probe, covered with GS was inserted via a working bronchoscope channel and advanced to the PPL in order to produce an EBUS image. The probe with the GS was confirmed to reach the lesion by EBUS imaging and X-ray fluoroscopy. When the lesion was not identified on the EBUS image, the probe was removed and a curette was used to lead the GS to the lesion. After localising the lesion, the probe was removed, and TBB and bronchial brushing were performed via the GS. Nineteen peripheral lesions (79.2%) were visualised by EBUS. All patients whose PPLs were visible on EBUS images subsequently underwent an EBUS-GS-guided diagnostic procedure. A total of 14 lesions (58.3%) were diagnosed. Even when restricted to PPLs <20 mm in diameter, the diagnostic sensitivity was 53%. In conclusion, endobronchial ultrasonography with guide sheath-guided transbronchial biopsy was feasible and effective for diagnosing peripheral pulmonary lesions.

Adult↗

Systemic lupus erythematosus with hepatic aneurysm, valsalva sinus aneurysm and associated polyangiitis: aneurysmal wall remodeling with dense fibrosis and calcification mediated by residual smooth muscle cells.

Isolated case reports have demonstrated the combination of hepatic aneurysm and systemic lupus erythematosus (SLE). The author experienced a rare autopsy case of a 36-year old Japanese male with SLE, a hepatic aneurysm, a Valsalva sinus aneurysm and associated polyangiitis. In the histopathological and ultrastructural examinations of the postmortem organs, most of the angiitis lesions were in the scar phase associated with the histopathological features of collagenous fibrosis with the diminution of the smooth muscle cells and elastic fibre layers. Massive calcification was seen on the wall and obliterative changes in the lumen with calcified thrombi. In the dense collagenous stroma of the aneurysmal wall, residual atrophic smooth muscle cells with immunohistochemical (HHF35+, alpha-smooth-muscle actin+, vimentin+ and desmin almost-) and ultrastructural features (spindle-shaped cells with a few rER and rich intracytoplasmic filaments with peripheral dense patches) were assumed to be involved in the histogenesis of the aneurysmal wall. Massive calcification and degeneration or dynamic remodeling of the extracellular matrices in the aneurysmal wall might be mediated by the residual smooth muscle cells. It was suspected that the generalized polyangiitis as a complication of SLE might have involved the intrahepatic arteries and Valsalva sinus wall and subsequently generated the aneurysm.

Adult↗

[Pleuropneumonectomy for pulmonary tuberculosis and chronic tuberculous empyema].

Surgical intervention is often necessary to treat either the active pulmonary tuberculosis or its sequelae such as destroyed lung, tuberculous empyema, and bronchial stenosis. Pleuropneumonectomy, which has been reported to be associated with high mortality and morbidity, is performed when the pleural space is completely obliterated due to previous inflammation or when preoperative empyema is complicated. In this article we report 3 patients receiving pleuropneumonectomy for pulmonary tuberculosis or chronic tuberculous empyema in recent 3 years. The mean operation time is 5 hours and 56 minutes, and the mean volume of intraoperative bleeding amounted to 1,417 ml. Autologous blood transfusion was prepared for all 3 patients and transfused them during the operation. No mortality, but postoperative complications were seen in 2 patients (67%) with a history of diabetes mellitus. They were sternum infection and bronchopleural fistula, which were successfully treated conservatively. Diabetes mellitus is thought to be a major risk factor for not only tuberculous disease itself, but also postoperative morbidity. As for surgical techniques, wide skin incision with multiple thoracotomy is necessary to keep a good operative field, and special care must be taken for great vessels and nerves at extrapleural dissection. We conclude that pleuropneumonectomy is relatively risky but effective surgical procedure for adequately selected patients with destroyed lung and tuberculous empyema.

Aged↗