Biomedical subjects

M Tokuda

Publications and source records attributed to M Tokuda.

At least 73 records · Page 4Linked to original sources

PubMed1999

Vectorcardiographic evaluation of myocardial infarct size: comparisons with thallium myocardial scintigraphy.

OBJECTIVE: To determine the usefulness of vectorcardiography (VCG) in assessing myocardial infarct size. METHODS: The correlation of spatial and scalar parameters of VCG with the percent defect volume (%DV) of thallium myocardial single photon emission computed tomography (SPECT) was investigated in 63 patients with first-onset myocardial infarction (MI). VCG parameters included: (1) spatial parameters: magnitude, azimuth and elevation of the maximal vector, vectors at 20 ms and 30 ms, and (2) scalar parameters: amplitudes of 20 ms and 30 ms vectors at X, Y, and Z scalar leads abbreviated as X20, Y20, Z20, X30, Y30 and Z30, respectively. RESULTS: For anteroseptal MI, the azimuth of 30 ms vector and Z20 showed a significant correlation with %DV (r = 0.572, P < 0.05 and r = 0.832, P < 0.001) while in anteroseptal MI with involvement of lateral wall, the azimuth of 30 ms vector and X30 were correlated with %DV significantly (r = 0.775, and r = 0.780, P < 0.01). For inferior and inferoposterior MI, the elevation of 30 ms vector and Y30 were correlated well with %DV (r = 0.871, P < 0.01, r = 0.928, P < 0.001 for inferior MI and r = 0.678, P < 0.01, r = 0.760, P < 0.001 for inferoposterior MI). CONCLUSION: VCG parameters, especially scalar parameters, can be used to evaluate myocardial infarct size easily and non-invasively with remarkable accuracy.

Aged↗

PubMed1998

Osteoblastic differentiation is enhanced by rapamycin in rat osteoblast-like osteosarcoma (ROS 17/2.8) cells.

The effects of three immunosuppressants (rapamycin, FK506 and cyclosporin A) on the proliferation and differentiation of rat osteoblasts-like osteosarcoma cell line, ROS 17/2.8 (ROS) cells were examined in vitro. All immunosuppressants showed a direct inhibition on the proliferation of ROS cells with different potencies. Growth inhibition by rapamycin was stronger than that by FK506 or cyclosporin A. Rapamycin caused a significant increase in alkaline phosphatase (ALP) activity and in the expression of osteopontin and osteocalcin mRNAs. FK506 caused a moderate increase in ALP activity and a decreased expression of osteopontin mRNA. Cyclosporin A caused a decrease in ALP activity and in the expression of type 1 alpha 1 collagen mRNA. Our study indicates that rapamycin directly acts on ROS cells and induces osteoblastic differentiation, however, the effect of FK506 and cyclosporin A is weak. Rapamycin significantly enhances the differentiation induced by 1,25(OH)2-vitaminD3.

Animals↗

PubMed1998

Liquid chromatography of guanidino compounds using a porous graphite carbon column and application to their analysis in serum.

The retention mechanism of guanidino compounds on a porous graphitic carbon seemed to be mainly hydrophobic interaction, according to the retention factors in buffer solutions and the results of an analysis by computational chemical calculation using molecular mechanics (MM2). The baseline separation of ten guanidino compounds was achieved by the addition of a hydrophobic counterion. The retention mechanism may be dynamic ion-exchange. The stable system was applied to the analysis of guanidino compounds in serum from nephritic patients. The effluent was monitored by a post-column labeling detection method using ninhydrin. The detection limit of guanidino compounds was a few picomoles; however, that of creatinine was one hundredth of those of the other compounds. The reproducibilities of the peak height and area of the ten guanidino compounds using gradient elution were quite high, and the standard deviations were within a few percent (n=5), except for creatinine. The recovery of the compounds from serum was more than 90% (n=5). The reproducibility of retention times was within 1% (n=5).

Buffers↗

PubMed1998

Regulation of neuronal plasticity in the central nervous system by phosphorylation and dephosphorylation.

Neuronal plasticity can be defined as adaptive changes in structure and function of the nervous system, an obvious example of which is the capacity to remember and learn. Long-term potentiation and long-term depression are the experimental models of memory in the central nervous system (CNS), and have been frequently utilized for the analysis of the molecular mechanisms of memory formation. Extensive studies have demonstrated that various kinases and phosphatases regulate neuronal plasticity by phosphorylating and dephosphorylating proteins essential to the basic processes of adaptive changes in the CNS. These proteins include receptors, ion channels, synaptic vesicle proteins, and nuclear proteins. Multifunctional kinases (cAMP-dependent protein kinase, Ca2+/phospholipid-dependent protein kinase, and Ca2+/calmodulin-dependent protein kinases) and phosphatases (calcineurin, protein phosphatases 1, and 2A) that specifically modulate the phosphorylation status of neuronal-signaling proteins have been shown to be required for neuronal plasticity. In general, kinases are involved in upregulation of the activity of target substrates, and phosphatases downregulate them. Although this rule is applicable in most of the cases studied, there are also a number of exceptions. A variety of regulation mechanisms via phosphorylation and dephosphorylation mediated by multiple kinases and phosphatases are discussed.

Animals↗

PubMed1998

A calcineurin inhibitor, FK506, blocks voltage-gated calcium channel-dependent LTP in the hippocampus.

The effects of FK506, an immunosuppressant and protein phosphatase 2B (calcineurin) inhibitor, on the voltage-gated calcium channel (VGCC)-dependent long-term potentiation (LTP) were investigated in the CA1 region of mice hippocampal slices. VGCC-dependent LTP was induced either by a brief application of a potassium channel blocker tetraethyleneanmonium (TEA), or by a strong tetanic stimulation under the blockade of NMDA-receptors. FK506 (1-50 microM) produced dose-dependent inhibition on TEA-induced LTP. Cyclosporin A (CysA 50 microM), another calcineurin inhibitor, showed a similar inhibitory effect on TEA-induced LTP. FK506 (10 microM) also blocked the strong tetanus-induced LTP, but had no effect on the post-tetanic potentiation. By using a subthreshold weak tetanic stimulation protocol, we also found that low concentration of FK506 (1 microM) produced neither inhibition nor potentiation on VGCC-dependent LTP. These results showed FK506 and CysA exerted inhibitory effects on VGCC-dependent LTP, and suggest that calcineurin is involved in the processes of this kind of synaptic plasticity.

Animals↗

PubMed1998

Computed tomography measurements of overinflation in chronic obstructive pulmonary disease: evaluation of various radiographic signs.

Using computed tomography (CT), the authors determined significant signs of overinflation. Both the pulmonary function tests (PFT) and CT of 74 patients who underwent thoracic surgery for lung cancer (44 with normal lung function, 30 with chronic obstructive pulmonary disease) were reviewed. The following were correlated with forced expiratory volume in 1 second/forced vital capacity (FEV1/FVC): tracheal index (transverse/anteroposterior diameter), sterno-aortic distance, thoracic cage ratios (anteroposterior/transverse diameters) at the tracheal carina (TC1) and 5 cm below (TC2); and depth of the azygoesophageal recess and the presence of intercostal lung bulging (ILB). Significant correlations were observed between FEV1/FVC and tracheal index (r = 0.578, p < 0.0001), TC1 (r = -0.523, p < 0.0001), TC2 (r = -0.533, p < 0.0001), and ILB (r = -0.462, p < 0.0001). Correlations were significant but weak between FEV1/FVC and sterno-aortic distance (r = -0.351, p = 0.0027) and depth of the azygoesophageal recess (r = -0.308, p = 0.0085). Reduced tracheal index and increased anteroposterior diameter of the thoracic cage correlated most significantly with a pulmonary function index of chronic airway obstruction.

Adult↗

PubMed1998

The Pasteurella haemolytica 35 kDa iron-regulated protein is an FbpA homologue.

In a previous investigation, a 35 kDa iron-regulated protein was identified from total cellular proteins of Pasteurella haemolytica grown under iron-depleted conditions. This study reports identification of the gene (fbpA) encoding the 35 kDa protein based on complementation of an entA Escherichia coli strain transformed with a plasmid derived from a P. haemolytica lambda ZAP II library. Cross-reactivity was demonstrated between an anti-35 kDa mAb and a 35 kDa protein expressed in this strain. Furthermore, a translated ORF identified on the recombinant plasmid corresponded with the N-terminal amino acid sequence of the intact and a CNBr-cleaved fragment of the 35 kDa iron-regulated protein. Nucleotide sequence analysis of the gene encoding the 35 kDa protein demonstrated homology with the cluster 1 group of extracellular solute-binding proteins, especially to the iron-binding proteins of this family. Complete sequence analysis of the recombinant plasmid insert identified three other predominant ORFs, two of which appeared to be in an operonic organization with fbpA. These latter components (fbpB and fbpC) showed homology to the transmembrane and ATPase components of ATP-binding cassette (ABC)-type uptake systems, respectively. Based on amino acid/DNA sequencing, citrate competition assay of iron affinity and visible wavelength spectra, it was concluded that the P. haemolytica 35 kDa protein functions as an FbpA homologue (referred to as PFbpA) and that the gene encoding this protein is part of an operon comprising a member of the FbpABC family of iron uptake systems. Primary sequence analysis revealed rather surprisingly that PFbpA is more closely related to the intracellular Mn/Fe-binding protein IdiA found in cyanobacteria than to any of the homologous FbpA proteins currently known in commensal or pathogenic members of the Pasteurellaceae or Neisseriaceae.

Amino Acid Sequence↗

PubMed1998

Regulation of protease expression in Porphyromonas gingivalis.

Although the strong protease activity of Porphyromonas gingivalis appears to be an important virulence property of these organisms, little information is currently available regarding the regulation of expression of the multiple protease genes. Utilizing the lacZ reporter gene strategy, the environmental factors which regulate the expression of the Arg-gingipain gene rgpA and the prtT protease gene were investigated. These two genes are reciprocally regulated since factors which retarded growth (iron depletion and nutrient limitation) appeared to upregulate rgpA expression while down-regulating prtT expression. However, inactivation of the major rgpA gene resulted in increased transcription of the prtT and tpr protease genes while decreasing expression of the Lys-gingipain kgp gene as detected by Northern blot analysis. By contrast, inactivation of the prtT gene did not significantly affect kgp expression but moderately decreased rgpA mRNA levels. These results indicate that the protease genes of P. gingivalis are not coordinately regulated and suggest that some of these enzymes play specific roles in the physiology and/or virulence of these organisms.

Adhesins, Bacterial↗

PubMed1998

Role of Arg-gingipain A in virulence of Porphyromonas gingivalis.

In order to access the role of the Porphyromonas gingivalis Arg-gingipain proteases in the virulence of this organism, a mutant defective in the rgpA gene was constructed in strain 381. This mutant, MT10, displayed only 40% of the Arg-specific cysteine protease activity of the wild-type strain. In addition, MT10, as well as the recently characterized protease mutant G-102, which is defective in the rgpB gene, displayed reduced self-aggregation, hemagglutination, and the ability to bind to immobilized type I collagen compared to levels of the wild-type parent. However, unlike mutant G-102, the rgpA mutant displayed increased binding to epithelial cells relative to that of the parental organism. Mutant MT10 also did not express detectable levels of the FimA protein as assessed by both Western and Northern blotting or fimbriae visible by electron microscopy of the cells. Furthermore, the ability of MT10 to degrade rat tail collagen fibers when it was cultured at 37 degrees C was markedly attenuated compared to that of strain 381. These results suggest that Arg-gingipain A may play a significant role in the pathogenicity of P. gingivalis by altering the colonization and toxic properties of the organism.

Adhesins, Bacterial↗

PubMed1998

Detection of anti-cytokeratin 8 antibody in the serum of patients with cryptogenic fibrosing alveolitis and pulmonary fibrosis associated with collagen vascular disorders.

BACKGROUND: It has been suggested that the humoral immune system plays a role in the pathogenesis of cryptogenic fibrosing alveolitis (CFA). Although circulating autoantibodies to lung protein(s) have been suggested, none of the lung proteins have been characterised. The purpose of this study was to determine the antigen to which the serum from patients with pulmonary fibrosis reacted. METHODS: The anti-A549 cell antibody was characterised in a patient with CFA using Western immunoblotting and immunohistochemical staining of A549 cells. As we identified that one of the antibodies against A549 cells was anti-cytokeratin 8, the expression of mRNA of cytokeratin 8 in A549 cells was evaluated. In addition, we attempted to establish an enzyme linked immunosorbent assay to measure the levels of anti-cytokeratin 8 antibody in the serum of patients with CFA and pulmonary fibrosis associated with collagen vascular disorders (PF-CVD). RESULTS: Initially two anti-A549 cell antibodies were detected in the serum of patients with pulmonary fibrosis, one of which was characterised as anticytokeratin 8 antibody by Western immunoblotting. We were able to establish an ELISA to measure anti-cytokeratin 8 antibody and found significantly higher levels in patients with CFA and PF-CVD than in normal volunteers, patients with sarcoidosis, pneumonia, and pulmonary emphysema. CONCLUSIONS: One of the anti-A549 cell antibodies in the serum of patients with CFA was against cytokeratin 8. The serum levels of anti-cytokeratin 8 antibody were increased in patients with CFA and PF-CVD. These results suggest that anticytokeratin 8 antibody may be involved in the process of lung injury in pulmonary fibrosis.

Adult↗

PubMed1998

Dual regulatory effects of interferon-alpha, -beta, and -gamma on interleukin-8 gene expression by human gingival fibroblasts in culture upon stimulation with lipopolysaccharide from Prevotella intermedia, interleukin-1alpha, or tumor necrosis factor-alpha.

In a previous study, we demonstrated that the amount of interleukin (IL)-8 mRNA expressed by human gingival fibroblasts stimulated with lipopolysaccharide (LPS) from Prevotella intermedia ATCC 25611 is increased by pre-treatment with beta or gamma interferon (IFN-beta or -gamma). In the present study, we identified the regulatory effects of these IFNs on IL-8 mRNA expression and IL-8 production by human gingival fibroblasts. Priming with IFN-alpha (alpha), -beta, or -gamma upregulated the IL-8 mRNA expression in response to P. intermedia LPS, whereas co-stimulation with these IFNs reduced the amount of mRNA expressed by the cells. The regulation of IL-8 mRNA expression induced by recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) or rHuIL-1alpha was similar to that induced by LPS. The IL-8 mRNA expression in response to P. intermedia LPS was enhanced by IFN-gamma independently of de novo protein synthesis, and was regulated, at least in part, at the transcriptional level. The IL-8 mRNA accumulation in response to P. intermedia LPS was inhibited by tosylphenyl-alanyl chloromethyl-ketone, an inhibitor of NF-kappaB activation, although the NF-kappaB activation itself was not altered by IFN-gamma. These findings suggest that IFNs might be capable of both enhancing and inhibiting inflammatory responses in periodontal tissues through the dual regulation of IL-8 production by gingival fibroblasts in response to bacterial components and cytokines.

Blotting, Northern↗

PubMed1998

Vectorcardiographic evaluation of myocardial infarct size: departure parameters are superior to conventional spatial parameters.

To determine whether the departure parameters derived from a "departure loop" of a vectorcardiogram are more accurate than conventional spatial parameters in evaluating myocardial infarct size, 74 patients with first-onset myocardial infarction (MI) were studied. The correlation between the departure parameters (amplitudes in scalar leads of the departure loop) and the percent defect volume of thallium myocardial scintigrams (%DV) was compared with that of the spatial parameters (magnitude, azimuth, and elevation of the original QRS loop). In anteroseptal MI, the amplitude of a 20-msec vector in the z-axis and the azimuth of a 30-msec vector (H30) were significantly correlated with %DV (r=0.783, p<0.001 and r=0.572, p<0.05). In anteroseptal MI with involvement of the lateral wall, the amplitude of a 30-msec vector in the x-axis and H30 showed significant correlation with %DV (r=0.802, p<0.001 and r=0.772, p<0.01). In inferior and inferoposterior MI, the amplitude of a 30-msec vector in the y-axis and the elevation of a 30-msec vector were significantly correlated with %DV (r=0.920, 0.891, p<0.001 and r=0.871, 0.678, p<0.01, respectively). In conclusion, the departure parameters are more accurate than the spatial parameters for evaluation of myocardial infarct size.

Aged↗

PubMed1998

[A case of systemic lupus erythematosus with pure red cell aplasia possibly caused by persistent infection of human parvovirus B 19].

We reported a 68-old female who was diagnosed to have systemic lupus erythematosus 18 years ago. She had been well under 5 mg of prednisolone until 1995, when she felt severe shortness of breath. Laboratory examinations disclosed severe anemia accompanying mild thrombocytopenia. Bone marrow aspiration revealed a complete absence of erythroid progenitor cells. She was diagnosed to have pure red cell aplasia (PRCA) as well as antiphospholipid syndrome. A judicious use of methylprednisolone including pulse therapy resulted in a prompt resolution of anemia as well as thrombocytopenia and the dose of corticosteroid was tapered successfully thereafter. Persistent infection of HPV B 19 in the patient with inactive SLE was considered as a main cause of PRCA because not only IgG-HPV B 19 antibody but viral DNA was demonstrated in her serum at the time of admission. The relationship between PRCA and anti-phospholipid syndrome in this patient was also noted.

Aged↗

PubMed1998

Plasma beta-carotene, retinol, and alpha-tocopherol levels in relation to glycemic control of children with insulin-dependent diabetes mellitus.

Plasma beta-carotene, alpha-tocopherol and retinol were measured in 15 female and 5 male children with insulin-dependent diabetes mellitus (IDDM), and the correlations with plasma hemoglobin A1c (HbA1c) and fructosamine were analyzed. Twelve female and 8 male children served as age-matched controls. The plasma beta-carotene and alpha-tocopherol levels of the IDDM children were significantly higher than those of the control children, but there were no differences in plasma retinol or total lipid levels. The plasma beta-carotene level, beta-carotene/retinol ratio and beta-carotene/total lipids ratio each showed significant correlations with serum HbA1c and fructosamine in all subjects studied. Similarly, the plasma alpha-tocopherol level and alpha-tocopherol/total lipids ratio were correlated with these indexes of glycemic control. These findings suggest certain mechanisms may exist to prevent lipid peroxidation and vascular complications in IDDM patients.

Adolescent↗

PubMed1997

Activation of R-Ras by Ras-guanine nucleotide-releasing factor.

Ras-GRF/CDC25(Mm), mSos, and C3G have been identified as guanine nucleotide-releasing factors for Ras family proteins. We investigated in this study the guanine nucleotide-releasing activities of Ras-GRF, mSos, and C3G toward R-Ras, which shows high sequence similarity to Ras. Ras-GRF markedly stimulated the dissociation of GDP from R-Ras, and C3G also promoted the release of R-Ras-bound GDP. Under the same conditions, mSos little affected the reaction. When Ras-GRF and R-Ras were coexpressed in COS7 cells, the remarkable accumulation of the active GTP-bound form of R-Ras was observed. C3G also increased active R-Ras in COS7 cells, while mSos did not give any effect. These results indicated that Ras-GRF and C3G could activate R-Ras. Furthermore, the activation of R-Ras by Ras-GRF was enhanced when cells were treated with ionomycin, which is known to increase the intracellular calcium concentration. The examination of tissue distribution of R-Ras, Ras-GRF, and mSos by the reverse transcription-polymerase chain reaction revealed that Ras-GRF was expressed only in brain and testis, whereas R-Ras, C3G, and mSos were expressed rather ubiquitously. These findings raise the possibility that R-Ras is activated by Ras-GRF in brain and testis, and by C3G in other tissues, respectively.

Animals↗

PubMed1997

Involvement of calmodulin-dependent protein kinases-I and -IV in long-term potentiation.

Multifunctional Ca2+/calmodulin-dependent protein kinases (CaMKs) are thought to be involved in the induction of long-term potentiation (LTP). In the present study, LTP was induced by theta burst stimulation in the Schaffer collateral area of the stratum radiatum in the hippocampal CA1 region of the rat hippocampus. LTP-induced and control hippocampal slices were studied by Western blot and immunohistochemical analyses using CaMK-I, -II and -IV antibodies. Increased amounts of all three CaMKs were found in LTP-induced hippocampal slices as indicated by Western blot as well as by the density of their immunoreactivity. Our data clearly shows that not only CaMK-II but also CaMK-I and -IV contribute to synaptic plasticity formed in LTP.

Animals↗

PubMed1997

Reticalmin: a novel calcium/calmodulin-dependent protein kinase IV-like protein in rat retina.

Western blot analysis of 100,000 g supernatant of rat retina using a polyclonal anti-Ca2+/ calmodulin-dependent protein kinase IV (CaM-kinase IV) antibody revealed an immunoreactive mass of 35 kDa, termed reticalmin. Lower amount of a isoform of CaM-kinase IV was also expressed in rat retina. Reticalmin did not react with anti-CaM-kinase IV C-terminal peptide antibody which recognized alpha and beta isoforms of CaM-kinase IV and calspermin. Immunohistochemically reticalmin was shown to be localized mainly in the outer segment of photo-receptor cells, and in dendrites of inner plexiform layers and may be in nuclei of ganglion cells and some inner nuclear layer cells.

Animals↗

PubMed1997

Changes in the expression of novel Cdk5 activator messenger RNA (p39nck5ai mRNA) during rat brain development.

We previously reported that a neuron-specific Cdk5 activator, p35nck5ai, was most prominent in the newborn rat brain. In the adult brain, the expression decreased in most regions except hippocampus and primary olfactory cortex. A novel neuron-specific Cdk5 activator, p39nck5ai, has been recently cloned. To clarify whether two activators were differentially distributed throughout brain development, in this study, we examined the spatial and temporal expression of p39nck5ai in the development rat brain. Northern blot analysis showed that p39nck5ai expression was low in 15-day old fetuses and newborn, and was most prominent in the 1-3 week-old rat brains. In the adult rat brain, expression declined to the same level as in newborn rat brain. In situ hybridization showed that p39nck5ai mRNA was weakly expressed in all neurons of all regions in the newborn rat brain and the transcriptional level was highest in all regions in the 3 week-old rat brain. In the adult, expression was decreased in most neurons except Purkinje and granule cells in the cerebellum which retained high levels. These results suggest that p35nck5a and p39nck5ai may have different functional roles in distinct brain regions during different states of the rat brain development.

Aging↗