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Biomedical subjects

Koji Miyazaki

Publications and source records attributed to Koji Miyazaki.

At least 19 recordsLinked to original sources
PubMed2006

A possible association between coronary plaque instability and complex plaques in abdominal aorta.

OBJECTIVE: Coronary plaque instability causes myocardial infarction (MI). Angiographic lesions with such instability are complex lesions. Complex carotid plaques were reported to be prevalent in unstable angina. We investigated associations between coronary plaque instability, such as MI and angiographic complex coronary lesions, and aortic plaques. METHODS AND RESULTS: Aortic MRI was performed in 146 patients undergoing coronary angiography, of whom 108 had coronary artery disease (CAD) and 44 also had MI. Prevalence of plaques in thoracic and abdominal aortas was higher in patients with than without CAD (73% and 94% versus 32% and 79%), but it was similar in CAD patients with and without MI. Notably, complex plaques in abdominal aorta were more prevalent in CAD patients with than without MI (36% versus 14%; P<0.025). In multivariate analysis, abdominal complex plaques were associated with MI (odds ratio [OR], 4.5; 95% CI, 1.5 to 13.8). Among patients without MI, thoracic and abdominal complex plaques were more prevalent in patients with than without complex coronary lesions (22% and 33% versus 2% and 7%; P<0.05). Abdominal complex plaques were also associated with complex coronary lesions (OR, 9.8; 95% CI, 1.1 to 85.9). CONCLUSIONS: Complex plaques in abdominal aorta were associated with MI and complex coronary lesions, suggesting a link between coronary and aortic plaque instability.

Adult↗
PubMed2006

Buffer solution can control the porosity of DNA-chitosan complexes.

The current studies examine the pore properties and biological effects of DNA-chitosan complexes, which may be useful as scaffolds for tissue engineering. The porosity of the DNA-chitosan complexes was controlled by rinsing them with several different pH 7.2 buffer solutions, including phosphate-buffered saline (PBS), Tris-HCl, boric acid, and N-(2-hydroxyethyl)piperazine-N'-(2-ethanesufonic acid) (HEPES). Rinsing with PBS resulted in 84% porosity, whereas rinsing with Tris-HCl produced 94% porosity. It was further found that daunorubicin hydrochloride complex intercalated with and bound to the groove of the DNA-chitosan complexes, indicating that DNA in the complexes maintains its double-stranded helical structure. The DNA-chitosan complexes were not toxic to MG-63 osteoblast-like cells and caused only a mild tissue response when implanted subcutaneously in the backs of rats. These results suggest that buffer-rinsed DNA-chitosan complexes may be useful as a scaffold material in tissue engineering.

Buffers↗
PubMed2006

Dynamics of RhoA and ROKalpha translocation in single living cells.

The RhoA-binding kinase (ROK) is one of the target kinases of RhoA and is known to play a critical role in regulating cytoskeletal rearrangement in cells. ROK translocates to the plasma membrane fraction; however, the mechanism of the translocation of ROK still remains obscure. To clarify the molecular mechanisms of the translocation of ROK, we co-transfected MDCK cells with cyan fluorescent protein-tagged RhoA and yellow fluorescent protein-tagged ROKalpha, or their variants, and monitored the localization and translocation of the two different fluorescent tagged-molecules in single living cells during epithelial growth factor (EGF) stimulation. Both RhoA (wild-type) and ROKalpha (wild-type) translocated to ruffling membrane with EGF stimulation in several minutes. A ROKalpha mutant, in which Rho-binding ability is disrupted, is unable to translocate to the membrane with RhoA. However, RhoA mutant Q63L/C190R, an active form lacking membrane localization activity, abolished the translocation of wild-type ROKalpha, suggesting that the translocation of RhoA is critical for ROK translocation to the membrane. Another mutant lacking the pleckstrin homology domain failed in translocation as well. On the other hand, it was surprising that the kinase dead mutant succeeded in translocation to the membrane after EGF stimulation. Based on these results, we propose the following ROKalpha translocation mechanism. ROKalpha binds to RhoA in cytosol and translocates to the membrane based on the membrane-targeting ability of active RhoA. After ROKalpha associates with the membrane, the pleckstrin homology domain provides the stability of ROKalpha on the membrane. The activation of enzymatic activity or adenosine triphosphate binding, however, is not directly related to the translocation mechanism, although we found that the membrane association is critical for the activation of the kinase activity.

Animals↗
PubMed2006

Degradation of methacrylate monomers in human saliva.

This study assessed the effect of the molecular structure of newly synthesized methacrylate monomers on their chemical stability in human saliva, whereby these monomers can be used as dental composite resins. Six model monomethacrylates and two urethane-modified BisGMA monomers were added to human saliva, and their change in concentration after 24, 48, and 72 hours were measured by high-performance liquid chromatography. Degradation of the six model monomethacrylate monomers was found to be influenced by the molecular structure, such as steric hindrance and presence of urethane bond in chemical backbone. Based on the degradation test results of these six monomers, urethane-modified BisGMA derivatives--in which the hydroxyl groups in original BisGMA monomer were substituted with alkyl isocyanate--were synthesized and subjected to degradation test. The urethane-modified BisGMA monomers showed a particular resistance to salivary hydrolysis. Results of this study thus suggested that urethane groups should be considered when designing new monomers for dental composite systems as they demonstrated improved resistance to hydrolysis.

Bisphenol A-Glycidyl Methacrylate↗
PubMed2005

Production of a transgenic pig expressing human albumin and enhanced green fluorescent protein.

We introduced a fusion gene of human albumin and enhanced green fluorescent protein (EGFP) into porcine oocytes using the sperm vector method, and produced a piglet that showed clear expression of GFP in the hooves and skin. PCR and Southern blotting analysis of genomic DNA extracted from the piglet's tissues, including the liver, showed that the tissues carried the transgene. RT-PCR analysis demonstrated that both the human albumin and EGFP genes were expressed in the tissues. The fact that human albumin gene was integrated and expressed in the liver of the transgenic pig opened a way for us to achieve our goal, which was the use of transgenic pigs for the bioartificial liver support system.

Albumins↗
PubMed2005

Flow-independent myocardial ischemia induced by endothelin-1: an NADH fluorescence analysis.

The endothelin-1 (ET-1) is known to cause myocardial ischemia; however, whether this effect is entirely dependent on vasoconstriction is uncertain. The aim of this study was to characterize the myocardial ischemia after the intracoronary administration of endothelin-1, and compare it with that induced by coronary stenosis. In the left anterior descending coronary artery of 15 dogs, a mild inflow reduction (30%) was produced for 10 minutes using intracoronary endothelin-1 (46 +/- 33 pmol/min) or coronary stenosis. The hearts were rapidly cross-sectioned at short axial plane and freeze-clamped within 120 milliseconds using a specially developed device to visualize and quantify the area of ischemia (%IA) with NADH fluorescence photography. The %IA was larger in the endothelin-1 group than in the stenosis group (66 +/- 23 versus 18 +/- 18, P = 0.0005); furthermore, the ischemia was transmural in the ET-1 group, but limited to subendocardium in the stenosis group. ET-1 increased the coronary arterial resistance especially in subepicardial region and produced smaller ischemic foci in microcirculation. The mechanism of larger ischemia produced by ET-1 might depend on pro-ischemic effects on myocytes and vasoconstriction of the coronary microcirculation, predominantly in the subepicardium in vivo.

Animals↗
PubMed2005

Evaluation of the quality of porcine somatic cell nuclear transfer embryo by gene transcription profiles.

This study aimed to evaluate the quality of porcine somatic cell nuclear transfer (SCNT) embryos by examining its gene transcription patterns. Embryos were produced by SCNT, intracytoplasmic sperm injection (ICSI) or under different conditions, and transcripts of genes for fibroblast growth factor receptor (FGFr) 2IIIc, FGFr72IIIb, X inactive-specific transcript (Xist), interleukin 6 (IL6), IL6 receptor (IL6r) alpha and c-kit ligand, were detected by real-time RT-PCR. The percentages of embryos in which these transcripts were detected were similar in SCNT and ICSI embryos. On the other hand, the transcriptional levels of the FGFr72IIIb and IL6ralpha genes were 0.5 times less and 2 times more, respectively, in SCNT blastocysts than those of ICSI blastocysts (p<0.05). When nuclear transfer was performed before or after activation of oocytes, embryos in the latter case showed significantly lower frequencies of having FGFr72IIIb (74% vs. 90%) and Xist (3% vs. 33%) transcripts compared to the former case embryos (p<0.05). When two lines of nuclear donor cells with different developmental potencies were used, the transcriptional profiles in the reconstructed embryos did not show any significant differences. Our study suggests that expression profiles of FGFr72IIIb, IL6ralpha, and Xist can be used as markers for the diagnosis of the developmental potency of porcine nuclear transfer embryos.

Animals↗
PubMed2005

Antibacterial activities and bonding of MMSA/TBB resin containing amphiphilic lipids.

The purpose of this study was to investigate the antibacterial activity of MMA/TBB resin containing newly developed amphiphilic lipids. The amphiphilic lipids, C10-L-Ala/pts and C12-L-Ala/pts, synthesized from the reaction of n-alkyl alcohol and L-alanine were dissolved in MMA at concentrations of 0.5, 1.0, 1.5, and 2.0 mol%. Resin mixtures of PMMA powder and each MMA liquid containing lipid and TBB were prepared for all tests. Both lipids gave antibacterial effect to MMA/ TBB resin. The addition of C12-L-Ala/pts to MMA resulted in a significantly higher antibacterial activity than the addition of C10-L-Ala/pts. In terms of bond strength, the bond strength of MMA/TBB resin to bovine dentin was significantly decreased by the addition of amphiphilic lipids. But for enamel, the bond strength of MMA/TBB resin with amphiphilic lipids was clinically acceptable for orthodontic brackets. In conclusion, amphiphilic lipids will be useful as a component of adhesive resin to give the latter an antibacterial effect.

Acrylic Resins↗
PubMed2005

PBS buffer solutions with different pH values can change porosity of DNA-chitosan complexes.

We examined the effect of phosphate-buffered saline (PBS) solution with different pH values on the formation of porosity in the DNA/chitosan complexes, and evaluated the intercalation behavior of the complexes. Four different PBS solutions with pH = 6.0, 7.0, 7.4, and 7.8 were used for rinsing water-insoluble DNA-chitosan complexes. All complexes showed high porosities ranging from 83 to 95%. Rinsing with PBS at pH 7.0, 7.4, and 7.8 reduced the porosity of the DNA-chitosan complexes. Re-rinsing with PBS at pH 7.4 reduced the porosity of the DNA-chitosan complex rinsed with PBS at pH 6.0. The appearances for porous formation were influenced by the differences in pH of PBS. Daunorubicin hydrochloride intercalated and bound in the grooves of DNA within all of the DNA-chitosan complexes, indicating that DNA in the complexes maintained its double-stranded helical structure. These results suggested that PBS-rinsed DNA-chitosan complex is promising as a scaffold material in tissue engineering.

Biopolymers↗
PubMed2005

Characterization of the expression of TLR2 (toll-like receptor 2) and TLR4 on circulating monocytes in coronary artery disease.

TLRs are receptors involved in the recognition of pathogens by the innate immune system, and TLR2 and TLR4 play important roles in the activation of monocytes. A total of 105 consecutive patients who underwent coronary angiography comprised of 46 with stable effort angina (SA), 41 with unstable angina (UA), and 18 with no significant CAD (CNT) were enrolled. The baseline expression levels of TLR2 and TLR4 on monocytes in peripheral blood mononuclear cells (PBMCs) were determined by flow-cytometric analysis. Since TLR2 expression has been reported to be regulated by TLR4 signaling, we cultured PBMCs with or without lipopolysaccharide (LPS, 1 microg/ml). At baseline, TLR4 levels (mean of fluorescence intensity ) in SA (145 +/- 58, p < 0.05) and UA (164 +/- 65, p < 0.01) were higher than those in CNT (107 +/- 37). As for TLR2, levels were higher in UA (108 +/- 36, p < 0.05) than in SA (94 +/- 18) and CNT (87 +/- 22). After stimulation with LPS, TLR2 levels increased in SA but decreased in UA. In conclusions, TLR4 levels increased in both SA and UA. Monocytes in UA were characterized by elevated TLR2 levels and unresponsiveness of the TLR2 levels to TLR4 stimulation.

Aged↗
PubMed2004

Deficiency of interleukin-1 receptor antagonist deteriorates fatty liver and cholesterol metabolism in hypercholesterolemic mice.

Although the anti-inflammatory effect of interleukin-1 (IL-1) receptor antagonist (IL-1Ra) has been described, the contribution of this cytokine to cholesterol metabolism remains unclear. Our aim was to ascertain whether deficiency of IL-1Ra deteriorates cholesterol metabolism upon consumption of an atherogenic diet. IL-1Ra-deficient mice (IL-1Ra(-/-)) showed severe fatty liver and portal fibrosis containing many inflammatory cells following 20 weeks of an atherogenic diet when compared with wild type (WT) mice. Expectedly, the levels of total cholesterol in IL-1Ra(-/-) mice were significantly increased, and the start of lipid accumulation in liver was observed earlier when compared with WT mice. Real-time PCR analysis revealed that IL-1Ra(-/-) mice failed to induce mRNA expression of cholesterol 7alpha-hydroxylase, which is the rate-limiting enzyme in bile acid synthesis, with concurrent up-regulation of small heterodimer partner 1 mRNA expression. Indeed, IL-1Ra(-/-) mice showed markedly decreased bile acid excretion, which is elevated in WT mice to maintain cholesterol level under atherogenic diet feeding. Therefore, we conclude that the lack of IL-1Ra deteriorates cholesterol homeostasis under atherogenic diet-induced inflammation.

Animals↗
PubMed2004

Protein kinase Cepsilon and the antiadrenergic action of adenosine in rat ventricular myocytes.

Adenosine-induced antiadrenergic effects in the heart are mediated by adenosine A(1) receptors (A(1)R). The role of PKCepsilon in the antiadrenergic action of adenosine was explored with adult rat ventricular myocytes in which PKCepsilon was overexpressed. Myocytes were transfected with a pEGFP-N1 vector in the presence or absence of a PKCepsilon construct and compared with normal myocytes. The extent of myocyte shortening elicited by electrical stimulation of quiescent normal and transfected myocytes was recorded with video imaging. PKCepsilon was found localized primarily in transverse tubules. The A(1)R agonist chlorocyclopentyladenosine (CCPA) at 1 microM rendered an enhanced localization of PKCepsilon in the t-tubular system. The beta-adrenergic agonist isoproterenol (Iso; 0.4 microM) elicited a 29-36% increase in myocyte shortening in all three groups. Although CCPA significantly reduced the Iso-produced increase in shortening in all three groups, the reduction caused by CCPA was greatest with PKCepsilon overexpression. The CCPA reduction of the Iso-elicited shortening was eliminated in the presence of a PKCepsilon inhibitory peptide. These results suggest that the translocation of PKCepsilon to the t-tubular system plays an important role in A(1)R-mediated antiadrenergic actions in the heart.

Adenosine↗
PubMed2004

Lack of interleukin-1 receptor antagonist modulates plaque composition in apolipoprotein E-deficient mice.

OBJECTIVE: Interleukin (IL)-1 plays an important role in atherosclerosis. IL-1 receptor antagonist (IL-1Ra) is an endogenous inhibitor of IL-1. However, the role of IL-1Ra in the development of atherosclerosis is poorly understood. METHODS AND RESULTS: Mice that lacked IL-1Ra (IL-1Ra-/-) were crossed with apolipoprotein E-deficient (E-/-) mice and formation of atherosclerotic lesions was analyzed after 16 weeks or 32 weeks consumption of a normal chow diet. This study focused on the comparison of atherosclerotic lesion between IL-1Ra+/+/apoE-/- (n=12) and IL-1Ra(+/-)/apoE-/- mice (n=12), because of the significantly leaner phenotype in IL-1Ra-/-/apoE-/- mice compared with the others. Interestingly, atherosclerotic lesion size in IL-1Ra+/-/apoE-/- mice at age 16 weeks was significantly increased (30%) compared with IL-1Ra+/+/apoE-/- mice (P<0.05). At 32 weeks, the differences of lesion size between these mice failed to achieve statistical significance. However, immunostaining demonstrated an 86% (P<0.0001) increase in the MOMA-2-stained lesion area of IL-1Ra+/-/apoE-/- mice. In addition, alpha-actin staining in these lesions was significantly decreased (-15%) compared with those in IL-1Ra+/+/apoE-/- mice (P<0.05). CONCLUSIONS: These results suggest an important role of IL-1Ra in the suppression of lesion development during early atherogenesis and furthermore indicate its role in the modulation of plaque composition.

Animals↗
PubMed2004

Intercalation behavior and tensile strength of DNA-lipid films for the dental application.

In this study, we prepared DNA-lipid films and examined their intercalation behavior and tensile strength as an indicator for usefulness as a dental material. The lipids were synthesized from the reaction of glycine, l-alanine, or l-glutamic acid with n-alkyl alcohol in the presence of p-toluenesulfonic acid. The self-standing, water-insoluble DNA-lipid films were prepared by casting the DNA-lipid complex from chloroform/ethanol solution. The DNA-lipid films formed intercalation complexes with ethidium bromide. This indicates that DNA-lipid films maintain a double helical structure. The tensile strengths of DNA films were 0.8-2.4MPa and were compatible with a commercially available material (Membrane) for guided tissue regeneration in dental use. We conclude that DNA-lipid films have potential for use as a material for the surface treatment of implanted materials or as a bone-guiding scaffold for dental application.

DNA↗
PubMed2004

Study on rat subcutaneous reaction to experimental polyurethane elastomers.

The purpose of the study was to investigate the biocompatibility of experimental elastomers, E580 and E590. The experimental elastomers and the control--a clinically used elastomer--were implanted into the subcutaneous tissue of rats. The tissue reactions were examined histologically on the 3rd, 7th, 14th, 28th, and 56th day after implantation. It was found that there were some irritant responses in the tissues adjacent to the implanted elastomers during the first week. However, the inflammatory tissue reaction subsided substantially from the second week onwards. The stable fibrous capsule surrounding the elastomer was formed after eight weeks. The tissue responses of the control, E580, and E590 were similar. The results suggested that the long-term tissue irritation of the experimental elastomers was so low such that they have the potential to be applied clinically.

Animals↗
PubMed2003

Evaluation of a soybean product fujiflavone P40 as an antiosteoporotic agent in rats.

The preventive effects of Fujiflavone P40 (a soybean isoflavone product) against both bone loss and periodontal alteration were evaluated using an ovariectomized rat model. Rats were divided into five groups: sham-operated (Sham), ovariectomized (OVX), OVX given Fujiflavone P40, OVX given 17beta-oestradiol, and OVX given the vehicle for 17beta-oestradiol, respectively. Fujiflavone P40 contains 46.6% isoflavones which consist of 24.1% daidzin, 16.5% glycitin and 5.9% genistin. Administration of Fujiflavone P40 to OVX rats suppressed the body weight gain until 5 weeks. Fujiflavone P40 also decreased total and high-density lipoprotein (HDL) cholesterols and triglyceride level of OVX rats, significantly. After 7 weeks, Fujiflavone P40 did not recover the coarsened fibre of the periodontal ligament. The ovariectomy decreased the uterine weight by 78%. The administration of 17beta -oestradiol recovered the weight loss by 99%, while Fujiflavone P40 restored it by 33%. The ovariectomy decreased the tibial bone mineral density (BMD) by 22%. The administration of 17beta-oestradiol to OVX rats recovered the tibial BMD decrease by 100%, while Fujiflavone P40 recovered it by 78%. The results suggest that Fujiflavone P40 may be useful as a preventive agent for osteoporosis.

Animals↗
PubMed2003

Estrogenic activity of tissue conditioners in vitro.

OBJECTIVES: In order to assess the estrogenic activities of plasticizers used in tissue conditioners and four commercial tissue conditioners, we carried out in vitro tests. METHODS: Seven plasticizers and two metabolites were diluted to concentrations ranging from 10(-9) to 10(-4)M. Four commercial tissue conditioners were also diluted to concentrations ranging from 2 x 10(-8) to 2 x 10(-3)g/ml. Estrogenic activities were tested by the E-screen test using MCF-7 cells. When estrogen is present, the cells proliferate. Instead of counting the cells or nuclei directly, cell numbers were assessed by measurement of the total protein content using the sulforhodamine B assay. The liquid compositions of the four commercial tissue conditioners were examined by high-performance liquid chromatography. RESULTS: n-Butyl benzyl phthalate, dibutyl phthalate, n-butyl phthalyl n-butyl glycolate, di-2-ethylhexyl phthalate and benzyl salicylate significantly increased proliferation of MCF-7 cells. The remaining two plasticizers, di-2-ethylhexyl adipate and benzyl benzoate, as well as two metabolites of dibutyl phthalate and di-2-ethylhexyl phthalate, i.e. monobutyl phthalate and mono-2-ethylhexyl phthalate, respectively, did not increase proliferation of MCF-7 cells at the concentrations tested. Four commercial tissue conditioners, Coe comfort (CC), Tissue Conditioner (TC), Hydro Cast (HC) and Denture Soft (DS) II, significantly increased proliferation of MCF-7 cells. HPLC data revealed the commercial products contained plasticizers: benzyl benzoate and dibutyl phthalate in CC, dibutyl phthalate in TC, n-butyl benzyl phthalate in HC and n-butyl phthalyl n-butyl glycolate in DS II. SIGNIFICANCE: Except for benzyl benzoate and di-2-ethylhexyl adipate, the plasticizers tested showed estrogenic activity. The four commercial tissue conditioners tested also showed estrogenic activity, and HC showed especially strong estrogenicity.

Breast Neoplasms↗