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Biomedical subjects

Y Miyamoto

Publications and source records attributed to Y Miyamoto.

At least 253 records · Page 14Linked to original sources

Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy: evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes.

BACKGROUND: In cardiac hypertrophy, both excessive enlargement of cardiac myocytes and progressive interstitial fibrosis are well known to occur simultaneously. In the present study, to investigate the interaction between ventricular myocytes (MCs) and cardiac nonmyocytes (NMCs), mostly fibroblasts, during cardiocytes hypertrophy, we examined the change in cell size and gene expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in cultured MCs as markers for hypertrophy in the neonatal rat ventricular cardiac cell culture system. METHODS AND RESULTS: The size of cultured MCs significantly increased in the MC-NMC coculture. Concomitantly, secretions of ANP and BNP into culture media were significantly increased in the MC-NMC coculture compared with in the MC culture (with the possible contamination of NMC <1% of MC). Moreover, in the MC culture, enlargement of MC and an increase in ANP and BNP secretions were induced by treatment with conditioned media of the NMC culture. A considerable amount of endothelin (ET)-1 production was detected in the NMC-conditioned media. BQ-123, an ET-A receptor antagonist, and bosentan, a nonselective ET receptor antagonist, significantly blocked the hypertrophic response of MCs induced by treatment with NMC-conditioned media. Angiotensin II (Ang II) (10(-10) to 10(-6) mol/L) and transforming growth factor-beta1 (TGF-beta1) (10(-13) to 10(-9) mol/L), both of which are known to be cardiac hypertrophic factors, did not induce hypertrophy in MC culture, but both Ang II and TGF-beta1 increased the size of MCs and augmented ANP and BNP productions in the MC-NMC coculture. This hypertrophic activity of Ang II and TGF-beta1 was associated with the potentiation of ET-1 production in the MC-NMC coculture, and the effect of Ang II or TGF-beta1 on the secretions of ANP and BNP in the coculture was significantly suppressed by pretreatment with BQ-123. CONCLUSIONS: These results demonstrate that NMCs regulate MC hypertrophy at least partially via ET-1 secretion and that the interaction between MCs and NMCs plays a critical role during the process of Ang II- or TGF-beta1-induced cardiocyte hypertrophy.

Angiotensin II↗

Differential modes of nuclear localization signal (NLS) recognition by three distinct classes of NLS receptors.

The targeting of karyophilic proteins to nuclear pores is mediated via the formation of a nuclear pore-targeting complex, through the interaction of nuclear localization signal (NLS) with its NLS receptor. Recently, a novel human protein, Qip1, was identified from a yeast two-hybrid system with DNA helicase Q1. This study demonstrates that Qip1 is a novel third class of NLS receptor that efficiently recognizes the NLS of the helicase Q1. Moreover, the data obtained in this study show that the specific interaction between Qip1 and the NLS of the helicase Q1 requires its upstream sequence of the minimal essential NLS. By using purified recombinant proteins alone in the digitonin-permeabilized cell-free transport system, it was demonstrated that the two known human NLS receptors, Rch1 and NPI-1, are able to transport all the tested NLS substrates into the nucleus, while Qip1 most efficiently transports the helicase Q1-NLS substrates, which contain its upstream sequence in so far as we have examined the system. Furthermore, in HeLa cell crude cytosol, it was found that endogenous Rch1 binds to all the tested NLS substrates, while the binding of endogenous NPI-1 is restricted to only some NLSs, despite the fact that NPI-1 itself shows binding activity to a variety of NLSs. These results indicate that at least three structurally and functionally distinct NLS receptors exist in the human single cell population, and suggest that the nuclear import of karyophilic proteins may be controlled in a complex manner at the NLS recognition step by the existence of a variety of NLS receptors with various specificities to each NLS.

Adenosine Triphosphatases↗

Non-decay type fast-setting calcium phosphate cement: hydroxyapatite putty containing an increased amount of sodium alginate.

A hydroxyapatite [(HAP) Ca10(PO4)6(OH)2] putty that behaves like a putty or self-curing resin was made by increasing the amount of sodium alginate in non-decay type fast-setting calcium phosphate cement (nd-FSCPC). nd-FSCPC became viscous as the sodium alginate concentration was increased. The best handling properties were obtained when nd-FSCPC contained 8% sodium alginate in its liquid phase. When a 2-kg glass plate was placed on the paste, HAP putty spread to form an area three times that of FSCPC paste. Thus, HAP putty is expected to be easier to use than FSCPC in the filling of bone defects. HAP putty did not decay; in fact, it set within approximately 20 min when immersed in distilled water immediately after mixing. The wet diametral tensile strength value of HAP putty was approximately 12 MPa after 24 h, the same as that for nd-FSCPC containing 0.5% sodium alginate in its liquid phase, or FSCPC that is free from sodium alginate. The elements constituting set HAP putty were examined using powder X-ray diffraction and found to be predominantly apatitic minerals after 24 h. Since the handling properties of a putty or self-curing resin-like cement are very useful in certain surgical procedures, HAP putty made by increasing the sodium alginate concentration in nd-FSCPC is potentially a valuable new biomaterial for use in plastic and reconstructive surgery, as well as oral and maxillofacial surgery.

Alginates↗

1alpha,25-dihydroxyvitamin D3-24-hydroxylase (CYP24) hydroxylates the carbon at the end of the side chain (C-26) of the C-24-fluorinated analog of 1alpha,25-dihydroxyvitamin D3.

The sequential oxidation and cleavage of the side chain of 1alpha, 25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) initiated by the hydroxylation at C-24 is considered to be the major pathway of this hormone in the target cell metabolism. In this study, we examined renal metabolism of a synthetic analog of 1alpha,25(OH)2D3, 24, 24-difluoro-1alpha,25-dihydroxyvitamin D3 (F2-1alpha,25(OH)2D3), C-24 of which was designed to resist metabolic hydroxylation. When kidney homogenates prepared from 1alpha,25(OH)2D3-supplemented rats were incubated with F2-1alpha,25(OH)2D3, it was mainly converted to a more polar metabolite. We isolated and unequivocally identified the metabolite as 24,24-difluoro-1alpha,25,26-trihydroxyvitamin D3 (F2-1alpha,25,26(OH)3D3) by ultraviolet absorption spectrometry, frit-fast atom bombardment liquid chromatography/mass spectroscopy analysis, and direct comparison with chemically synthesized F2-1alpha,25,26(OH)3D3. Metabolism of F2-1alpha,25(OH)2D3 into F2-1alpha,25,26(OH)3D3 by kidney homogenates was induced by the prior administration of 1alpha,25(OH)2D3 into rats. The C-24 oxidation of 1alpha,25(OH)2D3 in renal homogenates was inhibited by F2-1alpha,25(OH)2D3 in a concentration-dependent manner. Moreover, F2-1alpha,25,26(OH)3D3 was formed in ROS17/2.8 cells transfected with a plasmid expressing 1alpha,25(OH)2D3-24-hydroxylase (CYP24) but not in the cells transfected with that expressing vitamin D3-25-hydroxylase (CYP27) or containing inverted CYP27 cDNA. These results show that CYP24 catalyzes not only hydroxylation at C-24 and C-23 of 1alpha,25(OH)2D3 but also at C-26 of F2-1alpha,25(OH)2D3, indicating that this enzyme has a broader substrate specificity of the hydroxylation sites than previously considered.

Animals↗

Surface modifications of titanium in calcium-ion-containing solutions.

The surface of titanium was modified in calcium-ion-containing solutions to improve bone conductivity. Three kinds of aqueous solutions for the modification were prepared using calcium nitrate, calcium chloride, and calcium oxide. The pH values of the solutions were 3.9, 7.4, and 12.6, respectively. At first, anatase powder was immersed in the solutions of 7 days to confirm the effect of the solutions on the modification. As a result, calcium titanate was formed on the anatase powder, indicating that the titanium surface was possibly modified by the solutions. Titanium plates were then immersed in the solutions at ambient temperature for 7 days, washed by deionized water, and dried. X-ray photo-electron spectroscopy of surface-modified titanium revealed that the surface-modified layer contained calcium hydroxide and/or calcium titanate. The surface-modified layer in which titanium was modified was thickest in this order: calcium oxide, calcium chloride, and calcium nitrate solutions. Apatite was formed on the surface-modified titanium in Hanks' solution while apatite was undetected on unmodified titanium. Therefore, the surface modification of titanium may improve bone conductivity. The amount of apatite corresponded to the thickness of the surface-modified layer and the amount of calcium in the layer.

Apatites↗

Blast coating method: new method of coating titanium surface with hydroxyapatite at room temperature.

When a titanium plate was blasted with hydroxyapatite [HAP; Ca10[x](HPO4)[x](PO4)[6-x](OH)[2-x]] powder at room temperature using an ordinary sandblaster, the surface of the titanium plate was found to be coated with HAP homogeneously and completely. The coated layer was examined with energy dispersive X-ray spectroscopy and X-ray diffraction and was found to be the same as the HAP powder used with respect to composition and crystallographic structure. The coated HAP layer was tightly attached to the surface of the titanium plate, at least at the level of scanning electron microscopy. Interestingly, the HAP particles stuck together at room temperature as if they were sintered. The coating was stable against ultrasonication in water for at least 5 min, and it was difficult to remove by nail scratching. Thus, the bonding strength between the HAP powder and the titanium plate was much higher than that achieved with currently employed room temperature coating processes such as dipping, electrophoretic deposition, and electrochemical deposition. Therefore, the blast coating method is potentially valuable for the fabrication of useful biomaterials for hard tissue replacement.

Biocompatible Materials↗

Ischemia-reperfusion injury on the pancreas in rats: identification of acinar cell apoptosis.

An ischemia-reperfusion injury on the pancreas is involved in the pathophysiology of acute pancreatitis or tissue injuries after pancreas transplantation. On the other hand, recent studies have demonstrated that ischemia-reperfusion induces apoptosis in several organs such as kidney, heart, and brain. In the present study, we sought to characterize a pattern of injury during ischemia-reperfusion on the pancreas and determined whether ischemia-reperfusion on the pancreas causes the apoptotic process. Ischemia-reperfusion was induced by blocking the inferior splenic artery and removing the clamp in pentobarbital-anesthetized rats. Rats were sacrificed at 0-72 hr following a 60-min ischemia. Evans blue extravasation showed 3.5-fold increase at 2 hr after reperfusion, indicating a rapid increase of vascular permeability. Tissue myeloperoxidase activity, an index of neutrophil accumulation, significantly increased in a time-dependent manner until 48 hr after reperfusion. Histological analysis revealed the existences of interstitial cell infiltration and edema. DNA breaks of acinar cells were detected by gel electrophoresis and in situ nick end-labeling, and the numbers strikingly increased at 48 hr after reperfusion. Furthermore, Bax protein, an effector of apoptotic cell death, was expressed in acinar cells. The results indicate that an ischemia-reperfusion injury on the pancreas in rats resembles many features of acute pancreatitis. Apoptosis in acinar cells may be one of the specific features of the ischemia-reperfusion injury on the pancreas.

Amylases↗

Hyperthermia after cardiac surgery.

PURPOSE: To describe two cases of hyperthermia occurring after cardiac surgery. CLINICAL FEATURES: At the end of cardiopulmonary bypass, protamine was injected to reverse heparin. Following protamine, hypotension, pulmonary hypertension and hypoxia developed, but protamine induced hypotension recovered after administration of methyl predonisolone and catecholamines. Rectal temperature increased to 41 degrees C in the first case and 40 degrees C in the second after recovery from protamine induced hypotension. Arterial blood gas analysis, breathing 100% oxygen, showed severe acidosis and hypercapnia (pH 6.96, PaCO2 73 mmHg, PaO2 45 mmHg. BE -18 in the first case, and pH 7.13, PaCO2 59.9, PaO2 52.8, BE -11 in the second). At the same time, creatine phosphokinase levels showed 8400 u.L-1 in the first case and 4369 u.L-1 in the second. Serum and urine myoglobin concentrations were 334.2 ng.ml-1 and 1085 ng.ml-1, and 468 ng.ml-1 and 885 ng.ml-1, respectively. Efforts to cool the patients using alcohol were made. Following dantrolene (400 mg and 200 mg), the hyperthermia, acidosis, hypoxaemia and hypercarbia subsided (temperature 37.5 and 37.5 degrees C, PaO2 220 mmHg and 180 mmHg with 100% oxygen, PaCO2 35 mmHg and 41 mmHg respectively). Postoperatively, 20 mg.hr-1 dantrolene infusion were administered for 24 hr. In the first case, an anaphylactoid reaction was confirmed by increased plasma tryptase. CONCLUSION: We report two cases of hyperthermia after cardiac surgery in whom dantrolene was very effective in reducing the high rectal temperature.

Aged↗

Plasma levels of brain and atrial natriuretic peptides elevate in proportion to left ventricular end-systolic wall stress in patients with aortic stenosis.

Brain natriuretic peptide (BNP) is a novel cardiac hormone secreted predominantly from the ventricle. We examined the plasma levels of BNP and atrial natriuretic peptide (ANP) in 13 patients with aortic stenosis undergoing corrective surgery. Preoperative plasma BNP and ANP levels correlated highly with preoperative left ventricular end-systolic wall stress (ESS) (r = 0.96, p < 0.0001 and r = 0.95, p < 0.0001, respectively). Moreover, between preoperative and late postoperative states, the difference of the plasma levels of BNP and ANP correlated with the difference of ESS. In two patients with elevated ESS and quite high preoperative plasma BNP (> 1000 pg/ml), rapid decrease of the plasma level after operation was observed. These results suggest that synthesis and secretion of BNP and ANP are stimulated by the increase of left ventricular end-systolic wall stress in patients with aortic stenosis.

Aged↗

Carbohydrate-binding properties of the hemolytic lectin CEL-III from the holothuroidea Cucumaria echinata as analyzed using carbohydrate-coated microplate.

The carbohydrate-binding properties of the hemolytic lectin CEL-III from the Holothuroidea Cucumaria echinata were studied using the microplate assay system which we have recently developed [Hatakeyama et al. (1996) Anal. Biochem. 237, 188-192]. When the binding of CEL-III to lactose covalently immobilized on a microplate was examined using colloidal gold solution, the binding was detected with as little as 1 microgram/ml protein. Affinity of several carbohydrates to CEL-III was assessed by means of an inhibition experiment using the lactose-coated plate and it was found that N-acetylgalactosamine has the highest affinity for CEL-III, followed by lactose and lactulose. Examination of the binding of CEL-III to the lactose-coated plate at various pH values and temperatures revealed that the affinity is higher in the acidic pH region and at lower temperatures. From the Ca(2+)-dependence profile for the binding of CEL-III to the lactose-coated plate, the apparent dissociation constant for Ca2+ was estimated to be 2.3 mM. These results suggested that the carbohydrate-binding properties of CEL-III are closely related to its hemolytic activity, although an additional interaction between the protein and the lipid bilayer, which is enhanced in the alkaline pH region, also seems to be necessary for its hemolytic action.

Animals↗

Expression of Bcl-2 and PCNA in duct cells after pancreatic duct ligation in rats.

Obstruction of the pancreatic duct induces acinar cell deletion followed by duct proliferation and interstitial fibrosis. Apoptosis has been reported to be involved in the induction of acinar cell deletion after pancreatic duct ligation (PDL) in rats, however, the mechanism of pancreatic duct cell proliferation is still unknown. We hypothesized that Bcl-2 (antiapoptosis protein) and PCNA (cell cycle-related protein) could be involved in the mechanism of pancreatic duct cell proliferation after PDL. In PDL, rats, acinar cells decreased in number and disappeared completely after duct ligation and duct-lining cells increased in number and formed duct-tubular complexes. Immunohistochemical study showed that PCNA expression appeared in the ductules and centroacinar cells from early stages after duct ligation and that Bcl-2 expression in duct cells, which was faint in normal pancreas, increased significantly when acinar cells were diminishing. Western blotting demonstrated that Bcl-2 was detected as a single band at 26 kDa, and the intensity of Bcl-2 in PDL rats was approximately ninefold stronger than in normal pancreas. Expression of Bcl-2 and PCNA after pancreatic duct ligation may be related to the prevention of apoptosis and cell proliferation of pancreatic duct cells in rats.

Animals↗

Nitric oxide generation from hydroxyurea via copper-catalyzed peroxidation and implications for pharmacological actions of hydroxyurea.

We investigated the generation of nitric oxide (NO) by H2O2-dependent peroxidation of hydroxyurea in the presence of copper-containing proteins such as Cu,Zn-superoxide dismutase (Cu,Zn-SOD) or ceruloplasmin as a catalyst. In the reaction mixture of hydroxyurea, CuZn-SOD, and H2O2, NO generation was identified by measuring the specific electron spin resonance (ESR) signal of 2-phenyl-4, 4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO). The ESR signal of the NO-hemoglobin adduct was also detected in human red blood cells during copper-catalyzed peroxidation of hydroxyurea. The NO production during peroxidation of hydroxyurea was quantified as NO2- formation, measured by using the Griess assay, the amount of NO2- was dependent on the concentrating of hydroxyurea of the reaction mixture. ESR spin trapping with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) showed hydroxy radical (OH) generation in the reaction of H2O2 with either Cu,Zn-SOD or ceruloplasmin. Several OH scavengers, such as ethanol, thiourea, DMPO, and dimethylsulfoxide, and the metalchelating agent diethylenetriaminepentaacetic acid significantly inhibited NO generation from hydroxyurea. This indicates that NO release from hydroxyurea may be mediated by OH derived from the copper-catalyzed Fenton-like reaction. Incubation of hydroxyurea and Cu,Zn-SOD with xanthine oxidase and hypoxanthine in a system forming O2- -->H2O2 also resulted in appreciable NO production. These results suggest that NO production from hydroxyurea catalyzed by copper-containing proteins may be the molecular basis of the pharmacological and antitumor action of hydroxyurea.

Ceruloplasmin↗

Role of neutrophils in cerulein-induced pancreatitis in rats: possible involvement of apoptosis.

We investigated the role of neutrophils and the involvement of apoptosis in cerulein-induced acute pancreatitis. Male Sprague-Dawley rats were divided into 2 groups. In the control group, acute pancreatitis was induced by subcutaneous injections of cerulein. In methotrexate-treated group, the rats received intraperitoneal injections of methotrexate to produce neutrophil depletion before the injections of cerulein. The rats were sacrificed at the indicated time points until 72 h after the first injection of cerulein. Neutrophil depletion ameliorated pancreatic edema and vacuole formation in acinar cells during the early stages of cerulein-induced acute pancreatitis. Electron microscopy, DNA gel electrophoresis and in situ nick end-labeling revealed the involvement of apoptosis in acinar cells in cerulein-induced acute pancreatitis. Furthermore, the number of apoptotic acinar cells in neutrophil-depleted rats showed an about 2-fold increase during the late stages when compared with those in the control rats. Our results suggest that neutrophil depletion in cerulein-induced pancreatitis leads to amelioration of pancreatic injury during the early stage, and enhancement of apoptosis by neutrophil depletion occurs during the late stage.

Acute Disease↗

Respiratory responses to passive and active recovery from exercise.

To investigate the effect of the neural components associated with leg movements on the control of ventilation during recovery from exercise, we recorded the minute ventilation (VE), oxygen uptake (VO2), and carbon dioxide output (VCO2) of eight normal volunteers during recovery from moderate, steady-state cycle exercise (170 W). The recovery phases were undergone separately under two different conditions: 5 min of rest (passive recovery) on a bicycle ergometer and 3 min of pedaling at a work rate of 0W (active recovery) followed by 2 min of rest. The phase-1 responses were observed in all the variables studied at the transition of passive recovery but not in the active recovery phase. The kinetics of VCO2, during the off-transition were significantly faster than those of VE in both recoveries, indicating that the decreases in VCO2 could precede the decreases in VE. Although the levels of VE and VCO2 during active recovery were significantly higher than those during passive recovery, the decline in VE was closely proportional to that of VCO2 under both recovery conditions, with resultant indications of similar VE-VCO2 regression lines. These findings suggest that the flux of CO2 to the lungs is an important determinant of ventilatory drive during recovery, and that neither central command nor neural afferents from contracting muscles are requisite for the control of ventilation during recovery from exercise.

Adult↗

Semicircular suture annuloplasty for mitral regurgitation: appraisal of the Paneth-Burr method.

BACKGROUND AND AIMS OF THE STUDY: Mitral annuloplasty has long been considered as the basic procedure for mitral regurgitation (MR). However, it is increasingly advisable to avoid foreign material and to use a method that provides greater adjustment for control of annular size. In this study, the semicircular suture annuloplasty (Paneth-Burr method) was evaluated for its efficacy and durability; the optimum size of the mitral annulus was also assessed. METHODS: The Paneth-Burr method was modified using Gore-Tex suture, a crossing suture technique, and obturator for temporary annulus size adjustment. Between 1992 and 1994, 21 patients with an average age of 45 years (range: 8 months to 67 years) underwent such annuloplasty combined with various valvuloplasties and chordal reconstruction. The etiologies were degenerative in 11 patients, congenital in five, ischemic in three and endocarditis in two. RESULTS: Mean MR grades (0-4) decreased from 3.3 +/- 0.6 to 0.2 +/- 0.4 after surgery. At an average 24 months follow up (range: 12 to 41 months), 16 patients showed no increase in MR, but five showed increased MR grades (all grade 3 or less: three grade 3 and two grade 2). Only one patient with ischemic MR required reoperation. Patients with increased MR grade during follow up had a larger intraoperative annular size (> 90% of normal), while those with < 90% of normal mitral annular size showed no increase in MR beyond grade 2, except one with ischemic etiology. CONCLUSIONS: A measured semicircular suture annuloplasty appears to be useful for MR, with acceptable efficacy and durability. In addition, the annular size to be adjusted at surgery is proposed to be < or = 90% of normal mitral annular diameter.

Adolescent↗

[Two patients with mediastinal parathyroid cysts].

We encountered two patients with mediastinal parathyroid cysts. Patients 1 was a 44-year-old man with no complaints who had an abnormal shadow on a roentgenogram of the chest. An iperation was performed because a computed tomogram and a magnetic resonance image showed a cyst in the upper right part of the mediastinum, which was suspected to be a mediastinal thyroid cyst. Patient 2 was a 77-year-old woman with no complaints who had an abnormal shadow on a roentgenogram of the chest. Therapeutic thoracoscopy was performed because a mediastinal cyst was suspected based on the results of computed tomography and magnetic resonance image. Both patients had thin-walled that they were mediastinal parathyroid cysts. Mediastinal parathyroid cysts are very rare. We searched the medical literature but found previously published reports of only 39 cases.

Adult↗