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Biomedical subjects

K Tsuchida

Publications and source records attributed to K Tsuchida.

At least 55 records · Page 3Linked to original sources

Cytogenetic analysis shows that the unusually large chromosome in the sex-limited pB silkworm (Bombyx mori) strain consists of three chromosomes.

We have discovered an inordinately large chromosome pair at the pachytene stage in the oocyte of the sex-limited pB (black larval marking) silkworm (Bombyx mori) strain (TWPB). We have analyzed the composition and arrangement of this large chromosome. A genetic linkage analysis shows that the large chromosome is made up of the W chromosome, the second chromosome fragment (pB fragment), and the fifth chromosome (linkage group) containing at least the region from map position 0.0 to 40.8. We also observed a sex heterochromatin body (SB) that we deduced to be made up of condensed W chromosomes. The number of SBs in each female nucleus among the sucking stomach cells of the TWPB strain was variable. Evidently, the W chromosome of the TWPB strain is attached to another chromosome. The composition of the W chromosome, the second chromosome fragment, and the fifth chromosome was studied through linkage analysis for these three chromosomes. We used two strains derived from the TWPB strain, the sex-limited pM (moricaud larval marking)-like (TWPML) and the autosomal pM-like (T5PML). The results show that the TWPML strain originates through a detachment of the fifth chromosome from the large chromosome of the TWPB strain, and the T5PML strain originates through a detachment of the W chromosome from that. Accordingly, the large chromosome of the TWPB strain is arranged in the order W chromosome--second chromosome fragment--fifth chromosome.

Animals↗

Low density lipoproteins develop resistance to oxidative modification due to inhibition of cholesteryl ester transfer protein by a monoclonal antibody.

Although numerous studies have investigated the relationship between cholesteryl ester transfer protein (CETP) and high density lipoprotein (HDL) remodeling, the relationship between CETP and low density lipoproteins (LDL) is still not fully understood. In the present study, we examined the effect of the inhibition of CETP on both LDL oxidation and the uptake of the oxidized LDL, which were made from LDL under condition of CETP inhibition, by macrophages using a monoclonal antibody (mAb) to CETP in incubated plasma. The 6-h incubation of plasma derived from healthy, fasting human subjects led to the transfer of cholesteryl ester (CE) from HDL to VLDL and LDL, and of triglycerides (TG) from VLDL to HDL and LDL. These net mass transfers of neutral lipids among the lipoproteins were eliminated by the mAb. The incubation of plasma either with or without the mAb did not affect the phospholipid compositions in any lipoproteins. As a result, the LDL fractionated from the plasma incubated with the mAb contained significantly less CE and TG in comparison to the LDL fractionated from the plasma incubated without the mAb. The percentage of fatty acid composition of LDL did not differ among the unincubated plasma, the plasma incubated with the mAb, and that incubated without the mAb. When LDL were oxidized with CuSO4, the LDL fractionated from the plasma incubated with the mAb were significantly resistant to the oxidative modification determined by measuring the amount of TBARS and by continuously monitoring the formation of the conjugated dienes, in comparison to the LDL fractionated from the plasma incubated without the mAb. The accumulation of cholesteryl ester of oxidized LDL, which had been oxidized for 2 h with CuSO4, in J774.1 cells also decreased significantly in the LDL fractionated from the plasma incubated with mAb in comparison to the LDL fractionated from the plasma incubated without the mAb. These results indicate that CETP inhibition reduces the composition of CE and TG in LDL and makes the LDL resistant to oxidation. In addition, the uptake of the oxidized LDL, which was made from the LDL under condition of CETP inhibition, by macrophages also decreased.

Antibodies, Monoclonal↗

Pharmacokinetics of NS-49, a phenethylamine class alpha(1A)-adrenoceptor agonist, at therapeutic doses in several animal species and interspecies scaling of its pharmacokinetic parameters.

The pharmacokinetics of NS-49, a newly developed phenethylamine class alpha(1A)-adrenoceptor agonist, was investigated in rats, rabbits, and dogs given intravenous and oral doses that have little effect on the renal blood flow rate (approximating the range of clinical doses). A three-compartment open model adequately described the plasma NS-49 profiles with respective elimination half-lives of 18, 19, and 13 h after intravenous administration of NS-49 to rats, rabbits and dogs. After oral administration, the NS-49 plasma concentrations reached their maximums within 1.5 h in all the species tested, then decreased as in intravenous administration. The systemic availability was 80% for the rats, 70% for the rabbits, and 101% for the dogs. From the pharmacokinetic parameter values for these three species, we predicted human pharmacokinetics of NS-49 after oral administration with an animal scale-up approach. The area under the plasma concentration-time curve (AUC) after oral administration, as well as the total body clearance showed an excellent allometric relationship to body weight across the three species. The oral AUC value for humans therefore could be predicted from this correlation. The predicted value agreed well with the observed value in the clinical phase I study. It was difficult to predict the plasma concentration profile of NS-49 for humans after oral administration because the absorption rate constant (k(a)) that is essential for estimation of the maximum concentration exhibited no correlation across the species tested. But we approximately could simulate the plasma concentration profile of NS-49 for humans by using the k(a) value for the rats or rabbits.

Adrenergic alpha-1 Receptor Agonists↗

Experimental charge density and electrostatic potential in nicotinamide.

The accurate crystal structure of nicotinamide, 3-pyridinecarboxamide, was determined from X-ray and neutron diffraction experiments: C(6)H(6)N(2)O, M(r) = 122.13, monoclinic, P2(1)/c, Z = 4. The electron distribution at 150 K was determined by the maximum entropy method and the electrostatic potential in the crystal was calculated by Fourier convolution of the electron distribution. The electrostatic properties of the nicotinamide molecule depend on the molecular conformation. The asymmetric electrostatic potential field observed above and below the pyridine-ring plane is related to the rotation of the carboxamide group with respect to the pyridine plane. The positive potential peak at the C4 atom of the pyridine ring extends to the C=O-group side of the plane. The asymmetry of the potential on the C4 atom is consistent with the stereospecificity of hydride transfer in NAD(+)/NADH oxidoreduction.

Journal Article↗

A novel tripartite structure comprising a mariner-like element and two additional retrotransposons found in the Bombyx mori genome.

A clone of a DNA-mediated mobile element (transposon) corresponding to a mariner-like element (MLE) was obtained by carrying out the polymerase chain reaction with genomic DNA of Bombyx mori using a Hyalophora cecropia MLE sequence as a primer. This clone had a size of about 4.2 kb and, after sequencing, was found to contain an RNA-mediated, shorter retrotransposon named L1Bm, which was in turn integrated with a much longer retrotransposon named BMC1. Thus, the mobile elements made a novel tripartite structure. The BMC1 and L1Bm moieties of the composite structure each contained a 63-bp conserved sequence which was subsequently found to be highly conserved in all BMC1 and L1Bm elements registered so far. We propose that the 63-bp stretch may be a recognition site for a retrotransposition mechanism conducted by a reverse transcriptase and an endonuclease complex. On the basis of this inference, we propose a model that predicts how different types of BMC1 and L1Bm elements are dispersed in the genome. In addition, a phylogenetic tree made from the current and extant BMC1 and L1Bm sequences indicated that these elements can be classified into Subfamilies I and II.

Animals↗

Suppression of transforming growth factor beta and vascular endothelial growth factor in diabetic nephropathy in rats by a novel advanced glycation end product inhibitor, OPB-9195.

AIMS/HYPOTHESIS: Advanced glycation end products (AGEs) participate in the pathogenesis of diabetic nephropathy. We reported earlier that OPB-9195, a synthetic thiazolidine derivative and novel inhibitor of advanced glycation, prevented progression of diabetic glomerulosclerosis by lowering serum concentrations of advanced glycation end products and reducing their deposition in the glomeruli. Here, we examined their contribution and that of growth factors, such as transforming growth factor-beta (TGF-beta) and vascular endothelial growth factor (VEGF), to the progression of diabetic nephropathy. We also investigated the expression of type IV collagen in the kidneys of Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats, a Type II (non-insulin-dependent) diabetes mellitus model, after treatment with OPB-9195. METHODS: Using northern blots and immunohistochemical techniques, we determined the renal expression of TGF-beta and type IV collagen mRNAs and proteins in OLETF rats. We also examined OPB-9195's effects on renal expression of VEGF mRNA and protein. RESULTS: Concomitant increases in TGF-beta and type IV collagen expression were observed at each point in time in OLETF rats not given OPB-9195. In contrast, OPB-9195 treatment greatly suppressed the renal expression of TGF-beta, VEGF and type IV collagen mRNAs and proteins to that seen in non-diabetic rats. CONCLUSION/INTERPRETATION: Since OPB-9195, an AGE-inhibitor, prevented the progression of diabetic nephropathy by blocking type IV collagen production and suppressing overproduction of two growth factors, TGF-beta and VEGF, in diabetic rats, this compound warrants further investigation.

Animals↗

Inhibitory effect of TS-962 on the formation of early atherosclerotic lesions in high fat-fed hyperlipidemic hamsters.

The hypocholesterolemic and anti-atherosclerotic effect of TS-962, a specific ACAT inhibitor, was investigated in a hamster model fed a high fat diet containing 10% coconut oil and 0.05% cholesterol. Lipid accumulated atherosclerotic lesions were detected by using oil red O staining in the lesion-prone aortic arch. A high dose, estimated to be 15 mg/kg, of TS-962 decreased serum cholesterol to normal levels with reduction of liver cholesterol contents below normal levels, and as a consequence, entirely inhibited the lipid accumulation in the aortic arch. Furthermore, a low dose, estimated to be 1.5 mg/kg, of TS-962 remarkably inhibited aortic lipid accumulation by 73% compared with the control group, without changing either serum cholesterol level or liver cholesterol content. These findings suggest that TS-962 is effective in the primary prevention of atherosclerosis by directly suppressing the formation of foam cells in arteries.

Acetamides↗

Secretion of prebeta HDL increases with the suppression of cholesteryl ester transfer protein in Hep G2 cells.

Prebeta HDL are small, protein rich lipoproteins that are predominantly composed of apo A-I, without apo A-II. Prebeta HDL are secreted from the liver as nascent HDL and/or are produced in the incubated plasma by cholesteryl ester transfer protein (CETP). However, the role of CETP in the secretion of HDL from the liver has yet to be determined. In the present study, we examined the effect of the suppression of hepatic CETP by antisense oligodeoxynucleotides (ODNs) against CETP targeted to the liver on the secretion of apo A-I using a Hep G2 cell culture. The ODNs against CETP were coupled to asialoglycoprotein (ASOR) carrier molecules, which serve as an important method for the regulation of liver gene expression. Hep G2 cells were cultured in DMEM supplemented with 10 FBS. After 2 days, the medium was changed to DMEM with EGF and the cells were divided into three groups. The control group received saline, while the sense group was mixed with the sense ODNs complex and the antisense group was mixed with the antisense ODNs complex, respectively, for 2 days. Both the hepatic CETP mRNA and the CETP mass in the medium in the antisense group decreased significantly more than in the sense and the control groups (CETP mass: 1.697 + /- 0.410 ng/mg cell protein vs. 2.367 + /- 0.22 and 2.360 + /- 0.139, n = 3 in each determination). In contrast, both the hepatic apo A-I mRNA and the apo A-I mass in the medium in the antisense group were significantly higher than those in the sense and the control groups (apo A-I mass; 1.877 + /- 0.215 micro/mg cell protein vs. 1.213 + /- 0.282 and 1.097 + /- 0.144, n = 3 in each determination). The increase in apo A-I was mainly due to the increase in prebeta apo A-I. These findings may partly explain why HDL and apo A-I increase in patients with CETP deficiency, while also indicating the possibility that the original level of prebeta HDL is sufficient in such patients.

Apolipoprotein A-I↗

Pharmacological actions of monovalent ionophores on spontaneously beating rabbit sino-atrial nodal cells.

The effects of sodium (monensin) and potassium (nigericin and lonomycin A) ionophores on the spontaneous activity in rabbit sino-atrial (SA) nodal cells were investigated using microelectrode and whole-cell voltage-clamp techniques. In the multicellular preparations, the ionophores produced a negative chronotropic effect in a concentration-dependent manner, and at 3x10(-5) M significantly decreased the amplitude and duration of action potentials and enhanced the maximum rate of depolarization. The ionophores elicited dysrhythmias and then a sinus arrest often occurred. These responses were reversible. In whole-cell clamp experiments, monensin enhanced the L-type Ca2+ current (ICa), whereas lonomycin A and nigericin inhibited ICa. The fast component of the inactivation phase for ICa was decreased by the ionophores, but the slow component was unaffected. The activation and inactivation kinetics (d infinity and f infinity) were not altered. The ionophores did not affect the hyperpolarization-activated inward current. Monensin inhibited the delayed rectifier K+ current (I(K)), but lonomycin A and nigericin increased I(K). Its activation kinetics shifted in the depolarizing direction. The effects on the ionic currents were irreversible. Monensin (30 microM) increased cellular Ca2+ concentration ([Ca2+]i), using Ca2+ -sensitive fluorescent dye (fura-2). These results indicate that the monovalent ionophores depress the action potentials and produce a negative chronotropic effect due to direct and indirect modulations of the ionic currents and the [Ca2+]i level in rabbit SA nodal cells.

Action Potentials↗

Purification and characterization of fetal bovine serum beta-N-acetyl-D-galactosaminyltransferase and beta-D-glucuronyltransferase involved in chondroitin sulfate biosynthesis.

beta-N-Acetylgalactosaminyltransferase II and beta-glucuronyltransferase II, involved in chondroitin sulfate biosynthesis, transfer an N-acetylgalactosamine (GalNAc) and glucuronic acid (GlcA) residue, respectively, through beta-linkages to an acceptor chondroitin oligosaccharide derived from the repeating disaccharide region of chondroitin sulfate. They were copurified from fetal bovine serum approximately 2500-fold and 850-fold, respectively, by sequential chromatographies on Red A-agarose, phenyl-Sepharose, S-Sepharose and wheat germ agglutinin-agarose. Identical and inseparable chromatographic profiles of both glycosyltransferase activities obtained through the above chromatographic steps and gel filtration suggest that the purified enzyme activities are tightly coupled, which could imply a single enzyme with dual transferase activities; beta-N-acetylgalactosaminyltransferase and beta-glucuronyltransferase, reminiscent of the heparan sulfate polymerase reaction. However, when a polymerization reaction was performed in vitro with the purified serum enzyme preparation under the polymerization conditions recently developed for the chondroitin-synthesizing system, derived from human melanoma cells, each monosaccharide transfer took place, but no polymerization occurred. These results may suggest that the purified serum enzyme preparation contains both beta-N-acetylgalactosaminyltransferase II and beta-glucuronyltransferase II activities on a single polypeptide or on the respective polypeptides forming an enzyme complex, but is different from that obtained from melanoma cells in that it transfers a single GalNAc or GlcA residue but does not polymerize chondroitin.

Animals↗

Preventive nutritional factors in epidemiology: interaction between sodium and calcium.

1. It is generally believed, though difficult to prove, that diet plays a role in the risk of various diseases. Components of difficulties include several issues such as dietary assessment method, regression dilution bias, multicolinearity and interaction among nutrients. 2. The present study focuses on colinearity and interaction between sodium and calcium, which should be cautiously examined in nutritional epidemiological studies in relation to blood pressure and bone mineral density. 3. The World Health Organization's International Cooperative Cardiovascular Diseases and Alimentary Comparison study showed significant multicolinearity among urinary sodium, calcium and urea nitrogen as well as urinary calcium and magnesium. Urinary sodium and calcium had significant correlation (r = 0.438, P < 0.05, n = 48) by cross-centre analysis. 4. Interaction between sodium and calcium on bone mineral density is studied using the data set from bone mineral density screening for 1658 females, aged 20-40 years, in Yokohama, Japan. Among those who have lower calcium intake (< 600 mg/day), higher calcium intake (%) from small fish, which is likely to be associated with a high salt diet, related to significantly lower mineral bone density. 5. Interaction between sodium and calcium on bone mineral density among young Japanese females is suggested. Moderate sodium restriction is needed for prevention of not only cardiovascular diseases but also osteoporosis.

Blood Pressure↗

Arterial anastomosis using the telescoping method and an elastomeric adhesive.

We experimentally studied a new technique for anastomosis of small arteries which involves the telescoping method and a surgical adhesive with the objective of examining its clinical potential. This technique was applied to the unilateral femoral arteries of 27 mongrel dogs. After division of the artery, the distal artery was incised longitudinally, and the proximal end was invaginated into the opened artery. One stay suture and the elastomeric adhesive PUP201 were placed in the anastomotic site. The bursting and tensile strength and the patency were examined, and a histopathological study was performed at various intervals up to 1 year after the operation. The bursting strength exceeded 500 mm Hg. The patency rate of the anastomosed arteries was 100%. The mean percentage of stenosis to diameter of the proximal normal lumen was 20% one year after operation. The luminal surface of the anastomosed line was healed smoothly, and thrombi between the telescoped arteries were replaced by elastofibrotic union. In conclusion, small arterial anastomosis using the telescoping method and an elastomeric adhesive is easy and safe and provides good patency.

Anastomosis, Surgical↗

Embryonal renal remnant in the gubernaculum testis.

We present a rare case of embryonal renal remnant found by chance as a mass in the gubernaculum testis during an operation on a right undescended testis. We discuss the relationship between this tissue and paradidymis or supernumerary kidney.

Choristoma↗

AV reentrant and idiopathic ventricular double tachycardias: complicated interactions between two tachycardias.

An electrophysiological study was performed in a 61 year old man with Wolff- Parkinson-White (WPW) syndrome. At baseline, neither ventricular nor supraventricular tachycardias could be induced. During isoprenaline infusion, ventricular tachycardia originating from the right ventricular outflow tract (RVOT) with a cycle length of 280 ms was induced and subsequently atrioventricular reentrant tachycardia (AVRT) with a cycle length of 300 ms using an accessory pathway in the left free wall appeared. During these tachycardias, AVRT was entrained by ventricular tachycardia. The earliest ventricular activation site during the ventricular tachycardia was determined to be the RVOT site and a radiofrequency current at 30 W successfully ablated the ventricular tachycardia at this site. The left free wall accessory pathway was also successfully ablated during right ventricular pacing. The coexistence of WPW syndrome and cathecolamine sensitive ventricular tachycardia originating from the RVOT has rarely been reported. Furthermore, the tachycardias were triggered by previous tachycardias.

Adrenergic beta-Antagonists↗

The therapeutic effect of proton pump inhibitors on Helicobacter pylori-positive gastric ulcers.

The aim of the present study was to elucidate the risk factors that could delay gastric ulcer healing when either a proton pump inhibitor or an H2-receptor antagonist is used for gastric ulcer treatment. Endoscopically-diagnosed gastric ulcer patients (216 men and 96 women, mean age: 57+/-13 years) were investigated. All patients were consecutively recruited and randomly assigned to receive H2-receptor antagonist (n = 196) or proton pump inhibitor (n = 116) treatment for eight weeks. Chi-squared tests and multivariate analysis to determine factors influencing ulcer healing were used to analyse the patients profiles, endoscopic findings, and Helicobacter pylori-infection status. In the H2-receptor antagonist group, the most important risk factor was a large ulcer size (> 2 cm diam.), followed by a linear shape of the ulcer, undermining tendency of ulcer, previous history of gastric ulcer, and H. pylori infection. In the proton pump inhibitor group, linear shape of the ulcer was the only significant risk factor for slow ulcer healing; other factors, including H. pylori infection, were insignificant. These results indicate that ulcer morphology may be the most important information for predicting ulcer healing, and that H. pylori infection does not delay gastric ulcer healing when proton pump inhibitor treatment is used.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Dietary soybeans intake and bone mineral density among 995 middle-aged women in Yokohama.

To investigate relationship of dietary factors, especially source of calcium intake, to bone mineral density (BMD) among Japanese middle-aged women, a total of 995 healthy women age of 40 to 49 (mean +/- SD, 45 +/- 3), who lives in Yokohama-city, were recruited through convenience sampling by the municipal information paper and health announcement at each 18 public health center in 18 wards for the three-day course on prevention of osteoporosis from October 1996 to March 1998. The BMD of the 2nd metacarpal bone was measured using Computed X-ray Densitometry (CXD) method, by a trained radiologist. Dietary intake of calcium was assessed by self-reporting food frequency questionnaire on calcium dietary sources such as milk, dairy products, small fish, vegetables, and soybeans and carefully checked by trained dietician. An independent gradient of non-adjusted and adjusted BMD for age and weekly calcium intake, through soybeans intake frequency (p = 0.03) was noted. This study suggest soybeans, through possible beneficial effects of vitamin-K, soyprotein, and isoflavonoid, may affect BMD of middle aged women.

Adult↗