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Biomedical subjects

K Takeda

Publications and source records attributed to K Takeda.

At least 757 records · Page 42Linked to original sources

Effects of tumor necrosis factor-alpha on connective tissue metabolism in normal and scleroderma fibroblast cultures.

Recent studies have demonstrated that tumor necrosis factor-alpha (TNF-alpha) selectively decreases production of collagens I and III, the major types of collagen in the dermis, and increases production of collagenase in cultured dermal fibroblasts. The effects of TNF-alpha on collagens I, III and VI, fibronectin and collagenase gene expression by fibroblasts derived from normal individuals and patients with systemic sclerosis (SSc) were studied. SSc is characterized by excessive accumulation of collagen in the skin and in certain organs. TNF-alpha inhibited collagen production and mRNA levels of collagens I and III and of fibronectin, and stimulated collagenase activity and collagenase mRNA levels in SSs fibroblasts. Levels of mRNA for alpha 1 (VI) and alpha 3 (VI) collagen and for beta-actin were unaltered in SSc fibroblasts incubated with TNF-alpha. Similar results were observed for mRNA levels in normal fibroblasts incubated with TNF-alpha. These results suggest that TNF-alpha could be expected to be beneficial in the treatment of SSc. In addition, our results indicated that collagen-VI expression is regulated independently from expression of collagens I and III, and expression of fibronectin and collagens I and III are regulated in parallel in fibroblasts treated with TNF-alpha.

Adult↗

Conformational changes of alpha-lactalbumin and its fragment, Phe31-Ile59, induced by sodium dodecyl sulfate.

Conformational changes of bovine alpha-lactalbumin in sodium dodecyl sulfate (SDS) solution were studied with the circular dichroism (CD) method using a dilute phosphate buffer of pH 7.0 and ionic strength 0.014. The proportions of alpha-helix and beta-structure in alpha-lactalbumin were 34% and 12%, respectively, in the absence of SDS. In the SDS solution, the helicity increased to 44%, while the beta-structure disappeared. In order to verify the structural change from beta-structure to alpha-helix, the moiety, assuming the beta-structure in the alpha-lactalbumin, was isolated by a chymotryptic digestion. The structure of this alpha-lactalbumin fragment, Phe31-Ile59, was almost disordered. However, the fragment adopted a considerable amount of alpha-helical structure in the SDS solution. On the other hand, the tertiary structure of alpha-lactalbumin, detected by changes of CD in the near-ultraviolet region, began to be disrupted before the secondary structural change in the surfactant solution. Dodecyl sulfate ions of 80 mol were cooperatively bound to alpha-lactalbumin. Although the removal of the bound dodecyl sulfate ions was tried by the dialysis against the phosphate buffer for 5 days, 4 mol dodecyl sulfates remained per mole of the protein. The remaining amount agreed with the number of stoichiometric binding site, determined by the Scatchard plot, indicating that the stoichiometric binding was so tight.

Circular Dichroism↗

Secondary structural changes of large and small fragments of bovine serum albumin in thermal denaturation and in sodium dodecyl sulfate denaturation.

The helicities in various fragments of bovine serum albumin (BSA) were examined in the thermal denaturation and in sodium dodecyl sulfate (SDS) denaturation. The thermal denaturation was examined in a temperature range between 2 and 65 degrees C. The helicity decreased with a rise of temperature and it recovered to some degree upon cooling temperature. A rather high reversibility was observed in the BSA fragments, which were located in the N-terminal of the parent protein and then contained the first large loop with no disulfide bridge. The high reversibility was available also for the helicity in the first large loop of the fragment, disulfide bridges of which were reduced. The fragments, which were smaller than one domain, became unstable in the SDS denaturation. The helicities of such fragments decreased in lower SDS concentrations compared with those of the intact BSA and the large fragments, which contained one or more domains. A resistance to the SDS denaturation appeared in the helices of every large loop even after the fragmentation. On the other hand, helicities of the fragments decreased to 20-25% upon the reduction of disulfide bridges. However, the helicities of these fragments increased to 35-40% in the SDS denaturation.

Animals↗

Plasmalemmal, voltage-dependent ionic currents from excitable pulvinar motor cells of Mimosa pudica.

Plasmalemmal ionic currents from excitable motor cells of the primary pulvinus of Mimosa pudica were investigated by patch-clamp techniques. In almost all of the enzymatically isolated protoplasts, a delayed rectifier potassium current was activated by depolarization, while no currents were detected upon hyperpolarization. This sustained outward current was reversibly blocked by Ba and TEA and serves to repolarize the membrane potential. Outward single channel currents that very likely underly the macroscopic outward potassium current had an elementary conductance of approximately 20 pS. In addition, in a few protoplasts held at hyperpolarized potentials, depolarization-activated transient inward currents were observed, and under current clamp, action potential-like responses were triggered by depolarizing current injections or by mechanical perturbations. The activation characteristics of both inward currents and spikes showed striking similarities compared to those of action potentials in situ.

Barium↗

31P MR spectroscopy in hypertrophic cardiomyopathy: comparison with Tl-201 myocardial perfusion imaging.

Abnormal phosphate metabolism of the myocardium was evaluated in patients with hypertrophic cardiomyopathy (HCM) using 31P magnetic resonance (MR) spectroscopy. The results were compared with those from left ventricular function and thallium 201 (Tl-201) perfusion scintigraphy. Six normal volunteers and 19 patients with HCM were studied with a 1.5 T MR system. The spectra were localized to the myocardium using volume selection with the depth-resolved surface coil spectroscopy (DRESS) technique. Peak areas of 2,3-diphosphoglycerate (DPG), phosphodiesters (PDE), phosphocreatine (PCr), and beta-ATP were determined by fitting Gaussian functions to the phased spectra. The peak areas were corrected for contamination of blood adenosine triphosphate (ATP) and PDE. The corrected PCr/beta-ATP ratio in patients (1.07 +/- 0.10, mean +/- SE) was significantly lower compared with that in normal volunteers (1.71 +/- 0.13, p < .01). The PCr/beta-ATP ratio showed an abnormal decrease (< mean -2 SD of the controls) in 11 (58%) of 19 patients. The averaged PCr/beta-ATP ratio in 15 patients with normal left ventricular ejection fraction (LVEF) was 1.14 +/- 0.10, significantly lower than in healthy subjects. By contrast, the corrected PDE/PCr ratio in HCM did not differ significantly compared with that in healthy subjects (0.46 +/- 0.09 vs 0.36 +/- 0.09). The PDE/PCr ratio was abnormally elevated (> mean + 2 SD of the controls) in only four (21%) of the patients. On Tl-201 myocardial single-photon emission computed tomography (SPECT) imaging, the perfusion of the left ventricular wall looked normal in 6 and abnormal in 5 of 11 HCM patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

Effect of sterol biosynthesis inhibitor, SSF-109, on cholesterol synthesis in isolated rat hepatocytes.

The inhibitory effect of SSF-109 on cholesterol synthesis in isolated hepatocytes was studied using a radio-high-performance liquid chromatography system, and the results were compared with those of other inhibitors, triparanol and AMO-1618. SSF-109 caused accumulation of two trimethylsterols: lanosterol and 24-dihydrolanosterol. The distribution profile of [2-14C]mevalonate-originated radioactivity in cholesterol, lanosterol, dihydrolanosterol, 2,3-oxidosqualene, and squalene indicates that SSF-109 inhibits the 14 alpha-methyl demethylase activity. Triparanol accumulated the radioactivity of [2-14C]mevalonate in desmosterol and 2,3-oxidosqualene suggesting that triparanol inhibits sterol delta 24-reductase and 2,3-oxidosqualene cyclase. AMO-1618 caused enrichment of the radioisotope from [2-14C]mevalonate in 2,3-oxidosqualene but reduced it in squalene, suggesting that AMO-1618 acts on 2,3-oxidosqualene cyclase and some enzyme that catalizes a metabolic pathway between mevalonate and squalene.

Animals↗

Postprandial change in canine blood viscosity.

1. Postprandial variation in blood viscosity was studied in beagle dogs. 2. Blood viscosity increased following feeding. This change was caused by haematocrit elevation, which resulted mainly from splenic contraction. 3. Haemoconcentration, plasma viscosity and erythrocyte deformability did not contribute to the postprandial increment in blood viscosity.

Animals↗

Differences in levels of erythrocyte glutathione and its metabolizing enzyme activities among primates.

1. The levels of erythrocyte glutathione and the activities of its metabolizing enzymes--glutathione peroxidase (GSH-Px), glutathione S-transferase (GST) and glutathione reductase (GR)--were measured in four species of primates: human, rhesus monkey, common marmoset and common tree shrew. 2. There were marked differences in GSH-Px and GST activities among the primates, while GR activity and glutathione level were much less variable.

Animals↗

Stizolobic acid on frog spinal cord; possible species dependent activation of excitatory amino acid receptors.

1. We examined the effects of stizolobic acid, an amino acid isolated from a plant, Stizolobium hassjoo, on the binding of [3H]glutamic acid and [3H]kainic acid to synaptosomes from frog spinal cords and on the depolarization at the ventral roots of frog spinal cords. 2. Stizolobic acid inhibited the binding of [3H]kainic acid more potently than that of [3H]glutamic acid. 3. Among stizolobic acid derivatives, 3-Br-stizolobic acid was the most potent inhibitor of the binding of [3H]kainic acid, but the inhibitory potency was 100 times weaker than that of kainic acid. 4. Stizolobic acid and its derivatives could cause depolarization of the ventral root of frog spinal cord in a dose dependent manner, and 3-Br-stizolobic acid was a more potent inducer of depolarization than kainic acid, but the dose dependency of 3-Br-stizolobic acid was a little different from that of kainic acid. 5. The results suggest that stizolobic acid and its analogues act as a kainic acid agonist in frog spinal cord. 6. The present results and others indicate that stizolobic acid may interact with the different types of excitatory amino acid receptors dependent on the species of animals.

Amino Acids, Dicarboxylic↗

Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation.

We cloned a cDNA coding for a putative human heme oxygenase isozyme, designated type 2 (HO-2), and analyzed its function by transient expression assays. HeLa cells transfected with either HO-2 cDNAs or a cDNA coding for authentic heme oxygenase (HO-1) expressed the activity of heme oxygenase, although no activity was detected in the mock transfected cells. Using specific anti-HO-1 antibody, we showed that expression of a HO-1 cDNA resulted in the increase in its protein levels, but HO-1 protein was not detectable in the cells expressing HO-2 cDNAs. We thus confirmed the functional identity of HO-1 and HO-2. Then, we analyzed their expression in an erythroid cell line, YN-1-0-A. Treatment with hemin or by heat shock (42 degrees C) led to a remarkable increase in the HO-1 mRNA levels, while HO-2 mRNA expression was not induced at all, suggesting that they are under separate regulation.

Cell Line↗

Effects of emergency coronary angioplasty on left ventricular volume after acute myocardial infarction compared with intracoronary thrombolysis.

BACKGROUND: The effects of emergency coronary angioplasty on left ventricular remodeling have not been fully evaluated. We compared the effects of emergency coronary angioplasty on left ventricular volume with those of intracoronary thrombolysis in patients suffering from their first acute myocardial infarction (AMI). METHODS: A total of 51 patients who had a culprit lesion at site No 6 of the American Heart Association (AHA) classification were analyzed. Patients with re-obstruction of the culprit lesion after reperfusion were excluded from the study. The coronary angioplasty and the thrombolysis groups consisted of 28 and 23 patients, respectively. Left ventricular volume was measured by left ventriculography at 4 weeks after infarction. RESULTS: Clinical and angiographic background factors showed no significant differences between these two groups. The left ventricular end-systolic volume index was significantly smaller in the angioplasty group than in the thrombolysis group (31 +/- 14 versus 45 +/- 14 ml/m2, P < 0.01). Similarly, the left ventricular end-diastolic volume index was significantly smaller in the angioplasty group (64 +/- 13 versus 82 +/- 13 ml/m2, P < 0.01). Conversely, the left ventricular ejection fraction was larger in the angioplasty group than in the thrombolysis group (54 +/- 13 versus 45 +/- 13%, P < 0.05). Multivariate analysis revealed that the selection of coronary angioplasty was one of the factors significantly associated with a decrease in left ventricular end-systolic and end-diastolic volume. CONCLUSION: These results suggest that emergency coronary angioplasty produces better left ventricular remodeling than intracoronary thrombolysis after AMI.

Adult↗

Decreased type VI collagen gene expression in cultured Werner's syndrome fibroblasts.

Gene expression of collagens VI, I, and III in Werner's syndrome was studied by measuring messenger RNA (mRNA) and protein production levels in four fibroblast strains from patients with Werner's syndrome and comparing them with age-matched healthy subjects. Levels of type VI collagen mRNA were decreased in all Werner's syndrome fibroblast strains and the decreases were in parallel in all three chains (alpha 1, alpha 2, and alpha 3) of type VI collagen. A coordinate increase of the alpha 1(I) and alpha 1(III) collagen mRNA levels was observed in three of the four Werner's syndrome fibroblast strains. However, no qualitative abnormality of these mRNA transcripts in Werner's syndrome fibroblasts were found by Northern blot analysis. Changes in type VI and type I collagen mRNA correlated well with production levels of corresponding proteins, as determined by immunologic assays. These data suggest that there are changes in expression of multiple connective tissue constituents in Werner's syndrome fibroblasts.

Adult↗

Relation between bronchial responsiveness to methacholine and levels of IgE antibody against Dermatophagoides farinae and serum IgE in asthmatic children.

The relation between non-specific bronchial responsiveness and allergic sensitivity was evaluated in children with asthma. Bronchial responsiveness was determined by methacholine inhalation challenge test, and was expressed as a provocative concentration of methacholine causing a 20% fall in FEV1 (PC20). RAST titre for Dermatophagoides farinae and serum IgE level were evaluated as parameters of allergic sensitivity. When the PC20 values, RAST titres for D. farinae, and serum IgE levels of 47 asthmatic children and 16 normal controls were compared, the asthmatic children had significantly lower PC20 values and higher D. farinae-RAST titres and serum IgE levels than the normal controls. The correlation analyses in 47 asthmatic children have shown that there is no significant correlation between PC20 values and RAST titres for D. farinae (r = 0.04, P > 0.1) or between PC20 values and serum IgE levels (r = 0.03, P > 0.1). These results suggest that although both bronchial hyperresponsiveness and allergic sensitization may be in some way related to one another in children with asthma, the magnitude of allergic sensitization does not influence the degree of the non-specific bronchial responsiveness.

Adolescent↗

Effects of ATP on cultured smooth muscle cells from rat aorta.

1. Membrane ionic currents provoked by externally applied ATP were studied by patch-clamp techniques in cultured aortic smooth muscle cells of the rat. 2. Using standard bath and pipette solutions and whole-cell voltage-clamp, ATP evoked an inward current when the cell membrane potential was held at -50 mV and an outward current when the potential was held at 30 mV, with a reversal potential near -10 mV. 3. Application of ATP gamma S gave results similar to those obtained with ATP, while adenosine, AMP and alpha,beta-methylene ATP were ineffective. The ATP-activated current was inhibited by suramin, 100 microM. 4. ATP also induced a biphasic rise in internal free Ca levels as shown directly by Fura-2 measurements and by the increase in Ca-dependent K single-channel activity in cell-attached patches. 5. With outward current through K channels blocked by internal Cs and TEA, modification of the ionic composition of bath and pipette solutions revealed that the reversal potential for the ATP-induced whole-cell current closely followed ECl, the chloride equilibrium potential, and was insensitive to manipulations of the monovalent cation gradient. 6. These results indicate that in rat cultured aortic smooth muscle cells, ATP binding to P2-purinoceptors produces increases of internal free Ca levels and subsequent activation of both Ca-dependent K and Cl currents.

Adenosine Triphosphate↗

Effects of new quinolones on Mycoplasma pneumoniae-infected hamsters.

The efficacies of the new quinolones temafloxacin, ofloxacin, and ciprofloxacin were investigated against Mycoplasma pneumoniae in an experimental hamster pneumonia model. Hamsters were infected intratracheally with M. pneumoniae and sacrificed 18 h after the final medication, and their lungs were aseptically removed, homogenized, and cultured quantitatively. The efficacies of these drugs were determined by the CFU of M. pneumoniae in lungs. Temafloxacin and ofloxacin, but not ciprofloxacin, were active when the oral administration of 200 mg/kg of body weight per day (once per day) for 5 days was initiated 24 h after infection. Although no effect on the elimination of M. pneumoniae was observed after the administration of these drugs at 200 mg/kg/day at 5 days after infection, the continuous administration for 15 days of temafloxacin, but not ofloxacin or ciprofloxacin, significantly reduced viable M. pneumoniae in the lungs. These results suggest that temafloxacin and ofloxacin are effective in the acute phase of infection and, moreover, that temafloxacin is effective in the late stage of infection during which progressive lung alterations and continuous increases in mycoplasmal growth occurred. The peak levels of temafloxacin in sera and lungs after oral administration were similar to those of ofloxacin and higher than those of ciprofloxacin. The areas under the curve of temafloxacin in the lung tissue, however, were higher than those of ofloxacin and ciprofloxacin. On the basis of these results, temafloxacin and ofloxacin might be promising antimicrobial agents for the treatment of mycoplasmal infection.

Animals↗

Pure progressive autonomic failure: a clinicopathological study.

Two cases of pure progressive autonomic failure (PAF) are presented. A postmortem study of one case (case 2) showed a pathology resembling that of Parkinson's disease. Marked cell loss was noted in the substantia nigra, nucleus ceruleus, and intermediolateral column of the spinal cord, while cell loss in the sympathetic ganglion was not remarkable. This case may be an exceptionally rare case of late-onset PAF in which autonomic failure was mainly ascribed to preganglionic (and central) pathology, although autonomic function tests suggested postganglionic sympathetic disorder in both cases.

Aged↗