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Biomedical subjects

T Shimada

Publications and source records attributed to T Shimada.

At least 793 records · Page 44Linked to original sources

[Clinical significance of red cell distribution width in polycythemia vera].

We evaluated changes in red cell distribution width-standard deviation (RDW-SD) measured using a multiple parameter automated hematology analyzer E 4000 in patients with polycythemia vera (PV). Patients with iron deficiency anemia, those with chronic myelogenous leukemia, those with primary thrombocythemia, and normal subjects were examined as controls. In the patients with PV, as in those with the other 3 diseases, RDW-SD tended to be higher than in the normal controls when red blood cell counts were high. The RDW-SD in patients with PV transiently increased following administration of a myelosuppressive, which corresponded to the transition period from microcytes to normal blood cells. It was even higher during the polycythemic period than during the myelofibrotic period. This may be associated with hematopoietic abnormality due to extramedullary hematopoiesis. RDW-SD seems to well reflect the pathologic status of PV.

Adolescent↗

[A case of bilateral coronary artery to pulmonary artery fistulas associated with hepatic arteriovenous fistula].

A 55-year-old man, previously treated for primary hypothyroidism, was admitted for evaluation of atypical chest pain. On physical examination, the pulse rate was 60 and blood pressure was 132/84 mmHg. Heart sounds were normal and no murmur was heard. Abdominal palpation showed no abnormal finding, and bruit was not heard. The electrocardiogram was normal and stress-induced ischemia was not found. The chest X-ray showed no sign of pulmonary hyperaemia or mass lesion, and central shadow was normal (CTR = 44%). During right heart catheterization, the pressures were normal in the cardiac chambers, pulmonary artery, and wedge position. A significant increase in oxygen saturation was disclosed at pulmonary artery level (10%) and the upper site of the inferior vena cava (22%). Coronary arteriography found coronary artery fistulas from the left main trunk, the left anterior descending artery, the left circumflex, and the right coronary artery to the pulmonary trunk. The left and right ventriculograms were normal. The digital subtraction angiography of celiac artery showed hepatic arteriovenous fistula. Ultrasonography and computed tomography found no mass lesion of the liver. Since combination of the bilateral coronary artery with pulmonary artery fistula, and hepatic arteriovenous fistula is very rare, the present case is worth noting for the investigation of the pathogenesis of congenital arteriovenous fistulas.

Arterio-Arterial Fistula↗

[Anatomical and clinical studies on peroneal vascularized composite tissue transfer].

An anatomical study of the peroneal artery and vein and their branches was carried out on 86 cadaver legs. The number of cutaneous branches was 4.8 +/- 1.4 per leg. The length of the cutaneous branches was 5.5 +/- 1.6 cm. The external diameters of cutaneous branches at the skin distribution site were 0.6 +/- 0.2 mm for the artery and 0.8 +/- 0.3mm for the vein. The peroneal vessels gave off communicating branches to anterior or posterior tibial vessels 6.1 +/- 2.4 cm proximal from the lateral malleolus. The range of rotation of island flap when transposed proximally was 14.3 +/- 3.3 cm proximal from the head of the fibula, and when transposed distally, the range of rotation was 16.9 +/- 5.3 cm distal from the lateral malleolus. Based on the results obtained from these experimental studies, we have been performing peroneal vascularized tissue transfer since 1982. A total of 72 cases have been performed, and the results have been encouraging.

Adolescent↗

Potentiation by a biscoclaurine alkaloid, cepharanthine, of the toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin in HeLa cells.

Cepharanthine, a biscoclaurine alkaloid, causes an 8-fold enhancement of the cytotoxic effect of a conjugate of epidermal growth factor (EGF) coupled with Pseudomonas exotoxin in HeLa cells. Cepharanthine also potentiates the effect of Pseudomonas exotoxin. Cepharanthine does not affect the binding and uptake of 125I-EGF by HeLa cells, but it delays the release of radioactivity associated with 125I-EGF into the medium. Analysis by colloidal silica gradients using cell homogenates suggests that 125I-EGF accumulates in the lysosomes of cells treated with cepharanthine and that [3H]cepharanthine accumulates in lysosomes. The pH in HeLa cell lysosomes is 5.2, and cepharanthine does not significantly increase the pH. Electron microscopy shows an increased number of electron-dense bodies and dilated Golgi apparatus after cepharanthine treatment. Cepharanthine appears to accumulate in lysosomes, and it may delay degradation of EGF-Pseudomonas exotoxin in the cells as does 125I-EGF.

ADP Ribose Transferases↗

Transfer by pro gene transfection of tumor promoter-sensitive phenotype to promotion-insensitive JB6 cells.

Transfection of activated promotion sensitivity genes (pro genes) confers on insensitive (P-) cells susceptibility to induction of anchorage-independent growth by tumor-promoting phorbol esters. Promotion-sensitive (P+) JB6 cell variants, from which activated pro-1 and pro-2 were cloned, respond to 12-O-tetradecanoylphorbol-13-acetate (TPA) and various nonphorbol tumor promoters with anchorage-independent transformation that is irreversible 60-80% of the time. Anchorage-independent (Tx) clonal lines derived from these TPA-induced agar colonies were also tumorigenic in nude mice. This report has addressed the question of whether the phenotypes associated with parental P+ cells are transferred by transfection of activated pro-1 and pro-2. Clonal lines were established after transfection of JB6 P- cells with activated pro-1 or pro-2, induction of anchorage-independent colony formation by TPA, and growth of individual agar colonies to yield clonal transfectant lines. The lines so derived from transfected populations included Tx, P+, and P- lines, reflecting irreversible neoplastic transformation and greater and lesser degrees of preneoplastic progression, respectively. The anchorage-independent transfectants were found to be tumorigenic. Since untransfected P- cells subjected to the same single-cycle TPA treatment and cloning in agar yielded no anchorage-independent and few P+ transfectants, the appearance of P+ and Tx transfectants after pro-1 and pro-2 transfection is therefore likely to be due to the transfected pro genes. Indirect assay of pro gene uptake by quick-blot hybridization of transfectant cell DNA with the vectors into which pro genes had been cloned confirmed the association of transferred P+ and Tx phenotypes with the presence of the transfected DNA. Finally, assay of the sensitivity of P+ pro-1 and pro-2 transfectants to transformation by TPA at various concentrations showed that transfection with pro-1 or pro-2 conferred about equal responses that were somewhat lower than those observed with parental P+ controls. Taken together these data indicate that promotion-insensitive JB6 cells need only an activated pro gene and TPA exposure to become neoplastically transformed.

Animals↗

Metabolic activation of environmental carcinogens and mutagens by human liver microsomes. Role of cytochrome P-450 homologous to a 3-methylcholanthrene-inducible isozyme in rat liver.

The metabolic activation of procarcinogens and promutagens by human liver microsomal cytochrome P-450 has been investigated by means of a newly developed method measuring the induction of umu gene in Salmonella typhimurium TA1535/pSK1002 [T. Shimada and S. Nakamura, Biochem. Pharmac. 36, 1979 (1987)]. The chemicals examined were aflatoxin B1 (AFB1), eight carcinogenic heterocyclic aromatic amines isolated from protein and amino acid pyrolysates, and 2-aminoanthracene. Liver microsomes from six patients catalyzed the metabolic activation of these chemicals; 2-amino-3,5-dimethylimidazo[4,5-f]quinoline (MeIQ) and AFB1 were most actively bioactivated, followed by 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-aminoanthracene (2-AA) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline. At least two forms of human cytochrome P-450 may be involved in the activation of these procarcinogens. This suggestion was supported by the following lines of evidence: (a) addition of non-ionic detergent Emulgen 913 to the incubation mixture caused a more profound inhibition of microsome-catalyzed activation of AFB1 than of MeIQ, IQ and 2AA, (b) 7,8-benzoflavone stimulated the activation of AFB1 by about 2.5-fold, whereas it inhibited significantly the reactions with MeIQ, IQ and 2AA, and (c) polyclonal antibodies against a 3-methylcholanthrene-inducible form of rat cytochrome P-450 (P-450d) caused a marked inhibition of the metabolic activation of MeIQ, IQ and 2-AA by human liver microsomes though they did not show any effects on the microsomal activation of AFB1. Data are also presented showing that none of the reactions catalyzed by human liver microsomes were inhibited by antibodies to a phenobarbital-inducible form of rat cytochrome P-450 (P-450b). These results suggest that the human cytochrome P-450 isozyme that is immunochemically similar and, thus, homologous to rat P-450d plays a major role in the metabolic activation of several procarcinogens examined, and that the activation of AFB1 is catalyzed by another and, possibly, not phenobarbital-inducible form(s) of human cytochrome P-450.

Adult↗

Na+-dependent elevation of the acidic cell surface pH (microclimate pH) of rat jejunal villus cells induced by cyclic nucleotides and phorbol ester: possible mediators of the regulation of the Na+/H+ antiporter.

The effects of cyclic nucleotides and phorbol ester on the acidic cell surface pH of rat jejunal villi were studied by using single-barrelled pH-sensitive microelectrodes. Addition of dibutyryl cAMP (1 mM) to the mucosal bathing solution caused an elevation of the cell surface pH from 6.19 +/- 0.04 (n = 12 measurements from three animals) to 6.53 +/- 0.03 (12) in the presence of Na+ in the medium. However, dibutyryl cAMP had no significant effect in the absence of Na+ and presence of 1 mM amiloride. Dibutyryl cGMP (1 mM) also had an Na+-dependent inhibitory effect on the cell surface pH. A phorbol ester, phorbol 12-myristate 13-acetate, caused an elevation of the cell surface pH only in the presence of Na+ from 6.14 +/- 0.07 (12) to 6.46 +/- 0.08 (12). Phorbol and phorbol 13-acetate, which do not stimulate protein kinase C, were without significant effects. These results suggest that increased levels of the intracellular cyclic nucleotides and activation of protein kinase C raise the acidic cell surface pH by inhibiting the activity of the brush-border Na+/H+ antiporter in the rat jejunal villus cells.

Amiloride↗

Structure and distribution of lymphatic capillaries and fenestrated blood capillaries in the conduction system of the rabbit heart.

The distribution and structure of lymphatic and blood capillaries in the rabbit heart conduction system were investigated by transmission electron microscopy. The sinuatrial node, atrioventricular node, and atrioventricular bundle possessed a rich network of lymphatic capillaries, which were situated not only at the periphery but also in the interior of the conduction system. The fine structure of these lymphatic capillaries was essentially similar to those within the atria and the ventricles. Although blood capillaries within working myocardium were nonfenestrated, the heart conduction system was often supplied by fenestrated blood capillaries. In the atrioventricular node and bundle especially, fenestrated blood capillaries and lymphatic capillaries were topographically associated, forming an extensive microcirculatory system. The presence of fenestrated capillaries suggests that a fast transcapillary passage of metabolites occurs in these regions, while the lymphatic capillaries may play an important role in the removal of macromolecules and excess intercellular fluid.

Animals↗

Effects of L-propionylcarnitine on electrical and mechanical alterations induced by amphiphilic lipids in isolated guinea pig ventricular muscle.

We examined the effects of L-propionylcarnitine (Prop. C), a short-chain acylcarnitine, on amphiphile (L-lysophosphatidylcholine or L-palmitoylcarnitine)-induced electrophysiological and ultrastructural changes in isolated guinea pig ventricular papillary muscles, under acidic conditions (pH 6.9). Conventional microelectrode, tension-recording, and electron microscope techniques were used. Both amphiphiles, at a concentration of 10(-4) M, significantly decreased the resting membrane potential, action potential amplitude, and action potential duration, but increased the developed and resting tension. Such amphiphile-induced electrical changes were not observed in muscles pretreated with the beta-blocker, atenolol, although the mechanical changes remained unaffected. The application of Prop. C (10(-2) M), in the continued presence of the amphiphiles caused a return of the action potential duration and the developed tension to the control level. However, the resting potential and action potential amplitude remained unaffected; in fact, the maximum upstroke velocity (Vmax) of the action potential tended to decrease further. Pretreatment with Prop. C prevented all the amphiphile-induced electrophysiological and mechanical changes, except for Vmax. Electron microscopic studies revealed that amphiphile-induced ultrastructural changes were prevented, at least in part, in the presence of Prop. C. Thus, Prop. C antagonizes some of deleterious effects of amphiphiles, such as lysophosphatidylcholine and palmitoylcarnitine, upon the electrical and mechanical activities of the ventricular muscle, under acidic conditions.

Animals↗

Extra- and intracellular calcium in vanadate-induced contraction of vascular smooth muscle.

The effects of extracellular Ca and Ca antagonists on vanadate-induced contractions of vascular smooth muscles of aortae and mesenteric arteries from rabbits, guinea pigs, and Wistar-Kyoto (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) were studied. Vanadate-induced contractions of aortae were greatly diminished by extracellular Ca removal; the size of the remaining contraction was variable. Vanadate-induced contractions of mesenteric arteries, which were only observed in the presence of elevated K, were suppressed by the removal of Ca. Verapamil and nifedipine depressed vanadate-induced contractions of aortae from WKY and SHRSP, whereas they produced no or only slight inhibition of responses in guinea pig and rabbit aortae. Ca uptake into smooth muscle cell increased in the presence of vanadate, but the increase was much less than that induced by high K. In saponin-skinned smooth muscle, vanadate depressed the Ca-induced contraction. It is concluded that the vanadate-induced contraction utilizes both extracellular and intracellularly bound Ca, the relative contribution of which varies from preparation to preparation.

Animals↗

Dome formation of keratin-containing agranular cells from rat anterior pituitary gland in vitro.

A certain kind of cell in the pituitary gland exhibited immunoreactive keratin and dome formations in vitro. We obtained epithelial cells, which were able to subculture, from the outgrowth of anterior pituitary organ cultures. These cells lacked hormone secretory granules and exhibited immunoreactive keratin. Furthermore, they produced dome formations or cystic structures in monolayer culture and under three-dimensional culture condition using type I collagen gel. Dome formation was stimulated by dibutyryl cyclic AMP (dbcAMP, 10(-3) to 10(-5) M). Their responsiveness to dbcAMP is similar to that of several other epithelial cells that possess transport functions in vivo and in vitro. Although the origin of our cultured cells is unknown, these cells formed dome formations that possessed transport function and were related to cystic structures in the pituitary gland in vivo.

Animals↗

Cloning of the mitochondrial genome of Rana catesbeiana and the nucleotide sequences of the ND2 and five tRNA genes.

The entire mitochondrial genome of Rana catesbeiana was cloned into a plasmid vector pBR322 at the unique BamHI site and the nucleotide sequences of the ND2 gene and of its flanking genes were determined. The ND2 gene was encoded by 1,033 base pairs and, as deduced from the nucleotide sequence, the ND2 product consisted of 344 amino acids with a molecular weight of 37,561. This gene was flanked on the 5' side by the tRNA genes for isoleucine, glutamine, and methionine and on the 3' side by those for tryptophan and alanine. These genes were the same in their organization as those found in the mammalian and Xenopus laevis mitochondrial genomes. A comparison of the putative amino acid sequences of the ND2 proteins of different animal species revealed that six regions in the sequence were well conserved during evolution, suggesting that some of these conserved sequences are crucial for biological activity of the ND2 protein. The nucleotide sequence homologies between the five tRNA genes of R. catesbeiana and their counterparts of mammals and X. laevis were in the range of 55 to 85%, depending on the tRNA and animal species.

Amino Acid Sequence↗

Molecular interaction of bovine kininogen and its derivatives with papain.

The molecular interaction of bovine kininogen and its derivatives with papain was investigated. High-molecular-weight kininogen (HMWK) or low-molecular-weight kininogen (LMWK) and inactive papain treated with N-[N-(L-3-trans-carboxyoxiran-2-carbonyl)-L-leucyl]agmatine (E-64) formed, respectively, a complex, which was dissociable on sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE). The densitometric determination of the bands separated on SDS-PAGE and amino acid analysis of the samples extracted from the electrophoresis gel revealed that the complex between kininogen and papain is formed in a molar ratio of one to one. Moreover, analysis of the inhibition of the caseinolytic activity of papain by these kininogens indicated that HMWK, LMWK, and kinin-free derivatives obtained from both kininogens inhibit active papain with a stoichiometry of 1:1. On the other hand, the papain activity was inhibited by two kinds of cyanogen bromide fragments isolated from the heavy chain of HMWK. These two fragments with Ki values of 38 and 0.64 nM corresponded, respectively, to residue Nos. 47 to 243 and Nos. 244-360 of the HMWK heavy chain. These results suggest that in the intact HMWK and LMWK, one of the two potential reactive sites interacts with papain to form a complex and that the other reactive site becomes active only after separation of the two sites.

Amino Acids↗

Surface properties of autoagglutinating mesophilic aeromonads.

The surface characteristics of 24 autoagglutinating (AA+) mesophilic aeromonads were investigated. One group of 16 was found to be highly related serologically by their reactive pattern against O antisera generated against three reference strains. Subsequent characterization of 11 of these isolates (group 1) indicated that they had the following properties in common: precipitation after boiling (PAB+), membership of serogroup O:11 (typing scheme of Sakazaki and Shimada), resistance to lysis by bacteriophage Aeh1, and possession of a surface layer (S layer) as determined by transmission electron microscopy. Strains not exhibiting the same serologic reactivity pattern belonged to diverse serogroups (other than O:11), were generally susceptible to lysis by Aeh1, and were S layer negative by transmission electron microscopy (group 2). Analysis of selected isolates representing both groups indicated that group 2 strains were usually more hydrophobic than group 1 isolates in several different assays; both groups, however, possessed high surface charge as determined by binding to DEAE-cellulose. Group 1 isolates were more virulent than group 2 strains tested as determined by lower 50% lethal doses for mice. On the basis of the results of the kinetics of autoagglutination in broth, relative surface hydrophobicity, uptake of Congo red, agglutination of yeast cells, and electrophoretic protein profiles of whole-cell extracts, the surface layer associated with O:11 mesophilic aeromonads appears to be distinct from that of Aeromonas salmonicida. The results suggest that a new pathogenic group of mesophilic aeromonads linked through a common AA phenotype, serogroup, and S layer cause serious infections in both humans and animals (fish).

Aeromonas↗

The gene encoding the nonstructural protein of B19 (human) parvovirus may be lethal in transfected cells.

The B19 parvovirus is a cause of bone marrow failure in humans. B19 is toxic to erythroid progenitor cells in vitro. Viral products possibly responsible for toxicity were explored by transfection of cloned B19 genome into HeLa cells. The nonstructural (NS) protein was detected in cells 30 h after transfection. Plasmids containing the B19 genome were transfected with selectable marker genes in stable transformation assays. Plasmids that contained the left side of the B19 genome, which encodes the NS protein of the virus, inhibited antibiotic-resistant colony formation. Transformation occurred when NS protein expression was blocked by mutation. Suppression of transformation by NS protein was not tissue specific, suggesting a role for NS protein in toxicity for nonpermissive cells without parvovirus replication or virion accumulation.

Capsid↗

Alteration of left ventricular geometry during preload reduction and afterload increment.

To ascertain whether or not left ventricular geometry changes during preload reduction and afterload increment, the shortening characteristics of small segments in the left ventricular free wall were examined using 4 pairs of ultrasonic crystals in 10 dogs. Three pairs of ultrasonic crystals were circumferentially implanted in the basal, the midventricular and the apical portion of the left ventricle. Another pair of crystals were longitudinally placed in the midventricle. In the control state, the shortening at the apex was largest of all segments. During preload reduction, the end-diastolic length decreased significantly in each segment. The percent shortening decreased significantly at the apical and the longitudinal segment, but it remained unchanged at the midventricular and the basal segment. During afterload increment, the end-diastolic length increased significantly, but the percent shortening remained unchanged in each segment. We concluded that left ventricular geometry was altered during preload reduction and that the apical part is more responsive to preload change than the other portion.

Animals↗