Origin of patterning in neural tubes.
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Animals↗
Biomedical subjects
H Wada
Publications and source records attributed to H Wada.
[Differentiation of hematopoietic stem cell and expression of GPI-anchor protein in paroxysmal nocturnal hemoglobinuria].
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CD59 Antigens↗
Ultrastructural changes in canine lung preserved in newly developed solutions.
The present study was undertaken to clarify the effect of differences in the ionic composition of a preservation solution by investigating ultrastructures of pulmonary endothelial cells and oxygenation ability of preserved lungs after reperfusion. The relation between the ultrastructural changes and the oxygenation ability was also examined. Canine lungs flushed with an extracellular-type (ET)-Kyoto solution (group A, n = 6), with an intracellular-type-Kyoto solution and prostaglandin El (group B, n=6), or with Euro-Collins solution and prostaglandin E1 (group C, n=6) were stored at 4 degrees C for 20 hr. In transmission electron microscopic findings, the frequency of ultrastructural changes (protrusion of endothelial cells and cellular vacuolization) was significantly less in group A (23.3 +/- 9.1 and 13.3 +/- 5.2%, respectively) than in group B (64.4 +/- 6.1 and 42.2 +/- 8.8%, respectively). In group A, PaO2 after 40, 70, and 130 min of reperfusion was uniformly excellent (289.4 +/- 5.7, 303.3 +/- 7.0, and 303.0 +/- 19.6 mm Hg, respectively) and significantly higher than in groups B and C (202.6 +/- 32.0 and 185.9 +/- 23.0 mm Hg, respectively) after 70 min and higher than in group C (155.7 +/- 36.3 mm Hg) after 130 min of reperfusion. A negative correlation was noted between the frequency of cellular vacuolization and the PaO2 after reperfusion. These results indicated that extracellular ion composition has significantly better effect on pulmonary vascular ultrastructures than intracellular ion composition. This may be a factor making ET-Kyoto solution superior to the other two solutions in oxygenation ability after reperfusion. When attempting to develop better lung preservation solution, the ability to preserve the ultrastructures of preserved lung may be an important consideration in the evaluation.
Animals↗
Effect of inorganic phosphate and ADP on the myofilament sliding induced by laser flash photolysis of caged ATP.
Using the technique of laser flash photolysis of caged ATP, we have suggested that, under nearly isometric conditions, the unitary distance of myofilament sliding per ATP molecule (myosin head powerstroke) is about 10 nm. To give further information about the mechanism of myofilament sliding, we studied the effect of inorganic phosphate (Pi) and ADP on the photoreleased ATP-induced shortening of single glycerinated muscle fibers under very small external loads. Both the velocity and the distance of the myofilament sliding induced by 150 microM ATP increased by Pi (20 mM), and decreased by ADP (0.4 mM). On the other hand, Pi and ADP showed no significant effect on the myofilament sliding induced by 100 and 75 microM ATP. The potentiating effect of Pi on the myofilament sliding with 150 microM ATP can be explained as being due to the increase in population of AM.ADP.Pi with corresponding decrease in population of AM.ADP, and also the increase in population of M.ADP.Pi and M.ATP. Meanwhile, the inhibitory effect of ADP can be simply accounted for to be due to an accumulation of AM.ADP that already finished their force generating process. The ineffectiveness of Pi and ADP on the myofilament sliding with 100 and 75 microM ATP is consistent with the view that it is caused by almost synchronized single myosin head powerstrokes.
Actin Cytoskeleton↗
Translational initiation factor eIF-4E. A link between cardiac load and protein synthesis.
To define the coupling mechanism between cardiac load and the rate of protein synthesis, changes in the extent of eIF-4E phosphorylation were measured after imposition of a load. Electrically stimulated contraction of adult feline cardiocytes increased eIF-4E phosphorylation to 34% after 4 h, as compared with 8% phosphorylation in quiescent controls. However, eIF-4E phosphorylation did not increase upon electrical stimulation in the presence of 7.5 mM 2,3-butanedione monoxime, an inhibitor of actin-myosin cross-bridge cycling and active tension development. Treatment of adult cardiocytes with either 0.1 microM insulin or 0.1 microM phorbol 12-myristate 13-acetate increased eIF-4E phosphorylation to 23 and 64%, respectively, but these increases were not blocked by 2,3-butanedione monoxime. In canine models of acute hemodynamic overload in vivo, eIF-4E phosphorylation increased to 23% in response to left ventricular pressure overload as compared with 7% phosphorylation in controls. Acute volume overload had no effect on eIF-4E phosphorylation. These changes in eIF-4E phosphorylation account for differences in anabolic responses to acute pressure versus acute volume overload. These data suggest that eIF-4E phosphorylation is a mechanism by which increased cardiac load is coupled to accelerated rates of protein synthesis.
Animals↗
The coxD gene for heme O synthase in Synechocystis.
The cyanobacterial coxD gene for heme O synthase was cloned from Synechocystis sp. PCC 6803 and its nucleotide sequence was determined. The deduced amino-acid sequence of the gene was homologous to the amino-acid sequences of bacterial heme O synthesis. In contrast to the genes for heme O synthases in other prokaryotes, which are clustered together with genes for the structural subunit(s) of cytochrome oxidase, the coxD gene is not linked to such genes on the chromosome of Synechocystis.
Amino Acid Sequence↗
Direct high-performance liquid chromatographic and high-performance liquid chromatographic-thermospray-mass spectrometric determination of enantiomers of methamphetamine and its main metabolites amphetamine and p-hydroxymethamphetamine in human urine.
For the identification of drug abuse, a simple and rapid method which allows us to distinguish enantiomers of methamphetamine (MA) and its metabolites amphetamine (AP) and p-hydroxymethamphetamine (p-OHMA) in human urine was explored by coupling direct HPLC and HPLC-thermospray-mass spectrometry (HPLC-TSP-MS) both of which employ a beta-cyclodextrin phenylcarbamate-bonded silica column. HPLC analysis was performed after the solid-phase extraction from the urine sample with Bond Elut SCX, and D- and L-enantiomers of MA, AP and p-OHMA could be separated well. The proposed conditions are as follows: eluent, acetonitrile-methanol-50 mM potassium phosphate buffer (pH 6.0) (10:30:60, v/v) flow-rate, 1.0 ml/min temperature, 25 degrees C. The linear calibration curves were obtained for D- and L- MA and AP in the concentration range from 0.2 to 20 micrograms/ml; the relative standard deviation for D- and L-AP and D- and, L-MA ranged from 1.67 to 2.35% at 2 micrograms/ml and the detection limits were 50 ng/ml for D- and L-AP and D-MA and 100 ng/ml for L-MA. For the verification of the direct HPLC identification, HPLC-TSP-MS was also carried out under the same conditions except that acetonitrile-methanol-100 mM ammonium acetate (pH 6.0) (10:30:60, v/v) was used as an eluent. Upon applying the scan mode, 10 ng/ml for D- and L-AP and D-MA and 20 ng/ml for L-MA were the detection limits. Using the selected ion monitoring mode, 0.5 ng/ml, 0.8 ng/ml and 1 ng/ml could be detected for D- and L-AP, D-MA and L-MA, respectively.
Amphetamine↗
Characterization of the Fad12 mutant of Synechocystis that is defective in delta 12 acyl-lipid desaturase activity.
The Fad12 mutant of Synechocystis sp. PCC 6803 has a defect in the desA gene for delta 12 acyl-lipid desaturase. We identified a change in the nucleotide sequence of the structural gene for the desaturase, in which a leucine codon has been converted to a stop codon. Western blot analysis revealed that the delta 12 acyl-lipid desaturase was localized in both plasma membranes and thylakoid membranes of wild-type cells but was absent from both types of membrane in Fad12 cells. These findings suggest that the desaturation of fatty acids takes place in both types of membrane in Synechocystis sp. PCC 6803. The mutation in the delta 12 desaturase did not affect the lipid composition of thylakoid and plasma membranes, but it changed the fatty acid composition of lipids in similar ways in both types of membrane.
Amino Acid Sequence↗
Increased plasma-soluble fibrin monomer levels in patients with disseminated intravascular coagulation.
Plasma-soluble fibrin monomer (SFM) level in patients with disseminated intravascular coagulation (DIC) was significantly higher than the level in patients with pre-DIC or in non-DIC patients, and the level in patients with pre-DIC was significantly higher than that in non-DIC patients. There was no significant difference in plasma SFM levels among various diseases underlying DIC. Plasma SFM level in patients with good outcome was significantly decreased after treatment for DIC. The sensitivity of fibrin degradation products and platelet number was high for DIC, but not for pre-DIC. The sensitivity of thrombin-antithrombin III complex, plasmin-plasmin inhibitor complex, and SFM was high for both DIC and pre-DIC. The specificity of these markers was also high. Receiver operating characteristic analysis suggests that plasma SFM level could be the most useful marker for the diagnosis of both DIC and pre-DIC.
Amino Acid Sequence↗
Plasma tissue factor and tissue factor pathway inhibitor levels in patients with disseminated intravascular coagulation.
We measured the plasma levels of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in patients with disseminated intravascular coagulation (DIC) to examine the relationship between TFPI and vascular endothelial cell injury. Plasma TF (273 +/- 90 pg/ml) and TFPI (252 +/- 125 ng/ml) levels were significantly increased in patients with DIC compared with non-DIC patients. Plasma TF antigen level was significantly increased in pre-DIC patients (285 +/- 85 pg/ml), while the plasma TFPI level (152 +/- 54 ng/ml) was not markedly increased in such a state. The plasma TF/TFPI ratio was high in the pre-DIC patients (2.10 +/- 0.90), and low in the DIC patients (1.40 +/- 0.87) and healthy volunteers (0.84 +/- 0.26). There was no significant difference between the DIC patients with a good outcome and those with a poor outcome in terms of plasma TF levels, although the plasma TFPI level in the DIC patients with a good outcome (289 +/- 133 ng/ml) was significantly higher than that in those with a poor outcome (187 +/- 75 ng/ml). During the clinical course of DIC, plasma TF antigen was increased first, and an increase of the plasma TFPI level followed the increase in plasma TF level. These findings suggest that plasma TFPI is released from vascular endothelial cells and it may reflect vascular endothelial cell injury. It is conceivable that TF and TFPI may play an important role in the onset of DIC.
Anticoagulants↗
Lymphoblastic transformation of chronic myelomonocytic leukemia in an infant.
A 10-month-old infant with chronic myelomonocytic leukemia (CMML) of 5 months' duration, who had been treated only with transfusion, displayed leukemic transformation characterized by lymphoid morphology, PAS positivity, and myeloperoxidase negativity. Surface marker analysis of blast cells revealed expression of lymphoid-associated antigens (CD10 and CD19) but not myeloid-associated antigens (CD13, CD14, and CD33). These findings suggest that some cases of infantile CMML are clonal disorders arising in a pluripotent stem cell that can also differentiate along the lymphoid cell lineage.
Antigens, CD↗
Refractory pancreatitis associated with graft-versus-host disease in Fanconi anemia.
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Bone Marrow Transplantation↗
Experimental anti-GBM glomerulonephritis induced in rats by immunization with synthetic peptides based on six alpha chains of human type IV collagen.
The anti-glomerular basement membrane (GBM)-nephritis-inducing activity of six synthetic peptides having an amino acid sequence consisting of the six alpha chains of human type IV collagen was examined by injecting the peptides into rats. The peptides consisted of 27 amino acid residues from the non-collagenous domain (NC1) of the alpha 1 to alpha 6 chains and were non-consensus sequences sandwiched between two consensus sequences near the carboxyl terminus. Each peptide was coupled to keyhole limpet haemocyanin and injected with adjuvant into the footpads of 20 female WKY/NCrj rats. The number of rats with proteinuria (over 10.0 mg of urinary protein/15 h) and haematuria was 2 with the alpha 3 peptide, 8 with the alpha 4 peptide, and 1 with the alpha 5 peptide. Histological changes seen in the glomeruli were characteristic of those in anti-GBM nephritis. Linear deposition of rat IgG along the GBM was observed in five rats injected with the alpha 4 peptide. A nephritogenic monoclonal antibody against the alpha 4 peptide was established using lymph node cells from a rat injected with the alpha 4 peptide. The results indicate that alpha 4(IV)NC1 is a potent nephritogenic antigen like alpha 3(IV)NC1, which has already been recognized as a primary target antigen in Goodpasture's syndrome.
Amino Acid Sequence↗
Promotion and control of selective adhesion and proliferation of endothelial cells on polymer surface by carbon deposition.
The adhesion and proliferation of endothelial cells can be drastically improved when cultivated on a carbon-deposited polymer surface. When the surface of segmented polyurethane, in which endothelial cells are not capable of proliferating, is modified by carbon deposition, cell adhesion and proliferation occurred selectively on the carbon-deposited region. Carbon deposition to a tissue culture-treated polystyrene surface on which cells are capable of proliferating further promoted cell spreading and proliferation. Cell patterning on carbon-deposited segmented polyurethane was also attempted by controlling the direction or region of cell proliferation. Carbon deposition onto the polymer surface brought about a decrease in the water contact angle of the surface. Precoating fibronectin or laminin on the carbon-deposited polymer surface brought about cell adhesion and proliferation in the carbon-deposited region, but not in the non-carbon-deposited region. Our data suggest that promotion of cell proliferation on a carbon-deposited surface is probably due to selective adhesion of adhesive proteins to the surface.
Animals↗
Preservation of skin free-flap using trehalose.
In this study, we looked for a solution able to preserve traumatically amputated tissues such as nose tip, ear, and scalp for at least 48 hr that was easy to handle and low in price. Trehalose is a nonreducing disaccharide that can protect many prokaryotes, fungi, and yeasts against adverse conditions by stabilizing cell membranes. In order to study its protective effect, 60 skin free-flaps of the ears of rabbits were preserved in Euro-Collins (EC) solution or in modified Euro-Collins solution in which 7% trehalose replaced glucose (EC + 7%T) for 24, 48, and 72 hr at 4 degrees C. After completion of preservation, these flaps were transferred to the other ear with microvascular anastomosis. After 7 days, we obtained a survival rate of 100% of flaps after preservation for 24 hr in both solutions. But, after preservation for 48 hr or longer, this decreased to 60% (6 of 10 preserved flaps survived) in EC solution and 90% (9 of 10 flaps) in EC + 7%T solution. This difference became clearer after 72 hr of preservation, when the survival rate was higher and light microscopic observation showed less damage for flaps preserved in EC + 7%T solution (20% in EC and 70% in EC + 7%T) (P < 0.05). Thus, EC + 7%T solution was superior to EC solution in the preservation of skin free-flaps of the ear of rabbits. Trehalose shows great promise in the preservation of many organs, including storage of traumatically amputated tissues such as nose tip, ear, and scalp.
Adipose Tissue↗
Early evolution of the Metazoa and phylogenetic status of diploblasts as inferred from amino acid sequence of elongation factor-1 alpha.
To understand the early evolution of the Metazoa, it is necessary to determine the correct phylogenetic status of diploblastic animals (poriferan, cnidaria, and ctenophora). Despite clasdistic studies of morphological characters and recent molecular phylogenetic studies, it remains uncertain whether diploblasts are monophyletic or paraphyletic, and how these three phyla of diploblasts are phylogenetically related. To obtain insight into these phylogenetic problems, we sequenced almost the entire nucleic acid sequence of elongation factor-1 alpha from a sponge, two cnidarians, a ctenophora, and a turbellarian. We then investigated the phylogenetic status of the diploblasts. We compared the amino acid sequences, nucleotide sequences at the first and second codon positions, and those at the second positions. Phylogenetic trees were inferred by neighbor-joining, maximum likelihood, and maximum parsimony, and they supported the monophyly of the Metazoa. However, the phylogenetic relationships of the diploblast groups were not significantly resolved, although the trees preferred the monopoly of the diploblasts.
Amino Acid Sequence↗
Mitochondrial rDNA phylogeny of the asteroidea suggests the primitiveness of the paxillosida.
Asteroids display four distinct modes of developmental patterns: the indirect mode, the nonbrachiolarian mode, the direct mode, and the mode with a barrel-shaped larva. Among them the former two are planktotrophic, whereas the latter two are lecithotrophic. The direct mode and the mode with a barrel-shaped larva are thought to have evolved from the more primitive planktotrophic mode, the nonbrachiolarian and the indirect mode, respectively. However, whether the nonbrachiolarian mode or the indirect mode is the more primitive in asteroids is unresolved, despite discussion since early this century. A key aspect of this problem is the phylogenetic status of paxillosidans, since the nonbrachiolarian mode and the mode with a barrel-shaped larva are seen only in paxillosidans. To resolve this problem, we performed a molecular phylogenetic study of asteroids, based on the nucleotide sequences of mitochondrial rDNAs. Phylogenetic trees support a close relationship between the Asterinidae and the Solasteridae. We suggest that the paxillosidans are not a monophyletic group; rather, the Luidiidae (one family of Paxillosida) is a sister group to the rest of the asteroids. Although some aspects of our results contradict a recent study by Lafay et al. (1995, Syst. Biol. 44: 190-208) based on 28S rRNA sequences, both studies agree on a paraphyletic nature for the paxillosidans. We conclude that characters shared by paxillosidans are primitive; hence the primitive mode of development in asteroids is the nonbrachiolarian mode.
Animals↗
Resection of primary pulmonary hemangiopericytoma: a report of two cases.
We report herein the cases of two patients who underwent resection of a primary pulmonary hemangiopericytoma. The first patient was a 58-year-old man found to have a mass-like shadow of about 5 cm in diameter in the right hilum by a routine chest X-ray. Subsequent magnetic resonance imaging (MRI) showed a probable mediastinal tumor, and surgery was performed. Postoperative pathological examinations confirmed the diagnosis of pulmonary hemangiopericytoma. The second patient was a 21-year-old woman found to have a mass-like shadow of about 2 cm in diameter in the left middle lung field. As a preoperative diagnosis could not be made, exploratory surgery was performed and the left S6 segment was excised. A definitive diagnosis of pulmonary hemangiopericytoma was established postoperatively by pathological examination. Primary pulmonary hemangiopericytoma is an extremely rare type of tumor, with only 36 cases having been reported in the Japanese literature to date, including out 2 cases. A discussion following the case reports examines the sex, age, initial manifestations, sites, and methods of surgery employed in these 36 cases.
Adult↗