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Biomedical subjects

N Maeda

Publications and source records attributed to N Maeda.

At least 451 records · Page 25Linked to original sources

Purification of D2 dopamine receptor by photoaffinity labelling, high-performance liquid chromatography and preparative sodium dodecyl sulphate polyacrylamide gel electrophoresis.

[125I]N-azidophenethylspiperone ([125I]azido-NAPS) was used as a photoaffinity ligand for bovine D2 dopamine receptor. On photolysis, [125I]azido-NAPS was covalently incorporated into a major band of 94 kDa in bovine striatal membrane as assessed by autoradiography after sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) (10% acrylamide gel). The labelled D2 receptor protein from striatal membrane was solubilized and subjected to HPLC using gel filtration (TSK G3000SW) and hydroxyapatite gel (Pentax SH2010C), followed by two steps of preparative SDS-PAGE. The D2 receptor protein could be obtained as a single major polypeptide on SDS-PAGE by either silver staining or autoradiography.

Affinity Labels↗

Cell surface galactosyltransferase acts as a general modulator of rat acinar cell proliferation.

Several physiological parameters were examined for inducing acinar cell proliferation and corresponding increased expression of beta 1-4 galactosyltransferase. In this study, dietary changes causing acinar cell proliferation included the following: the introduction of animals to a liquid diet (causing gland atrophy) followed by reintroduction of solid chow, gustatory stimulation provided by the introduction of 0.5% citric acid to animal drinking water, and removal of the submandibular gland with subsequent reliance on the parotid gland for salivary protein. Alterations in growth factor levels were produced by injecting animals with a chronic (3 day) regimen of either nerve growth factor or epidermal growth factor. Parotid gland proliferation could be blocked in all cases except EGF by the injection of propranolol, a beta-adrenoceptor antagonist, or the galactosyltransferase specific modifier protein, alpha-lactalbumin. EGF-induced proliferation could, however, be prevented by treating the animals with monoclonal antibody to EGF receptor or galactosyltransferase modifier protein alpha-lactalbumin. These results for normal acinar cell proliferation suggest a direct role for cell surface beta 1-4 galactosyltransferase in signalling and maintaining active cell growth.

Animals↗

Contribution of glycoproteins to fibrinogen-induced aggregation of erythrocytes.

The contribution of membrane glycoproteins to the velocity of fibrinogen-induced erythrocyte aggregation was examined using a rheoscope combined with a video camera, an image analyzer and a computer. The structure of glycoproteins was modified with proteolytic enzymes, trypsin or alpha-chymotrypsin. (1) Mild enzymatic treatment of erythrocytes decreased the velocity of erythrocyte aggregation, but more intense treatment increased the velocity remarkably. (2) The erythrocyte aggregation was affected not only by the density of surface negative charge of erythrocytes, but also by the structural changes of glycoproteins. (3) Erythrocyte deformability and the morphological characteristics were not altered by these enzymatic treatments. The physiological significance of glycoproteins of erythrocyte surface for the survival of erythrocytes and for the suspension stability of blood was discussed.

Adult↗

A cerebellar Purkinje cell marker P400 protein is an inositol 1,4,5-trisphosphate (InsP3) receptor protein. Purification and characterization of InsP3 receptor complex.

P400 protein is a 250 kd glycoprotein, characteristic of the cerebellum, which is accumulated at the endoplasmic reticulum, at the plasma membrane and at the post-synaptic density of Purkinje cells. In this study, we purified inositol 1,4,5-trisphosphate (InsP3) receptor from mouse cerebellum and examined the possibility that P400 protein is identical with cerebellar InsP3 receptor protein. InsP3 receptor was solubilized with Triton X-100 from a post-nuclear fraction of ddY mouse cerebellum and was purified with high yield by sequential column chromatography on DE52, heparin-agarose, lentil lectin-Sepharose and hydroxylapatite. In these chromatographies, P400 protein co-migrated completely with the InsP3 binding activity. The purified receptor is a 250 kd protein with a Bmax of 2.1 pmol/microgram and a KD of 83 nM. It reacted with three different monoclonal antibodies against P400 protein, indicating that P400 protein is the same substance as the InsP3 receptor (P400/InsP3 receptor protein). Electron microscopy of the purified receptor showed a square shape with sides approximately 25 nm long. Binding assays of the cerebella of Purkinje cell-degeneration (pcd) mice with [3H]InsP3 demonstrated that the InsP3 binding sites in the cerebellum are distributed exclusively on the Purkinje cells. Immunohistochemical analysis indicated that P400/InsP3 receptor is present at the dendrites, cell bodies, axons and synaptic boutons of the Purkinje cells.

Animals↗

Selection of variants of avian infectious bronchitis virus showing tropism for different organs.

Avian infectious bronchitis virus strain Kagoshima-34 isolated from the kidneys of a chicken that died of nephrosis/nephritis lost its nephropathogenicity during intratracheal passage in SPF chickens. The resultant virus acquired stronger respirotropism but reduced tropism for kidneys. On the other hand strain Tottori-2 isolated from the trachea of a chicken suffering from severe respiratory disease did not lose its respirotropism after serial intravenous passage in SPF chickens. The serological properties of the passaged virus were investigated by virus neutralisation test. The antibody titres of both strains of virus fluctuated with progressive passage. The serological properties of the virus isolated from respiratory organs were not necessarily the same as those of the isolates made from the kidneys.

Animals↗

Temperature effect on oxygenation and metabolism of perfused rat hindlimb muscle.

The effects of temperature on oxygenation and metabolism in perfused rat hindlimb was studied at 35 degrees C and 15 degrees C. Oxygenation of myoglobin and oxidation of cytochrome aa3 in the thigh (quadriceps) muscle were estimated from the difference spectra measured with a rapid-scanning spectrophotometer. Simultaneously, oxygen uptake and release of lactate and pyruvate were measured. (1) In hypothermia, glycolysis played a major role in energy metabolism even though Cyt aa3 was maintained in a more oxidized state than in normothermia. (2) P50 of myoglobin in perfused rat hindlimb was 5.0 mmHg at 35 degrees C, 2.3 mmHg at 25 degrees C and 1.1 mmHg at 15 degrees C. The delta H degree was -13.0 kcal/mol. (3) When about 30% of myoglobin was deoxygenated at both 35 degrees C and 15 degrees C, the oxygen uptake started to decrease and lactate release increased. (4) At 35 degrees C, the oxidation level of cytochrome aa3 was same as the oxygenation level of myoglobin. At 15 degrees C, however, the oxidation level of cytochrome aa3 was clearly higher than the oxygenation level of myoglobin. The oxygen uptake at 15 degrees C was about one third that at 35 degrees C. In conclusion, in order to maintain the aerobic condition of cytochrome aa3 in mitochondria of rat skeletal muscle, a tissue oxygen tension higher than 12 mmHg at 35 degrees C, and higher than 3 mmHg at 15 degrees C is required.

Animals↗

Effects of high alpha-linolenate and linoleate diets on erythrocyte deformability and hematological indices in rats.

Rats were fed either a high alpha-linolenate diet or a high linoleate diet from weaning to 4 mon of age. Soybean oil was used as a control. Phospholipid compositions of erythrocytes from the three dietary groups were not significantly different. However, the difference in the alpha-linolenate (18:3n-3)/linoleate (18:2n-6) ratio of the diets was reflected in the n-3/n-6 ratios of the 20 and 22 carbon highly unsaturated fatty acids except for docosahexaenoic acid (22:6n-3) in the phospholipids. Despite the significant differences in the fatty acid compositions of phospholipids, no measurable differences were detectable in erythrocyte deformability, whole blood viscosity and hematological indices of the three dietary groups. These results indicate that the beneficial effects of the high alpha-linolenate diet, as compared with the high linoleate diet, are exerted without significant changes in these parameters.

Animals↗

Resurfacing of skin defects of the hand using the arterialised venous flap.

In 22 patients an arterialised venous flap, with arterial blood only flowing through the vein of the flap, was used to cover skin defects of the hand. There were 17 complete successes, four partial successes and one complete failure. The main indications for use of the arterialised venous flap are to cover extensive skin loss with exposed bone or tendon where conventional flaps cannot be applied and to reconstruct the digital arterial defect where both revascularisation and skin coverage are needed.

Adolescent↗

Architecture of Purkinje cells of the reeler mutant mouse observed by immunohistochemistry for the inositol 1,4,5-trisphosphate receptor protein P400.

P400 protein, which is identical to the inositol 1,4,5-trisphosphate receptor protein, is a glycoprotein closely associated with the membranes of Purkinje cells. Three types of monoclonal antibodies against P400 protein were employed for the immunohistochemical detection of Purkinje cells in the cerebellum and brainstem of the normal and reeler mouse. Purkinje cells in both types of mice were immunoreactive against anti-P400 antibodies, and the soma, dendrites, axon and even terminal boutons in the cerebellar and vestibular nuclei could be clearly visualized. In the cerebellum of the reeler mutant, the heterotopic Purkinje cells both within and below the granule cell layer were also immunopositive and could be clearly differentiated from the deep cerebellar nuclei, in which neurons were immunonegative. The molecular layer of the reeler cerebellum varied in thickness and certain parts were completely defective. The dendrites within the molecular layer extended from Purkinje cells whose cell bodies were located in the normal position, abnormally in the granule cell layer, or at the surface of the central mass. Outside the cortex of the cerebellum, ectopic Purkinje cells were demonstrated in 3 cerebellar nuclei, the cerebellar medulla and peduncle, and brainstem of the normal and reeler mouse.

Animals↗

Three independent insertions of retrovirus-like sequences in the haptoglobin gene cluster of primates.

The human haptoglobin two-gene cluster (HP-HPR) contains two retrovirus-like elements. One (RTVL-Ia) is in the first intron of the HPR gene, and the second (RTVL-Ic) is at the 3'-end of the gene cluster. The chimpanzee three-gene cluster (HP-HPR-HPP) contains an additional, third copy (RTVL-Ib) in the intergenic region between HPR and HPP. RTVL-Ia and RTVL-Ib are essentially full size and have the general structure, 5'-LTR-gag-pol-env-3'-LTR, while RTVL-Ic lacks about one-third of its 5'-part. Although none of the elements has retained long open reading frames, we could detect stretches having amino acids identical to various parts of Moloney murine leukemia virus (Mo-MuLV) proteins. We conclude that the RTVL-I elements were derived from a virus very similar in structure to Mo-MuLV. The DNA sequences surrounding the insertion points of the three RTVL-I elements are not alike and allow the inference that they integrated into the haptoglobin gene cluster independently at some time after the initial formation of the triplicated gene cluster in primates. Comparison of the nucleotide sequences of the three elements leads to the hypothesis that foreign DNA introduced into the genome can initially accumulate mutations more rapidly than the genomic sequences surrounding it.

Amino Acid Sequence↗

A physical map of the human salivary proline-rich protein gene cluster covers over 700 kbp of DNA.

By using a linking library, we have experimentally linked, ordered, and spaced four of the six loci that constitute the human salivary proline-rich protein (PRP) multigene family. The methods used for mapping these four PRP genes may be useful in other multigene systems in which no probes unique to each member of genes are available, but in which some enzyme site that occurs only once in each member of the family can be found. The remaining two PRP loci have been provisionally mapped and linked within the gene cluster primarily on the basis of the resulting order giving a simple map. The order of the six loci that most simply accounts for our data is PRB2, PRB1, PRB4, PRH2, PRB3, and PRH1. The PRP gene cluster spans at least 700 kbp on chromosome 12 at p13.2. A scheme for the evolution of the cluster that requires an initial gene duplication followed by three unequal but homologous crossovers is given.

Blotting, Southern↗

Expressed cerebellar-type inositol 1,4,5-trisphosphate receptor, P400, has calcium release activity in a fibroblast L cell line.

P400, inositol 1,4,5-trisphosphate receptor (InsP3-R), is a key protein to understanding the mechanisms of inositol 1,4,5-trisphosphate (InsP3)-mediated Ca2+ mobilization. We obtained the cerebellar-type P400/InsP3-R cDNA and generated an L cell transfectant (L15) that produces cDNA-derived P400/InsP3-R. In membranes, this protein displays high affinity, specificity, and capacity for InsP3, as does the cerebellar P400/InsP3-R. InsP3 can also induce greater 45Ca2+ release from the membrane vesicles of L15 cells than from those of control L cells. These results provide direct evidence that the cDNA-derived P400/InsP3-R protein is actually involved in physiological Ca2+ mobilization, through binding to InsP3 molecules in the same manner as the cerebellar P400/InsP3-R.

Animals↗

Erythrocyte rheology.

Two main subjects of erythrocyte rheology, deformation and aggregation, are discussed in detail, on the basis of biochemical structure. The close relationship between the life span (or cell aging) and the rheology of individual erythrocytes is also briefly described. A currently important problem is emphasized, that is, the molecular aspect of the dynamic cytoskeletal structure and the mechanism of its regulation. This concerns not only the rheological function and the survival of circulating erythrocytes, but also the pathophysiology of abnormal erythrocytes.

Blood Viscosity↗

Nail abrasion: a new treatment for ingrown toe-nails.

A simple new method for the treatment of ingrown toe-nails was devised and applied to 33 toe-nails in 22 patients. The entire nail surface, except for the margin, was abraded until the nail became flexible with a Schreu's skin grinder equipped with a steel bar. When there was infection or granulation of the nail wall, the imbedded nail corner was trimmed before abrasion. The patients underwent additional treatments when one treatment was not enough to achieve cure or when discomfort appeared, which was the first symptom of recurrence. The average number and duration of treatments was 2.9 times and 16 months, respectively. Relief from pain occurred in all patients soon after abrasion was performed. Recurrence of infection and granulation was observed in only one patient. Five patients (23%) remained problem-free for more than one year without any additional treatment, because their incurved nail was gradually improved in shape by repetition of this procedure. This therapy was particularly effective in patients with severely incurved nails.

Adult↗