Search PubMed⌕ Search

Biomedical subjects

Y Yano

Publications and source records attributed to Y Yano.

At least 361 records · Page 20Linked to original sources

ATP-driven tubule extrusion from axonemes without outer dynein arms of sea-urchin sperm flagella.

We have prepared axonemes without outer dynein arms from sea-urchin (Pseudocentrotus depressus, Hemicentrotus pulcherrimus) sperm flagella by selective solubilization with NaCl. Electron microscopy revealed that the axonemes gradually lost their outer arms in 0.5 M NaCl during 10 min. Such axonemes retained 42.8 +/- 7.3% of their total axonemal ATPase activity and showed C, A, D and B bands in the dynein region of 4% SDS-gel, while a solubilized fraction of the outer arms consisted almost entirely of A band polypeptide. We have succeeded in causing extrusion of the outer doublets from such axonemes by addition of ATP and trypsin. A bundle of outer doublets was sometimes observed to be extruded first from an axoneme and to show bending motion for a while, subsequently followed by a sliding of separate doublets past each other. The speed of the tubule extrusion process was slower and around 60% of that of intact axonemes having both types of arm. These observations indicate that the inner arms have a function equivalent to that of the outer arms, of sliding on adjacent doublets, although the inner arms seem to be constituted from polypeptide(s) different from that of the outer arms.

Adenosine Triphosphatases↗

Sliding velocity between outer doublet microtubules of sea-urchin sperm axonemes.

Using a dark-field microscope equipped with a high-efficiency TV camera including a video tape-recorder, we recorded the sliding movement between outer doublet microtubules of the demembranated axonemes of sea-urchin (Pseudocentrotus depressus and Hemicentrotus pulcherrimus) sperm flagella by adding ATP and trypsin at 25 degrees C. The time and length of the sliding doublet microtubules from axonemes were measured directly from the image on the picture monitor from the video tape. The sliding velocity was almost constant in the range from 0 to 2% polyethylene glycol concentration in the reactivation medium and decreased a little at more than 2%. We prepared various lengths of axoneme fragments by homogenizing whole axonemes and found that the shorter fragments showed similar sliding velocity to that of longer ones at less than 200 microM ATP, but slightly decreased speed at more than 500 microM. ATP. The sliding movement sometimes stopped and the percentage of sliding axonemes was lower below 2 micrograms/ml trypsin. Above 3 micrograms/ml, the process appeared to be more like disintegration than sliding movement, which may be due to excess digestion by trypsin. Sliding speed was therefore measured in a reactivation medium containing 2% polyethylene glycol with the addition of ATP and 2 micrograms/ml trypsin. The velocity increased in proportion to the increase in ATP concentration. Vmax was approximately 14 micrograms/s at 2 mM ATP. In order to compare the Km for the sliding velocity with that of the ATPase activity of the axonemes, we measured ATPase activity of axonemes prepared and assayed under conditions in which sliding movement in the axonemes could be induced. Neither the curve of ATPase activity nor the curve of sliding velocity plotted against ATP concentration obeyed Michaelis-Menten kinetics. The close relationship between ATPase activity and sliding velocity suggested that 'sliding-movement-coupled ATPase activity' may well be reflected in the axoneme ATPase reported here.

Adenosine Triphosphatases↗

Thermodynamic analysis of the nonspecific interaction of sodium dodecyl sulfate with swollen Bio-Gel beads.

In frontal gel chromatography on Bio-Gel P-2, sodium dodecyl sulfate (SDS) at the concentrations below its critical micelle concentration (cmc) showed anomalously high partition coefficients (Kav obs) indicative of strong interactions with the swollen gel phase; further, Kav obs was found to increase with concentration and temperature. This preferential partition of SDS in the Bio-Gel phase was analyzed in terms of the transfer free energy of SDS from the mobile phase (0.1 M NaCl) to the swollen Bio-Gel phase. The results showed that the overall transfer process is primarily governed by hydrophobic free energy arising from the anomalous nature of hydrated water in the gel matrix; that is, in highly hydrated water "iceberg" formation is evidently limited and the hydrophobic free energy is accordingly lowered, resulting in the preferential partition of SDS in the swollen Bio-Gel phase. The increase in the negativity of transfer free energy with concentration, though relatively small, indicated a definite tendency for the formation of SDS clusters in the gel phase. Finally, a model illustrating the states of SDS molecules in the gel matrix is presented, which may also be pertinent to SDS-protein and SDS-amylose complexes.

Chromatography, Gel↗

Biological activity of macromomycin.

Macromomycin (MCR) is a polypeptide antimuor antibiotic isolated from the culture broth of Streptomyces macromomyceticus. Antitumor activities of MCR were examined against three different tumor system, i.e., EHRLICH ascites carcinoma, L1210 leukemia and LEWIS lung carcinoma. Daily intraperitoneal treatment with MCR for 5 days showed a strong inhibition against EHRLICH ascites carcinoma. Both single and repeated intraperitoneal injections of MCR were effective over a wide dose range against intraperitoneally inoculated L1210 leukemia and MCR intravenously administered was also active against intravenously inoculated L1210 leukemia. Daily local subcutaneous injections of MCR produced the prolongation of life span of mice to which LEWIS lung carcinoma was subcutaneously inoculated with some cured mice, but daily intraperitoneal injections of MCR showed no activity. Single intravenous administration of MCR inhibited early LEWIS lung carcinoma, but not advanced LEWIS lung carcinoma. The combination of MCR with aracytidine, or cyclophosphamide showed a synergistic activity against L1210 leukemia. MCR was not inactivated by treatment with serum, although neocarzinostatin was markedly inactivated by the same treatment.

Animals↗

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of bovine serum albumin oligomers produced by lipid peroxidation.

The oligomers of bovine serum albumin were produced by controlled reaction with peroxidizing linoleic acid to examine their possible utility as calibration proteins insodium dodecyl sulfate-polyacrylamide gel electrophoresis. The polymerization was effected in reaction mixtures containing linoleic acid undergoing peroxidation in the presence of ascorbic acid, and conditions that yield soluble oligomers with a wide molecular weight distribution were established. The interaction of these soluble oligomers with sodium dodecyl sulfate exhibited a binding isotherm indistinguishable from that obtained with bovine serum albumin. Furthermore, sodium dodecy sulfate-polyacrylamide gel electrophoresis of the albumin oligomers conformed to the empirical relation of molecular weight to mobility that pertains to the use of these oligomers as standard molecular weight markers.

Electrophoresis, Polyacrylamide Gel↗

Evaluation and application of alumina-based Rb-82 generators charged with high levels of Sr-82/85.

Generator-produced Rb-82, a 75-sec positron emitter with potential for myocardial blood-flow imaging, was studied with various ion-exchange columns to evaluate the characteristics of alumina as an adsorber for the 25-day Sr-82 parent. Test columns of alumina, Bio Rex 70, and Chelex 100 were loaded with multimillicurie amounts of no-carrier-added Sr-82/Sr-85 (Sr-85 is a production contaminant). The breakthrough of Sr-82/Sr-85, and the yield of Rb-82, were determined for long-term elutions from each column with up to 4 liter of 2% NaCl solution at pH 8 to 9. The breakthrough of Sr-82/85 was 10(-6)-10(-5) from aluminal 10(-6)-10(-4) from Chelex 100 and Bio Rex 70. The effects of eluent flow rate and concentration, and of alumina volume, on the breakthrough and yield were also studied. An improved and automated Rb-82 generator was used for myocardial and brain blood-flow studies in experimental animals and in man; it was equipped with solenoid flow-control valves and five in. of lead shielding for the alumina columns, which were charged with 25-50 mCi Sr-82 (100-150 mCi Sr-85). The Rb-82 generator with alumina column provided up to 20-40 mCi of Rb-82 as often as every 5-10 min with less than 10(-5) breakthrough of Sr-82/85 over the 2- to 3-mo, useful life of the generator.

Aluminum Oxide↗

An osmotic effect operative in frontal gel chromatography.

The osmotic effect operative in frontal gel chromatography was quantitatively studied. When mixtures of a non-penetrating solute (Kav = 0) and a partially penetrating solute (0 less than Kav less than 1) were subjected to frontal gel chromatography, the latter formed a coextensive concentration gradient across the trailing boundary of the former, leading to the formation of a second plateau where the concentration exceeded that of the original solution plateau. It was shown that this anomaly, which we have previously predicted, was a direct consequence of osmotic perturbation of the bead size of the Sephadex gel and could be satisfactorily described by an equation based solely on the osmotic distention of the gel beads. Finally, the implications of the osmotic effect in the frontal chromatographic analysis of acceptor-ligand interactions is discussed and a method for correcting this effect is presented.

Chromatography, Gel↗

Morphogenesis of pancreatoblastoma, infantile carcinoma of the pancreas: report of two cases.

Two cases of infantile carcinoma of the pancreas were diagnosed as pancreatoblastoma based on the morphogenesis of the tumors. These encapsulated tumors adhered to the head of the pancreas and to the descending portion of the duodenum. Histologic examination revealed an organoid structure made up of cords or nodules of squamoid cells with elongated nuclei arranged in a parallel fasciculating pattern (squamoid corpuscles), surrounding tubular structures of columnar epithelial cells and intermediate light cell masses with little differentiation. Electron microscopy revealed zymogen-like granules and well developed granular endoplasmic reticulum in the cytoplasm. There were no detectable islet cells in tumor tissue. Both of these tumors could be derived from the ventral pancreas and be isolated by the lack of communication with the duct of Wirsung. As the duct of Santorini was patent, extirpation of these organoid tumors would not influence secretion of pancreatic juice. Considering the favorable prognosis after extirpation of these tumors, they should be differentiated from the usual adenocarcinoma of the pancreas occurring in adults.

Adolescent↗

Three dimensional imaging of the myocardium with radionuclides.

Transverse sections of the distribution of 129Cs and 201Ti in the human myocardium were obtained using 36 and 72 views of the thorax with a large field of view Anger camera. The cardiac cycle was divided into 100-msec intervals to obtain motion images of an average cycle of the beating heart. At least 300,000 events must be detected for each cardiac phase of each section for quantitative work. Over 8 million events from the upper thorax must be accumulated in gated studies if three or more sections are obtained for 8 intervals of 100 msec to 150 msec.

Animals↗