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Biomedical subjects

M Hoshi

Publications and source records attributed to M Hoshi.

At least 199 records · Page 11Linked to original sources

Properties of 1-methyladenine receptors in starfish oocyte membranes: involvement of pertussis toxin-sensitive GTP-binding protein in the receptor-mediated signal transduction.

In response to a meiosis-inducing hormone, 1-methyladenine (1-MA), starfish oocytes undergo reinitiation of meiosis with germinal vesicle breakdown. The 1-MA-initiated signal is, however, inhibited by prior microinjection of pertussis toxin into the oocytes, suggesting that a guanine nucleotide-binding protein (G protein) serving as the substrate of pertussis toxin is involved in the 1-MA receptor-mediated signal. We thus investigated properties of 1-MA receptors by means of binding of the radiolabeled ligand to the oocyte membranes. There were apparently two forms of 1-MA receptors with high and low affinities in the membranes. The high-affinity form was converted into the low-affinity one in the presence of a non-hydrolyzable analogue of GTP. A 39-kDa protein, which had been identified as the alpha-subunit of the major substrate G protein for pertussis toxin, was also ADP-ribosylated by cholera toxin only when 1-MA was added to the membranes. The ADP-ribosylated 39-kDa alpha-subunit could be immunoprecipitated with antibodies raised against the carboxy-terminal site of mammalian inhibitory G-alpha. These results indicate that 1-MA receptors are functionally coupled with the 39-kDa pertussis toxin-substrate G protein in starfish oocyte membranes.

Adenine↗

A novel saccharide structure, Xyl 1----3 Gal 1----(SO3-)3,4 Fuc----, is present in acrosome reaction-inducing substance of the starfish, Asterias amurensis.

The jelly coat of echinoderm eggs contains a glycoconjugate, acrosome reaction-inducing substance (ARIS), that is essential for triggering the acrosome reaction in homologous spermatozoa. In the starfish, Asterias amurensis, ARIS is a sulfated glycoprotein of an apparent molecular size of greater than 10(7). Since its biological activity is dependent mostly on its sugar moiety, oligosaccharides liberated by hydrolysis with 10 mM H2SO4 for 60 min at 100 degrees C from pronase digests of ARIS (P-ARIS) were chemically analyzed. The main oligosaccharide purified by high-performance anion-exchange chromatography was determined to be Xyl1----3Gal1----(SO3-)3,4Fuc by compositional analysis and FAB mass spectrometry. This structure indicates that ARIS possesses a novel saccharide chain having sulfated fucose as an internal residue.

Acrosome↗

Senile cataract-related accumulation of Lewis(x) glycolipid in human lens.

A glycosphingolipid that reacted positively to anti-stage-specific embryonic antigen-1 (SSEA-1) antiserum accumulated in human lens in association with aging and senile cataract formation. Since this antiserum recognizes Lewis(x) (Le(x)) structure, Gal beta 1-4(Fuc alpha 1-3)GlcNAc-, which is a typical tumor-associated and differentiation-related saccharide chain, the lens glycolipid was predicted to be a Lex antigen. The glycolipid purified from cataractous lens tissues was indeed a Lex glycolipid, Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1- 4Glc beta 1-1 ceramide. Enhanced expression of the Lex glycolipid may affect the organization of lens plasma membranes through Le(x)-Le(x) interactions, as suggested for compaction in mouse preimplantation embryos and embryonic teratocarcinomas, resulting in lens opacification, namely cataract.

Cataract↗

Mitogen-activated-protein-kinase-catalyzed phosphorylation of microtubule-associated proteins, microtubule-associated protein 2 and microtubule-associated protein 4, induces an alteration in their function.

Mitogen-activated protein kinase (MAPK), a serine/threonine-specific protein kinase which is generally activated by stimulation with various growth factors and phorbol esters, utilizes microtubule-associated protein (MAP) 2 as a good substrate in vitro. We have found that MAPK-catalyzed phosphorylation of MAP2 resulted in a significant loss in its ability to induce tubulin polymerization. The chymotryptic fragments, containing a microtubule-binding domain of MAP2, were phosphorylated by MAPK and the ability of the fragments to induce tubulin polymerization was also greatly decreased by the phosphorylation, suggesting that phosphorylation of the microtubule-binding domain is important for functional alteration of MAP2. In addition to MAP2, a 190-kDa heat-stable MAP (MAP4) found in various tissues and cells, was a good substrate for MAPK in vitro. Phosphorylation of MAP4 inactivated tubulin polymerization. We examined the effect of phosphorylation of MAP2 and MAP4 on the dynamics of microtubules nucleated by purified centrosomes in vitro. The data showed that MAPK-catalyzed phosphorylation of MAP2 and MAP4 reduced their ability to increase the apparent elongation rate and the number of microtubules nucleated by the centrosome. Thus, MAPK is capable of phosphorylating MAPs and negatively regulating their microtubule-stabilizing function.

Animals↗

Purification and characterization of a vitelline coat lysin from Ciona intestinalis spermatozoa.

In Ciona intestinalis a chymotrypsin-like activity is involved in sperm penetration of the egg vitelline coat. A chymotrypsin-like enzyme has been purified from spermatozoa by a protocol including ion exchange chromatography, gel filtration, and native polyacrylamide gel electrophoresis. The purified enzyme resulted homogeneous when analyzed by SDS-PAGE. The molecular weight of the chymotrypsin-like enzyme was estimated to be 35 kDa by gel filtration and 24 KDa by SDS-PAGE in nonreducing conditions. The pH optimum of the enzyme is 8.4 and its activity is enhanced by Ca2+. It shows the highest activity towards the synthetic substrate Suc-Ala-Ala-Pro-Phe-AMC. Furthermore, by electron microscopy, the purified enzyme affects the structure of egg vitelline coat, and thus it fulfills one of the criteria of a lysin.

Amino Acid Sequence↗

Evidence that metalloendoproteases are involved in gamete fusion of Ciona intestinalis, ascidia.

The use of specific inhibitors and substrates of metalloendoproteases provides evidence that in many systems these enzymes are involved in membrane fusion events. In this study, we investigated whether metalloendoproteases are involved in Ciona sperm-egg fusion. In vitro fertilization assays with the metal chelator 1,10-phenanthroline, specific metalloendoprotease substrates, and the vital stain Hoechst 33342 suggested that a Zn(2+)-dependent metalloendoprotease(s) takes part in Ciona sperm-egg fusion. Furthermore, electrophysiological recordings showed that insemination carried out in the presence of either 1,10-phenanthroline or the substrate CBZ-Gly-Phe-NH2 fails to induce fertilization potential or any other change in membrane potential. These results support the hypothesis that in Ciona intestinalis, a metalloendoprotease(s) is functional in gamete fusion.

Animals↗

Participation of 650-kDa protease (20 S proteasome) in starfish oocyte maturation.

A protease involved in oocyte maturation of a starfish, Asterina pectinifera, was explored. Trypsin-like and chymotrypsin-like activities of the 650-kDa protease in oocyte extract were revealed to increase more than twice under the influence of 1-methyladenine before germinal vesicle breakdown (GVBD) during maturation. The inhibitory potencies of leupeptin and its five analogs against the chymotrypsin-like activity, but not the trypsin-like activity, of this protease was well in accord with those against GVBD (Takagi Sawada et al. (1989). Dev. Biol. 133, 609-612). These results indicate that the chymotrypsin-like activity of the 650-kDa protease (most probably 20 S proteasome) plays a key role in starfish oocyte maturation.

Amino Acid Sequence↗

Iron(II) sulphate (Fricke solution) oxidation yields for 8.9 and 13.6 keV X-rays from synchrotron radiation.

The oxidation yields (G) for 8.86 and 13.55 keV X-rays produced by synchrotron radiation were measured using an iron(II) sulphate (Fricke) solution. Monoenergetic X-rays were produced using a silicon crystal monochromator. The X-rays were absorbed in 0.4 M sulphuric acid-iron(II) sulphate solution and FeIII ion yields were measured and corrected for escape fractions resulting from scattering using Monte Carlo calculations. Doses in the solution were determined using a thin window, parallel plate chamber calibrated against a primary standard free-air chamber at the Electrotechnical Laboratory (Osaka, Japan). Yields (G) of 1.50 +/- 0.06 and 1.43 +/- 0.06 mumol J-1 were obtained for 8.86 and 13.55 keV X-rays respectively.

Oxidation-Reduction↗

A novel ceramide trihexoside from the eggs of the sea urchin, Hemicentrotus pulcherrimus.

Glucosylceramide (Glc beta 1-1Cer) and a novel ceramide trihexoside (Gal beta 1-6Gal beta 1-6Glc beta 1-1Cer) were purified from the eggs of the sea urchin, Hemicentrotus pulcherrimus. Their chemical structures were determined by gas-liquid chromatography, methylation analysis, chromic acid oxidation, enzymatic hydrolysis, enzyme-linked immunosorbent assay, fast atom bombardment mass spectrometry, and proton nuclear magnetic resonance spectroscopy. The ceramide trihexoside has a novel carbohydrate structure, and its core structure, Gal beta 1-6Glc, is also novel. The ceramide moieties of these glycolipids are almost identical. Two fatty acids, 22:1 and 22h:1, constitute more than 80% of the total acids. Long-chain bases are all phytosphingosine, approximately 90% of which is n-t18:0. The finding of melibiosylceramide (Gal alpha 1-6Glc beta 1-1Cer) from the eggs of another sea urchin species [Kubo, H. et al. (1988) J. Biochem. 104, 755-760] and the present finding of the novel ceramide trihexoside suggest that there are a variety of unique sugar structures in sea urchin glycosphingolipids.

Animals↗

A novel difucosylated neutral glycosphingolipid from the eggs of the sea urchin, Hemicentrotus pulcherrimus: I. Purification and structural determination of the glycolipid.

A novel fucose-containing neutral glycosphingolipid (GL-5) was purified from the eggs of the sea urchin, Hemicentrotus pulcherrimus. The chemical structure was determined to be Fuc alpha 1-3GalNAc beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-4Glc beta 1-1Cer by methylation analysis, partial acid hydrolysis, fast atom bombardment mass spectrometry, and proton nuclear magnetic resonance spectroscopy. The unique characteristics of GL-5 are that: the reducing terminal disaccharide portion is not Gal beta 1-4Glc but GlcNAc beta 1-4Glc; it includes a GalNAc beta 1-4GlcNAc sequence and a Fuc-GalNAc linkage; the defucosylated core is a novel trisaccharide chain; and the sugar structure is one of the smallest ever characterized for a difucosylated glycolipid. The major fatty acids were 22:1 and 22h:1, and about 30% of the total acids was 2-hydroxylated. All the long-chain bases were phytosphingosines, of which about 90% was n-t18:0. The similarity of the ceramide moiety to that of glucosylceramide from the same eggs [Kubo, H. et al. (1992) J. Biochem. 111, 726-731] suggests a close biosynthetic relationship between GL-5 and the glucosylceramide.

Animals↗

A novel difucosylated neutral glycosphingolipid from the eggs of the sea urchin, Hemicentrotus pulcherrimus: II. Structural determination by two-dimensional NMR.

A novel fucosylglycolipid from the eggs of the sea urchin, Hemicentrotus pulcherrimus, was determined by using two-dimensional NMR methods. Subspectra extraction by the homonuclear Hartmann-Hahn method was useful for identification of the individual sugar components. The homonuclear Hartmann-Hahn and double-quantum-filtered correlated spectra were analyzed to establish the assignments of sugar proton resonances. On the basis of the resonance assignments, the linkages of the individual sugar components were determined to be as follows. [formula: see text] This glycolipid contains a novel skeletal structure with the linkages of GalNAc beta 1-4GlcNAc beta 1-4Glc beta. We also observed that 2-hydroxylation of the fatty acids induced appreciable chemical shift changes of the proton resonances of the phytosphingosine moiety as well as the anomeric proton resonance of the reducing terminal glucose.

Animals↗

Neutron discrepancies in the DS86 Hiroshima dosimetry system.

More than a decade has passed since a complete revision was initiated of the radiation doses received by survivors of the Hiroshima and Nagasaki atomic bombings. The new dosimetry system (DS86) was completed in 1986 and adopted shortly thereafter. Overall, DS86 was noted to be a clear improvement over the old dosimetry system. However, based on limited validation measurements, troublesome inconsistencies were suggested for neutrons. Since 1986, a substantial number of additional neutron activation measurements have been made in mineral and metal samples from Hiroshima. Importantly, a large number of measurements have now been made at distances beyond 1 km. Here, inconsistencies between neutron activation measurements and DS86 calculations for Hiroshima are examined using all available measurement data, including new measurements for 36Cl which extend the measurement range to more than 1.7 km from the epicenter, and Monte Carlo modeling calculations for each sample measured. Results show that thermal neutron activation measured beyond approximately 1 km in Hiroshima (at distances most relevant for radiation-risk evaluation) is two to 10, or more, times higher than that calculated based on DS86. Similar trends observed when comparing results by several independent measurement laboratories, using different analytical methods, suggest that the DS86 calculations for low-energy neutrons are in error. Because of the importance of the Hiroshima data in radiation risk evaluation, this large discrepancy is in need of resolution.

Japan↗

Benchmark test of transport calculations of gold and nickel activation with implications for neutron kerma at Hiroshima.

A benchmark test of the Monte Carlo neutron and photon transport code system (MCNP) was performed using a 252Cf fission neutron source to validate the use of the code for the energy spectrum analyses of Hiroshima atomic bomb neutrons. Nuclear data libraries used in the Monte Carlo neutron and photon transport code calculation were ENDF/B-III, ENDF/B-IV, LASL-SUB, and ENDL-73. The neutron moderators used were granite (the main component of which is SiO2, with a small fraction of hydrogen), Newlight [polyethylene with 3.7% boron (natural)], ammonium chloride (NH4Cl), and water (H2O). Each moderator was 65 cm thick. The neutron detectors were gold and nickel foils, which were used to detect thermal and epithermal neutrons (4.9 eV) and fast neutrons (> 0.5 MeV), respectively. Measured activity data from neutron-irradiated gold and nickel foils in these moderators decreased to about 1/1,000th or 1/10,000th, which correspond to about 1,500 m ground distance from the hypocenter in Hiroshima. For both gold and nickel detectors, the measured activities and the calculated values agreed within 10%. The slopes of the depth-yield relations in each moderator, except granite, were similar for neutrons detected by the gold and nickel foils. From the results of these studies, the Monte Carlo neutron and photon transport code was verified to be accurate enough for use with the elements hydrogen, carbon, nitrogen, oxygen, silicon, chlorine, and cadmium, and for the incident 252Cf fission spectrum neutrons.

Californium↗

Meteorological observations at Hiroshima on days with weather similar to that of the atomic bombing.

An attempt to explain discrepancies between measured neutron-induced radioactivities (i.e., 152Eu, 60Co) and calculated yields based on the Dosimetry System 1986 was made by considering moist air densities at different altitudes over Hiroshima, Japan. The investigation checked the validity of moist air density estimates used in the Dosimetry System 1986 computer codes. Meteorological observations were conducted to obtain atmospheric temperature and pressure profiles for the Hiroshima area. By coupling these observations with surface measurements taken on 6 August 1945 at Ebayama Park (3.6 km from the hypocenter), estimates of temperature, pressure, and humidity at the time of detonation were derived. This allowed the calculation of densities for dry air, moist air, and water vapor. The results proved similar to the Dosimetry System 1986 estimates. Water vapor density calculations showed the largest difference (at most 7%). These results imply that the 152Eu yield contradiction in the Dosimetry System 1986 calculations vs. real measurements for large ground distances (> 900 m) is not the result of an erroneous estimate of moist air density in Hiroshima.

Cobalt Radioisotopes↗

Specific activities of 60Co and 152Eu in samples collected from the Atomic-Bomb Dome in Hiroshima.

Neutron-induced activities 60Co and 152Eu have been measured for samples collected from the Atomic-Bomb Dome locating at 161 m from the hypocenter of the Hiroshima Bomb. Specific activities 60Co/Co and 152Eu/Eu at the time of the detonation have been determined as 10.0 +/- 1.0 Bq mg-1 (steel sample S4) and 80 +/- 9 Bq mg-1 (granite sample G1), respectively. Detailed measurements of 60Co and 152Eu activities for samples collected from various locations of the Dome show almost no directional dependence whether the sample faced to the epicenter or not, nor vertical height dependence between 17 m height and the ground level. In addition, 152Eu was not detected in the sample collected from the basement. It has been shown that the present 60Co activity value, the nearest steel one to the hypocenter, as well as other short distance data are systematically lower than the calculated values based on the neutron fluence of the DS86.

Cobalt Radioisotopes↗

Comparison of the measured gamma ray dose and the DS86 estimate at 2.05 km ground distance in Hiroshima.

The gamma ray dose at 2.05 km ground distance from the Hiroshima atomic bomb hypocenter was measured from roof tile samples by a thermoluminescence technique. Two tile samples collected at 2.45 km also were analyzed to check the reliability of the background estimates. The result for the 2.05 km distance was 129 +/- 23 mGy for five-tile sample average. This value is 2.2 times larger than the corresponding DS86 estimate. These results and those in the literature show the DS86 estimate is 50% or less of the measured value 2.05 km from the Hiroshima hypocenter.

Gamma Rays↗

Histopathological evaluation on the effect of induced hypertension chemotherapy presurgically performed in patients with advanced carcinoma of the stomach.

Stomachs resected from 13 patients with an advanced gastric carcinoma were examined histopathologically to evaluate the effect of induced hypertension chemotherapy (IHC) using angiotensin II. Eight of the 13 patients, randomly chosen, were treated presurgically by IHC with a regimen of 5-fluorouracil, adriamycin and mitomycin C; the same regimen was used in the remaining five patients but without inducing hypertension. Clinical evaluation of the effect gave rise to the confirmation, presurgically, of complete response in three and partial response (PR) in two patients in the IHC group, whereas in the non-IHC group, the highest rating was PR, which was attained in only one patient. Also in the histopathological assessment based on a five-step grading, the IHC group earned in average a higher score than the non-IHC group, with a difference proved significant by Wilcoxon's test. A histological rating of Grade 3, the highest effectiveness, was given to three patients in the IHC group, in one of whom the resected stomach disclosed no viable carcinoma cells but only fibrotic areas replacing carcinoma. There was also a correlation between the clinical and histological ratings as proved to be significant by Spearman's test. We conclude that in gastric carcinoma, the effect of chemotherapy is enhanced by angiotensin II-IHC.

Adenocarcinoma↗