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Biomedical subjects

K Mochizuki

Publications and source records attributed to K Mochizuki.

At least 127 records · Page 7Linked to original sources

Characterization of a plant SINE, p-SINE1, in rice genomes.

We have previously found that a short interspersed element (SINE), named p-SINE1, is present in the Waxy gene of Oryza sativa in two copies. Here, we cloned five members of p-SINE1 located at other loci in O. sativa and determined their nucleotide sequences. These sequences had a T-rich pyrimidine tract at their defined 3' end and were flanked by direct repeats of a sequence of mostly 14-15 bp long like p-SINE1s in the Waxy gene. The consensus sequence derived from total seven members of p-SINE1 was 123 bp in length and had an internal promoter region for RNA polymerase III. The 5'-half region of the sequence was partially homologous to the tRNA-related block of rabbit C family, one of SINEs in the animal system. Two of the seven p-SINE1 members were not present in the corresponding loci in African rice, Oryza glaberrima, and may thus be available for classification of some rice strains. Comparison of the nucleotide sequences of the Waxy gene between O. sativa and O. glaberrima showed that base substitutions have frequently occurred in a p-SINE1 member (p-SINE1-r1) and a transposable element Tnr1 also present in the Waxy gene, suggesting that these elements, which appear as repetitive sequences in the rice chromosome, tend to acquire base substitutions at a higher frequency than do unique sequences.

Base Sequence↗

Fine structure of atrial natriuretic peptide (ANP)-granules in the atrial cardiocytes in the hamster, guinea pig, rabbit, cat and dog.

In the hamster, guinea pig, rabbit, dog and cat, the right and left atria and ventricles were examined by immunohistochemistry, and the right auricular cardiocytes were studied by transmission electron microscopy. Moreover, ANP-granules in the cardiocytes were analyzed by ultrastructural morphometry. Immunohistochemically, the most intensely ANP-reactive cardiocytes were localized in the right auricle, particularly more prominent in the hamster and guinea pig than in the rabbit, dog and cat. The immunoreaction in the dog and cat was weaker than that in the rabbit. ANP-immunoreactivity was not detected in the ventricular myocardium of any of all species examined, but was occasionally observed in the subendocardium of the ventricular septum. Ultrastructurally, ANP-granules were localized principally in the perinuclear region associated with the Golgi apparatus and scattered throughout the sarcoplasmic layers. The Golgi apparatus of the cardiocytes was better developed in the hamster and guinea pig than in the rabbit, dog and cat. It was poorly-developed in the dog and cat. By ultrastructural morphometry, the number of granules was greatest in the hamster followed by the guinea pig, rabbit and dog or cat, in this order. On the other hand, the diameter of granules was largest in the guinea pig and reduced via the hamster to the rabbit. The diameter was significantly smaller in the dog than in the rabbit. The diameter of granules of the cat was lay between the rabbit and dog.

Animals↗

Effect of intravitreal injection of norfloxacin on the retina in pigmented rabbits.

The effect of an intravitreal injection of norfloxacin on the retina was evaluated by in-vivo electroretinogram (ERG) and histological examination in pigmented rabbits. The intraocular pharmacokinetics after an intravitreal injection of norfloxacin were also investigated. An intravitreal injection of 50 micrograms norfloxacin produced no significant change in the ERG. An injection of 500 micrograms norfloxacin decreased the amplitude of the oscillatory potentials, and delayed their peak latencies 3 hours after the injection, but these changes recovered within 7 days. A marked suppression of the c-wave was noted in one pigmented rabbit. Neither 50 micrograms nor 500 micrograms norfloxacin caused any apparent changes in the visual evoked potential and in the retinal histology 7 days after the injection. The intraocular pharmacokinetic study showed that the concentration of norfloxacin in the choroid-retina was almost the same level as that in the vitreous body 3 hours after the injection. However, the former was higher than the latter 7 days after the injection, suggesting the persistency of norfloxacin in the choroid-retina. An intravitreal injection of 50 micrograms norfloxacin could be used without retinal toxicity. A high dose-intravitreal use of norfloxacin needs careful attention with respects to its persistency in the pigmented ocular tissues.

Animals↗

Intraocular penetration and effect on the retina of fluconazole.

The intraocular concentration of fluconazole was measured in nonvitrectomized and vitrectomized eyes after an intravenous administration of 5 or 25 mg/kg fluconazole in albino rabbits. Respective fluconazole concentrations in the aqueous, vitreous and serum 1 hour after administration were 2.87, 1.72 and 4.60 micrograms/ml at 5 mg/kg administration, and 14.93, 7.05 and 20.63 micrograms/ml at 25 mg/kg administration, indicating high and dose-dependent intraocular penetration of fluconazole. Intraocular penetration of intravenously administered fluconazole was moderately, not very much, enhanced by vitrectomy. The in-vitro electroretinogram (ERG) remained unchanged after perfusion with 20 micrograms/ml of fluconazole. The in-vivo ERG and the visual evoked potential was unchanged after the daily administration of 25 mg/kg fluconazole for 8 days. The toxicity of fluconazole on the retina would be low and within safety limits so far as it is used at clinical dosage. Fluconazole may have a place in the treatment of fungal ocular infections.

Animals↗

Fine structure of atrial natriuretic peptide(ANP)-granules in the atrial cardiocytes in the pig, cattle and horse.

In the pig, cattle and horse, the right and left atria and ventricles were examined by immunohistochemistry, and the right atrial and auricular cardiocytes were studied by transmission electron microscopy. Moreover, ANP-granules in the cardiocytes were analyzed by ultrastructural morphometry. Immunohistochemically, the most intensely ANP-reacted cardiocytes were localized in the right auricle, particularly more prominent in the pig and cattle than in the horse. Ultrastructurally, ANP-granules were located principally in the perinuclear region associated with the Golgi apparatus and throughout the sarcoplasmic layers. They were classified into 2 types, A and B. A-granule had a conspicuous electron-dense core enveloped by a membrane. B-granule had a fibrillogranular core enveloped by an indistinct membrane. By TEM equipped with a goniometer stage, a part of the limiting membrane of B-granules became visible as the stage tilt progressed. By ultrastructural morphometry, the number of each type of granules was significantly greater in the pig and cattle than in the horse. In the pig and cattle, the total number of both types of granules in the auricular cardiocytes was significantly greater than that in the atrial ones. The diameter of each type of granules was significantly larger in the pig and cattle than in the horse. The diameter of A-granules was significantly larger than that of B-granules.

Animals↗

Fine structure of atrial natriuretic peptide (ANP)-granules in the atrial cardiocytes of the mouse, rat and Mongolian gerbil.

Mouse, rat and Mongolian gerbil atrial and ventricular cardiocytes were examined by immunohistochemistry, and the right atrium including the auricle was examined by transmission electron microscopy. In addition the ANP granules of both right atrial and auricular cardiocytes were analyzed by ultrastructural morphometry. ANP immunoreactivity was detected in the atria of all three species, and the most intensely reacting cardiocytes were localized in the right auricular part of the atrium. These reactions were more prominent in the mouse and rat than in the Mongolian gerbil. ANP immunoreactivity was not detected in the ventricular myocardium of any of the three species, but was occasionally seen in the subendocardium of the ventricular septum. Ultrastructurally, the ANP granules in the auricular and atrial cardiocytes were observed to be variable in size and number, and these granules were located principally in the paranuclear region in association with the Golgi apparatus, and found throughout the sarcoplasmic layers in all three species. The ANP granules were classified into two types: A-granules containing a conspicuous electron-dense core possessing a membrane, and B-granules having profiles with a fibrillogranular, less electron-dense core than the A-granules and an indistinct membrane. The features of these granules were similar in all three species. When examined by ultrastructural morphometry, the number of each type granule and the total number of granules in the right auricular and atrial cardiocytes of the mouse and rat were significantly greater than in the Mongolian gerbil. The total number of granules in the right auricular cardiocytes was significantly greater than in the cardiocytes of the right atrium exclusive of the auricle, however, there was no significant difference between the number of A-granules and B-granules in the three species. The diameter of each type of granule in the right auricular and atrial cardiocytes of the mouse and rat was significantly greater than in the Mongolian gerbil, and the diameter of the A-granules was significantly greater than the diameter of the B-granules in all three species.

Animals↗

Fine structure of the atrial cardiocytes in the house musk shrew (Suncus murinus).

The atrial and ventricular cardiocytes of the house musk shrew were examined by immunohistochemistry, and the right atrium containing the auricle was studied by transmission electron microscopy. The atrial natriuretic peptide (ANP)-granules of the cardiocytes in the auricle and the rest of the atrium were also analyzed by ultrastructural morphometry. On immunohistochemistry, ANP immunoreactivity was detected in the atria, with the most intensely reacting cardiocytes being localized in the right auricular part of the atrium. ANP immunoreactivity was not detected in the ventricular muscles. On ultrastructure, in most of the atrial cardiocytes including the auricle, ANP-granules, well-developed Golgi apparatus and rough endoplasmic reticulum were observed, and the nuclei were characteristically situated in the periphery of the cardiocytes, being different from many other mammalian hearts. The ANP-granules were classified into two types (A and B), with most of these granules being located in the paranuclear region in association with the Golgi apparatus, and a few ANP granules being observed throughout the sarcoplasmic layers intervening between the myofibrilar bundles. On ultrastructural morphometry, the total number of granules in the right auricular cardiocytes was significantly greater than those in the atrial cardiocytes, and the diameter of the A-granules was significantly greater than that of the B-granules in both the atrial and auricular cardiocytes.

Animals↗

Tissue culture study on Mongolian gerbil's (Meriones unguiculatus) malignant melanoma.

We tried to culture melanoma cells from a Mongolian gerbil's (Meriones unguiculatus) malignant tumor. In primary culture, most of cells have abundant melanin granules in their cytoplasm. Melanin granules decreased through 5 to 15 serial passages and disappeared after 15 th passage. The morphology of the cells varied from spindle to large polydendritic cells. Although typical melanin granules were not seen when the cells were stained by Masson-Fontana method, the cells were positive for DOPA reaction. Electron microscopically, most of the cells have well-developed Golgi apparatus and rough-surfaced endoplasmic reticulum with dilated cistern, and premelanosome-like granules were frequently observed in their cytoplasm.

Animals↗

Atrial natriuretic peptide (ANP)-granules of auricular cardiocytes in aging Mongolian gerbils.

The atrial natriuretic peptide (ANP)-granules of right auricular cardiocytes were studied in aging Mongolian gerbils by immunohistochemistry and transmission electron microscopy, and analyzed by ultrastructural morphometry. Immunohistochemically, the ANP immunoreactions were weaker in 3 and 4-year-old animals than in 90-day-old, 1- and 2-year-old animals. There was no difference in the reaction among 90-day-old, 1 and 2-year-old animals, or between 3 and 4-year-old animals. Ultrastructurally, the morphological features in 1 and 2-year-old animals were similar to those of 90-day-old animals, but distinct in 3 and 4-year old animals by the presence of a few lysosomal structures in the cell. By ultrastructural morphometry, number and diameter of ANP-granules of 3 and 4-year-old animals were significantly smaller than those in the 90-day-old, 1 and 2-year-old. There was no significant difference in number and diameter among the 90-day-old, 1 and 2-year-old, or between the 3 and 4-year-old.

Aging↗

[Separation of high density lipoprotein (HDL) using anti-apo A-I, apo A-II immuno-affinity chromatography to evaluate the effects of probucol].

An assessment has been made regarding the changes of the particles of lipoprotein A-I without A-II (Lp A-I) and lipoprotein A-I with A-II (LpA-I/A-II) which correspond to HDL subfraction isolated by the use of anti-apo A-I and A-II antibody affinity columns in order to quantitatively and qualitatively investigate the change of HDL caused by administration of probucol and pravastatin which are therapeutic drugs for hypercholesterolemia. Probucol caused significant decreases of HDL-cholesterol, plasma apo A-I/apo A-II ratio and particles larger in diameter than 10.4 nm. Comparing Lp A-I and A-I/A-II ratios with those in normolipidemic controls and the ratios before and after administration of probucol, the decrease of LpA-I ratio was found to be remarkable after prolonged administration of probucol, and it was presumed that the decrease of HDL cholesterol by prolonged administration reflects the decrease of LpA-I particles more than the decrease of LpA-I/A-II. On the other hand, no significant change was seen in HDL cholesterol, plasma apo A-I/apo A-II ratio or HDL particle size in the pravastatin group. It is considered essential to observe HDL from the aspect of apoprotein, which plays an important role in the metabolism of lipoprotein, in the assessment of the anti-atherogenic activity of HDL cholesterol in future. In other words, it is necessary to analyze the change of HDL from the aspect of Lp A-I and Lp A-I/AII and investigate their respective metabolisms and roles.

Adult↗

Serodiagnosis of cancer by using Candida cytochrome c recognized by human monoclonal antibody HB4C5.

Cytochrome c from various sources, such as Candida krusei, yeast, horse, and cattle, was found to be recognized by human monoclonal antibody HB4C5 specific to lung cancer. Therefore, the cytochrome c was applied to the measurement of antibody amount in patient sera with a similar reactivity to the antibody HB4C5 for serodiagnosis of cancer. The cytochrome c from Candida krusei was most valuable for the serodiagnosis of various cancers, and the yeast cytochrome c was also useful. However, horse and bovine cytochrome c did not react with antibody of the cancer patients. By using Candida cytochrome c lung, bile duct, esophagus, and liver cancers were detected at high rates of more than 50%. In the case of lung cancer, the detection rates of small-cell, squamous, large-cell and adenocarcinoma were 78%, 63%, 100%, and 34%, respectively. The rate for small-cell carcinoma was higher than that with the currently used NSE assay system, and the rate for squamous carcinoma was comparable to that with the SCC assay system, although the system using cytochrome c did not show similar reactivity to that with the SCC system. Furthermore, lung cancer was detected at early stages by using cytochrome c, and even in the case of adenocarcinoma, the rate at early stages with the cytochrome c system was higher than that with the CEA assay system. On the other hand, false positive rates of benign diseases and normal were low--8% and 2%, respectively.

Antibodies, Monoclonal↗

Immune response of the Mongolian gerbil (Meriones unguiculatus) to sheep red blood cells.

The immune responses of Mongolian gerbils, Meriones unguiculatus, to sheep red blood cells (SRBC) were studied as compared to those of mice. After a single injection of SRBC, hemagglutinin titers in gerbils were significantly lower and hemolytic plaque-forming cells (PFC) in the spleen were less in number as compared to the response of mice. In gerbils the PFC response to a higher dose of bacterial lipopolysaccharide (LPS) was rather higher than in mice. The delayed-type hypersensitivity (DTH) assay on the foot-pad revealed that the responsiveness was considerably lower in gerbils than in mice.

Animals↗

Effects of vancomycin and ofloxacin on rabbit ERG in vivo.

The retinal toxicity of vancomycin and ofloxacin was studied by electroretinogram (ERG) recorded before and after intravitreal injection in rabbits. A dose of 1.0 mg vancomycin caused no change in the ERG for at least 8 weeks after the injection. The ERG was nonrecordable during one to four weeks after an intravitreal injection of 10 mg vancomycin, with recovery of only the c-wave. A dose of 200 micrograms ofloxacin did not cause deterioration of the b-wave, the c-wave or the oscillatory potentials throughout the follow-up period of 8 weeks. Considering the individual susceptibility of the ERG components to these antimicrobial agents and taking into account the difference in vitreous volume between the rabbit and man, the results of this study indicate the recommended intravitreal doses of these antimicrobial agents for clinical use are as follows: vancomycin 1.0 mg/0.1 ml, and ofloxacin 200 micrograms/0.1 ml.

Animals↗

Serodiagnosis of cancer using porcine carboxypeptidase A as an animal antigen recognized by human monoclonal antibody HB4C5.

Protein from hog which is recognized by human monoclonal antibody (HB4C5), generated from a patient with large cell lung carcinoma, was identified as carboxypeptidase A by comparison of the protein with carboxypeptidase A in enzymatic activity, immunologic reactivity, and amino acid sequence. Carboxypeptidase A activity was also found in human cancer tissue, and purified antigen from cancer tissue recognized by the antibody HB4C5 was reacted with rabbit anti-carboxypeptidase A serum, indicating that carboxypeptidase A is an antigen of HB4C5. Since large amounts of carboxypeptidase A can be obtained from porcine sources, a simple method for its purification was established. The fraction which was most reactive with HB4C5 was obtained from acetone powder of porcine pancreas by successive applications of water extraction, ammonium sulfate precipitation, trypsin treatment, and Mono Q column chromatography. Its apparent molecular weight was 40,000, according to SDS polyacrylamide gel electrophoresis. When the reactivity of IgG in sera with the purified carboxypeptidase A was measured, the detection rates for lung, ovary, larynx, uterus, and liver cancer were more than 50%, while the rates for stomach and breast cancer were around 30%, and pancreatic cancer, benign diseases, and normal controls were minimally detected.

Amino Acid Sequence↗

Histone H2B as an antigen recognized by lung cancer-specific human monoclonal antibody HB4C5.

Histone H2B was demonstrated to be an immunoreactive material recognized by the human monoclonal antibody HB4C5, which had been already established to be specific for lung cancers. The inhibitory effect of histone H2B on the activity of HB4C5 antibody to immunostain the cytoplasmic antigen in lung adenocarcinoma tissue indicated that histone H2B at least had antigenic determinant comparable to the cytoplasmic antigen. A mouse anti-histone H2B monoclonal antibody could immunostain the cytoplasm of lung adenocarcinoma cells in sliced tissue sections in the same manner as the human monoclonal antibody HB4C5. Enzyme-linked immunosorbent assay of HB4C5 antibody on plastic immunoplates coated with histone H2B also showed specific reactivity of this antibody with histone H2B, and the reaction was effectively inhibited when extra histone H2B or mouse anti-histone H2B monoclonal antibody was added to the reaction mixture. These results consistently lead us to a conclusion that histone H2B possesses antigenicity to the human monoclonal antibody HB4C5.

Adenocarcinoma↗

Characterization of a lung cancer-associated human monoclonal antibody HB4C5.

A human monoclonal antibody HB4C5, which is known to be associated with lung cancers, was characterized. Two kinds of light chains with 30-kD and 32-kD molecular mass were found to comprise the molecule of this IgM antibody. Various combinations of the two light chains could be contained in this IgM antibody molecule, which might have resulted in the heterogeneity of this antibody with different reactivities. The presence of the 30-kD light chain was indispensable for the antibody activity of cancer recognition. Monomeric 180-kD units of the antibody, having 32-kD light chains exclusively, which showed marginal activity of cancer recognition, could be removed by immunosorbent column chromatography with histone H2B-Sepharose 4B. F(ab')2 of the HB4C5 antibody containing the active 30-kD component as the dominant light-chain constituent could be purified by hydroxyapatite chromatography by HPLC, facilitating clinical applications of this lung cancer-associated monoclonal antibody of human origin.

Antibodies, Monoclonal↗