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Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 451 records · Page 25Linked to original sources
PubMed1985

Characterization of N-glycolylneuraminic acid-containing gangliosides as tumor-associated Hanganutziu-Deicher antigen in human colon cancer.

Hanganutziu-Deicher (HD) antigen-active N-glycolylneuraminic acid (NeuGc)-containing gangliosides were isolated and characterized from human colon cancer tissues. The antigenic gangliosides were detected by thin-layer chromatography by our newly developed method (H. Higashi, Y. Fukui, S. Ueda, S. Kato, Y. Hirabayashi, M. Matsumoto, and M. Naiki. J. Biochem., 95: 1517-1520, 1984) of enzyme immunostaining using affinity-purified chicken antibody against hematoside containing NeuGc (II3NeuGc-LacCer) and horseradish peroxidase-conjugated rabbit anti-chicken lgG. One to six species of the antigenic gangliosides were isolated from seven of 16 cases of colon cancer, whereas no antigenic compound was detected in all apparently normal colorectal tissues from 17 individuals without colorectal cancer. Tissues from different patients showed different patterns of molecular species of the antigenic gangliosides. Densitometric determination indicated that HD antigenic sialic acid, NeuGc, accounted for about 1% or less of the total lipid-bound sialic acids. Four species of antigenic gangliosides were identified as hematoside and hematoside-containing O-acyl ester (II3NeuGc-LacCer and II3 4- or 7-O-acyl-NeuGc-LacCer), GM2-containing NeuGc (II3NeuGc-GgOse3Cer), and sialylparagloboside (IV3NeuGc-nLcOse4Cer) by their behaviors on 2-dimensional thin-layer chromatography, and the effects of mild alkaline treatment, sialidase treatment, periodate oxidation, and endo-beta-galactosidase treatment.

Antigens, Heterophile↗
PubMed1984

In vivo and in vitro studies on cocaine metabolism: ecgonine methyl ester as a major metabolite of cocaine.

The levels of cocaine, benzoylecgonine and ecgonine methyl ester were measured periodically in the urine of dogs and rabbits by gas chromatography-chemical ionization selected ion monitoring mass spectrometry after subcutaneous injection of 2.9 mumol/kg (1.0 mg/kg) cocaine--HCl. Ecgonine methyl ester persisted much longer than benzoylecgonine. Urinary ecgonine methyl ester could be identified for 72 h in all the experimental dogs. The ester accounted for 6.6-27.1% of a dose of cocaine in dogs and 8.8-31.9% in rabbits 24 h after administration. Excretion of benzoylecgonine in urine was 6.3-19.2% in dogs and 7.5-32.9% in rabbits. This confirms that ecgonine methyl ester is one of the major metabolites of cocaine as well as benzoylecgonine. Excretion of the parent drug and its two hydrolyzed metabolites in faeces was very small, less than 1% of administered dose. Tri-o-tolylphosphate and eserine significantly inhibited cocaine hydrolysis to benzoylecgonine and ecgonine methyl ester, respectively. Parathion and EDTA, however, had no effects on cocaine hydrolysis in vivo. In vitro demethylation of cocaine to benzoylecgonine was demonstrated in the plasma from dogs and this production of benzoylecgonine was much more than that of non-enzymatic formation in buffer at physiological pH. It was concluded that a large part of the benzoylecgonine excreted in urine is an in vivo enzymatic product of cocaine. On the other hand, plasma cholinesterase and liver esterase mediated ecgonine methyl ester formation. This liver esterase was abundant in the soluble fraction and could be found in both of microsomal and mitochondrial fractions.

Animals↗
PubMed1984

Quantitation of cocaine, benzoylecgonine and ecgonine methyl ester by GC-CI-SIM after Extrelut extraction.

A method of gas chromatography-chemical ionization selected ion monitoring (GC-CI-SIM) is described for the determination of cocaine, benzoylecgonine and ecgonine methyl ester in biological materials using an Extrelut extraction technique. Recoveries of cocaine, benzoylecgonine and ecgonine methyl ester by this technique were 95, 81 and 97%, respectively. The method uses cocaine-d5, benzoylecgonine-d5 and lidocaine as internal standards, and isobutane as reagent gas for chemical ionization. Sensitivity of the method proved to be 1 ng/ml for cocaine and benzoylecgonine, and 10 ng/ml for ecgonine methyl ester when used in a 10-ml urine sample. With animal experiments, ecgonine methyl ester as well as benzoylecgonine was confirmed as a major metabolite of cocaine.

Animals↗
PubMed1984

Localization of actin and myosin for the study of ameboid movement in Dictyostelium using improved immunofluorescence.

The distribution of actin and myosin in Dictyostelium amebae at different developmental stages was studied by improved immunofluorescence ("agar-overlay" technique). Both were localized at the cortical region of amebae in all early developmental stages. In amebae with polarized morphology, bright fluorescence with antiactin was seen in the anterior pseudopode. The cortex in the posterior end was also stained with antiactin. On the other hand, very specific crescent-shaped staining with antimyosin was seen at the posterior cortex. In cells in contact with each other, actin was concentrated at the contact region, whereas myosin was localized specifically in the cortex on the other side of the contact region. At the aggregation stage, when monopodial amebae migrate forming streams, actin staining was seen all around the cell periphery, with intense fluorescence in the anterior pseudopode. On the other hand, specific staining of myosin was seen only at the posterior cortex. The cleavage furrow of cells performing cytokinesis displayed distinct myosin staining, and this staining represented the filamentous structure aligned in parallel to the axis of constriction. These findings indicate that myosin staining reflects the portion of the cell cortex where contraction occurs and the motive force of ameboid movement is generated at the posterior cortex of a migrating cell.

Actins↗
PubMed1984

Sensitive enzyme-immunostaining and densitometric determination on thin-layer chromatography of N-glycolylneuraminic acid-containing glycosphingolipids, Hanganutziu-Deicher antigens.

A sensitive enzyme-immunochemical staining method was developed for detection of N-glycolylneuraminic acid (NeuGc)-containing glycosphingolipids (GSLs) on silica gel thin-layer chromatography. The procedure consists of immune reaction among NeuGc-containing GSLs, affinity-purified chicken anti-NeuGc-LacCer and horseradish peroxidase-conjugated rabbit anti-chicken IgG, and the peroxidase reaction using 4-chloro-1-naphthol as a chromogenic substrate. Quantitative determination was achieved by direct densitometric scanning of the enzyme-immunostained spots on the chromatogram. As little as 0.5 pmol of NeuGc-LacCer, NeuGc-nLcOse4Cer, and NeuGc-nLcOse6Cer (0.64-1.0 ng) could be detected with a good signal-to-noise ratio. A semi-linear detector response was observed up to 50 pmol of each GSL. This procedure can be applied easily to other glycolipid antigen systems.

Antigens, Heterophile↗
PubMed1984

Isolation of transformation-defective, replication-nondefective early region 1B mutants of adenovirus 12.

We isolated three adenovirus 12 early region 1B mutants (in205B, in205C, and dl205) by ligation of the cleaved DNA-protein complex and transfection of human embryo kidney cells with the ligation products. These mutants could replicate efficiently in human embryo kidney or KB cells but showed markedly reduced transforming capacities both in vitro and in vivo. In cells infected with the mutants, the early region 1B gene was transcribed efficiently. In cells infected with in205B, the products corresponding to the early region 1B-coded 19,000-molecular-weight polypeptide was detected by in vitro translation but not immunoprecipitated extract of labeled cells. In cells infected with in205C or dl205, the products corresponding to the same polypeptide were not detected by either in vitro translation or immunoprecipitation of labeled cell extracts. The results suggest that the 19,000-molecular-weight polypeptide encoded by early region 1B is required for cell transformation but not for viral propagation.

Adenoviruses, Human↗
PubMed1984

Characterization of a host range mutant of human adenovirus 12 defective in early region 1B.

We isolated an adenovirus 12 early region 1B mutant (in206B) by ligation of the cleaved DNA-protein complex and transfection of human embryo kidney cells with the ligation products. By deduction from the DNA sequencing analysis, the two polypeptides (with molecular weights of 19,000 and 54,000) coded in the early region 1B were fused in this mutant to produce a large polypeptide. This mutant could replicate efficiently in 293 cells but not efficiently in KB or human embryo kidney cells. In KB cells, viral DNA replication could not be detected after infection with in206B. In human embryo kidney cells, viral DNA replication did occur, but the transition of viral mRNA patterns from early to late did not occur even after DNA replication, resulting in failure to produce the late polypeptides. These results indicate that the early region 1B products may be involved in both viral DNA replication and regulation of transcription.

Adenoviruses, Human↗
PubMed1984

Expression of the E4 gene is required for establishment of soft-agar colony-forming rat cell lines transformed by the adenovirus 12 E1 gene.

Rat 3Y1 cells were transfected with recombinant gARC ( pSV2gpt carrying the adenovirus 12 early region 1 [E1] gene), and focus formation was observed in monolayer cultures after culture of cells in gpt-selective medium (Eagle medium containing 10% fetal calf serum, xanthine, thymidine, aminopterin, and mycophenolic acid) for 10 days, followed by focus formation. Transformed E1Y cell lines were then established from these foci. The E1Y cells were transformed morphologically similarly to cells transformed with intact adenovirus 12 DNA but formed no colonies in soft-agar culture and induced tumors in transplanted rats only after a long incubation period. For the establishment of completely transformed cells, 3Y1 cells were transformed with combinations of gARC , pE3 (pBR322 carrying the adenovirus 12 E3 gene), and gE4 ( pSV2gpt carrying the adenovirus 12 E4 gene) DNA. E1- 3Y cells (3Y1 cells transformed with gARC and pE3 DNA), E1- 4Y cells (3Y1 cells transformed with gARC and gE4 DNA), and E1-3- 4Y cells (3Y1 cells transformed with gARC , pE3 , and gE4 DNA) were established. These transformed cell lines were compared for growth in Eagle medium with 2 or 10% fetal calf serum, colony formation in soft-agar culture, and tumor growth in rats transplanted with the transformed cells. Several transformed cell lines of E1- 4Y and E1-3- 4Y cells showed colony formation in soft-agar culture and abundant expression of the E1B gene. T antigen f was seen by immunofluorescence as flecks in these cells, in which the E4 gene was transcribed, but was not seen in E1Y cells, suggesting that T antigen f was encoded by the E4 gene. The suggestion was confirmed by the observation that T antigen f was detected in COS-1 cells transfected singly with gE4 DNA by immunofluorescence with polyclonal and monoclonal antibodies. Transcription of the E4 gene was confirmed in gE4 -transfected COS-1 cells. T antigen f, one of the E4 gene products, was identified as a polypeptide of molecular weight 11,000 (E4- 11K ) by immunoprecipitation with monoclonal antibodies. The above results also suggest that expression of the E4 gene gives cells the advantage of forming colonies in soft-agar culture. A tendency was noticed for E1B gene expression to be enhanced by E4 gene expression. The relationship between enhancement of colony formation in soft-agar culture and enhancement of E1B gene expression is discussed.

Adenoviruses, Human↗
PubMed1984

Tumor-associated expression of glycosphingolipid Hanganutziu-Deicher antigen in human cancers.

Expression of heterophile Hanganutziu-Deicher (HD) antigen on the cell surface of various human malignant tumor tissues was studied by membrane immunofluorescence staining with chicken antiserum against N-glycolylneuraminyllactosylceramide (HD3) and fluorescein-conjugated rabbit anti-chicken IgG. The HD antigen was demonstrated in samples from 38 of 77 (38/77, 49%) cases, consisting of gastric (9/16), breast (8/14), colorectal (3/12), nasopharyngeal (4/7), and uterine (2/3) carcinomas, leukemias (5/10), malignant lymphomas (2/5), and other cancers (5/10). Histological examination indicated that expression of the antigen by the gastric, colorectal and breast carcinomas and leukemias was not related to their histological types. The cytoplasm and the surface of the malignant glandular epithelial cells were both specifically stained by immunofluorescence staining of frozen sections in one colon adenocarcinoma. Glycosphingolipids (GSLs) were extracted from the colon cancer tissue and two HD antigen-active GSLs were detected on thin-layer chromatography (TLC) by enzyme-immunostaining using affinity-purified chicken anti-HD3 and horseradish peroxidase-conjugated rabbit anti-chicken IgG. One migrated to the same position as HD3 on TLC, and the other to a position similar to that of authentic N-glycolylneuraminylneolactotetraosylceramide.

Antigens, Heterophile↗
PubMed1984

Automatic control in anesthesia: a comparison in performance between the anesthetist and the machine.

This report is divided into two parts. First, we developed two new servo control systems by modifying an existing one. The original system was designed to control inspired halothane concentration using mean arterial pressure; the two new systems were designed to control inspired halothane concentration using end-tidal concentration or to control mean arterial pressure using the automated infusion of nitroprusside. Second, we compared the performance of experienced physician and nurse anesthetists (nine, six, and six experiments, respectively). The experiments incorporated a standardized testing sequence of two changes in desired blood pressure (set point) and two pharmacologically induced disturbances in blood pressure (perturbations). The scoring was designed to examine how fast blood pressure changed (90% response time), how far past the set point it went (overshoot), how long it took to eliminate most of the fluctuations in blood pressure (settling time), and the degree of fluctuation of blood pressure after settling (stability). Given three systems to be tested, there were (3 X 14) 42 possible mean scores for the machine and 42 for the anesthetists. The machine scored better than the anesthetists in 38 out of 42 of the mean scores; the differences were statistically significant in 19 out of 42 scores. The wide scatter in performances of the anesthetists prevented the achievement of significance in nine cases with large differences between means. Thus when the scores from the three systems were combined to achieve a larger n value, the machine outperformed the anesthetist in 12 out of 14 scores.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia, Inhalation↗