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Biomedical subjects

M Kojima

Publications and source records attributed to M Kojima.

At least 559 records · Page 31Linked to original sources

Purification and characterization of p-coumaroyl-D-glucose hydroxylase of sweet potato (Ipomoea batatas) roots.

p-Coumaroyl-D-glucose hydroxylase in sweet potato (Ipomoea batatas Lam.) has been purified to apparent electrophoretic homogeneity using a combination of anion-and cation-exchange, hydrophobic and gel filtration chromatography. The purified enzyme was a monomer with a molecular weight of 33,000 and pI of 8.3. The purified enzyme showed not only hydroxylase activity but also polyphenol oxidase activity. L-Ascorbic acid was the best electron donor for the hydroxylation reaction, which had an optimum pH of 7.0. The enzyme hydroxylated p-coumaroyl-D-glucose, p-coumaric acid, and p-cresol but did not act on o-coumaric acid, m-coumaric acid, 4-hydroxy-3-methoxycinnamic acid, p-hydroxybenzoic acid or L-tyrosine. While the enzyme utilized p-coumaroyl-D-glucose and p-coumaric acid equally at pH 7.0, it hydroxylated only p-coumaroyl-D-glucose at pH 5.5. The enzyme oxidized diphenols such as D,L-(3,4-dihydroxyphenyl) alanine and caffeic acid, but exhibited no clear pH optimum in this reaction characteristic of polyphenol oxidase. Both the hydroxylase and the polyphenol oxidase activities were strongly inhibited by beta-mercaptoethanol, diethyldithiocarbamate, KCN, and p-coumaric acid (in concentrations higher than 5 mM). Ammonium sulfate and sodium chloride activated the hydroxylase activity but not the polyphenol oxidase activity of the enzyme. The enzyme activity and L-ascorbic acid contents changed in a manner suggesting their involvements in chlorogenic acid biosynthesis during incubation of sliced sweet potato root tissues.

Ammonium Sulfate↗

Fulminant hepatitis B: induction by hepatitis B virus mutants defective in the precore region and incapable of encoding e antigen.

Clones of hepatitis B virus were propagated from 10 cases of fulminant hepatitis B after amplification by polymerase chain reaction and their nucleotide sequences of the precore region were determined. All 113 clones from 9 cases had a point mutation from guanine to adenine at nucleotide 83 in the precore region, which converted codon 28 for tryptophan (TGG) to a stop codon (TAG) and prohibited the synthesis and secretion of hepatitis B e antigen. Precore-region defects were not detected in any of 23 clones from the remaining 1 case. By contrast, precore-region defects were not found in any of 180 clones from 8 cases of acute hepatitis B without hepatic failure serving as controls. The source of infection was traceable in 3 cases. The same precore-region defect, along with the sequence identity of 435 nucleotides, was observed in clones from the case of a baby and his grandmother, who carried the virus and was implicated in the transmission, and also in clones from two pediatricians and the carrier patients they attended. These findings support the hypothesis that precore-defective mutants have stronger activity to induce fulminant hepatitis than nondefective viruses.

Adolescent↗

Interaction between phospholipids and biocompatible polymers containing a phosphorylcholine moiety.

Random and block copolymers containing a phospholipid polar group in their side chain were synthesized by the copolymerization between 2-methacryloyloxyethyl phosphorylcholine and styrene. These copolymers showed amphiphilic character, especially poly(methacryloyloxyethyl phosphorylcholine-block-styrene) formed stable polymer micelles in water. The interaction between natural phospholipid, dipalmitoylphosphatidylcholine and methacryloyloxyethyl phosphorylcholine copolymers was investigated. The amount absorbed of dipalmitoylphosphatidylcholine from its liposomal solution on to the poly(methacryloyloxyethyl phosphorylcholine-co-styrene) surface increased with increase of methacryloyloxyethyl phosphorylcholine composition. Moreover, when poly(methacryloyloxyethyl phosphorylcholine-block-styrene) was added to dipalmitoylphosphatidylcholine solution, organization of dipalmitoylphosphatidylcholine molecules and stabilization of bilayer structure of dipalmitoylphosphatidylcholine liposome were found. This means that methacryloyloxyethyl phosphorylcholine moieties in the copolymer have a strong affinity to dipalmitoylphosphatidylcholine molecules. The blood compatibility of methacryloyloxyethyl phosphorylcholine copolymers was also investigated with particular attention to the aggregation ability of platelets after contacting methacryloyloxyethyl phosphorylcholine copolymers; this ability decreased when platelets were put in contact with polymers without a methacryloyloxyethyl phosphorylcholine moiety. On the other hand, aggregation ability remained at almost the same level to that of original platelets after contact with methacryloyloxyethyl phosphorylcholine copolymers. From these findings, we concluded that methacryloyloxyethyl phosphorylcholine copolymers show excellent blood compatibility due to adsorption of lipids from plasma and the formation of an organized adsorption layer of lipids on the surface of the methacryloyloxyethyl phosphorylcholine copolymers.

1,2-Dipalmitoylphosphatidylcholine↗

Distribution and characterization of diglycosyldiacylglycerol isomers with different anomeric configuration in higher plants.

The novel diglycosyldiacylglycerol (DGDG), galactosyl(beta 1----6)-galactosyl(beta 1----3')-diacylglycerol (B isomer), present in Adzuki beans was found to be distributed together with the well-known galactosyl(alpha 1----6)-galactosyl(beta 1----3')-diacylglycerol (A isomer), in all (10) of the higher plants examined. The highest levels were found in leguminous seeds were the amounts were always less than 33% of the total DGDG of mature seeds. The highest proportion of the B isomer was found in Adzuki bean seed DGDG (26-33%), with the lowest in pea seed DGDG (2%). The amounts of the B isomer in DGDG of Adzuki and kidney beans cotyledons were almost equal to those in mature seeds. Immature seeds and hypocotyls of three kinds of beans also contained the B isomer in small amounts compared with the mature beans, while only trace amounts of the isomer was found in other organs such as leaves, stems, pods, roots and generative organs of plants, except for root from kidney beans. The molecular species composition of the principal diacylglycerol moieties in the A and B isomers of DGDG were found to be significantly different among several plant seeds, although the component diacylglycerol species were qualitatively similar to each other.

Carbohydrate Sequence↗

Oscillatory potentials and permeability of the blood-retinal barrier in noninsulin-dependent diabetic patients without retinopathy.

Electroretinography and vitreous fluorophotometry were performed in 36 eyes of 36 noninsulin-dependent diabetic patients and in 32 eyes of 32 healthy control subjects between the ages of 30 and 59 years. Fluorescein fundus angiograms showed no abnormalities in either group. Peak implicit time of the first deflection of the oscillatory potential, interpeak interval between the first and second deflections, and the sum of the amplitudes of the upward deflections were analyzed. Inward permeability of the blood-retinal barrier was calculated by vitreous fluorophotometry and computer simulation. The peak implicit time of the first deflection and the interpeak interval between the first and second deflections of the oscillatory potential were significantly longer in diabetic patients than in control subjects (P less than 0.01). No significant difference in inward permeability of the blood-retinal barrier existed between the two groups. These results indicate that a selective delay in the peak implicit time of the oscillatory potential (neurosensory retinal abnormality) may be present in noninsulin-dependent diabetic patients, representing retinal functional changes before changes in blood-retinal barrier permeability are apparent.

Adult↗

Malfunction of arterial sarcoplasmic reticulum leading to faster and greater contraction induced by high-potassium depolarization in young spontaneously hypertensive rats.

The contribution of sarcoplasmic reticulum was studied with regard to the increase in arterial contraction induced by a high-potassium depolarization in spontaneously hypertensive rats (SHR). The 20 mmol/l potassium-induced contraction of femoral arteries was faster and greater in 6-week-old SHR than in age-matched normotensive Wistar-Kyoto (WKY) rats. Relaxation after washing the arteries with a Krebs solution was slower in SHR than in WKY rats. When the sarcoplasmic reticulum of SHR arteries had been depleted of calcium by caffeine in a calcium-free solution, the rate of high-potassium-induced contraction of the calcium-depleted SHR arteries was slowed, the same result as that with non-calcium-depleted WKY arteries. In ryanodine-treated arteries, the rate and magnitude of high-potassium-induced contraction were enhanced slightly in SHR and greatly in WKY rats, resulting in no final difference between SHR and WKY rats. Ryanodine slowed the relaxation rate in WKY rats but not in SHR. These results suggest that the diminution in ability of sarcoplasmic reticulum to sequester calcium may be responsible for the faster rate and greater magnitude of high-potassium-induced contraction with the slower relaxation in SHR arteries. We postulated that genetic malfunction of sarcoplasmic reticulum causes the increased contraction of arterial smooth muscle leading to the enhanced vasoconstriction and elevated blood pressure in SHR.

Animals↗

Posttransfusion fulminant hepatitis B associated with precore-defective HBV mutants.

Fulminant hepatitis B developed in 8 recipients of blood units without detectable hepatitis B surface antigen on routine screening. All 124 hepatitis B virus (HBV) DNA clones propagated from their sera possessed defects in the precore region. A point mutation from guanine to adenine at nucleotide 83, converting codon 28 for tryptophan (TGG) to a stop codon (TAG), was the commonest, and it was found in all 113 clones from 7 cases. The remaining case displayed 1 clone with this point mutation and 10 clones with an insertion of 2 base pairs after nucleotide 26. Antibody to hepatitis B core antigen (anti-HBc) was detected in a high titer in 1 of 10 pilot plasma samples of blood units transfused to this case. HBV DNA clones propagated from it exhibited the same precore-region defects as those from the recipient. On the basis of these results HBV mutants, defective in the precore region, would appear to be responsible for posttransfusion fulminant hepatitis B, and the exclusion of blood units with high-titered anti-HBc would be efficacious in preventing it.

Amino Acid Sequence↗

Monocytoid B lymphocytes and epithelioid cell clusters in abscess-forming granulomatous lymphadenitis. With special reference to cat scratch disease.

In order to clarify the appearance of monocytoid B lymphocytes (MBLs) in abscess-forming granulomatous lymphadenitis (AGL) and the relation between AGL and cat-scratch disease (CSD), 48 cases of AGL were studied histologically. MBLs were present in about 50% of AGL cases. Warthin-Starry (WS) silver stain-positive bacteria, which are the causative agent of CSD, were present in 52.4% of AGL cases with MBLs and 59.2% of AGL cases without MBLs. The appearance of MBLs in AGL was not related to various clinical features, including disease interval from initial lymphadenopathy to lymph node biopsy. Histologically, epithelioid cell clusters appeared in about 70% of MBL-positive AGL cases, but were not observed in MBL-negative AGL. Therefore, a close interaction between MBLs and epithelioid cells in AGL is suggested, and we emphasize that the histological features of some AGL cases resemble those of toxoplasmic lymphadenitis.

Abscess↗

Abnormality in sarcoplasmic reticulum-dependent arterial contraction in responses to caffeine and noradrenaline in spontaneously hypertensive rats.

We studied caffeine- and noradrenaline-induced contraction in tail arteries from 4-week-old male SHR and age- and sex-matched WKY. After the sarcoplasmic reticulum Ca2+ had been depleted by the Ca(2+)-free EGTA (0.1 mmol/l) solution, the caffeine (10 mmol/l)-induced contractions in a low-Ca2+ (0.5 mmol/l) solution were smaller in SHR than in WKY. After the sarcoplasmic reticulum had been loaded with Ca2+ in physiological Ca2+ (2.5 mmol/l) solution, caffeine- and noradrenaline (10(-5) mol/l)-induced contractions in a Ca(2+)-free EGTA solution were smaller in SHR than in WKY. The Ca2+ concentration-tension relationship in skinned arterial fibres was similar in WKY and SHR. These data suggest that the ability of the sarcoplasmic reticulum to take up Ca2+ and store Ca2+ is decreased in SHR. The decreased take up and store of Ca2+ may increase cytosolic Ca2+, which elevates arterial resistance and develops hypertension in gene hypertension.

Animals↗

Hepatitis B virus mutants with precore-region defects in two babies with fulminant hepatitis and their mothers positive for antibody to hepatitis B e antigen.

Clones of hepatitis B virus (HBV) DNA were propagated from sera of two babies who developed neonatal fulminant hepatitis B, as well as from sera of their mothers who carried HBV with antibody to hepatitis B e antigen, and the precore-region sequences were determined. A point mutation from guanine to adenine, converting codon 28 for tryptophan (TGG) to a stop codon (TAG), was detected in 18 of 20 HBV DNA clones from mother and all 31 clones from baby in one family, and invariably in 55 clones from mother and three clones from baby in the other family. These results indicate that HBV mutants defective in the precore region in some carrier mothers with antibody to hepatitis B e antigen may transmit fulminant hepatitis B to their babies.

Amino Acid Sequence↗

Accumulation of tri-n-butyltin chloride and triphenyltin chloride by oral and via gill intake of goldfish (Carassius auratus).

1. The bioconcentration factors (BCF) of both chemicals via gill intake of goldfish reached plateaux after 21 days of exposure. The average BCF values of Bu3SnCl and Ph3SnCl (N = 2) were 1976 and 1384, respectively, after 21-28 days of exposure. 2. The BCF values by oral intake of goldfish reached plateaux after 28 days for Bu3SnCl but did not for Ph3SnCl. The BCF values of Bu3SnCl and Ph3SnCl were 0.04 and 0.10, respectively, after 35 days. 3. The order of the BCF values via gill intake was reversed compared with that by oral intake. Further, the contamination of freshwater fish by these compounds was discussed using the results of these experiments.

Administration, Oral↗

[Establishment and characterization of a human undifferentiated carcinoma cell line (HMG)].

The undifferentiated carcinoma cell line (HMG) was established from a nude mouse tumor which had been produced by transplantation of a intraperitoneal tumor of 27-year-old woman. The HMG cell line has the following biological properties. 1. The HMG cells are round to oval in shape and grow as floating cell aggregates like a rouleau or a cluster of grapes. 2. 100 passages have been carried out over a year, and the population doubling time is about 17 hrs. 3. In the original tumor, keratin and vimentin were expressed simultaneously, in HMG cells, however, only localization of vimentin was confirmed. 4. By chromosomal analysis, over 90% of the cells revealed 46, XX, with no karyological abnormalities, at passage 82. 5. When heterotransplanted into the subcutis of a nude mouse, HMG cells produced a undifferentiated carcinoma resembling the original tumor.

Adult↗

DNA polymerase action blocked by adenine adducts induced by 5-hydroxymethylchrysene sulfate.

Modification of M13mp10 single-stranded DNA with 5-hydroxymethylchrysene (5HCR) sulfate, the ultimate carcinogenic metabolite of 5-methylchrysene, resulted in formation of N6[(chrysen-5-yl)methyl]adenine and N2[(chrysen-5-yl)methyl]-guanine at the ratio of 2.7:1. Measurement of DNA synthesis using this modified template and E.coli DNA polymerase I (Klenow fragment) demonstrated that increasing levels of adducts caused a progressive decline in replication. Analysis of reaction products on DNA-sequence gels revealed DNA elongation to be arrested exclusively at adenine adducts in -AAAGGA- and -AACA- sequences.

Adenine↗

Effect of phosphoenolpyruvate on energy metabolism of ischemic liver in anesthetized rats--31P-MRS study.

The effect of phosphoenolpyruvate (PEP) on energy metabolism of ischemic liver was examined in anesthetized rats. In vivo 31P-NMR spectroscopy (31P-MRS) was used to monitor cellular energy metabolism. Hepatic ischemia was induced by temporarily clamping the portal vein for 60 minutes. The liver adenosine triphosphate (ATP) levels decreased remarkably during ischemia, and they gradually increased after ischemia but did not return to pre-operative levels. PEP effectively increased the levels of ATP. The ATP levels of the PEP-treated rats were significantly higher than those of the control rats, and also intracellular acidosis was improved during post-ischemic reperfusion. These findings suggest that PEP may have a cytoprotective effect and improve the energy metabolism in the ischemic liver.

Adenosine Triphosphate↗

[Pharmacokinetic and clinical evaluations of flomoxef in neonates].

To evaluate pharmacokinetics and clinical efficacy of flomoxef (6315-S, FMOX) in neonates, FMOX was administered to 21 neonates. With 20 mg/kg and 40 mg/kg of intravenous drip-infusion of FMOX 60 minutes, half lives (T 1/2's) was 64.9 minutes and 130.3 minutes, respectively, and when 20 mg/kg of FMOX was infused intravenously to 2 cases, half lives were 70.8 minutes and 110.1 minutes, respectively. When 45-100 mg/kg of FMOX was administered to 17 neonates with infections (pneumonia 8, sepsis 1, sepsis suspected 2, intrauterine infection 2, urinary tract infection 2, omphalitis 2), the efficacy rate was 88.2% (15 of 17). No adverse reactions were observed clinically in the 21 neonates. Transient elevation of eosinophilia was observed in 1 case and transient elevation of S-GOT and S-GPT 1 in another. These results suggest that FMOX is an effective and safe antibiotic to use in neonates.

Alanine Transaminase↗

[Inhibition of cholesterol synthesis and cataract].

The cholesterol requirements of the lens for the formation of plasma membranes are met by self-synthesis only until early postnatal life. This ability subsequently decreases with increasing age and probably ceases altogether. The cholesterol supply then has to come from exogenous sources. These findings are of great importance for the qualitative assessment of extrahepatic side effects of HMG-CoA-reductase inhibitors given for disturbances of lens transparency. The observation of cataracts in chronic toxicity tests with high doses in beagle dogs suggested that the lens enzyme was also affected by the inhibitor, thus causing cataracts. In contrast to other workers' results, we did not find any activity of HMG-CoA-reductase in our experiments with calf, bovine, dog and rat lenses. Preliminary studies with bovine eyes demonstrate a direct correlation between the cholesterol serum level and the content of the aqueous humor. This shows the importance of the exogenous supply for the lens requirements. A further strong argument is that the use of HMG-CoA-reductase inhibitors in patients with pathologically increased cholesterol level is safe as far as the lens transparency is concerned, as demonstrated in clinical trials.

Animals↗