Separation of oligonucleotides by high-performance ion-exchange chromatography on a non-porous ion exchanger.
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Chromatography, DEAE-Cellulose↗
Biomedical subjects
K Matsubara
Publications and source records attributed to K Matsubara.
Microdeletion associated with the integration process of hepatitis B virus DNA.
Hepatitis B virus (HBV) DNA is often found in integrated form in hepatocellular carcinomas (HCC) and in non-cancerous liver cells of chronic carriers of HBV. However, the process of integration has not been well understood. Analyses of integrant DNA was expected to give clues. However, the majority of the integrants are products of multistep rearrangements following integrations, and analysis of randomly selected samples do not give clues for understanding the process of primary integrant formation. Therefore, one must select an appropriate integrant(s) that has a simple structure. We surveyed a collection of integrants prepared from many HCC's, and found one integrant that has the simplest structure so far studied: The viral genome is almost complete, is joined to cellular DNA using the cohesive end of the viral DNA, and furthermore, the "left" and "right" flanking cellular DNA's are almost contiguous. Analysis of the unoccupied sites in cellular DNA showed that, although almost contiguous, it has generated a microdeletion (15 base pairs) in the target sequence. This target sequence has a short region of homology to the sequence in the viral genome located close to the junction. One integrant with strikingly similar features has been reported independently. Two similar, but not identical cases from literatures could be added to this category. Therefore, the integrants with these properties may represent a unique category among those prepared from hepatocellular carcinomas. Based on these findings, we propose that this integrant represents the primary product of integration, and discuss the intermediate acting in the process of integration.
Base Sequence↗
Expression of alpha-amylase in human lung cancers.
Thirty-three human lung tumors were studied for the expression of alpha-amylase by immunohistochemical and Northern blot analyses. Twenty of them were adenocarcinomas, among which 17 proved to be adequate for mRNA analyses and were, except for two, amylase mRNA producers. Seven were squamous cell carcinomas, none of which produced amylase. The remaining six consisted of two undifferentiated small cell carcinomas, and one each of undifferentiated large cell carcinoma, carcinoid tumor, mucoepidermoid carcinoma, and metastatic lung cancer; the mucoepidermoid carcinoma proved to be an amylase producer. These observations strongly suggest that among lung cancers, the production of alpha-amylase is a property commonly associated with adenocarcinomas and can be used for distinguishing cell types. Histogenesis and carcinogenesis in lung cells are discussed in connection with the cells that produce amylase.
Adenocarcinoma↗
The production of active human thyroid-stimulating hormone from alpha and beta mRNAs in Xenopus laevis oocytes.
Active human thyroid-stimulating hormone (hTSH) was produced by Xenopus laevis oocytes following injection of an mRNA mixture of hTSH beta and alpha subunits synthesized by T3 RNA polymerase. Some of the hTSH molecules were secreted into the medium, while others remained in the cells. The active molecules consisted of alpha and beta subunits and were in highly glycosylated form. The Xenopus laevis oocyte-produced hTSH stimulated the rat thyroid cell line FRTL-5 to produce and secrete the cyclic AMP as does authentic hTSH.
Animals↗
Gas chromatographic determination for forensic purposes of petroleum fuel inhaled just before fatal burning.
The determination of petroleum fuel in the blood of burned bodies was carried out by three different gas chromatographic procedures. Seven components of gasoline (isopentane, n-pentane, 2-methylpentane, benzene, 2-methylhexane, 3-methylhexane and toluene) and five of kerosene (xylene, C9H20, mesitylene, pseudocumene and C11H24) were chosen as indicators with a coefficient of variation of 5-24%. The methods were applied to four autopsy cases with a relatively low carboxyhaemoglobin (HbCO) content. When gasoline exposure had occurred, the blood concentrations determined were almost identical whatever the components selected. Great variations in the components determined were found after kerosene exposure, and hydrocarbons greater than or equal to C14 were hardly inhaled by the victims. A higher content of fuel in the left than in the right ventricular blood observed in the autopsy cases suggests fuel inhalation just before death. The same phenomenon was also observed in the content of blood HbCO. Determinations of petroleum fuel and HbCO in both the right and left ventricular blood would be useful for the forensic diagnosis on burned bodies with a low HbCO content.
Aged↗
Swallowing-induced arrhythmia.
Deglutition tachyarrhythmia, first reported in 1926 by Sakai and Mori, is a rare clinical entity. Vagal reflex has been supposed to be responsible for the induction of the arrhythmia. We have recently studied a patient with swallowing-induced premature atrial contractions. Atropine sulfate and catecholamine infusion increased the frequency of the arrhythmia, while beta blockade suppressed it. It is suggested that the mechanism of the swallowing-induced arrhythmia might be related to a sympathetic reflex. As far as we know, there have been few reports in which the sympathetic reflex was proposed to be responsible for the arrhythmia.
Adult↗
Pancreatic secretory trypsin inhibitor in cancer.
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Humans↗
Cholesterol-lowering effect of N-(alpha-methylbenzyl)linoleamide (melinamide) in cholesterol-fed diabetic rats.
Cholesterol loading of diabetic rats is known to induce marked hyperlipoproteinaemia, and we have reported that enhancement of the activity of intestinal acyl-CoA:cholesterol acyltransferase (ACAT), one of the key enzymes involved in cholesterol absorption, might play an important role in the development of hypercholesterolaemia in these animals. In the present study, we have shown that treatment with N-(alpha-methylbenzyl)linoleamide (melinamide), a new hypocholesterolaemic drug, caused a substantial decrease of the enhanced intestinal ACAT activity in diabetic rats, but did not affect intestinal cholesterol esterase activity. Furthermore, marked improvement of hypercholesterolaemia in cholesterol-fed diabetic rats occurred concomitantly with the drug treatment. These results suggest that intestinal ACAT activity is closely related to the serum cholesterol level in diabetic rats, and show that melinamide lowers intestinal ACAT activity.
Animals↗
Activation of brain function by S-135, a benzodiazepine receptor inverse agonist.
1. S-135, 2-(5-methylthien-3-yl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinoline-3- one, bind binds to benzodiazepine receptors with a high affinity and shows pharmacological actions opposite to those of conventional benzodiazepine drugs. 2. S-135 induced no convulsion in mice by itself, but selectively potentiated the effect of subconvulsive dose of pentylenetetrazole. 3. S-135 potentiated rat crossed extensor reflex and Ro 15-1788 completely antagonized this potentiation. 4. S-135 antagonized pentobarbital-induced anesthesia, tetrabenazine-induced ptosis and reserpine-induced hypoactivity and shortened immobilization time in the despair test in mice, indicating that this compound possesses antidepressive properties. 5. S-135 antagonized amnesia in mice and rats in passive avoidance tasks. 6. Glucose utilization in brain areas relating to memory and arousal functions was enhanced following S-135 treatment. 7. These results indicate that S-135 can be a useful drug for activating depressed brain function.
Animals↗
Overlapping two genes in human DNA: a salivary amylase gene overlaps with a gamma-actin pseudogene that carries an integrated human endogenous retroviral DNA.
The human salivary amylase gene (amy1), consisting of eleven exons, is expressed in the salivary gland and in some amylase-producing tumors. Its uppermost exon and the following intron, along with the 5'-flanking region of this gene, are shown to be superimposed with a gamma-actin pseudogene sequence, a portion of which is transcribed into salivary amylase mRNA and another portion of which serves as a promoter for the amy1 gene. In the further upstream region, the gamma-actin pseudogene sequence is interrupted by a human endogenous retroviral nucleotide sequence.
Actins↗
Nucleotide sequence and chromosomal mapping of the human c-yes-2 gene.
We molecularly characterized the second gene, c-yes-2, of two copies of yes-related genes which we previously found to contain in the human genome. First, nucleotide sequence analysis revealed that the c-yes-2 gene is a pseudogene of the c-yes-1 gene. Second, by using two independent methods, hybridization of both DNAs from sorted chromosomes and metaphase spreads with c-yes-2 DNA, we assigned the c-yes-2 gene to chromosome 22q11.2. This chromosomal localization is consistent with that given in our previous report. The failure of proper mapping in our experiment might have been caused by instability of hybrid cell clones.
Base Sequence↗
Regional localization of the LCA oncogene to human chromosome region 2q14----q21.
A novel human oncogene, LCA, was assigned to region 2q14----q21 by in situ molecular hybridization. The present regional mapping substantiates the previous assignment that was performed by Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids.
Carcinoma, Hepatocellular↗
Cloning and nucleotide sequence of the brnQ gene, the structural gene for a membrane-associated component of the LIV-II transport system for branched-chain amino acids in Salmonella typhimurium.
The genetically defined gleR-brnQ region responsible for the branched-chain amino acid transport in Salmonella typhimurium was mapped in the 3.3-kilobase SalI-PstI segment of plasmid pOH56 by complementation analysis. By subcloning and genetic recombination analysis, the gleR and brnQ3 mutational sites were localized within the 0.85-kilobase SalI-EcoRV segment, and brnQ4 within the 0.8-kilobase EcoRV-HindIII segment. The nucleotide sequence of the brnQ gene and its flanking regions was determined. The brnQ gene is encoded by the sequence starting 24 base pairs upstream from the EcoRV site. Transcription of the brnQ gene starts at three sites separated by 171, 173 and 174 nucleotides, respectively, from the initiation codon. The promoter sequences can be seen in the immediate upstream region of the transcription initiation sites. There is a long silent region between the transcription initiation sites and a potential Shine-Dalgarno nucleotide sequence. The coding sequence of the brnQ gene, which is 1317 base pairs long, specifies a very hydrophobic protein of 439 amino acid residues.
Amino Acid Sequence↗
[The method for stress assessment of workers (Part 1). Stress score by self-rating method].
In order to evaluate the grade of reaction to stressors, especially those on occupational life of workers, the following stress survey was conducted. Eighteen new items of occupational life stressors were proposed in addition to the social readjustment rating scale (S.R.R.S.) prepared by Holmes. Furthermore, two items, "own stress tolerance" and "own stress at present," were designed in addition to the foregoing 65 stressors. Marriage is given a score of 50 in reference-standard of stress strength. These 67 items of 1,630 workers were evaluated by the self-rating method ranging in score from 0 to 100. The results can be summarized as follows: 1. As for each item, we examined the average value of the total sample and analyzed them by sex, age, kind of occupation, position, and length of service. We named these stress scores and analyzed the ranking of 65 stress scores. The item with the highest score was "death of spouse (score 82.7)" and the item with the lowest one was "income increase (score 24.7)." The scores of 27 out of 65 items were higher than 50. Next, 65 items were classified into 4 groups, that is, individual life, family life, occupational life, and social life. 2. Scores of the newly designed items to examine the ability to adjust oneself to place of work were "own stress tolerance (score: 73.7)" and "own stress at present (48.8)," respectively. 3. We examined the difference by sex, age, kind of occupation, position, and length of service based on the average value of stress score by t-test and the following results were obtained. 1) Scores of those in the thirties were higher than those in the twenties. 2) Section heads and leaders were under greater stress than division heads. In the same way, the differences among the four groups were analyzed and the following results were observed. 1) The only significant difference was seen in the occupational life stress group. 2) In the occupational life stress group, the scores of those in the thirties, forties, and fifties were higher than those in the twenties. Section heads and leaders were under greater stress than the staff. Workers in service for more than 21 years had higher scores than workers in service for less than 10 years. 4. The practical uses of our stress survey are also discussed in this report.
Adult↗
Increased activity of intestinal acyl-CoA: cholesterol acyltransferase in rats with streptozocin-induced diabetes and restoration by insulin supplementation.
We examined the activities of intestinal acyl-CoA:cholesterol acyltransferase (ACAT) and cholesterol esterase, enzymes regulating cholesterol absorption, in rats with streptozocin-induced diabetes (STZ-D) to clarify the effect of diabetes on cholesterol absorption. Three weeks after the induction of diabetes, plasma cholesterol levels were slightly but significantly increased in diabetic rats compared with control animals, whereas a far more remarkable increase in plasma cholesterol was observed in diabetic rats when fed an atherogenic diet containing 1% cholesterol, 0.5% cholic acid, and 5% lard. Microsomal ACAT activity in intestinal mucosa was three times higher in diabetic than in control rats. However, no significant difference in the enzyme activity could be detected between diabetic animals fed control chow and those fed the atherogenic diet. Furthermore, insulin supplementation given to diabetic rats caused a reduction of enzyme activity to the levels found in control animals. In contrast, cholesterol esterase activity in rat intestinal mucosa was unaffected by either the induction of diabetes or the atherogenic diet feeding. In conclusion, we disclosed that apparent ACAT activity in intestinal mucosa is elevated in STZ-D rats. Therefore, we postulate that enhancement of CoA-dependent cholesterol esterification in the intestine might be one of the major factors responsible for hypercholesterolemia in diabetes.
Animals↗
Cryo-electron microscopy of hepatitis B virus core particles produced by transformed yeast: comparison with negative staining and ultrathin sectioning.
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Freezing↗
The lca as an onco-fetal gene: its expression in human fetal liver.
The lca-transforming DNA was isolated from human hepatocellular carcinomas. This gene has no homology with known transforming DNA from human sources, and its role in neoplastic tissue formation has been left unanswered. In this communication, we report that RNAs prepared from human fetal livers hybridize to the lca DNA probe. The RNA is 1.8 kilobase in size and appears in the fetal liver only for a limited period during its development, viz. 19 weeks through 24 weeks of gestation. No other tissues carry detectable levels of the lca messenger RNA. Fetal hepatocytes at 5 weeks of gestation showed no transcripts of lca, but upon culturing for 2 more weeks in vitro, the cells became producers of the lca messenger RNA. These results suggest that the lca plays some role in the proliferative stage of the liver.
Carcinoma, Hepatocellular↗
Co-amplification of integrated hepatitis B virus DNA and transforming gene hst-1 in a hepatocellular carcinoma.
Transforming activity was detected in a hepatocellular carcinoma (HCC) carrying four integrated hepatitis B virus (HBV) DNA. This transforming gene was identified as hst-1, that lies in chromosome 11, band q13.3. One of the integrated HBV DNAs was found to lie close to the hst-1, and the hst-1 and the integrated HBV DNA were found to be co-amplified. The region of the amplification was limited. A model has been proposed that correlates the viral integration, amplification and activation of an oncogene.
Carcinoma, Hepatocellular↗