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Biomedical subjects

T Ide

Publications and source records attributed to T Ide.

At least 307 records · Page 17Linked to original sources

Defect in prereplicative phase of G0-specific ts mutant, tsJT60.

A temperature-sensitive mutant, tsJT60, grew exponentially at both 34 degrees and 39.5 degrees C, but when stimulated from the resting state it entered S phase at 34 degrees but not at 39.5 degrees C. The mutated function appeared to be a prerequisite throughout from 0 to 9 h following the stimulation, in order that G0-arrested cells would enter S phase. When the arrested cells were stimulated with serum, the amount of and synthesis of protein increased at 34 degrees but not at 39.5 degrees C. The amount of polysome fraction was much smaller in stimulated and unstimulated cells at 39.5 degrees C than in those stimulated at 34 degrees C. Of the events reported to increase shortly after the stimulation, uridine transport increased at both temperatures. Mutation in tsJT60 cells may be concerned with the function prerequisite to induce protein synthesis following serum stimulation, resulting in the blocking of cell cycle progression toward S phase at 39.5 degrees C.

Animals↗

Isolation of ts mutant cells which arrest in G1/G0 phase at the non-permissive temperature in the presence of appropriate growth factors from a Fischer rat cell line, 3Y1.

Two types of cell-cycle-ts mutants were isolated from Fischer rat cell line, 3Y1, and characterized. Clones in one complementation group, tsJT51 and tsJT341, grew at 34 degrees C in the presence of 10% fetal bovine serum (FBS). When the cells growing at 34 degrees C were transferred to 39.5 degrees C, they were arrested alive in G1/G0 phase in the presence of both FBS and epidermal growth factor (EGF), but died in the presence of one of these growth factors. The cells in the other complementation group, tsJT59, tsJT308, tsJT314 and tsJT349, grew at 34 degrees C in the presence of 10% FBS. When the cells growing at 34 degrees C were transferred to 39.5 degrees C, they were arrested alive in G1/G0 phase in the simultaneous presence of FBS, EGF and insulin, but died quickly if one of these growth factors was lacking. Growth-arrested cells at 39.5 degrees C were viable at least one or two weeks and had a potency to resume growth following the shift-down of temperature. Those are assumed to be ts mutant cells which enter and stay in G1/G0 phase from the cell cycle at the non-permissive temperature only in the presence of appropriate growth factors.

Animals↗

Events blocked in prereplicative phase in senescent human diploid cells, TIG-1, following serum stimulation.

When senescent human diploid cells, TIG-1, were stimulated with serum at the end of their proliferative life span, such biochemical events as uptakes of 2-deoxyglucose and uridine, and expression of c-myc, were enhanced. However, RNA synthesis, polyamine accumulation, thymidine uptake and DNA synthesis were not enhanced at all. Protein synthesis increased only moderately as compared with that observed in younger cells. These results indicated that the events in prereplicative phase known to be independent on protein synthesis are induced in senescent cells after the stimulation with serum, whereas those required protein synthesis failed to increase to the same extent as seen in young cells.

Blood↗

Oxidation and esterification of geometrical and positional isomers of octadecenoic acids in perfused rat liver.

The metabolic fate of the geometrical isomers of 6-, 9-, and 11-octadecenoic acids was studied in isolated perfused rat livers. Although the ketogenicities of these monounsaturated fatty acids generally decreased as the double bond was moved away from the carboxyl group, a dependence on the geometrical difference was found only for 9-octadecenoate, the rate being significantly lower in the cis-isomer. Very low density lipoprotein lipid secretion was distinctly and specifically decreased when trans-9-octadecenoic acid was perfused, but no such difference was observed with other isomers. Various trans-isomers were actively incorporated into the hepatic lipids and secreted apparently at the expense of preexisting endogenous cis-octadecenoate. The trans-isomers were all incorporated exclusively at the 1-position of hepatic phosphatidylcholine. Concentrations of the cis-octadecenoate in the glycerolipids secreted and remaining in the liver were characteristically modified depending on the location of the ethylenic bond of the cis-octadecenoate. Thus, both the location of the double bond in the acyl-chain and the geometrical configuration specifically influence the rate of oxidation and esterification of octadecenoic acid in perfused rat liver.

Animals↗

Hypocholesterolemic effect of gamma-linolenic acid as evening primrose oil in rats.

The hypocholesterolemic effect of polyunsaturated fatty acids was compared in male rats given high-cholesterol diets containing either evening primrose oil (EPO, linoleic plus gamma-linolenic), safflower oil (SFO, linoleic) or olive oil (OLO, low linoleic) at the 10% level. EPO with a phytosterol content of 1.47% was found to be more hypocholesterolemic than SFO (phytosterols 0.34%), and rats given EPO excreted more neutral (cholesterol and its metabolites) but not acidic steroids during the first 2 weeks of the feeding. Even when the phytosterol content of EPO and SFO was adjusted to be the same (0.67%), EPO was still more hypocholesterolemic than SFO but to a lesser extent, although fecal neutral steroid excretion was comparable in these two dietary fat regimens. The results indicate a significant hypocholesterolemic efficacy of gamma-linolenic acid.

Adipose Tissue↗

Dietary manipulation of the disappearance of trans-octadecenoates in rat tissues.

The effects of dietary manipulations on the fate of trans-octadecenoates deposited in the tissues of rats were examined. Male rats were fed on a 15% fat diet containing trans-octadecenoic acids (46.6% of total fatty acids) for 35 days followed by various diets free of trans-fatty acids. After removal of trans-fatty acids from the diet, there were phased disappearances of trans-octadecenoates from the circulation; a rapid and broad reduction in one day and a slow and gradual reduction thereafter. The rate of the initial reduction in serum trans-octadecenoates was highest on a high fat (20%) diet in relation to low fat (1 or 5%) diets. However, the disappearance rate at the later stage was apparently the same among the various groups and trans-fatty acid contents in the serum declined to about 10% of the initial value in 2 weeks and thereafter. After 35 days, the concentration of trans-octadecenoates remaining in the adipose tissue was markedly lower in rats fed on a high protein (40%) diet. The effects of dietary fat type and cholesterol on the fate of serum trans-octadecenoates were virtually the same, but livers from rats fed on the cholesterol-free safflower oil diet contained more trans-fatty acids than those from rats fed on the corresponding olive oil diet. Thus, the amounts of trans-fatty acids stored in the tissues cannot be merely predicted from serum levels. It seems that both dietary fat and protein affect the metabolic rate of trans-octadecenoates in rats.

Adipose Tissue↗

Activities of liver mixed function oxidase system in rats fed trans fat.

Rats were fed diets containing either camellia oil or partially hydrogenated corn oil as a source of cis or trans octadecenoate, respectively, in the presence of adequate linoleic acid. After 35 days of feeding the diets, activities of several hepatic drug metabolizing enzymes as well as the content of hepatic microsomal cytochrome P-450 were determined. Geometrical difference in the dietary fat did not affect the amount of microsomal protein nor the content of cytochrome P-450. Also, activities of NADPH cytochrome C reductase, aminopyrine N-demethylase and biphenyl hydroxylase were approximately the same between two groups of rats. Aniline hydroxylase was slightly elevated in the rats fed trans fat. It was concluded that the difference in the geometry of dietary fatty acids had little effect in modulating the hepatic mixed function oxidase system.

Animals↗

Cytochalasin D inhibits the progression from the Go to S phase at the mid-prereplicative stage in GC-7 cells stimulated with serum.

When the growth of serum-arrested GC-7 cells, a clone from African green monkey kidney, was induced by the addition of 10% calf serum, they began to enter S phase after 15-16 h. When stimulated cells were cultured in the presence of 0.6 micrograms/ml of cytochalasin D, the entrance into S phase was inhibited. Treatment of cells with cytochalasin D during the period earlier than 8 h or later than 11 h after the serum stimulation showed no or little inhibitory effect on the entrance of cells into S phase. Inhibition of the entrance into S phase was observed only when stimulated cells were treated with cytochalasin D during the periods including 9-10 h after stimulation. A rapid increase in protein synthesis occurred 9-12 h after the serum stimulation and was inhibited in the presence of cytochalasin D. These and other results suggested that in the course of the prereplicative process from Go through S phase only the stage around 9-10 h after the start of the cell cycle was sensitive to cytochalasin D and that the block of the cycle was correlated with the inhibition of protein synthesis at this stage.

Animals↗

Responses of plasma cyclic AMP, serum immunoreactive insulin, C-peptide immunoreactivity and blood sugar levels to glucagon in patients with liver diseases.

Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI), serum c-peptide immunoreactivity (CPR) and blood sugar (BS) were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight) in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was the response suppressed. The peak response of BS was found significantly later in liver cirrhosis patients than in normal controls. IRI and CPR responses to glucagon were lower in acute hepatitis patients than in normal controls and liver cirrhosis patients. IRI levels and their sum were also lower in acute hepatitis patients, although CPR levels were not significantly different. Thus, the ratio of the sum of CPR from 0 to 60 min to that of IRI was significantly higher in acute hepatitis, indicating impaired pancreatic secretion of insulin to glucagon stimulation as well as increased uptake of insulin by the liver in acute hepatitis.

Adult↗

Sister chromatid exchanges induced by cancer chemotherapeutic agents in vitro and in vivo: consideration of the hazard of drugs as possible mutagens and carcinogens causing second malignancies.

Sister chromatid exchanges (SCEs) induced by chemotherapeutic agents currently used for treating various malignancies were studied in vitro and in vivo. Whether the agents are potentially mutagenic/carcinogenic is discussed on the basis of recent clinicopathological observations on secondary malignancy. In the in vitro studies, the induction of SCEs by various anticancer agents, including three anticancer antibiotics, six alkylating agents, and three antimetabolites, was examined and compared with induction by two known carcinogens, 4-nitroquinoline 1-oxide and N-ethyl-N'-nitronitrosoguanidine. In the in vivo studies, SCE frequencies were examined in the lymphocytes from 74 blood samples drawn from 40 patients with cancer or leukemia at various times after the agents had been administered alone or in combination. The SCE frequencies induced by the anticancer antibiotics were variable because the drugs had different modes of action: generally, the drugs have strong cytotoxicity that must affect the viability of cells. On the other hand, alkylating agents induced significantly high levels of SCE frequencies in vitro; the modes of SCE inducibility were similar to those of the two carcinogens. The fact that cells were viable up to relatively high molar concentrations of the agents and that high SCE levels persisted for a long period suggests that the lesions induced by alkylating agents are long-lived and may therefore be more frequently involved in mutagenesis. This finding may be compatible with the clinical observation that the vast majority of patients with second malignancy are found in the group treated with alkylating agents. Antimetabolites generally showed only a weak SCE induction or none in vivo and in vitro.

Alkylating Agents↗

Loss of responsiveness in senescent human TIG-1 cells to the DNA synthesis-inducing effect of various growth factors.

Responses of human diploid cells, TIG-1, were examined with respect to their ability to initiate DNA synthesis under the influence of various growth factors and their combinations. The following agents stimulated DNA synthesis in quiescent TIG-1 cells at 37-49 PDL (population doubling level) (66-79% of lifespan completed): fetal bovine serum; tumor-derived DNA synthesis factors such as those from rat rhodamine fibrosarcoma, human adenoma and from the conditioned medium of cultured human pituitary cells; human and mouse epidermal growth factors; tumor promotors such as 12-O-tetradecanoylphorbol 13-acetate and teleocidin; microtubule-disrupting agents as colchicine, vinblastine, podophyllotoxin and TN-16; melittin; and dexamethasone. Cells at 58-60 PDL (94-97% of lifespan completed) were stimulated to synthesize DNA by fetal bovine serum, tumor-derived DNA synthesis factors and epidermal growth factors, but not by other agents. Finally, in senescent cells at 62 PDL (100% of lifespan completed), any of these growth factors and of their combinations failed to induce DNA synthesis at all. These senescent cells, however, still retained the ability to initiate DNA synthesis following infection with SV40 as reported previously [Exp. Cell Res., 143 (1983) 343-349].

Carcinogens↗

Oxidation and esterification of cis- and trans-isomers of octadecenoic and octadecadienoic acids in isolated rat liver.

The metabolism of 9-octadecenoic and 9,12-octadecadienoic acids with different geometrical configurations was compared in isolated perfused rat liver. More ketone bodies were produced when the trans-isomers were infused. In contrast, only the cis-isomer augmented the triacylglycerol secretion almost entirely as very-low-density lipoprotein (VLDL). Although these responses were independent of the difference in the degree of unsaturation in both the cis- and trans-isomers, the trans-monoenic acid compared to the trans-dienic acid was incorporated more readily into perfusate and hepatic lipids. Quantitative information was obtained with radioactive tracer experiments. The hepatic uptakes of 9-[10-14C]octadecenoic acids were comparable in the cis- and trans-isomers. The trans-octadecenoic acid compared to the cis counterpart was oxidized more readily and incorporated more into liver phospholipid but less into perfusate and liver triacylglycerol. These reciprocal responses counterbalanced each other. The lower rates of triacylglycerol synthesis and secretion in the liver perfused with the trans-octadecenoic acid was confirmed using [2- 3H]glycerol as a tracer. The marked difference in the channelling of cis- and trans-fatty acids in the pathways of oxidation and esterification seems to modify the VLDL secretion in perfused rat liver. Present observations indicate a considerable difference in the fate of unsaturated fatty acids with different configurations. trans-Fatty acids are expected to be an efficient energy source in animal tissues and may not be hyperlipidemic.

Animals↗

Purification and characterization of alpha-glucosidases produced by Saccharomyces in response to three distinct maltose genes.

alpha-Glucosidases or maltases (EC 3.2.1.20) were purified to electrophoretic homogeneity from a respective strain of Saccharomyces cerevisiae which carries a single MAL gene, either MAL alpha, MAL beta, or MAL gamma, using gluconate-Sepharose affinity chromatography and isoelectrofocusing. Of these maltases, two types of maltase were obtained from the MAL gamma strain, the pI values of which were 5.6 and 5.9. From the MAL alpha and MAL beta strain was obtained only one type of maltase with the pI at 5.6 which was identical to one of the maltases from the MAL gamma strain. These four maltases possessed the same properties, except for pI. They were monomers with molecular weights of between 66 000 and 67 000. With regard to the substrate specificity, they hydrolyzed maltose and sucrose exclusively but not alpha-methylglucoside nor maltooligosaccharide. They did not differ in immunological properties.

Genes↗