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51.
Epidermal growth factor (EGF) produces inositol phosphates and increases cytoplasmic free calcium in cultured porcine thyroid cells 总被引:2,自引:0,他引:2
N Takasu M Takasu T Yamada Y Shimizu 《Biochemical and biophysical research communications》1988,151(1):530-534
The initial signal for thyroid cell proliferation is unknown. This is the first report to show that epidermal growth factor (EGF) produces inositol phosphates and increases cytoplasmic free calcium ([Ca2+]i) in the thyroid gland. In cultured porcine thyroid cells, 10 nM EGF produces a breakdown of phosphatidylinositol and stimulates inositol phosphate production. Ten nM EGF increases [Ca2+]i, measured using fura-2, a fluorescent Ca2+ indicator; the EGF-induced [Ca2+]i response occurs immediately, reaches a maximum within several seconds, and then slowly declines. EGF stimulates production of inositol phosphates, which seem to increase [Ca2+]i. Inositol phosphate production and an increase in [Ca2+]i after EGF-stimulation may function as an initial signal for thyroid cell proliferation. 相似文献
52.
Gelatinases of metastatic cell lines of murine colonic carcinoma as detected by substrate-gel electrophoresis 总被引:6,自引:0,他引:6
S Yamagata Y Ito R Tanaka S Shimizu 《Biochemical and biophysical research communications》1988,151(1):158-162
Gelatinases were detected in the conditioned medium of murine colonic carcinoma cells by SDS-polyacrylamide gel electrophoresis using gels copolymerized with gelatin. Several gelatinase activities differing in molecular weight were detected but the major activities migrated with molecular weights of 60,000 and 95,000. The enzymes did not hydrolyze bovine serum albumin or casein, and required calcium for activity. All of the gelatinase activities were inhibited by EDTA, 1,10-phenanthroline and dithiothreitol but not by N-ethylmaleimide and phenylmethylsulfonyl fluoride. The 95,000 dalton gelatinase was separated from the 60,000 dalton gelatinase by affinity chromatography on Ricinus communis agglutinin-agarose, and the former activity was markedly increased in highly metastatic cell lines as compared with its activity in poorly metastatic cell lines. 相似文献
53.
Human fibrinogen (concentration 8.4 mg/mL) was ligated (cross-linked) with factor XIIIa and dithiothreitol (DTT) at pH 8.5, ionic strength 0.45. With 7.5 μg/mL of factor XIIIa alone, there was almost no γ-γ ligation, but with 2 mM DTT added, oligomers appeared, and γ-γ and Aα-Aα ligation was nearly complete after 3 days. At 38 μg/mL of factor XIIIa, some γ-γ and Aα-Aα ligation occurred even without DTT. For fibrinogen concentrations of 4.0 and 8.4 mg/mL, 38 μ/mL factor XIIIa, 2.0 mM DTT, clot-like gels formed and the shear modulus of elasticity increased slowly over several days to a constant value. The final modulus was similar in magnitude to those of ligated clots of α-fibrin (clotted by thrombin) and α-fibrin (clotted by batroxobin) under the same conditions. However, the opacity was somewhat higher; whereas in fine fibrin clots there is minimal lateral association of the protofibrils, in fibrinogen gels at the same pH and ionic strength the protofibrils (which are presumably single chains of fibrinogen monomers joined end to end at their D domains) are evidently associated in bundles (although not to the degree seen in coarse fibrin clots). Creep and creep recovery measurements showed almost perfect elastic behavior, with essentially no creep under stress and complete recovery after removal of stress. The modulus was scarcely affected by introduction of lithium bromide by diffusion to a concentration of 0.6M, which in unligated fibrin clots causes substantial softening. Whereas in fine fibrin clots (both αβ-fibrin and α-fibrin) factor XIIIa causes only γ-γ ligation, addition of 2 mM DTT produced some α-α ligation in these also. 相似文献
54.
Calcium alginate gel stabilized with a polyelectrolyte complex (PEC) consisting of potassium poly(vinyl alcohol) sulfate (KPVS) and trimethylammonium glycol chitosan iodide (TGCI) was used for the immobilization of beta-amylase. The immobilization was made by gelling aqueous droplets of enzyme solution including both sodium alginate and KPVS in a CaCl(2) solution containing TGCI. The activity of the enzyme entrapped into the stabilized gel beads was evaluated by studying the batch reaction kinetics of enzyme-catalyzed hydrolysis of maltotetraose. Repeated kinetic measurements, totaling 18, were carried out at fixed time intervals. After each measurement the beads were stirred for 1 day in a freshly prepared 10 mM NaCl solution at 3 degrees C. It was found that the immobilized system remained stable without leading to a serious loss of the activity or to a large leakage of the enzyme from the support. This was explained as being due to a PEC-crosslinked contracted network structure of the stabilized gel matrix. 相似文献
55.
The effects of the addition and removal of glycerol on the metabolic activities of human platelets were studied. Platelet concentrates (PC) with 20 ml plasma were stored with 3-7% (v/w) glycerol in 150-ml polyvinylchloride plastic bags for 2 days at 22 degrees C with constant agitation. Incubation of glycerol with platelets produced a dose-dependent inhibition of oxygen consumption. The inhibitions of glucose utilization and lactate production had reached the plateau level at 3% glycerol. The rate of adenosine triphosphate (ATP) generation of control platelets was 9.8 nmol/min/10(9) platelets, in which over 90% ATP generation was derived from oxidative phosphorylation. There was a dose-dependent decrease (up to 20%) by glycerol in the rate of platelet ATP generation. Glycerol inhibited glycolysis more than oxidative phosphorylation. However, the inhibition potency diminished with increasing concentrations of glycerol. The energy metabolism of platelets after removal of 5% glycerol was examined. Deglycerolized platelets after 1 hr incubation facilitated energy metabolism more strongly than that of 24 hr incubation. The platelet aggregation response to collagen was not impaired by a cycle of the addition and removal of glycerol. The results indicate that glycerol lowered the rate of ATP generation of platelets stored at 22 degrees C. However, the removal of glycerol reversed the decreased energy metabolism. 相似文献
56.
The present study was designed to determine whether a bag made from ethylene-vinyl acetate copolymer (EV) with superior flexibility at subzero temperature is suitable for a storage container of single-donor apheresis platelets. Apheresis platelets were stored with 100 ml plasma in 1-liter bags made of EV or standard polyvinyl chloride (PVC) plastic at 22 degrees C with constant agitation. The oxygen permeability of the 1-liter EV bag averaged 1447 nmol/min/atm, which was about 1.5 times higher than that of PVC bags. The partial oxygen tension (PO2) of platelet concentrates (PC) has linearly decreased to 16 mm Hg with increasing platelet counts. The level of the partial carbon dioxide was always higher in EV bags than in PVC bags. Oxygen consumption rates of platelets stored in EV and PVC bags with a sufficient oxygen supply averaged 1.25 and 1.20 nmol/min/10(9) platelets, respectively. The rates of glucose consumption and lactate production were not changed in two bags. Ninety percent of the total ATP production of about 8 nmol/min/10(9) platelets were generated through the aerobic metabolism. The platelet counts in the 1-liter EV and PVC bags, at which PO2 is 16 mm Hg, were 2.2 and 1.5 x 10(11) platelets, respectively. The study indicates that apheresis platelets stored in EV bags at 22 degrees C have no different metabolic changes when compared with those of PVC bags. In addition, the number of platelets maintaining the aerobic metabolism is 1.5 times higher than that of PVC bags. 相似文献
57.
Expression of actin mRNAs in denervated chicken skeletal muscle 总被引:3,自引:0,他引:3
The expression of actin genes in chicken pectoralis muscle denervated 1 week after hatching was examined 1-8 weeks after the operation by RNA blot hybridization using a generic actin cDNA probe and DNA probes specific for alpha-skeletal and alpha-cardiac actin genes. Total and alpha-skeletal actin mRNAs/microgram total RNA decreased to about half of the levels found in contralateral control muscle, while the expression of alpha-cardiac actin mRNA was up-regulated. Consequently, alpha-cardiac actin mRNA formed about 15% of the total actin mRNA as compared to less than 1% found in control muscle. The expression of actin genes in the denervated muscle was similar to that in the late embryonic muscle. These results suggest that innervation is required to show the expression pattern of striated muscle actin genes found in mature muscle. 相似文献
58.
Summary Changes in glycoconjugates of the zona pellucida induced by maturation, ovulation and fertilization of mouse oocytes have been studied by means of light microscopic methods of cytochemistry. These methods consisted of periodic acid-Schiff (PAS), Alcian Blue pH 1.0 and pH 2.5, and peroxidase-labelled lectin diaminobenzidine (PO—LT—DAB) procedures in combination with the digestion technique with neuraminidase. According to the results obtained, glycoconjugates of the zona pellucida of fertilized eggs contained a smaller amount of sulphate groupings than that in ovarian oocytes, whereas their reactions for sialic acid and fucose residues were significantly stronger in intensity in the former, as compared with those in the latter. The cytophysiological significance of such cytochemical changes in glycoconjugates of the zona pellucida has been discussed with special reference to its functional alterations following maturation, ovulation and fertilization. 相似文献
59.
M Nagayoshi Y Hirata M Tamaru S Sugimoto J Shimizu K Hirabayashi T Matsutani 《Nihon seirigaku zasshi. Journal of the Physiological Society of Japan》1986,48(1):14-25
The regional levels of several cell marker proteins in the brain and the ability of operant discrimination learning on a multiple fixed ratio (FR), fixed interval (FI) schedule were determined in rats with microencephaly induced by prenatal treatment with methylazoxymethanol (MAM), an antimitotic agent, on the 11 th to 13 th days (Group A) or on the 15 th day (Group B) of gestation. The cell marker proteins were determined with a sensitive enzyme immunoassay. Neuron-specific enolase (NSE; gamma gamma-enolase) had a significantly lowered level in the neocortex anterior in Group A. Non-neuronal enolase (NNE; alpha alpha-enolase) was significantly reduced in the superior colliculus, lateral geniculate body and optic nerve, but increased 1.5 fold in the retina in Group A. S-100b protein, a marker of astroglial cells, showed no significant change. As for the learning performance, the Group B animals showed an elevated behavioral activity and made evident discrimination between the FI and FR schedule. But Group A animals had prolonged FR components requiring responses to light on, and their spontaneous activity counts recorded by Automex showed an inhibition of behavior in light environments. These findings suggest a causative role of some developmental abnormality in the central visual system, indicated by the aberrant cell marker levels, in the disturbed learning ability of the Group A animals. 相似文献
60.
N-Carbamoylsarcosine amidohydrolase, a novel enzyme involved in the microbial degradation of creatinine in Pseudomonas putida 77, was purified 27-fold to homogeneity with a 63% overall recovery through simple purification procedures including successive ammonium sulfate fractionation, DEAE-cellulose chromatography, and crystallization. The relative molecular mass of the native enzyme estimated by the ultracentrifugal equilibrium method is 102,000 +/- 5000, and the subunit Mr is 27,000. The Km and Vm values for N-carbamoylsarcosine are 3.2 mM and 1.75 units/mg protein, respectively. Ammonia, carbon dioxide, and sarcosine were formed stoichiometrically from N-carbamoylsarcosine through the action of the purified enzyme preparation. N-Carbamoyl amino acids with a methyl group or hydrogen atom on the amino-N atom and possessing glycine, D-alanine, or one of their derivatives as an amino acid moiety served well as substrates for N-carbamoylsarcosine amidohydrolase. N-Carbamoylsarcosine, N-methyl-N-carbamoyl-D-alanine, N-carbamoylglycine, and N-carbamoyl-D-alanine were hydrolyzed at relative rates of 100, 12.8, 9.8, and 7.3, respectively, by the enzyme. N-Carbamoyl derivatives of D-tryptophan, D-phenylalanine, and those of some other amino acids including D-phenylglycine and p-hydroxy-D-phenylglycine were also hydrolyzed by the enzyme. For the L-isomers of all N-carbamoyl amino acids tested there was no production of ammonia, carbon dioxide, or the corresponding amino acids due to the action of the enzyme. Cupric, mercuric, and silver ions inhibited the enzyme strongly, and some thiol reagents were also found to be inhibitory. 相似文献