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Biomedical subjects

G Weber

Publications and source records attributed to G Weber.

At least 559 records · Page 31Linked to original sources
PubMed1982

Rat hepatomas: chemotherapy with lycurim and pyrazofurin.

Hepatoma 8999 was sensitive to Lycurim [1,4-di-(methylsulfonyloxy-ethylamino)-1,4-dideoxy-ms-erythritol] with a mean lethal dose (LD50) of 8.1 X 10(-8) M for a 6-hour treatment in vitro. The drug dose lethal to 10% of the rats with Lycurim (10 mg/kg) injected ip 12 times into hepatoma 8999-bearing BUF rats at 10-day intervals provided a mean increase in life-span (ILS) of 156%. The more rapidly growing, less differentiated hepatoma 3924A was tenfold less sensitive to Lycurim in vitro, and three treatments in vivo (10 mg/kg given every 8 days) gave an ILS of only 18% in ACI/N rats. Because hepatoma 8999 had a high adenosine kinase activity, the effect of Pyrazofurin (PF; 3-beta-D-ribofuranosyl-4-hydroxypyrazole-5-carboxamide) was examined in vitro: The LD50 was 8.5 X 10(-8) M in a 6-hour exposure. In hepatoma 3924A, with a fifteenfold lower adenosine kinase, the LD50 was 22-fold higher. Three treatments with PF (4 mg/kg given every 2 days) in hepatoma 8999 caused an 18% ILS and no host toxicity, but in hepatoma 3924A no significant ILS was observed. Lycurim combined with PF (0.05 microM each) in hepatoma 8999 cells in vitro provided synergistic kill, but Lycurim and PF (0.3 and 1 microM, respectively) in hepatoma 3924A cells yielded summation. When 10 rats with hepatoma 8999 were treated 15 times with the optimal dose of Lycurim (7.5 mg/kg every 10 1/2 days), 1-year survivors numbered 7. Alternate doses of Lycurim (7.5 mg/kg) and PF (3 mg/kg) at 5-day intervals for 4 months to 10 rats gave an ILS of 152% with eight 1-year survivors and no host toxicity.

Adenosine Kinase↗
PubMed1982

Cytotoxic and cell kinetic effects of 3,4,5-trihydroxybenzohydroxamic acid (VF 122) in hepatoma 3924A cells.

3, 4, 5-Trihydroxybenzohydroxamic acid (VF 122), an inhibitor of ribonucleotide reductase, killed rat hepatoma 3924A cells in tissue cultured after 7 days of incubation. A concentration of 15 microM caused 50% inhibition of colony-forming ability (IC50). Under the same conditions, hydroxyurea, also an inhibitor of ribonucleotide reductase, had an IC50 of 52 microM. Treatment for 1 hr with VF 122 of exponentially growing culture resulted in a biphasic exponential dose-response curve (Do = 0.5 mM, IC50 = 0.75 mM, and Do = 1.8 mM, IP = 0.8 mM). In plateau-phase cells, a threshold exponential curve was obtained (Do = 2.2 mM, Dq = 1.7 mM, and IC50 = 2.7 mM). Exponentially growing hepatoma 3924A cells were more sensitive to VF 122 than were plateau-phase cultures. In contrast, hydroxyurea killed only exponentially growing 3924A hepatoma cells, exhibiting an exponential plateau dose-response curve without achieving achieving an IC50 value at concentrations from 1 to 200 mM. In synchronized cultures, VF 122 (1 MM for 1 hr) was toxic for cells in mid and late G1 phase, in early and mid S phase, and, to a lesser degree, in G2 phase, Hydroxyurea (10 mM for 1 hr) killed cells only in S phase. Proliferating and resting hepatoma 3924A cells recovered from sublethal and potentially lethal damage induced by VF 122.

Animals↗
PubMed1982

Decreased content of reduced and oxidized nicotinamide-adenine dinucleotide phosphate in rat hepatomas.

Tissue contents of NADPH and NADP+ were measured in freeze-clamped samples of normal rat liver and in four transplantable rat hepatomas covering a wide range of growth rates. Lowry cycling procedures were employed for analysis, using alkaline extracts for NADPH and acid extracts for NADP+. The mean NADPH content in 33 normal livers was 515 nmol/g wet weight, and mean NADP+ content was 311 nmol/g wet weight. In the four hepatomas, the amounts of both NADPH and NADP+ were low, and the extent of decrease correlated with tumor growth rate. In the slowly growing hepatoma 9618A, total NADP was slightly decreased (63% control) and more extensive decreases were observed in the medium growth rate tumors 47C and 8999 (38% and 19%, respectively, of control). In the rapidly growing hepatoma 3924A, total NADP was drastically decreased to 3% of the control liver value. Measurement of NADPH and NADP+ recovery from extracts of hepatoma 3924A showed that there were no inhibitors that might have blocked the activity of the assay enzymes. The NADPH/NADP+ ratio was close to the normal liver value in all four hepatomas. A 30-sec period of ischemia did not cause significant change in NADPH, but gave 33% decrease in liver NADP+. A 5-min period of ischemia decreased NADP+ to 50% of the zero-time value in liver, and to 71% in hepatoma 3924A, but was without effect on NADPH.

Animals↗
PubMed1981

[Preliminary data on the different endothelial reaction to Con A at different levels of the arterial tree in rabbits and in Cynomolgus monkeys].

Delayed appearance of atherosclerotic lesions in cerebral arteries has been observed not only in man but also in monkeys and rabbits submitted to atherogenic diets. Previous observations of ours had shown a Con A positive reaction ("glycocalyx" or "surface coat") at the luminal surface and in the plasmalemmal vesicles of aortic endothelial cells of rabbits an other laboratory animals. The "surface coat" is now reputed the site of lipoproteinlipase activity whose importance in atherogenesis has recently been stressed. In our present observations, the endothelial cells Con A reactivity after Bernhard and Avrameas which was not previously studied in the cerebral arteries of rabbits and monkeys has resulted always lacking in this arterial district. Those observations may help explaining delayed appearance of atherosclerotic lesions in cerebral arteries.

Animals↗
PubMed1981

Purification and properties of CTP synthetase from Ehrlich ascites tumor cells.

CTP synthetase (UTP: ammonia ligase (ADP-forming), EC 6.3.4.2) was purified 200-fold from Ehrlich ascites tumor cells with about 50% purity as judged by polyacrylamide gel electrophoresis. The molecular weight estimated to be 118 000 by gel filtration. The optimal pH was 8.6. 2-Mercaptoethanol was required for optimal activity and stabilization of the enzyme. Magnesium was essential for the reaction and other divalent cations were ineffective. Ammonia could replace glutamine as the amino donor. When other substrates were at saturating concentrations, Michaelis-Menten kinetics were observed yielding Km values for UTP, ATP and glutamine of 0.18, 0.8 and 0.13 mM, respectively. With ATP at subsaturating concentration, the double-reciprocal plot for UTP saturation was sinusoidal and the Hill plot showed an n value of 1.3. The double-reciprocal plot for ATP saturation, when UTP was at subsaturating concentration, departed from Michaelis-Menten kinetics with an n value of 1.4. These data suggest the existence of cooperative interactions between enzyme and substrates at subsaturating concentrations of ATP or UTP. GTP was not essential, but it acted as an activator on the glutamine reaction; optimal activation was observed at 1 mM GTP. The affinity for glutamine was not affected by GTP.

Amino Acids↗
PubMed1981

Effect of hydrostatic pressure on lysozyme and chymotrypsinogen detected by fluorescence polarization.

The effect of hydrostatic pressure upon solutions of chymotrypsinogen and lysozyme at room temperature has been followed by employing a new technique [Chryssomallis, G. S., Drickamer, H. G., & Weber, G. (1978) J. Appl. Phys. 49, 3084] that permits the measurement of fluorescence polarization at pressures of up to 10 kbar. Lysozyme shows a stable, reversible 60% increase in apparent volume when the pressure is raised to 9 kbar. This can be given a simple interpretation in terms of solvent penetration of the structure at higher pressures. In contrast, the results with chymotrypsinogen are time dependent and only partially reversible on release of the pressure. They involve conversion (tl/e = 5 min) to a form with a lower rotational rate at approximately 6 kbar and return to a fast-rotating form at higher pressure. This latter form persists on pressure release. The possibility of generating what are clearly metastable conformations, not only in chymotrypsinogen but also in flavodoxins [Visser, A. J. W. G., Li, T. M., Drickamer, H. G., & Weber, G. (1977) Biochemistry 16, 4879], indicates that there are unresolved questions about the relative stability of protein conformations which can be profitably investigated by high-pressure experiments.

Chymotrypsinogen↗
PubMed1981

Pressure-induced reversible dissociation of enolase.

A study of the polarization of the intrinsic fluorescence and the fluorescence of dansyl conjugates of enolase shows that an increase in hydrostatic pressure, in the range of 1 bar-3 kbar, promotes the dissociation of this protein into dimers. The dissociation of oligomeric proteins under pressure is predicted to be a general phenomenon by a model that assumes the existence of small "free volumes" at the intersubunit boundaries. The same model predicts a dependence of the standard volume change in the dissociation reaction upon the pressure, owing to the additional surface compressibility of the monomers, and numerical analysis of the results clearly shows that dependence for enolase. For a midpoint dissociation pressure of 1.5 kbar the standard volume change in the dissociation reaction is delta V p0 = -65 +/- 8 mL mol-1 and the dependence of the volume change upon pressure (dVp0/dp) is approximately -30 mL mol-1 kbar-1. The reversibility of the pressure effects is shown to be better than 95% by either polarization or fluorescence spectrum recovery. The pressure perturbation of the fluorescence polarization is a method of general applicability to studies of protein aggregation, and it can be also of value in characterizing the effect of ligands on the aggregation of oligomeric proteins.

Dansyl Compounds↗
PubMed1981

Carbamoyl phosphate synthetase (glutamine-hydrolyzing): increased activity in cancer cells.

The specific activity of carbamoyl phosphate synthetase (glutamine-hydrolyzing), the first and rate-limiting enzyme of de novo uridine 5'-triphosphate biosynthesis, was increased in 13 transplantable hepatomas, particularly in the rapidly growing tumors (5.7- to 9.5-fold), and the rise was correlated with tumor growth rates. Thus, synthetase activity was linked with both hepatic neoplastic transformation and progression. Synthetase specific activity was so elevated in a transplantable sarcoma (18-fold) and a kidney adenocarcinoma (5-fold). The increased activity should enhance the capacity of the pathway and should confer selective advantages to cancer cells.

Adenocarcinoma↗
PubMed1981

Guanosine-5'-phosphate synthetase and guanosine-5'-phosphate kinase in rat hepatomas and kidney tumors.

The behavior of the activities of GMP synthetase (xanthosine-5'-phosphate: L-glutamine amino-ligase(AMP-forming),EC 6.3.5.2) and GMP kinase (ATP: (d)GMP phosphotransferase,EC 2.7.4.8) was elucidated in cytosol preparations of rat tissues, including fetal, neonatal and regenerating liver, in a transplantable kidney tumor, and in a spectrum of 11 hepatomas of different growth rates. GMP kinase activity was 60-fold or more higher than GMP synthetase activity in all of the examined tissues. GMP synthetase activity was increased in all hepatomas and in the kidney tumor, compared to control tissues, reaching 5.5-fold the normal liver values in the most rapidly growing hepatoma. This increase correlated with the tumor growth rates. GMP kinase activity showed no consistent pattern of alteration in the tumors. In both fetal and neonatal rat liver the activity of GMP synthetase was 2.5-times higher than in livers of adult rats, but GMP kinase activity did not change markedly during liver development. After partial hepatectomy GMP synthetase activity was elevated, reaching a peak of 155% of the sham-operated control values by 36 h after the operation. GMP kinase activity was not affected by partial hepatectomy. After 3 days starvation hepatic GMP kinase activity decreased slightly faster than total cytosol protein, while GMP synthetase activity was preferentially maintained. These results indicate that GMP synthetase activity was linked with cellular proliferation in differentiating, regenerating and neoplastic tissues.

Animals↗
PubMed1981

Purification and properties of dihydrothymine dehydrogenase from rat liver.

Rat liver dihydrothymine dehydrogenase, the rate-limiting enzyme of thymidine and uridine degradation, was purified to homogeneity as judged by polyacrylamide disc gel electrophoresis, sedimentation velocity, and Ultrogel ACA-34 elution profile. The enzyme has a molecular weight of 220,000 +/- 5,000 as determined by Ultrogel ACA-34 and sedimentation equilibrium The s20,w value of the enzyme was 9.2 S. The isoelectric point was at pH 5.25. The enzyme is composed of two identical subunits of an approximate molecular weight of 110,000 +/- 3,000 as determined by sodium dodecyl sulfate disc gel electrophoresis. The enzyme contains 4 mol of FAD and 3 mol of iron per mol of enzyme. Flavin released from the enzyme by boiling was identified as FAD by absorption spectra and thin layer chromatography, indicating that the enzyme is a flavometal protein. During dialysis, the enzyme was stabilized by 2-mercaptoethanol, but neither NADPH nor thymine was effective. The relative rates of reduction of pyrimidine analogues substituted at position 5 were 5-fluorouracil > 5-bromouracil > 5-diazouracil > 5-iodouracil > 5-nitrouracil, with 5-fluorouracil and 5-diazouracil 70% faster than thymine. Uracil was reduced 25% faster than thymine. The pH optimum for the forward and reverse reactions was 7.4. In the presence of NADPH, the apparent Km was 2.6 microM for thymine and 1.8 microM for uracil. Apparent Km for NADPH was 15 microM with thymine as substrate and 11 microM with uracil. In the reverse reaction, apparent Km values were 43 microM for dihydrothymine and 193 microM for dihydrouracil; apparent Km for NADP+ was 3.8 microM with dihydrothymine as substrate and 2.9 microM with dihydrouracil.

Amino Acids↗
PubMed1981

Psoriasis and human growth hormone: aetiology and therapy.

First report on nine patients suffering from psoriasis who were treated with somatostatin or bromocriptin or both. These are inhibitors of HGH. The therapeutical effect on skin lesions and psoriatic alterations to joints is described.

Adult↗