Active site mutations in DNA topoisomerase I distinguish the cytotoxic activities of camptothecin and the indolocarbazole, rebeccamycin.

DNA topoisomerase I (Top1p) catalyzes topological changes in DNA and is the cellular target of the antitumor agent camptothecin (CPT). Non-CPT drugs that target Top1p, such as indolocarbazoles, are under clinical development. However, whether the cytotoxicity of indolocarbazoles derives from Top1p poisoning remains unclear. To further investigate indolocarbazole mechanism, rebeccamycin R-3 activity was examined in vitro and in yeast. Using a series of Top1p mutants, where substitution of residues around the active site tyrosine has well-defined effects on enzyme catalysis, we show that catalytically active, CPT-resistant enzymes remain sensitive to R-3. This indolocarbazole did not inhibit yeast Top1p activity, yet was effective in stabilizing Top1p-DNA complexes. Similar results were obtained with human Top1p, when Ser or His were substituted for Asn-722. The mutations altered enzyme function and sensitivity to CPT, yet R-3 poisoning of Top1p was unaffected. Moreover, top1delta, rad52delta yeast cells expressing human Top1p, but not catalytically inactive Top1Y723Fp, were sensitive to R-3. These data support hTop1p as the cellular target of R-3 and indicate that distinct drug-enzyme interactions at the active site are required for efficient poisoning by R-3 or CPT. Furthermore, resistance to one poison may potentiate cell sensitivity to structurally distinct compounds that also target Top1p.     

Effect of Zeitgeber Intensity Reduction on a Simulated Dual-Oscillator Human Circadian System: Classical and Dynamic Analysis

The two-oscillator model of human circadian rhythmicity was analyzed when a zeitgeber relative intensity of 1, 0.5, or 0.1 was introduced into the equations. Fourier analysis was compared with dynamic analysis such as attractor reconstruction or Liapunov exponent calculation. After a 50 or 90% reduction in zeitgeber intensity, the dynamics of the system became equivalent and differed significantly from those of a system with maximal zeitgeber intensity. When 10% aleatory noise was added to the data, the analysis was still applicable, and the results obtained were essentially the same as in the absence of noise. Dynamic analysis could thus provide a distinct classification for periodic data, based on the type of analysis.     

The binding of anionic and nonionic surfactants to collagen through the hydrophobic effect

The adsorption of nonionic surfactants on hide powder previously treated with anionic surfactants has been studied. The adsorption of nonionic surfactants takes place through hydrophobic interactions. A mechanism has been proposed for this interaction, assuming that the nonionic surfactant has been fixed by means of secondary adsorption (hydrophobic interaction) after the primary adsorption of the anionic surfactant (ionic and hydrophobic interaction) which makes it possible.     

Actinomycin D and cycloheximide actions on activity of some intestinal enzymes of adult hamster

Actinomycin D, at a dose of 0.25 micrograms/g body wt, produced slight increases in intestinal enzymatic activity on hamsters. At a high dose (1.5 micrograms/g body wt), actinomycin D produced inhibition of lactase activity, whereas maltase, sucrase and alkaline phosphatase activity decreased in males and increased in females. Cycloheximide (1.5 micrograms/g body wt), produced no changes in enzymatic activity. In the male and female hamster, the different actions of the antibiotic can be explained by the variations in the cortisol release produced by stress.     

A new function for overall optimization of yeast production

The influence of several operating variables on the productivity and nutritional quality of Candida utilis under carbon-limited growth conditions was studied. Four responses were analyzed: biomass yield (Y), in vitro dry matter digestibility (d), nucleic acid cotents, and protein content (P). Three operating variables were studied: dilution rate, temperature, and carbon-nitrogen ratio in the growth medium. Correlation describing the influence of operating variables on the studied response parameters were developed. From these correlations response surfaces were drawn. A new function, alpha = PdY, is proposed as a more comprehensive criterion for selecting the optimal conditions under the constraint of satisfying multiple objectives. It is demonstrated to be superior to the yield coefficient alone in this respect, since it accounts for factors that are important to the nutritional aspects of the process.     

Production of staphylococcal enterotoxins C1 and C2 and thermonuclease throughout the growth cycle

Synthesis of enterotoxins C1 and C2 and thermonuclease throughout the growth cycle was investigated with Staphylococcus aureus type strains FRI137 and FRI361 and S. aureus isolates M5 (C1) and L2 (C2) of animal origin. Both enterotoxins were produced during the exponential growth phase or at the beginning of the stationary phase. The minimal incubation time (7 to 12 h) and the lowest population (10(7) to 2 x 10(9) CFU/ml) associated with detectable enterotoxin (1 to 6.5 ng/ml) were related to the total amount of toxin produced after 24 h. Thermonuclease was detected in all samples whenever enterotoxins were detected. Furthermore, strain FRI137 produced thermonuclease earlier and at lower cell populations than it did enterotoxin C1. Patterns of enterotoxin and thermonuclease synthesis did not correlate. The concentration of toxins increased throughout the growth cycle, while the concentration of thermonuclease remained constant during the last hours of the growth cycle.     

Action of Thiobacillus thiooxidans on Sulphur in the Presence of a Surfactant Agent and its Application in the Indirect Dissolution of Phosphorus

The addition of a surfactant agent (Tween 80) to a medium containing sulphur and a culture of Thiobacillus thiooxidans increased the attachment of bacteria to sulphur, the rate of sulphur oxidation and sulphuric acid production. This acid was used to dissolve phosphorus from calcium phosphate. The yield was higher than reported for other microorganisms although dissolution was not increased significantly by Tween addition.