Membrane-bound calmodulin in Xenopus laevis oocytes as a novel binding site for melatonin

Melatonin has been suggested as a physiological antagonist of calmodulin. In this work, we have characterized melatonin binding sites in Xenopus laevis oocyte membranes. Binding of [125I]melatonin by X. laevis oocyte membranes fulfills all criteria for binding to a receptor site. Binding was dependent on time, temperature, and membrane concentration and was stable, reversible, saturable, and specific. The binding site was also pharmacologically characterized. Stoichiometric studies showed a high-affinity binding site with a Kd of 1.18 nM. These data are in close agreement with data obtained from kinetic studies (Kd=0.12 nM). In competition studies, we observed a low-affinity binding site (Kd=63.41 microM). Moreover, the binding site was characterized as calmodulin. Thus, binding was dependent on calcium and blocked by anti-CaM antibodies in a concentration-dependent manner. Calmodulin inhibitor chlorpromazine also inhibited binding of the tracer. From these results, it is suggested that membrane-bound calmodulin acts as a melatonin binding site in Xenopus laevis oocytes, where it might couple cellular activities to rhythmic circulating levels of melatonin. This hypothesis correlates with the previous findings describing melatonin as a physiological antagonist of calmodulin.     

Antimutagenic activity of natural phenolic compounds present in the common bean ( Phaseolus vulgaris ) against aflatoxin B 1

Polyphenols with antimutagenic and anticarcinogenic properties are present in fruits, vegetables and legumes. In this study, the Salmonella typhimurium tester strains TA98 and TA100 were used in the microsuspension assay to examine the antimutagenic effect of phenolic compounds extracted from the common bean (Phaseolus vulgaris) against mutagenicity induced by aflatoxin B 1 (AFB 1 ). A dose-response curve was constructed for AFB 1 ; from which a level of 40ng AFB 1 /tube was selected for all antimutagenicity assays. The AFB 1 and phenolic extract (PE) were not toxic to the bacteria at concentrations tested. In the case of PE, results were similar to the number of spontaneous revertants for TA98 and TA100. The inhibitory effect of PE against AFB 1 mutagenicity was dose-dependent at the lower concentrations tested (2.5, 5, 10, 12.5, 15 and 25 μg-equivalent ( + )-catechin/tube for TA98; 0.5, 1, 1.5, 2.5, 5, 10 and 25 μg-equivalent ( + )-catechin/ tube for TA100). Further, a two-stage incubation procedure was used to investigate the potential interaction between PE and AFB 1 . The greatest inhibitory effect of the PE on AFB 1 mutagenicity occurred when PE and AFB 1 were incubated together. When the bacteria were first incubated with PE followed by a second incubation with AFB 1 , lower inhibition was observed. Lower inhibition was also observed when the bacteria were first incubated with AFB 1 followed by a second incubation with PE. The results suggest that the mechanism of inhibition could involve the formation of a chemical complex between of PE and AFB 1 .     

Quantitative determination of non-defect unstable structures in PVC through selective nucleophilic substitution with C6H5SNa

Summary The nucleophilic substitution with C₆H₅SNa, at -30°C, is compared for five PVC samples with various contents of isotactic triads. The conversion curves consist of a very fast stage followed by a steady one. All the samples are found to behave in the same way except for the content of the structures involved in the fast period. This content is estimated by extrapolating the straight lines of the steady stages to zero time. The obtained values are found to be a linear function of the content of isotactic triads. The results, together with some earlier ones, allow for the content of the labile non-defect structures in PVC to be determined.     

Modified determination of 2-thiobarbituric acid value in fats and oils

Several methods for the determination of TBA-value were compared with a new direct modification (omitting the previous distillation), using 1-butanol as the sole solvent. The absorbance is measured at 530 nm after heating to 95 degrees C for 120 min, and cooling. The effect of variables was tested by the Youdens ruggedness test (reaction time, temperature, cooling, concentration of sample, thiobarbituric acid, water, heavy metals, antioxidants, chelating agents, bubbling with oxygen or nitrogen). The Lambert-Beers law is valid in the absorbance range between 0.05 and 0.8. The repeatability is better than that of the Dzikowskis procedure and comparable with those of more complicated distillation methods. The standard deviation is in a linear relation with the absorbance (range 0.1-1.0). The method was found satisfactory for evaluating lard, cooking fat, soybean, sunflower, and rapeseed oils in the stage of beginning rancidity.