Electrophysiologic correlates of habituation and conditioned reflex elaboration

The average correlation coefficient between the EEG of different parts of the cortex gradually decreases as the rabbit is habituated to the experimental surroundings. Elaboration of an electo-defensive conditioned reflex leads to its enhancement which is replaced by a phase of temporary decrease, to be followed again by an enhancement of EEG correlation. The pronouncedness of theta-frequencies in the spectral and coherent functions behaves in a similar way. Successive changes of delta-activity level reveal no regularity.     

Spectral manifestations of different types of acriflavine binding to DNA in ultraviolet and visible region]

Difference absorption spectra (complex-sum of the initial reagents) are obtained in the visible and longwave UV region for the system of actiflavine and DNA in a number of cases differing in initial and final degrees of DNA filling by the dye, in particular separately for two types of dye binding to DNA. For these binding types conventional absorption spectra are calculated. In the visible region for the first binding type ("strong" binding) red shift of the absorption band is observed; for the second type ("weak" binding) we observed splitting of the band, short wavelength component being highly prevailing, and hypochromism. In the UV region for both binding types the spectra changed in approximately similar way; a slight blue shift and a rather remarkable hypochromism are observed. It is shown that the dye brings the main contribution into the spectral changes in the UV region. If to take into account the spectral properties of molecular aggregates the data obtained are compatible with the intercalation model for "strong" binding and dye stacking on DNA for "weak" binding.     

The effect of acridine dyes on the molecular structure of DNA]

DNA--acriflavin complexes have been investigated by the methods of flow birefringence and viscometry. The intrinsic viscosity and the optic anysotropy of the complex increase with the increasing quantities of binding dye. Experimental data are treated on the basis of different models of binding. At high ionic strength (mu = 0,1) one type of binding takes place which is described by the intercalation model. In this case the thermodynamic rigidity of DNA-molecule within the complex is proportional to "r". In solutions of low ionic strength (mu = 0,001), two types of DNA-acriflavin binding occur: intercalation and external binding. At low ionic strength, the spectrophotometric titration technique is shown to give a reduced value of "r".     

Rhythmical processes in the cerebral cortex bioelectrical activity in arousal reaction: qualitative nonlinear analysis involving refractoriness

An integral equation that takes into account the absolute and relative refractory periods of cortical cells was added to the earlier obtained integral equations describing the cooperation of excitatory and inhibitory neurons of the cerebral cortex, and its substantiation was suggested. The steady states and the stable region of oscillations of the mean membrane potential of excitatory neurons were investigated depending on the mean afferent influx value on the assumption of the normal threshold distribution of cells. The existence of a parameter multitude in the region of stability was shown when a suppression of the delta-rhythm and an enhancement of the theta-rhythm up to its dominance take place increasing nonspecific afferent influx, which shows up a white noise, and decrease of the theta-rhythm during further increase in nonspecific activation. Simultaneously the frequencies of the major cerebral rhythms increase. The conformity of the theoretically calculated dependences of changes of the spectrum upon increasing nonspecific afferent influx to the experimental data obtained during the electrical stimulation of the rabbit midbrain reticular formation was demonstrated.     

Dynamics of protein ligand binding on multiple time scales: NADH binding to lactate dehydrogenase

Although the importance of atomic motion to how proteins function has been conjectured for several decades, the characterization of protein dynamics on multiple time scales is scant. This is because of severe experimental and theoretical difficulties, particularly characterizing the nanosecond to millisecond time scales. Here, we apply advanced laser-induced temperature-jump relaxation spectroscopic techniques to examine the kinetics of NADH binding to lactate dehydrogenase over this time scale. The bimolecular rate process, at about 290 micros, is easily observed as are multiple faster events (with relaxation times of 200 ns, 3.5 micros, and 24 micros), revealing a rich dynamical nature of the binding step. The results show that there are multiple structures of bound enzyme-ligand complexes, some of which are likely to be far from the catalytically productive structure. The results have important implications for interpretations of the binding thermodynamics of ligands to LDH and, by extension, to other proteins. The observed processes likely play a role in the dynamics of the chemistry that is catalyzed by lactate dehydrogenase.     

Frequency and amplitude windows in the combined action of DC and low frequency AC magnetic fields on ion thermal motion in a macromolecule: theoretical analysis

We solved the differential equation describing combined action of DC and AC magnetic fields on thermal motion of ions in a biological macromolecule. The solution showed the occurrence of a new set of resonant peaks for ion oscillations under the influence of magnetic fields. After establishment of steady ion oscillations in the macromolecule interior that is well shielded from the action of small particles of the medium surrounding this molecule, the change in energy of ion thermal motion could be sufficient to alter the conformation state of the macromolecule. On this basis, a diversity of biological phenomena can be explained, including the appearance of the known "frequency" and "amplitude" windows, without any resort to the ideas of participation of cyclotron or parametric resonances in these effects.     

Methyl Jasmonate Increases the Tropane Alkaloid Scopolamine and Reduces Natural Herbivory in Brugmansia suaveolens: Is Scopolamine Responsible for Plant Resistance?

The tropane alkaloid (TA) scopolamine is suggested to protect Brugmansia suaveolens (Solanaceae) against herbivorous insects. To test this prediction in a natural environment, scopolamine was induced by methyl jasmonate (MJ) in potted plants which were left 10?days in the field. MJ-treated plants increased their scopolamine concentration in leaves and herbivory decreased. These findings suggest a cause?Ceffect relationship. However, experiments in laboratory showed that scopolamine affect differently the performance of the specialist larvae of the ithomiine butterfly Placidina euryanassa (C. Felder & R. Felder) and the generalist fall armyworm Spodoptera frugiperda (J. E. Smith): the specialist that sequester this TA from B. suaveolens leaves was not negatively affected, but the generalist was. Therefore, scopolamine probably acts only against insects that are not adapted to TAs. Other compounds that are MJ elicited may also play a role in plant resistance against herbivory by generalist and specialist insects, and deserve future investigations.     

Tryptophan-free human PNP reveals catalytic site interactions

Human purine nucleoside phosphorylase (PNP) is a homotrimer, containing three nonconserved tryptophan residues at positions 16, 94, and 178, all remote from the catalytic site. The Trp residues were replaced with Tyr to produce Trp-free PNP (Leuko-PNP). Leuko-PNP showed near-normal kinetic properties. It was used (1) to determine the tautomeric form of guanine that produces strong fluorescence when bound to PNP, (2) for thermodynamic binding analysis of binary and ternary complexes with substrates, (3) in temperature-jump perturbation of complexes for evidence of multiple conformational complexes, and (4) to establish the ionization state of a catalytic site tyrosine involved in phosphate nucleophile activation. The (13)C NMR spectrum of guanine bound to Leuko-PNP, its fluorescent properties, and molecular orbital electronic transition analysis establish that its fluorescence originates from the lowest singlet excited state of the N1H, 6-keto, N7H guanine tautomer. Binding of guanine and phosphate to PNP and Leuko-PNP are random, with decreased affinity for formation of ternary complexes. Pre-steady-state kinetics and temperature-jump studies indicate that the ternary complex (enzyme-substrate-phosphate) forms in single binding steps without kinetically significant protein conformational changes as monitored by guanine fluorescence. Spectral changes of Leuko-PNP upon phosphate binding establish that the hydroxyl of Tyr88 is not ionized to the phenolate anion when phosphate is bound. A loop region (residues 243-266) near the purine base becomes highly ordered upon substrate/inhibitor binding. A single Trp residue was introduced into the catalytic loop of Leuko-PNP (Y249W-Leuko-PNP) to determine effects on catalysis and to introduce a fluorescence catalytic site probe. Although Y249W-Leuko-PNP is highly fluorescent and catalytically active, substrate binding did not perturb the fluorescence. Thermodynamic boxes, constructed to characterize the binding of phosphate, guanine, and hypoxanthine to native, Leuko-, and Y249W-Leuko-PNPs, establish that Leuko-PNP provides a versatile protein scaffold for introduction of specific Trp catalytic site probes.