On the reliability of quantitation of biological effect in drug–interceptor–DNA systems

Relative insensitivity of theoretical estimation of biological effect in drug–interceptor–DNA systems is found with respect to variation of parameters of quasiphysiological conditions. The “inertness” of the biological response, in part, justifies the use of parameters of intermolecular interaction, derived from independent physicochemical experiments, in estimation of relative biological effect in the theory of interceptor/protector action.     

A stochastic model for the NMR analysis of the heteroassociation of biologically active aromatic molecules

A stochastic model for the NMR analysis of the heteroassociation of two aromatic compounds was developed, which takes into account all physically possible reactions of association of molecules in solution. Expressions for calculating the experimentally observed proton chemical shift were obtained in the general form, and an algorithm for calculating the parameters of heteroassociation using the stochastic model was proposed. The effects of limitations of the basic and general models, as compared with the stochastic model, on the model parameters of the heteroassociation of various biologically active aromatic molecules was analyzed. It was shown that the basic model can be used with a sufficient degree of accuracy for systems with a relatively small contribution of heteroassociation reactions to the total dynamic equilibrium in solution, whereas the general model describes satisfactorily the parameters of heteroassociation practically for all systems studied.     

Ferredoxin reduction by polarographic methods]

The reduction of iron-sulphur protein of the higher plant ferrodoxin has been studied by polarographical methods. Ferredoxin initiates a reversible wave with E1/2=--0,61 v (N. C. E.) at pH 7. Protein absorption greatly influences the electrochemical reduction. The protons have been shown to take part in the electrode reaction. The potentiometrically obtained data about the difference between E1/2 and E0=--0.70 v and its causative factors are discussed. As a result of the experiments with modification of ferredoxin active centre it has been concluded that the active centre participates in the polarographical reduction.     

Assembly of the sarcoplasmic reticulum proteins in differentiating rat skeletal muscle cell cultures: localization by immunofluorescence of sarcoplasmic reticulum proteins in differentiating rat skeletal muscle cell cultures

Immunofluorescent staining techniques were used to study the distribution of the Ca(2) + Mg(2+)-dependent ATPase and calsequestrin in primary cultures of differentiating rat skeletal muscle cells, grown for different periods of time under various culture conditions. In mononucleated myoblasts calsequestrin was detected after 45 h in culture whereas the ATPase was not detected until 60 h. After cell fusion began, both proteins could be identified in all multinucleated cells. Myoblasts grown for longer than 60 h in low Ca(2+) medium contained calsequestrin and the ATPase, even though they were unable to fuse. These studies at the cellular level confirm biochemical findings on the biosynthesis of calsequestrin and the ATPase. Immunofluorescent staining of myoblasts showed that calsequestrin first appears in a well-defined region of the cell near one end of the nucleus. At later times, the staining occupied progressively larger regions adjacent to the nucleus and took on a fibrous appearance. This suggests that calsequestrin first accumulates in the Golgi region and then gradually spreads throughout the cell. In contrast, the ATPase appeared to be concentrated in many small patches or foci throughout the cytoplasm and was never confined to one particular region, although some parts of the cell often stained more intensely than others. In multinucleated cells, alternating dark and fluorescent strands parallel to the longitudinal axis of the cells were evident.     

Connection of outer membrane protein composition in Yersinia pestis cells with intrinsic plasmids]

A set of isogenic derivatives of Yersinia pestis EV strain was obtained including the variants harbouring the different compositions of Yersinia own plasmids. The protein profiles of outer membranes of the set of strains were defined. The polyacrylamide gel electrophoresis has shown the small 6.1 Md plasmid to code an outer membrane protein with mol mass 29 kDa, different from pesticin I, while the heavy 60.0 Md plasmid encodes the 15-16 kDa polypeptide different from monomers of F1 and T-antigens of plague microbe.     

A microscopic model of complex formation of intercalators with DNA: analysis of NMR spectroscopy data

A model of intercalation complex formation of ligands with an arbitrary oligonucleotide sequence has been proposed, which takes in the explicit form the microscopic constants and the parameters of the cooperativity of binding to particular sites of the oligomer into account. The model has been adapted for the analysis of NMR spectroscopy data.     

Quantitation of the molecular mechanisms of biological synergism in a mixture of DNA-acting aromatic drugs

It is suggested that the widely reported biological synergism of a mixture of DNA-targeting aromatic drug molecules both in vivo and in vitro can be explained, in part, at the molecular level by competition between two basic mechanisms: the 'interceptor' (hetero-association between Drug1 and Drug2) and 'protector' mechanisms (complexation of Drug1 and Drug2 on DNA-binding sites). In the present work a complete analytical methodology has been developed to quantify these processes, providing an estimate of the relative importance of the interceptor/protector mechanisms using just a set of equilibrium association constants. The general methodology may be applied to other molecules with receptors for aromatic drugs.     

Theoretical analysis of dynamic force spectroscopy experiments on ligand-receptor complexes

The forced rupture of single chemical bonds in biomolecular compounds (e.g. ligand-receptor systems) as observed in dynamic force spectroscopy experiments is addressed. Under the assumption that the probability of bond rupture depends only on the instantaneously acting force, a data collapse onto a single master curve is predicted. For rupture data obtained experimentally by dynamic AFM force spectroscopy of a ligand-receptor bond between a DNA and a regulatory protein we do not find such a collapse. We conclude that the above mentioned, generally accepted assumption is not satisfied and we discuss possible explanations.     

Evaluation of the effectiveness of antibacterial substances in treating an experimental form of bubonic plague in monkeys]

The modelling of glandular plague and selection of the conditions for estimating the efficacy of new antibacterials for the treatment of the infection were performed on hamadryads (baboons). The experiments showed that the average LD50 of the culture of a highly virulent strain of Yersinia pestis on its subcutaneous administration to the animals was 2089 viable microbes. In 18 per cent of the episodes the experimental glandular plague in the animals was complicated by secondary plague pneumonia. Subcutaneous administration of 2 x 10(7) viable microbial cell of the plague pathogen caused acute sepsis and the animal death. The treatment of the experimental glandular plague in the hamadryads demonstrated that new antibacterials such as amikacin, netilmicin, ceftriaxone, cefotaxime, ceftizoxime, doxycycline, rifampicin, ofloxacin and ciprofloxacin were not inferior in their efficacy to streptomycin and tetracycline successfully used in the therapy of patients with plague.