The Degradation and Aging of Biological Systems as a Process of Information Loss and Entropy Increase.

The problem of the degradation and aging of bioorganisms is herein considered from the viewpoint of statistical physics. Two typical timescales in biological systems-the time of metabolic processes and the time of the life cycle-are used. A kinetic equation describing the small timescales of the systems' characteristic processes in is proposed. Maintaining a biosystem in a time-stable state requires a constant inflow of negative entropy (negentropy). Ratios are proposed to evaluate the aging and degradation of systems in terms of entropy. As an example, the aging of the epithelium is studied. The connection of our approach to the information theory of aging is discussed, as well as theoretical constructions related to the concept of cooperon and its changing with time.

A New Model of Hemoglobin Oxygenation.

The study of hemoglobin oxygenation, starting from the classical works of Hill, has laid the foundation for molecular biophysics. The cooperative nature of oxygen binding to hemoglobin has been variously described in different models. In the Adair model, which better fits the experimental data, the constants of oxygen binding at various stages differ. However, the physical meaning of the parameters in this model remains unclear. In this work, we applied Hill's approach, extending its interpretation; we obtained a good agreement between the theory and the experiment. The equation in which the Hill coefficient is modulated by the Lorentz distribution for oxygen partial pressure approximates the experimental data better than not only the classical Hill equation, but also the Adair equation.

C60 Fullerene Clusters Stabilize the Biologically Inactive Form of Topotecan.

There is a range of experimental proofs that biologically relevant compounds change their activity in the presence of C60 fullerene clusters in aqueous solution, which most frequently act as a nanoplatform for drug delivery. Inspired by this evidence, we made an effort to investigate the interaction of fullerene clusters with the antibiotic topotecan (TPT). This study proceeded in three steps, namely, UV/vis titration to confirm complexation and in vitro assays on proliferating and nonproliferating cells to elucidate the role of C60 fullerene in the putative change in TPT activity. Surprisingly, although the nonproliferating cell assay is consistent with the titration data and confirms complex formation, it contradicted the results of the proliferating cell assay. The latter showed that the mixture of TPT and fullerene affects the cells in the same way as pure TPT, as if there were no fullerenes in solution at all, whereas the action of TPT was expected to be enhanced. We explained this contradiction by the specific stabilization of the biologically inactive carboxylate form of the antibiotic adsorbed in the alkaline shell of large fullerene clusters, which leads to neutralization of the drug delivery function and almost zero net biological effect of the antibiotic in vitro. The practical outcome of the work is that fullerene clusters can be used for the selective delivery of pH-sensitive drug forms.

Interceptor potential of C60 fullerene aqueous solution: a comparative analysis using the example of the antitumor antibiotic mitoxantrone.

We performed a qualitative and quantitative analysis of intermolecular interactions in aqueous solution between the antitumor antibiotic mitoxantrone and C60 fullerene in comparison with interactions between the antibiotic and well-known aromatic molecules such as caffeine and flavin mononucleotide, commonly referred to as interceptor molecules. For these purposes, we obtained equilibrium hetero-association constants of these interactions using a UV/Vis titration experiment. Special attention was paid to the interaction of C60 fullerene with mitoxantrone, which has been quantified for the first time. Based on the theory of interceptor-protector action and using a set of measured equilibrium constants we managed to estimate the relative biological effect of these mixtures in a model living system, taking human buccal epithelium cells as an example. We demonstrated that C60 fullerene is able to restore the functional activity of the buccal epithelium cell nucleus after exposure to mitoxantrone, which makes it possible to use C60 fullerene as regulator of medico-biological activity of the antibiotic.

The Use of the Statistical Entropy in Some New Approaches for the Description of Biosystems.

Some problems of describing biological systems with the use of entropy as a measure of the complexity of these systems are considered. Entropy is studied both for the organism as a whole and for its parts down to the molecular level. Correlation of actions of various parts of the whole organism, intercellular interactions and control, as well as cooperativity on the microlevel lead to a more complex structure and lower statistical entropy. For a multicellular organism, entropy is much lower than entropy for the same mass of a colony of unicellular organisms. Cooperativity always reduces the entropy of the system; a simple example of ligand binding to a macromolecule carrying two reaction centers shows how entropy is consistent with the ambiguity of the result in the Bernoulli test scheme. Particular attention is paid to the qualitative and quantitative relationship between the entropy of the system and the cooperativity of ligand binding to macromolecules. A kinetic model of metabolism. corresponding to Schrödinger's concept of the maintenance biosystems by "negentropy feeding", is proposed. This model allows calculating the nonequilibrium local entropy and comparing it with the local equilibrium entropy inherent in non-living matter.

Hetero-association models of non-covalent molecular complexation.

The present review discusses the current state-of-the-art in building models enabling the description of non-covalent equilibrium complexation of different types of molecules in solution, which results in the formation of supramolecular structures different in length and composition (hetero-association or supramolecular multicomponent co-polymerisation). The description is focused on standard physical and chemical quantities such as experimental observables and equilibrium parameters of interaction (equilibrium constants and concentrations). The major partial cases of the hetero-association models, such as finite and indefinite isodesmic and cooperative complexations, and Benesi-Hildebrand and Langmuir adsorption models are considered. Future challenges in the development of the hetero-association models are provided.

The theory of interceptor-protector action of DNA binding drugs.

The review discusses the theory of interceptor-protector action (the IPA theory) as the new self-consistent biophysical theory establishing a quantitative interrelation between parameters measured in independent physico-chemical experiment and in vitro biological experiment for the class of DNA binding drugs. The elements of the theory provide complete algorithm of analysis, which may potentially be applied to any system of DNA targeting aromatic drugs. Such analytical schemes, apart from extension of current scientific knowledge, are important in the context of rational drug design for managing drug's response by changing the physico-chemical parameters of molecular complexation.

Study of the properties of doxorubicin-resistant cells affected by acute leucosis.

The stiffness of cell membrane was found to be one of the factors determining resistance of a cell in vitro to antibiotic doxorubicin action. Membranes of surviving cells are negatively charged (-35 - -30 mV) and have high values of stiffness (2.2-5.1 µÐ Ð°) at the doxorubicin concentrations in the medium of 1-500 µg/ml. If the drug concentration and exposure time are being increased, only cells with 'soft' membrane (0.25-1 µÐ Ð°) and positive surface potential (15-29 mV) survive. The data obtained have important prognostic value in studying drug resistance of tumour blood cells and can be used as objective markers of efficiency of the antitumor therapy.

C60 fullerene affects elastic properties and osmoregulation reactions of human lymphocytes.

The influence of aqueous solution of pristine C60 fullerene (C60FAS) on functional activity of lymphocytes from a healthy person was studied for the first time. By means of atomic force microscopy, it was found that C60FAS in a concentration of 0.1 mg/ml increases the stiffness of the lymphocyte membrane by 41% (p < 0.05) and lowers the functional activity of the plasmalemma surface, thereby constraining the use of its membrane material in physiological reactions using a hypotonic model in vitro. However, a cell retains the ability to regulate its volume and demonstrates relative resistance to hypo-osmotic stress. The resistance of lymphocytes in hypo-osmotic medium is facilitated by activation of the nucleus by C60 fullerene particles, which regulates the implementation of two consistent phases of an increase and decrease of cell volume, thereby retaining cell viability. All these indicate the impact of C60 fullerene on the cellular nucleus.

Shape-independent model (SHIM) approach for studying aggregation by NMR diffusometry.

NMR diffusometry has been gaining wide popularity in various areas of applied chemistry for investigating diffusion and complexation processes in solid and aqueous phases. To date, the application of this method to study aggregation phenomena proceeding beyond the dimer stage of assembly has been restricted by the need for a priori knowledge of the aggregates' shape, commonly difficult to know in practice. We describe here a comprehensive analysis of aggregation parameter-dependency on the type and shape selected for modeling assembly processes, and report for the first time a shape-independent model (designated the SHIM approach), which may be used as an alternative in cases when information on aggregates' shapes is unavailable. The model can be used for determining equilibrium aggregation parameters from self-diffusion NMR data including equilibrium self-association constant and changes in enthalpy, ΔH, and entropy, ΔS.

Quantitative correlation of the in vitro biological effect with parameters of molecular complexation in mutagen-interceptor systems.

According to the theory of interceptor-protector action a quantitative link between the physico-chemical parameters of molecular complexation and in vitro biological effect in aromatic drug-interceptor systems must exist. In the present communication such link between relative change in mutagenicity of IQ-type aromatic mutagens on addition of aromatic interceptor molecules with equilibrium hetero-association constants of mutagen-interceptor complexation has been found using the published in vitro data in bacteria cell systems.

Interceptor effect of C60 fullerene on the in vitro action of aromatic drug molecules.

C60 fullerenes are spherical molecules composed purely of carbon atoms. They inspire a particularly strong scientific interest because of their specific physico-chemical properties and potential medical and nanotechnological applications. In this work we are focusing on studying the influence of the pristine C60 fullerene on biological activity of some aromatic drug molecules in human buccal epithelial cells. Assessment of the heterochromatin structure in the cell nucleus as well as the barrier function of the cell membrane was performed. The methods of cell microelectrophoresis and atomic force microscopy were also applied. A concentration-dependent restoration of the functional activity of the cellular nucleus after exposure to DNA-binding drugs (doxorubicin, proflavine and ethidium bromide) has been observed in human buccal epithelial cells upon addition of C60 fullerene at a concentration of ~10(-5 )M. The results were shown to follow the framework of interceptor/protector action theory, assuming that non-covalent complexation between C60 fullerene and the drugs (i.e., hetero-association) is the major process responsible for the observed biological effects. An independent confirmation of this hypothesis was obtained via investigation of the cellular response of buccal epithelium to the coadministration of the aromatic drugs and caffeine, and it is based on the well-established role of hetero-association in drug-caffeine systems. The results indicate that C60 fullerene may reverse the effects caused by the aromatic drugs, thereby pointing out the potential possibility of the use of aromatic drugs in combination with C60 fullerene for regulation of their medico-biological action.

Evidence of entropically driven C60 fullerene aggregation in aqueous solution.

In the present work, we report the first experimental evidence of entropically driven C60 fullerene aggregation in aqueous solution, occurring with nearly zero enthalpy change.

Spectroscopic study of proflavine adsorption on the carbon nanotube surface.

Despite the fact that non-covalent interactions between various aromatic compounds and carbon nanotubes are being extensively investigated now, there is still a lack of understanding about the nature of such interactions. The present paper sheds light on one of the possible mechanisms of interaction between the typical aromatic dye proflavine and the carbon nanotube surface, namely, π-stacking between aromatic rings of these compounds. To investigate such a complexation, a qualitative analysis was performed by means of ultraviolet visible, infrared, and nuclear magnetic resonance spectroscopy. The data obtained suggest that π-stacking brings the major contribution to the stabilization of the complex between proflavine and the carbon nanotube.

C60 fullerene aggregation in aqueous solution.

In the present work we develop a novel approach for quantification of the energetics of C60 fullerene aggregation in aqueous media in terms of equilibrium aggregation constant KF. In particular, it is shown that the experimental determination of the magnitude of KF is possible only within the framework of the 'up-scaled aggregation model', considering the C60 fullerene water solution as a solution of fullerene clusters. Using dynamic light scattering (DLS) data we report the value, K(F) = 56,000 M(-1), which is in good agreement with existing theoretical estimates and the results of energetic analyses. It is suggested that the proposed 'up-scaled model' may be used in any instances of non-specific aggregation resulting in formation of large spherical particles. The measurement of the translational diffusion coefficient and the dimensions of the light scattering particles using a DLS approach with respect to C60 fullerene aggregates is found to contain significant systematic errors originating from the interaction effect that is well-known for micellar solutions. As a result, corrections to the equations associated with DLS data are proposed.

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.

Complexation of C60 fullerene with aromatic drugs.

The contributions of various physical factors to the energetics of complexation of aromatic drug molecules with C(60) fullerene are investigated in terms of the calculated magnitudes of equilibrium complexation constants and the components of the net Gibbs free energy. Models of complexation are developed taking into account the polydisperse nature of fullerene solutions in terms of the continuous or discrete (fractal) aggregation of C(60) molecules. Analysis of the energetics has shown that stabilization of the ligand-fullerene complexes in aqueous solution is mainly determined by intermolecular van der Waals interactions and, to lesser extent, by hydrophobic interactions. The results provide a physicochemical basis for a potentially new biotechnological application of fullerenes as modulators of biological activity of aromatic drugs.

Development of an analytical approach to study a three-component hetero-association by means of spectrophotometry.

A case of 1:m:n complexation in a three-component system containing any possible heterocomplexes formed between the non-self-aggregating, absorbing ligand A and two self-aggregating, non-absorbing ligands B and C was considered for the first time in an application for molecular spectroscopy. All expressions necessary for full quantitative analysis of experimental data in three-component mixtures were obtained, viz., the law of conservation of mass and the expression for an experimentally observed parameter. These expressions can be directly utilized in mathematical software for performing standard curve-fitting procedures or solutions of specific tasks such as calculations of the concentration of various types of complexes. The numerical test of the 1:m:n model, accomplished with the aid of ultraviolet-visible light experimental data in a three-component system (proflavine-caffeine-nicotinamide), proved the validity of the developed approach.

Random versus sequential pathway of molecular self-assembly.

Two important assumptions are often made in the analysis of molecular self-assembly at equilibrium, viz., that sequential is preferred to random aggregation and that the equilibrium constants at each stage of aggregation are equal, though both assumptions have not been justified strictly. In the present work we show that molecular self-assembly leading to formation of linear polymers and proceeding in a random manner appears to be less entropically favored than sequential aggregation, which provides a physical background for assuming sequential aggregation when studying molecular self-assembly in solution. Exact equations for analysis of experimental data for molecular assembly proceeding in a sequential manner were derived by taking strict account of the profile of the equilibrium constant, which provides a physically more correct approach than that using the conventional indefinite equilibrium constant (EK) model.