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Biomedical subjects

I N Kurochkin

Publications and source records attributed to I N Kurochkin.

At least 19 recordsLinked to original sources
PubMed2001

Bioelectrochemical analysis of neuropathy target esterase activity in blood.

Bioelectrochemical analysis of neuropathy target esterase (NTE) and its inhibitors is based on the combination of the NTE-catalyzed hydrolysis of phenyl valerate and phenol detection by a tyrosinase carbon-paste electrode. The use of the tyrosinase electrode improves 10-fold the sensitivity of NTE detection in comparison with a spectrophotometric method. The tyrosinase electrode was found to be suitable for measurements in whole human blood where spectrophotometric detection is considerably restricted. The specificity of NTE in blood for mipafox and di-2-propyl phosphorofluoridate was close to that for neuronal NTE. The NTE-like activity in blood was determined to be 0.19 +/- 0.02 nmol/min/mg of protein.

Animals↗
PubMed2000

Structural and functional properties of Langmuir films of antibodies based on amphiphilic polyelectrolytes.

We optimized the procedure for the formation of Langmuir films of antibodies based on amphiphilic polyelectrolytes and studied the physicochemical and immunochemical properties of the films obtained. Their immunochemical properties were compared with the immunochemical activity of antibodies in Langmuir films without amphiphilic polyelectrolytes and with antibodies adsorbed on the surface of polystyrene and graphite. The efficiency of immune adsorption by the films based on amphiphilic polyelectrolytes was shown to be greater; the affinity of antibodies and surface concentration of their active conformation depended on the type of amphiphilic polyelectrolytes used to obtain the films. We investigated the structure of these films at the surface of highly oriented pyrolytic graphite using the method of atomic force microscopy. Changes in the structure of the films under study caused by the increase of surface pressure were demonstrated.

Antibodies↗
PubMed1999

A new approach for determination of neuropathy target esterase activity.

Neuropathy target esterase (NTE) was shown to be an excellent biochemical marker for screening of organophosphates (OPs) with respect to their ability to result in organophosphate induced delayed neurotoxicity (OPIDN). This paper describes a new biosensor approach to the analysis of NTE and its inhibitors. The method is based on the combination of NTE enzymatic hydrolysis of phenyl valerate (PV) with phenol detection by the Clark-type oxygen electrode modified by immobilized tyrosinase. The validity of this biosensor method is confirmed by the facts that the calibration curves for NTE obtained by colorimetric and flow-through electrochemical methods were nearly identical and the titration of NTE by test inhibitor mipafox was shown to yield the same pI50 values. The developed electrochemical methods can be considered as a promising approach both for serial express NTE analysis and for kinetic characteristics of NTE.

Animals↗
PubMed1998

Kinetic behavior of a receptor-enzyme system: a model for drug addiction.

A theoretical kinetic model describing the behavior of a receptor-enzyme system during formation of drug addiction is considered in this article. The model assumes concomitant action of narcotic on at least two targets with opposite effects. Theoretical kinetic principles that the system must satisfy for the development of drug addiction are formulated. These kinetic principles are the slow inactivation of receptor-enzyme system and the divergence of characteristic times of dynamic concentration of product and enzyme.

Enzymes↗
PubMed1998

STM studies of LB films of amphiphilic polyelectrolytes with antibodies and enzymes.

The structure of LB films of protein-polyelectrolyte complexes transferred onto the pyrographite surface was studied by STM. The images of the films obtained at various protein concentrations in the water subphase and different values of surface pressure were captured. The topology of the surface covered by one or three layers of the films studied was investigated. It is shown that at a protein concentration of 1 mg/ml in the water subphase, the films are composed of aggregated protein molecules, and their structure has an insular character. An increase in the number of transferred layers up to three results in a virtually complete covering of the surface. A decrease in the protein concentration in the water subphase to 1 microgram/ml enabled us to prepare films consisting of individual non-aggregated protein molecules.

Antibodies↗
PubMed1997

Langmuir films from antibodies based on amphiphilic polyelectrolytes.

A technique for forming Langmuir films from antibodies based on an amphiphilic polyelectrolyte was developed. The physicochemical and immunochemical properties of the Langmuir films obtained were studied. The interaction of HBsAg with the films was found to be described by a model with one binding site, whereas that of HBsAg with antibodies adsorbed on a polystyrene plate, by a model with a positive cooperativity. The use of the novel Langmuir films from antibodies increases the sensitivity of the immunoenzyme assay.

Animals↗
PubMed1996

Potentiometric biosensors for cholinesterase inhibitor analysis based on mediatorless bioelectrocatalysis.

A potentiometric method for cholinesterase inhibitor analysis based on mediatorless bioelectrocatalysis has been developed. The method includes coimmobilization of three enzymes, butyrylcholinesterase, choline oxidase and peroxidase, on composite carbon electrodes. Catalytic hydrolysis of butyrylcholine and subsequent catalytic oxidation of choline result in the formation of hydrogen peroxide leads to a shift in the electrode potential. The detection limit for trichlorfon analysis is 2 x 10(-13) M. Electrodes remain stable for at least 4 weeks when stored at 277 K.

Biosensing Techniques↗
PubMed1994

Glucose potentiometric electrodes based on mediatorless bioelectrocatalysis. A new approach.

A potentiometric method has been developed for analysis of glucose on the basis of glucose oxidase and peroxidase co-immobilized on the surface of an electrode made of composite carbonic material. Catalytic oxidation of glucose results in the formation of hydrogen peroxide. Mediatorless peroxidase catalysis of electroreduction of hydrogen peroxide leads to a shift in the electrode potential. The rate of the increase of the electrode potential is proportional to glucose concentration and calibration curve is linear at 0.025-2.0 mM. Electrodes permit at least 100 measurements without any loss in the activity. Electrodes remain stable for 90 days when stored at 277 K.

Biosensing Techniques↗
PubMed1993

Affinity biosensors based on preconcentration/voltammetric analysis. Detection of phenothiazine drugs at Langmuir-Blodgett films of tyrosine hydroxylase.

A new route for operating affinity biosensors based on the voltammetric monitoring of the accumulated guest (analyte) is described. High sensitivity and selectivity accrue from the coupling of the specific receptor binding process and the inherent sensitivity of the preconcentration/pulse-voltammetric scheme. The redox (measurement) process results in dissociation of the receptor-guest complex, thus allowing multiple analytical determinations. The receptor layer also serves as an effective barrier that excludes interfering species. The new concept of preconcentration/voltammetric affinity biosensors is illustrated in connection with the detection of phenothiazine drugs using Langmuir-Blodgett films of their receptor, the enzyme tyrosine hydroxylase. The effect of various experimental variables upon the sensor performance is described.

Biosensing Techniques↗
PubMed1992

Receptor binding on whole cells can oscillate.

The study of the kinetics of binding of the opiate receptor agonist [3H]DADLE with NG108-15 cell suspensions has revealed a new periodic biological phenomenon, i.e., oscillations of the cellular receptor activity. The absence of oscillations for binding of the receptor antagonist shows that oscillations occur as a result of the transformation of the receptor signal only.

Animals↗
PubMed1992

[Receptor binding oscillations].

A kinetic analysis of the delta-opiate receptor agonist, [3H] DADLE, binding to cell (NG108-15) suspensions has led to the discovery of a new periodic biological phenomenon, namely, receptor activity oscillations. The absence of oscillations for the antagonist binding to the receptors suggests that oscillations are generated only as a result of receptor signal transformation. The correlation between the quantitative characteristics of the kinetic curves and the experimental conditions points to the fact that receptor binding oscillations may be due either to the receptor binding to the G-protein or the interaction of the receptor (or G-protein) with microtubules.

Cells, Cultured↗
PubMed1991

The 65-kDa protein from pig heart. A new substrate for Clostridium botulinum ADP-ribosyltransferase (exoenzyme C3).

In the pig heart sarcolemma, a 65 kDa protein is found to be ADP-ribosylated by Clostridium botulinum ADP-ribosyltransferase (exoenzyme C3). ADP-ribosylation of this protein is regulated by guanyl nucleotides and cytosol factor in a fashion similar to that for other C3 substrates. The new exoenzyme C3 substrate was partially purified. This protein is supposed to be a GTP-binding one.

ADP Ribose Transferases↗
PubMed1991

[Tyrosine hydroxylase activity in two-dimensional monomolecular films].

The activity of a neurospecific enzyme of tyrosine hydroxylase (TH) in monomolecular films formed onto solid surface was studied. The data obtained show that the formation of two-dimensional films onto negative-charge surfaces by the Langmuir-Schafer technology does not lead to the inactivation of the enzyme. Neuroleptic trifluoperazine increased the activity of TH. Monomolecular films of TH may be used as a sensitive element of biosensors for primary monitoring of neuroleptic-like compounds.

Animals↗
PubMed1990

[The effect of delta-9-tetrahydrocannabinol on the receptor and physicochemical properties of the membranes in the rat brain].

It has been demonstrated that delta-9-THC does not affect the specific binding of 3H-IQNB, 3H-DAGO and 3H-dihydroalprenolol, decreases the level of specific binding 3H-LSD and 3H-spiperone, a 2-3-fold increase in the total and nonspecific binding being observed in this case, and also increases the microviscosity of the rat obtain membranes and disrupts lipid-protein interactions. Increasing the microviscosity of membranes by other method (lipid peroxidation) differently affects the binding of radioactively labeled ligands with the membranes from rat brain.

Animals↗
PubMed1990

[Kinetic characteristics of the effect of GTP on properties of opioid receptors].

Specific interactions of guanine nucleotides with rat brain membrane preparations were investigated; a two-step scheme of this interaction was proposed. The effect of GTP on the agonist binding to rat brain opioid receptors was studied. Nucleotides were shown to inhibit the ligand interactions with mu- and sigma-receptors. It was found that the GTP-induced inhibition differs from that of the GTP stable analog, GppNp, i.e., GTP does not initiate any noticeable dissociation of the receptor system component and its liberation from the membrane into solution as is the case with GppNp. A kinetic model of the GTP interactions with the opioid receptor system stipulating that the affinity of high affinity centers for mu- and delta-ligands changes after nucleotide hydrolysis on the N-protein, was proposed. The differences in the mechanisms of GTP effect on mu- and sigma-receptors were revealed, i.e., the inhibition of the ligand binding to mu-receptors may occur just after GTP binding to the N-protein. The ligand-receptor interactions in the presence of GTP were modelled according to the proposed scheme.

Adenine Nucleotides↗
PubMed1989

[Lyophilized membranes as a test system for analyzing the properties of neuromediator receptors and activity of neurotropic agents].

The effect of lyophilization and prolonged storage of rat brain membrane preparations on the properties of receptors of cholinergic and adrenergic neuromediator systems has been studied by the radioreceptor assay. Lyophilized membrane preparations have been shown to be highly stable as compared with fresh membranes and retain their binding properties during storage for 1 year.

Animals↗
PubMed1988

Analysis of ligand-receptor binding by the difference method.

A new method has been proposed for analysis of experimental data on ligand-receptor binding at equilibrium. This method makes it possible to detect heterogeneity of a receptor system in cases where the contribution of the high-affinity site to total binding is rather small and the problem of graphic discrimination of a model cannot be solved unambiguously by other methods. The difference method permits us to exclude experiments on measuring nonspecific binding. A computer program for analysis of ligand-receptor binding has been worked out in which the difference method and traditional methods of binding isotherm analysis are realized. Numerical modeling has shown that the best strategy in experimental data processing is the treatment of total binding isotherms by both the difference method and regression analysis, including the nonspecific binding constant as one of the regression parameters.

Kinetics↗