Search PubMed⌕ Search

Biomedical subjects

S V Konev

Publications and source records attributed to S V Konev.

At least 19 recordsLinked to original sources
PubMed2004

[Determination of tyrosine- and tryptophan-containing peptides in plasma by absorption in UV spectral region].

A method for determining the cumulative concentration of tyrosine- and tryptophan-containing peptides (TSP and TPP) in blood plasma: 0.2 ml of blood plasma is added 0.1 ml of 1.8 M chloric acid, the sample is centrifuged, the supernatant (0.2 ml) is added 1.8 ml of 0.7 M NaOH and the optic density is measured at 290 nm in the quarts or plastic flask. Equations were derived to calculate separately the concentrations of TSP and TPP, whose normal values are 1.28 +/- 0.31 and 0.25 +/- 0.06 mmol/l, respectively. The increasing total content of peptides in thyrotoxicosis was shown to be mainly conditioned by a higher TSP concentration (a 2.4-fold increase). Therefore, TSP is a more sensitive marker in case of endogenous intoxication versus the generally known index of the cumulative content of oligopeptides.

Humans↗
PubMed2002

Fluorescent probing of the ligand-binding ability of blood plasma in the acute-phase response.

The acute-phase response alters the composition of carrier proteins in plasma, which may affect the blood deposition and transport of biomediators and drugs. The effect of the acute-phase response on the ligand binding ability of plasma was studied in leukemic children with and without systemic inflammation (sepsis and septic shock). To target different transport proteins, differentially charged fluorescent dyes were used: anionic ANS (8-anilinonaphthalene-1-sulfonate), uncharged Nile red, and cationic Quinaldine red. Human serum albumin was a principal carrier for ANS and competed for Nile red binding with lipoproteins. The synchro-scan fluorescence spectra of Nile red in plasma distinguished two species of the dye bound to serum albumin and to low-density and/or very low-density lipoproteins. The binding of Quinaldine red did not correlate with albumin and lipoprotein levels, and was probably determined by alpha(1)-acid glycoprotein. Compared with the control group, leukemia increased Quinaldine red binding by 65% and did not significantly affect the binding of other probes. Sepsis and septic shock did not change the binding of Quinaldine red, but progressively decreased ANS binding, finally by about 33%, and shifted Nile red distribution from serum albumin toward lipoproteins. These changes reflected a modified composition of the three principal transport proteins in plasma in the acute-phase response. Simple and rapid fluorescent tests developed in this study can be used to evaluate the acute-phase response and to optimize drug administration protocols in clinical practice.

Acute-Phase Reaction↗
PubMed2000

Bioenergetic response of isolated nerve terminals of rat brain to osmotic swelling.

The swelling of nerve terminals of rat brain in a hypotonic medium (230 mOsm) induced the potential-independent entrance of 45Ca2+ into synaptosomes and intrasynaptosomal mitochondria that changed the energy status of synaptosomes, the rate of O2 consumption and the content of ATP being decreased. The ratio ATP/ADP decreased from 6.5 +/- 0.26 (310 mOsm medium) to 3.1 +/- 0.18 (the medium 230 mOsm). Studies on the equilibrium distribution of K+ (86Rb+) and [3H]TPP+ showed that contents of these cations in the nerve terminals were virtually the same on incubation in both iso- and hypotonic media. This indicated that the swelling did not damage intrasynaptosomal mitochondria and plasma membranes of the synaptosomes. The inhibition of oxidative phosphorylation increased twofold the rate of glycolysis. The incubation of synaptosomes in calcium-free medium (230 mOsm) in the presence of EGTA (1 mM) prevented the inhibition of oxidative phosphorylation and synthesis of ATP by the osmotic swelling. Ruthenium Red (10 microM) in the medium 230 mOsm inhibited the entrance of 45Ca2+ into the intrasynaptosomal mitochondria and normalized the oxidative phosphorylation to the control level (310 mOsm medium). The decrease in the energy potential of synaptosomes induced by the hypoosmotic shock is suggested to be associated with the increase in Ca2+ content in the cytoplasm, its transport into the mitochondria, and the inhibitory effect on oxidative phosphorylation.

Adenosine Diphosphate↗
PubMed2000

Effects of nitric oxide donors on Ca2+-dependent [14C]GABA release from brain synaptosomes: the role of SH-groups.

Nitric oxide (NO) modulates processes of synaptic transmission at pre- and postsynaptic levels. In the present work we studied the mechanisms of action of NO on [gamma-14C]amino-n-butyric acid ([14C]GABA) release in rat cortical synaptosomes. NO donors--S-nitroso-L-cysteine and hydroxylamine (but not sodium nitroprusside)--inhibited the neurotransmitter efflux in a concentration range from 10 microM to 1 mM. Nitrosocysteine completely and selectively suppressed the Ca2+-dependent (vesicular) [14C]GABA release, while not affecting the Ca2+-independent component of the [14C]GABA transport. The influence of NO donors was not related to activation of guanylyl cyclase, since the membrane-permeable cGMP analog dibutyryl-cGMP did not mimic and the guanylyl cyclase inhibitor methylene blue did not change the NO effects. In contrast, the membrane-permeable SH-reagent N-ethylmaleimide (NEM) resembled the effects of NO donors on the Ca2+-dependent [14C]GABA release. The degree of inhibition of the release by nitrosocysteine, hydroxylamine, and NEM correlated with their ability to oxidize intra-synaptosomal SH-groups. These data suggest that synaptosomal sulfhydryl groups are the target for NO action at the presynaptic level. The NO-induced oxidation of thiols may be involved in physiological and, especially, pathological effects of nitric oxide in the central nervous system.

Animals↗
PubMed1999

Influence of plasma membrane depolarization on cAMP level in rat brain synaptosomes.

We studied the influence of plasma membrane depolarization on cAMP content in presynaptic nerve endings (synaptosomes) isolated from brain hemispheres (HS) and cerebellum (CS). Depolarization by elevated [K+]o decreased basal cAMP level in both types of synaptosomes; reduced cAMP content in HS and increased cAMP in CS in the presence of IBMX; and lowered forskolin-stimulated cAMP accumulation in both the HS and the CS. Similar results were obtained when depolarization was induced by veratrine or when [Ca2+]i was elevated by treatment of the synaptosomes with the ionophore A23187. In Ca2+-free media, depolarization was not able to affect the synaptosomal cAMP levels. These data suggest that in brain synaptosomes intracellular cAMP pathway is modulated by alterations in [Ca2+]i.

1-Methyl-3-isobutylxanthine↗
PubMed1999

Different modes of ozone-induced lipid oxidation in Candida utilis yeast cells and isolated membrane preparations.

Significant differences in the development of ozonolysis of lipids in membrane preparations and intact cells of the Candida utilis yeast were revealed. First, unlike isolated membranes, in which lipid modifications can be initiated by low ozone doses (< 0.5 micromol O3/mg protein) and develop proportionally to the treatment dose, in intact yeast cells, even the most ozone-sensitive sterols and nitrogen-containing phospholipids (phosphatidylcholine and phosphatidylethanolamine) did not undergo oxidative destruction at doses up to 6.0 micromol O3/mg protein. Second, the peculiarity of the ozone-initiated lipid modification in intact cells was that different classes of lipids exhibited different sensitivity to ozone. With an increase in the ozone dose, neutral lipids (sterols) and nitrogen-containing phospholipids (phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin) were modified to a greater extent. Third, the accumulation of lipid peroxidation products upon ozone treatment of cells, in contrast to the isolated membranes, was absent at low ozone doses and was recorded only after the lethal damage. It is suggested that these differences are related to both the function of antioxidative enzymes (catalase, superoxide dismutase, peroxidase, etc.) and the difference between the structural states (i.e., stability and accessibility to oxidation) of lipids in the isolated membranes and the intact cells.

Candida↗
PubMed1999

[Assessment of body intoxication as a balance between accumulation and binding of toxin in plasma].

A novel approach to assessing endogenic intoxication (EI) as an imbalance between toxin accumulation and binding by albumin in blood plasma is proposed. The intoxication criterion (IC) is determined by the ratio of the content of medium-weight molecules (MWM) and effective albumin concentration (EAC): IC = MWN/EAC. In children with oncohematological diseases the development of EI is associated with a 4-fold drop of MWM content and a twofold decrease of EAC, and hence, the imbalance between these two values increases 9-10 times. The proposed approach notably improves the sensitivity of diagnosis of the early stages of EI. Application of a new fluorescent marker pirrone red for measuring albumin binding capacity is validated and an algorithm of EAC determination using calibration curve of probe binding to standard albumin solution is described.

Humans↗
PubMed1998

Photoreactivation of the cytochrome oxidase complex with cyanide: the reaction of heme a3 photoreduction.

Electron transfer activity of isolated cytochrome oxidase inhibited by low concentrations of cyanide by 93-95% was shown to rise no less than three times under exposure to visible light. Irradiation with visible light was found to increase the rate of reduction of cytochrome oxidase heme groups in the presence of sodium dithionite. Based on these results, it is suggested that the modification of the catalytic and spectral characteristic of the cytochrome oxidase-cyanide complex is due to the photostimulation of the intramolecular electron transport at the interheme (heme a heme a3) transfer stage, i.e., is caused by photoreduction of the enzyme's heme a3-CN complex.

Animals↗
PubMed1998

Effect of calcium on the energy status of rat brain synaptosomes under acidosis.

Incubation of rat brain synaptosomes at pH 6.0 in Ca2+-containing medium is associated with a decrease in the ATP content and the rate of oxygen consumption. ATP/ADP ratio decreased from 6.6 +/- 0.24 at pH 7.4 to 3.2 +/- 0.17 at pH 6.0. The content of 86Rb+ and [3H]tetraphenylphosphonium measured at pH 7.4 did not change after preincubation at pH 6.0, indicating the absence of lesion of synaptosomal plasma membranes and intrasynaptosomal mitochondria. Incubation with 1 mM EGTA in Ca2+-free medium as well as addition of 1 mM ouabain or 10 microM ruthenium red prevents the effect of acidosis. Similar results were obtained when 5 mM pyruvate was used as a mitochondrial substrate instead of glucose. It is suggested that acidosis-induced decrease in the ATP level is associated with the increase in Ca2+ concentration in the cytoplasm and its transport into mitochondria. Ouabain reverses this process due to activation of Na+/Ca2+ exchange.

Acidosis↗
PubMed1997

Hypoosmotic shock activates Ca2+ channels in isolated nerve terminals.

Influence of hypotonic swelling on Ca2+ (45Ca2+) uptake in rat brain synaptosomes was studied. A decrease in medium osmolality from 310 to 260-180 mOsm led to a progressive stimulation of 45Ca2+ accumulation. The effect was blocked by verapamil (IC50 = 5 microM), CoCl2 (IC50 = 58 microM) and retained at a fixed concentration of external sodium indicating the involvement of Ca2+ channels rather than Na+/Ca2+ exchange in swelling-induced Ca2+ influx. The populations of calcium channels observed in hypoosmotic and depolarizing conditions are different in three aspects: (i) kinetics of 45Ca2+ entry; (ii) insensitivity to dihydropyridines and omega-conotoxin GVIA; (iii) insensitivity to preliminary depolarization by high potassium. The effects of swelling and depolarization on Ca2+ uptake were additive. No change in membrane potential monitored with diS-C3-(5) was recorded during synaptosome hypotonic swelling. The results suggest the existence in synaptosomal plasma membrane of volume-dependent calcium-permeable channels with properties distinct from those of the voltage-dependent calcium channels. Activation of these channels may constitute an early event in volume regulation of nerve terminals in anisoosmotic conditions.

Animals↗
PubMed1997

[The regulation of cell volume: the mechanisms of intracellular signalling].

The mechanisms of intracellular [correction of intercellular] signalling responsible for cell volume regulation are elucidated in the 2nd part of the review. Data on a nature of the "volume" sensor and signals acting on it, the properties of structures and mechanisms regulating the cell volume, are discussed as well as genetic regulation of the cell volume responses.

Animals↗
PubMed1996

Swelling-induced activation of Na+,K+,2Cl- cotransport in C6 glioma cells: kinetic properties and intracellular signalling mechanisms.

Swelling of C6 glioma cells in hypotonic medium (180 mOsm) results in two- to three-fold activation of K+ (86Rb+) influx suppressed by 10 microM bumetanide. Bumetanide-sensitive transport of 86Rb+ is dependent on extracellular K+, Na+ and Cl- both in iso-osmotic conditions and under hypo-osmotic shock, supporting the notion that it is mediated by Na+,K+,2Cl- cotransport. Inhibitors of protein kinase C (10 microM polymyxin B and l microM staurosporine) had no significant effect on basal cotransport but reduced its hypotonic stimulation by 70-80%. Similar results were obtained with calmodulin antagonist R24571 (10 microM), indicating Ca2+/calmodulin-dependence of the process. Influence of polymyxin B and R24571 was not additive. Swelling-activated Na+,K+,2Cl- cotransport was also suppressed by protein kinase C activator PMA (l microM). By contrast, preincubation of cells with inhibitors of protein phosphatases (100 microM vanadate, 5 mM fluoride and 0.5 microM okadaic acid) activated greatly the bumetanide-sensitive 86Rb+ uptake in isotonic conditions, while a subsequent hypotonic swelling led to smaller or no increment. These results indicate the involvement of Ca2+/calmodulin-dependent staurosporine/polymyxin B-sensitive protein kinase other than protein kinase C in swelling-induced activation of Na+,K+,2Cl- cotransport in glial cells.

Bumetanide↗
PubMed1996

[The effect of cytoskeleton modification on the process of trypsin-induced cell aggregation].

Studies were performed on the influence of cytoskeleton modification (by colchicine, vinblastine, and cytochalasin B, and by neoplastic transformation) on cell aggregation induced by small trypsin concentrations (the final concentrations equal to 1-100 mg/ml) added to a suspension of chick embryo fibroblasts dispersed from a monolayer with EDTA. It was shown that alteration of microfilaments resulted in the loss of the ability of plasma membranes to structural rearrangements. It is suggested that aggregation of these cells is realized due to a gradual exposure of adhesive sites along with proteolytic splitting of protein components of plasma membranes.

Animals↗
PubMed1995

Effect of proteolysis on the state of lipid phase in rat brain synaptosomal membranes.

Enzymatic proteolysis of the proteins of synaptic membranes has been found to be accompanied by the promotion of lipid peroxidation probably mediated by the liberation of membrane-bound iron. As fluorescent probes pyrene and diphenylhexatriene show, the microviscosity and micropolarity of membrane lipid phase rise as a result of lipid peroxidation. Different structural changes induced by proteolysis are displayed under inhibition of lipid peroxidation. Thus, the microviscosity of the bulk lipid phase appears to be lowered and the annular lipid microviscosity raised. Another explanation of the fluorescent data for annular lipids is the exclusion of pyrene molecules from this lipid pool, leading to a reduction of the probe local concentration. The changes observed in membrane lipid phase are considered as primary structural effects of proteolysis, not mediated by a phospholipase activation.

Animals↗
PubMed1994

Swelling-induced K+ influx in cultured primary astrocytes.

The effect of swelling of cultured primary astrocytes from rat brain in hypotonic medium on K+ influx has been studied. A decrease in osmolality from 310 to 180 mOsm increased the activity of sodium pump (ouabain-inhibited 86Rb+ influx) and Na+,K+,2Cl- cotransport (ouabain-insensitive bumetanide-inhibited 86Rb+ influx) by 70 and 35%, respectively. It is suggested that activation of these transport systems makes it possible to retain a high potassium concentration in the cells under regulatory volume decrease.

Animals↗
PubMed1994

Osmotic regulation of sodium pump in rat brain synaptosomes: the role of cytoplasmic sodium.

The effect of hypoosmolality of incubation medium on the rat of ouabain-sensitive 86Rb+ transport in rat brain synaptosomes was studied. A decreased osmolality from 310 to 250 mOsm increased the rate of 86Rb+ uptake from 3.72 to 6.23 nmol/mg of protein min. To evaluate the involvement of cytoplasmic sodium in sodium pump stimulation inhibitors of ion channels and transport pathways able to increase [Na+]in were used. Tetrodotoxin (1 microM), amiloride (0.5 mM) and verapamil (0.1 mM) had no influence on the osmotic response of the sodium pump. The decrease of sodium concentration in incubation medium to 15 mM, leading to a practical loss of its transmembrane gradient, did not abolish stimulation of pump. No increase in 22Na+ influx or intrasynaptosomal sodium content was registered at hypotonic conditions. It is suggested that osmotic regulation of Na+,K(+)-ATPase is not connected with an increase of internal sodium through opening of sodium channels, or with activation of other membrane sodium-transporting systems.

Animals↗