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Biomedical subjects

O Vinogradova

Publications and source records attributed to O Vinogradova.

At least 19 recordsLinked to original sources
PubMed2004

Effect of two-week tail suspension on forelimb and hindlimb [correction of hindlimd] small arteries in rats.

This study was aimed at comparing the effect of 2-week tail suspension on the characteristics of median artery and saphenous artery isolated from forelimb and hindlimb, accordingly. The segments of arteries were mounted in two-channel myograph for recording their isometric wall tension. In post-suspension rats segments of saphenous artery showed augmented sensitivity to noradrenaline and serotonin. More than two-fold decreased of EC50 was observed both before and after endothelium removal. Along with that, the sensitivity of median artery segments to vasoconstrictors was similar in post-suspension and control rats. However, the diameter of relaxed median artery in post-suspension rats was increased. Relaxation to acetylcholine compared to controls was augmented in both vascular regions of post-suspension rats being more pronounced in saphenous artery preparations.

Acetylcholine↗
PubMed2004

Neurogenic constrictor response of isolated small renal arteries in rats after 2-week simulated microgravity.

The study was aimed at investigation of the effects of 2-week tail suspension upon the constrictor responses of isolated small renal arteries in rats. 1st-2nd-order branches of renal artery were perfused with saline under the constant flow conditions. Constrictor responses to electrical stimulation of periarterial nerves, noradrenaline and serotonin were investigated. In post-suspension rats as compared to controls the response to nerve stimulation was slightly reduced during 15-Hz stimulation, but similar at smaller frequencies. Thus, simulated microgravity has no prominent effect of neurogenic responses of renal vessels, in agreement with non-changed density of periarterial adrenergic nerve plexus. Along with that, in post-suspension rats impairment of prejunctional sympathetic mechanisms might be compensated by augmented sensitivity of vascular smooth muscle to vasoconstrictors.

Animals↗
PubMed2004

Hemodynamic changes in post-suspension rats during gradual hemorrhage.

We examined the changes of hemodynamic parameters in nembutal-anesthetized rats during gradual hemorrhage (2 ml/100 g body weight during 30 min). In control rats blood pressure began to decline starting from 3rd min of bleeding and from 5th min it was accompanied by cardiac deceleration. Hindlimb vascular resistance was only slightly increased up to 15th min (by 20-30%) and then began to grow drastically. Less prominent changes of hemodynamics were observed in post-suspension rats. The results indicate that when activity of sympathetic nervous system is blunted with anesthetic post-suspension rats demonstrate higher hemodynamic stability during acute hemorrhage.

Animals↗
PubMed2002

Long-term dynamics of blood pressure in intact and sympathectomized post-suspension rats.

Low-frequency dynamics of mean arterial pressure (MAP) and pulse interval (PI) was studied in intact and sympathectomized rats after 2-week tail suspension. Correlation properties of long-term (f<0.8 Hz) MAP fluctuations were analyzed using R/S Hurst method. It was shown that time series of successive increments in MAP level display power-law fractal anticorrelations in wide range of time scales. After sympathectomy and/or suspension averaged Hurst exponents were more close to the value for uncorrelated random process (H=0.5), indicating deterioration of regulatory feedbacks. Besides, a decrease of coherence between MAP and PI in low-frequency range was observed after suspension in both intact and sympathectomized rats, indicating alteration of non-sympathetic cardiovascular control mechanisms.

Journal Article↗
PubMed2001

Effect of tail suspension on haemodynamics in intact and sympathectomized rats.

The effect of a 2-week tail suspension on mean arterial pressure (MAP) and pulse interval (PI) was studied in conscious chronically instrumented intact and sympathectomized rats. Sympathectomy was induced by 6-hydroxydopamine (100 mg x kg(-1), i.v.), injected 1 day before, and on days 5 and 10 of suspension. During suspension the intact rats had the same levels of MAP and PI as the intact controls. After release from suspension their MAP did not change: PI decreased (by 9%), but only for 1 h. Sympathectomy augmented the haemodynamic changes after release from suspension: tachycardia (10%) and hypotension (19%) were observed during the entire recording period (2 h). Therefore, in rats, post-suspension hypotension becomes apparent only after elimination of sympathetic influences. In spite of unaltered systemic parameters, intact post-suspension rats showed diminished responses to intravenous administration of tyramine (100 microg x kg(-1)) and phenylephrine (1-2 microg x kg(-1)). In addition, they showed augmented haemodynamic changes associated with natural behaviour. The increase of MAP and the decrease of PI on transition from rest to movement were more pronounced in the intact post-suspension rats than in the control rats (MAP 8.3% vs 4.5%, PI 15% vs 9%). In control rats the spectrum power density of low-frequency (0.0195-0.25 Hz) MAP fluctuations depended only weakly on behavioural activity, whereas in post-suspension intact rats it was profoundly augmented during movements (by 170%). Since the rapid adjustment of haemodynamics to behavioural activity is controlled by the sympathetic nervous system, the unbalanced condition of the cardiovascular system after suspension may reflect an altered sympathetic nerve control of the circulation.

Animals↗
PubMed2001

Oscillatory model of novelty detection.

A model of novelty detection is developed which is based on an oscillatory mechanism of memory formation and information processing. The frequency encoding of the input information and adaptation of natural frequencies of network oscillators to the frequency of the input signal are used as the mechanism of information storage. The resonance amplification of network activity is used as a recognition principle for familiar stimuli. Application of the model to novelty detection in the hippocampus is discussed.

Animals↗
PubMed2001

The neurogenic constrictor response of isolated rat saphenous artery is reduced after 2-week tail suspension.

Under real or simulated microgravity conditions the control of arterial vascular tone is greatly disturbed. The low arterial vessel reactivity to sympathetic influences may be the cause of an increase in flow in hind limb skeletal muscles in tail-suspended (TS) rats. Our previous experiments with constant pressure perfusion of rat hind limb demonstrated the reduced vasoconstrictor responses to sympathetic nerve stimulation in TS rats. Responses to exogenous noradrenaline depended on the perfusion conditions. It is known that the vessels of various branching orders noticeably differ in nerve density and in sensitivity to vasoconstrictor agonists. So under neurogenic or exogenous noradrenaline influences the vascular resistance may be increased at different levels of vascular bed, thus making the data analysis seriously complicated. This uncertainty may be overcome by investigation of a single vessel isolated from hind limb vascular bed. The saphenous artery, a resistance artery with dense innervation, is a very convenient object for this purpose. Thus, this study was aimed at comparing the effects of 2-week tail suspension upon the constrictor responses of isolated saphenous artery to neurogenic and exogenous noradrenaline stimuli in rats.

Animals↗
PubMed2000

A structural basis for integrin activation by the cytoplasmic tail of the alpha IIb-subunit.

A key step in the activation of heterodimeric integrin adhesion receptors is the transmission of an agonist-induced cellular signal from the short alpha- and/or beta-cytoplasmic tails to the extracellular domains of the receptor. The structural details of how the cytoplasmic tails mediate such an inside-out signaling process remain unclear. We report herein the NMR structures of a membrane-anchored cytoplasmic tail of the alpha(IIb)-subunit and of a mutant alpha(IIb)-cytoplasmic tail that renders platelet integrin alpha(IIb)beta(3) constitutively active. The structure of the wild-type alpha(IIb)-cytoplasmic tail reveals a "closed" conformation where the highly conserved N-terminal membrane-proximal region forms an alpha-helix followed by a turn, and the acidic C-terminal loop interacts with the N-terminal helix. The structure of the active mutant is significantly different, having an "open" conformation where the interactions between the N-terminal helix and C-terminal region are abolished. Consistent with these structural differences, the two peptides differ in function: the wild-type peptide suppressed alpha(IIb)beta(3) activation, whereas the mutant peptide did not. These results provide an atomic explanation for extensive biochemical/mutational data and support a conformation-based "on/off switch" model for integrin activation.

Amino Acid Sequence↗
PubMed2000

NMR-Based amide hydrogen-deuterium exchange measurements for complex membrane proteins: development and critical evaluation.

A method for measuring site-specific amide hydrogen-deuterium exchange rates for membrane proteins in bilayers is reported and evaluated. This method represents an adaptation and extension of the approach of Dempsey and co-workers (Biophys. J. 70, 1777-1788 (1996)) and is based on reconstituting (15)N-labeled membrane proteins into phospholipid bilayers, followed by lyophilization and rehydration with D(2)O or H(2)O (control). Following incubation for a time t under hydrated conditions, samples are again lyophilized and then solubilized in an organic solvent system, where (1)H-(15)N HSQC spectra are recorded. Comparison of spectra from D(2)O-exposed samples to spectra from control samples yields the extent of the H-D exchange which occurred in the bilayers during time t. Measurements are site specific if specific (15)N labeling is used. The first part of this paper deals with the search for a suitable solvent system in which to solubilize complex membrane proteins in an amide "exchange-trapped" form for NMR quantitation of amide peak intensities. The second portion of the paper documents application of the overall procedure to measuring site-specific amide exchange rates in diacylglycerol kinase, a representative integral membrane protein. Both the potential usefulness and the significant limitations of the new method are documented.

Amides↗
PubMed2000

Regulation of hemodynamics in sympathectomized rats after adaptation to tail suspension.

The organism adapts to actual or simulated microgravity by complex interactions of nervous, hormonal and local control mechanisms. Sympathetic nervous system is believed to play the leading role in this adaptation (Robertson et al. 1994). However, this conclusion seems to be rather deductive, as it has not been proved directly. Chronic sympathectomy provides a straightforward approach to this problem. We have studied the role of sympathetic nervous system in adaptation of cardiovascular system to simulated microgravity by tail suspension in intact and sympathectomized rats.

Animals↗
PubMed1999

Dynamics of blood pressure and pulse interval duration in rats after 14 day suspension.

At present, the pronounced changes in cardiovascular system observed in microgravity and on returning to 1 g are mostly attributed to the regulatory disturbances. The tail suspension model is used to simulate the effects of microgravity on muscle, bone and, on cardiovascular system of rats. Having in mind all limitations of this method, we decided to use it for analysis of the acute effects of cessation of tail suspension on regulation of cardiovascular system in rats.

Animals↗
PubMed1998

A membrane setting for the sorting motifs present in the adenovirus E3-13.7 protein which down-regulates the epidermal growth factor receptor.

The adenovirus E3-13.7 protein interferes with endosomal protein sorting to down-regulate the epidermal growth factor receptor and related tyrosine kinase receptors. The cytoplasmic C terminus of this protein contains three protein sorting motifs which are related to the function of E3-13.7. In this study, the structure of a 23-residue polypeptide corresponding to this domain was examined using solution NMR and CD spectroscopic methods. The peptide was observed to exist in a mostly random structural state in aqueous solution but underwent high affinity association with dodecylphosphocholine micelles, where it adopted an ordered structure. The affinity of this peptide for the micellar surface and the structure of the bound peptide were independent of pH variation, surface charge, or attachment of a myristoyl anchor to the N-terminal. Studies with phospholipid vesicles suggested that the micellar structural results can be extrapolated to a true lipid bilayer. On the micellar surface all three sorting motifs are closely associated with the water/apolar interface: 72-YLRH and 87-LL lie within interfacial amphipathic helices, while 76-HPQY is non-helical and dimples just above the surface. These results contribute to the development of an understanding of the basis for specificity in recognition of sorting motifs by components of the cellular protein trafficking machinery.

Adenovirus E3 Proteins↗
PubMed1998

Structural characterization and immunochemical detection of a fluorophore derived from 4-hydroxy-2-nonenal and lysine.

Aging and the progression of certain degenerative diseases are accompanied by increases in intracellular fluorescent material, termed "lipofuscin" and ceroid, respectively. These pigments are observed within granules composed, in part, of damaged protein and lipid. Modification of various biomolecules by aldehyde products of lipid peroxidation is believed to contribute to lipofuscin and ceroid formation. However, little direct evidence currently exists because the structures responsible for the fluorescent, cross-linked nature of this material are not well characterized. In this study, we have identified a fluorescent product formed in the reaction of Nalpha-acetyllysine and 4-hydroxy-2-nonenal (HNE), a major product of lipid peroxidation and the most reactive of these compounds under physiological conditions [Esterbauer, H., Shaur, R. J. & Zollner, H. (1991) Free Radical Biol. Med. 11, 81-128]. This fluorescent compound, characterized as a 2-hydroxy-3-imino-1,2-dihydropyrrol derivative, appears to form upon oxidative cyclization of the nonfluorescent 2:1 lysine-HNE Michael adduct-Schiff base cross-link. Polyclonal antibody was raised to the Nalpha-acetyllysine-HNE fluorophore and found to be highly specific to the chromophore structure of the compound. This antibody has been used to conclusively demonstrate that the lysine-HNE derivative of this fluorophore forms on protein upon exposure to HNE. The results of this study therefore provide the basis for future investigations on the contribution(s) of HNE-derived fluorophore formation to lipofuscin and ceroid accumulation.

Aldehydes↗
PubMed1998

On choosing a detergent for solution NMR studies of membrane proteins.

Translational diffusion coefficients and catalytic activities were measured for the integral membrane protein diacylglycerol kinase (DAGK) in a variety of types of detergent micelles. Despite the structural diversity of the detergents examined, the translational diffusion coefficients observed for DAGK spanned a fairly limited range of values: 2.7 to 4.7 (x10(-7) cm2/s). No general correlation was observed between the diffusion coefficients for the detergent-DAGK aggregates and the sizes of the corresponding protein-free micelles. These results indicate that the effective molecular weights of the DAGK-detergent aggregates were determined more by the structural properties of the protein than by the properties of the detergents. The catalytic activity of DAGK in detergents having medium-length alkyl chains such as dodecylphosphocholine or decylmaltoside was usually observed to be substantially higher than in short-chain detergents such as octylphosphocholine or octylglucoside. Taken together, the diffusion and activity results indicate that medium-chain detergents are generally preferred for use in NMR studies of complex membrane proteins because they are no worse than short-chained detergents in terms of increasing the effective molecular weight of the protein of interest while they are considerably better at maintaining native-like protein conformation. Among the 10 detergents examined, only sodium dodecylsulfate was observed to be unable to support DAGK activity under any conditions examined, suggest that this well-known protein denaturant should be used with care in studies of complex membrane proteins.

Detergents↗
PubMed1997

Escherichia coli diacylglycerol kinase: a case study in the application of solution NMR methods to an integral membrane protein.

Diacylglycerol kinase (DAGK) is a 13-kDa integral membrane protein that spans the lipid bilayer three times and which is active in some micellar systems. In this work DAGK was purified using metal ion chelate chromatography, and its structural properties in micelles and organic solvent mixtures studies were examined, primarily to address the question of whether the structure of DAGK can be determined using solution NMR methods. Cross-linking studies established that DAGK is homotrimeric in decyl maltoside (DM) micelles and mixed micelles. The aggregate detergent-protein molecular mass of DAGK in both octyl glucoside and DM micelles was determined to be in the range of 100-110 kDa-much larger than the sum of the molecular weights of the DAGK trimers and the protein-free micelles. In acidic organic solvent mixtures, DAGK-DM complexes were highly soluble and yielded relatively well-resolved NMR spectra. NMR and circular dichroism studies indicated that in these mixtures the enzyme adopts a kinetically trapped monomeric structure in which it irreversibly binds several detergent molecules and is primarily alpha-helical, but in which its tertiary structure is largely disordered. Although these results provide new information regarding the native oligomeric state of DAGK and the structural properties of complex membrane proteins in micelles and organic solvent mixtures, the results discourage the notion that the structure of DAGK can be readily determined at high resolution with solution NMR methods.

Amino Acid Sequence↗
PubMed1997

Effect of 3-week suspension on sympathetic vasoconstrictor response in rat.

One of the main problems arising after gravitational unloading is an orthostatic intolerance leading to failure in supporting the upright posture and performing natural locomotion. Among a number of causes for the orthostatic intolerance the decreased circulating blood volume, increased venous distention, alterations in microcirculation, loss of muscular tonus, and regulatory disturbances could be mentioned. The later cause has been intensively studied recently. The aim of the present study is to examine the alterations induced by simulated gravitational unloading in the reaction of resistance vessels of isolated hind limb to the sympathetic stimuli in rats.

Animals↗
PubMed1996

Escherichia coli diacylglycerol kinase is an alpha-helical polytopic membrane protein and can spontaneously insert into preformed lipid vesicles.

Escherichia coli diacylglycerol kinase (DAGK) is a 13.2 kDa enzyme which spans the cytoplasmic membrane three times. Functional DAGK was purified to homogeneity using a polyhistidine tag and Ni(II)-chelate chromatography. Transmission Fourier transform infrared spectroscopy (FT-IR) of DAGK in phosphatidylcholine multilayers led to the conclusion that > or = 90 of DAGK's native 121 residues are alpha-helical, consistent with a model in which DAGK consists of two amphipathic alpha-helices and three transmembrane helices. Polarized attenuated total reflection FT-IR studies of DAGK in oriented multilamellae yielded data consistent with a topological arrangement in which the three transmembrane helices are well-aligned with the bilayer normal while the two amphipathic helices are approximately parallel with the membrane plane. The ability of DAGK to spontaneously insert into preformed lipid vesicles was examined using a novel assay system involving DAGK-catalyzed phosphorylation of a fluorescently tagged diacylglycerol. When micellar DAGK is diluted into L alpha-phase vesicles spontaneous insertion of the enzyme is fairly efficient (ca. 30%). DAGK refolding and insertion from delipidated urea-solubilized DAGK into lipid vesicles is also modestly efficient (3.8 +/- 2.1%) above the gel to liquid crystalline phase transition temperature. The insertion studies indicate that the difference in energy barriers (delta delta G++) between pathways leading to catalytically productive folding and insertion of DAGK relative to unproductive pathways is < 4 kcal/mol. However, additional studies carried out with mutant forms of DAGK indicated that the differences between refolding/insertion pathways for DAGK in vivo and in vitro can be significant.

Amino Acid Sequence↗