PubMed Health⌕ Search

Biomedical subjects

V A Bondarenko

Publications and source records attributed to V A Bondarenko.

At least 19 recordsLinked to original sources
PubMed2002

[The assessment of radiation hazards in the "MIR" and ISS orbits from the data of vehicle and personal dosimetric monitoring].

The paper describes the procedure of estimating total radiation risk to crewmembers during lifetime and possible lifetime reduction in consequence of participation in the Mir and ISS missions in different periods of the solar activity. The procedure includes analysis of data of vehicle and personal dosimetry, and calculations of radiation doses in various Mir compartments and accumulated by body tissues of cosmonauts. Calculated doses showed good consistency with the doses measured with R-16 on board Mir and personal dosimeters. To a first approximation, estimation of doses to cosmonauts and radiation risk as a result of participation in ISS missions took into account similarity of the Mir and ISS basal modules (geometry, dimensions and mass values) and was performed with the use of the space station shielding model that had been described elsewhere. The model of ISS radiation shielding will be updated as data of dosimetry of ISS compartments and phantom studies are available.

Astronauts↗
PubMed2001

Inhibition of retinal guanylyl cyclase by the RGS9-1 N-terminus.

Cyclic GMP plays a key role in retinal phototransduction and its photoreceptor concentration is precisely controlled by the cooperative action of cGMP phosphodiesterase (PDE) and retinal guanylyl cyclase (retGC). However, studies of the relationship between these two systems have focused only on a Ca(2+)-mediated, indirect connection. Using a retinal "regulator of G-protein signaling" (RGS9-1) and its fragments, we show that the N-terminus of RGS9-1 inhibits retGC activity. We also indicate that the GGL domain and/or the RGS domain function as an internal suppressor against the N-terminus, suggesting that proteins bound to these domains regulate the inhibitory activity of the N-terminus. Direct interaction of retGC with RGS9-1 and its N-terminus is also proved by immunoprecipitation and an overlay technique. Since RGS9-1 also controls the lifetime of transducin-activated PDE through regulating GTPase activity of transducin, this study strongly suggests that RGS9-1 mediates the direct interaction between PDE and retGC systems, and that this ingenious mechanism plays an important role in tuning of cGMP concentration in photoreceptors.

Amino Acid Sequence↗
PubMed2000

Phosphorylation by cyclin-dependent protein kinase 5 of the regulatory subunit of retinal cGMP phosphodiesterase. I. Identification of the kinase and its role in the turnoff of phosphodiesterase in vitro.

Cyclic GMP phosphodiesterase (PDE) is an essential component in retinal phototransduction. PDE is regulated by Pgamma, the regulatory subunit of PDE, and GTP/Talpha, the GTP-bound alpha subunit of transducin. In previous studies (Tsuboi, S., Matsumoto, H. , Jackson, K. W., Tsujimoto, K., Williamas, T., and Yamazaki, A. (1994) J. Biol. Chem. 269, 15016-15023; Tsuboi, S., Matsumoto, H., and Yamazaki, A. (1994) J. Biol. Chem. 269, 15024-15029), we showed that Pgamma is phosphorylated by a previously unknown kinase (Pgamma kinase) in a GTP-dependent manner in photoreceptor outer segment membranes. We also showed that phosphorylated Pgamma loses its ability to interact with GTP/Talpha, but gains a 10-15 times higher ability to inhibit GTP/Talpha-activated PDE than that of nonphosphorylated Pgamma. Thus, we propose that the Pgamma phosphorylation is probably involved in the recovery phase of phototransduction through shut off of GTP/Talpha-activated PDE. Here we demonstrate that all known Pgammas preserve a consensus motif for cyclin-dependent protein kinase 5 (Cdk5), a protein kinase believed to be involved in neuronal cell development, and that Pgamma kinase is Cdk5 complexed with p35, a neuronal Cdk5 activator. Mutational analysis of Pgamma indicates that all known Pgammas contain a P-X-T-P-R sequence and that this sequence is required for the Pgamma phosphorylation by Pgamma kinase. In three different column chromatographies of a cytosolic fraction of frog photoreceptor outer segments, the Pgamma kinase activity exactly coelutes with Cdk5 and p35. The Pgamma kinase activity ( approximately 85%) is also immunoprecipitated by a Cdk5-specific antibody, and the immunoprecipitate phosphorylates Pgamma. Finally, recombinant Cdk5/p35, which were expressed using clones from a bovine retina cDNA library, phosphorylates Pgamma in frog outer segment membranes in a GTP-dependent manner. These observations suggest that Cdk5 is probably involved in the recovery phase of phototransduction through phosphorylation of Pgamma complexed with GTP/Talpha in mature vertebrate retinal photoreceptors.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed2000

[Radiation environment of orbital complex "Mir" during minimum of the 22-nd solar cycle (1994-1996)].

Analysis of radiation dynamics on board the orbital complex MIR in the period of 1994 through to 1996 overlapping minimum of the 22nd solar cycle (SC) was performed. Radiation parameters in the working compartments during eight MIR missions (MIR 15-22) were evaluated. According to the solar dynamics during minimal activity with the lowest meanings of Wolf numbers (< 20), the daily average dose rate on MIR reached its peak over the whole 22nd SC and by a factor of more than three exceeded meanings typical for solar maximum. A statistical method was used to compute correlation coefficients and set up an equation of linear regression of the absorbed dose with cosmophysical parameters of the SC minimum. Meanings of the correlation coefficients for SC minimum were substantially lower as compared with SC maximum, that is there was no distinct correlation between dose rate and the parameters under study. Radiation doses to cosmonauts violated the admissible radiation limits for ground exposures but not radiation limits established for manned space missions. Reduction of dose loading on cosmonauts during SC minimum can be achieved by planning shorter missions or utilization of a program of radiation protection for traverse of SAA ERB.

Astronauts↗
PubMed1999

Suppression of GTP/T alpha-dependent activation of cGMP phosphodiesterase by ADP-ribosylation by its gamma subunit in amphibian rod photoreceptor membranes.

Our previous study has shown that P gamma, the regulatory subunit of cGMP phosphodiesterase (PDE), is ADP-ribosylated by endogenous ADP-ribosyltransferase when P gamma is free or complexed with the catalytic subunits of PDE in amphibian rod photoreceptor membranes. The P gamma domain containing ADP-ribosylated arginines was shown to be involved in its interaction with T alpha, a key interaction for PDE activation. In this study, we describe a possible function of the P gamma ADP-ribosylation in the GTP/T alpha-dependent PDE activation. When rod membranes were preincubated with or without NAD and washed with a buffer containing GTP, the PDE activity of NAD-preincubated membranes was increased by the GTP-washing only to approximately 50% of that of membranes preincubated without NAD. The P gamma release by the GTP-washing from these NAD-preincubated membranes was also suppressed to approximately 50% of that preincubated without NAD. Taking into consideration that approximately 50% of P gamma is ADP-ribosylated under these conditions, these observations suggest that the ADP-ribosylated P gamma cannot interact with GTP/T alpha. We have also shown that a soluble fraction of ROS contains an enzyme(s) to release the radioactivity of [32P]ADP-ribosylated P gamma in concentration- and time-dependent manners, suggesting that the P gamma ADP-ribosylation is reversible. Rod ADP-ribosyltransferase solubilized from membranes by phosphatidylinositol-specific phospholipase C was separated into two fractions by ion-exchange columns. Biochemical characterization of these two fractions, including measurement of the Km for NAD and P gamma, estimation of their molecular masses, ADP-ribosylation of P gamma arginine mutants, effects of ADP-ribosyltransferase inhibitors on the P gamma ADP-ribosylation, and effects of salts and pH on the P gamma ADP-ribosylation, indicates that rod ADP-ribosyltransferase contains two isozymes, and that these two isozymes have similar properties for the P gamma ADP-ribosylation. Our observations strongly suggest that the negative regulation of PDE through the reversible P gamma ADP-ribosylation may function in the phototransduction mechanism.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1999

Solar particle events observed on MIR station.

Radiation impact of the SPEs on board the MIR space station and in the interplanetary space is discussed in the report. The data of the on-board radiation dosimeter R-16 were used to measure the SPE absorbed doses. Some of SPEs (such as September-October 1989 series of very large SPEs) were measured in detail by Liulin active high sensitive dosimetric instrument installed on board MIR station. MIR station orbit measurements of the absorbed doses are compared with the interplanetary absorbed doses from SPEs estimated by the data obtained by the METEOR satellite spectrometer. The equivalent dose beyond the magnetosphere resulting from the September 29, 1989 solar flare in a spacecraft module with ordinary shielding thickness (approximately 10 g/cm2 of Al) is far higher than the maximum permissible dose of acute single exposure (50 cSv) and comparable with the maximum permissible dose of 2 year mission (118 cSv). Such large SPEs are a serious hazard in interplanetary missions and call forth of special administrative countermeasures.

Equipment Design↗
PubMed1998

A possible role of RGS9 in phototransduction. A bridge between the cGMP-phosphodiesterase system and the guanylyl cyclase system.

In the current concept of phototransduction, the concentration of cGMP in retinal rod outer segments is controlled by the balance of two enzyme activities: cGMP phosphodiesterase (PDE) and guanylyl cyclase (GC). However, no protein directly mediates these two enzyme systems. Here we show that RGS9, which is suggested to control PDE activity through regulation of transducin GTPase activity (He, W., Cowan, C. W., and Wensel, T. G. (1998) Neuron 20, 95-102), directly interacts with GC. When proteins in the Triton X-100-insoluble fraction of bovine rod outer segments were isolated by two-dimensional gel electrophoresis and binding of GC to these proteins was examined using a GC-specific antibody, proteins (55 and 32 kDa) were found to interact with GC. However, the activity of GC bound to the 55-kDa protein was not detected. This observation was elucidated by the finding that the 55-kDa protein inhibited GC activity in a dose-dependent manner. Amino acid sequence showed that five peptides derived from the 55-kDa protein were identical to corresponding peptides of RGS9. Together with other biochemical characterization of the 55-kDa protein, these observations indicate that the 55-kDa protein is RGS9 and that RGS9 inhibits GC. RGS9 may serve as a mediator between the PDE and GC systems.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1998

[Pancreatic function and the danger of the occurrence of acute pancreatitis during the use of early enteral tube feeding after an operation on the bile ducts].

The amylase and lipase activity dynamics was studied in experiment on 43 dogs using the method of loading with polysubstrate eatable mixtures while the jaundice simulation after a bile passage restoration. The amylase and lipase amplitude and yield increase as an answer to food introduction in the animals with the obturating jaundice simulation and after a bile diversion operation conduction was established. The fact that such possible dangerous complication as an acute pancreatitis did not occur witnesses the security of enteral probe nutrition.

Acute Disease↗
PubMed1997

Residues within the polycationic region of cGMP phosphodiesterase gamma subunit crucial for the interaction with transducin alpha subunit. Identification by endogenous ADP-ribosylation and site-directed mutagenesis.

Interaction between the gamma subunit (Pgamma) of cGMP phosphodiesterase and the alpha subunit (Talpha) of transducin is a key step for the regulation of cGMP phosphodiesterase in retinal rod outer segments. Here we have utilized a combination of specific modification by an endogenous enzyme and site-directed mutagenesis of the Pgamma polycationic region to identify residues required for the interaction with Talpha. Pgamma, free or complexed with the alphabeta subunit (Palphabeta) of cGMP phosphodiesterase, was specifically radiolabeled by prewashed rod membranes in the presence of [adenylate-32P]NAD. Identification of ADP-ribose in the radiolabeled Pgamma and radiolabeling of arginine-replaced mutant forms of Pgamma indicate that both arginine 33 and arginine 36 are similarly ADP-ribosylated by endogenous ADP-ribosyltransferase, but only one arginine is modified at a time. Pgamma complexed with Talpha (both GTP- and GDP-bound forms) was not ADP-ribosylated; however, agmatine, which cannot interact with Talpha, was ADP-ribosylated in the presence of Talpha, suggesting that a Pgamma domain containing these arginines is masked by Talpha. A Pgamma mutant (R33,36K), as well as wild type Pgamma, inhibited both GTP hydrolysis of Talpha and GTP binding to Talpha. Moreover, GTP-bound Talpha activated Palphabeta that had been inhibited by R33,36K. However, another Pgamma mutant (R33,36L) could not inhibit these Talpha functions. In addition, GTP-bound Talpha could not activate Palphabeta inhibited by R33,36L. These results indicate that a Pgamma domain containing these arginines is required for its interaction with Talpha, but not with Palphabeta, and that positive charges in these arginines are crucial for the interaction.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1996

Possible stimulation of retinal rod recovery to dark state by cGMP release from a cGMP phosphodiesterase noncatalytic site.

Cyclic GMP phosphodiesterase, a key enzyme for phototransduction, contains alpha, beta (Palphabeta), and two gamma (Pgamma) subunits. In addition to catalytic sites, Palphabeta has two classes of noncatalytic cGMP binding sites with different affinities (Kd values <100 nM and >1 microM). Pgamma regulates Palphabeta as an inhibitor of cGMP hydrolysis and as a stimulator of cGMP binding to the high affinity noncatalytic sites. Pgamma release from Palphabeta by the GTP-bound alpha subunit of transducin (GTP.Talpha) interrupts these two functions. Here we describe a novel regulation of the Pgamma release by [cGMP] and its physiological implication. We isolated Pgamma mutants that exhibit abnormally one of these two functions, indicating the distinct domains in Pgamma are involved to express these functions. When [cGMP] was high ( approximately 5 microM), Pgamma responsible for the inhibition of cGMP hydrolysis was preferentially released, and cGMP hydrolysis activity of Palphabeta was increased about 10 times. When [cGMP] was low (less than approximately 0.5 microM), Pgamma responsible for the stimulation of cGMP binding to the high affinity sites was released. The Pgamma release resulted in the decrease of relative affinity of cGMP for the high affinity sites to at least (null)/1;10, followed by the rapid release of cGMP from one of the high affinity sites (apparent t1/2 = 3.8 s). cGMP ( approximately 5 microM) inhibited the extraction of Palphabeta from rod membranes by a Mg2+-free hypotonic buffer. The inhibition of Palphabeta extraction was not affected by Pgamma, suggesting that Palphabeta detects on the order of micromolar [cGMP] using low affinity noncatalytic sites on Palphabeta. Because [cGMP] is approximately 5 microM in darkness and lowered by photoexcitation and phosphodiesterase concentration is approximately 30 microM in rod photoreceptors, it is possible that cGMP phosphodiesterase functions to increase cytoplasamic [cGMP] after [cGMP] is reduced to the illuminated level.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1996

Discrimination of two functions of photoreceptor cGMP phosphodiesterase gamma subunit.

cGMP phosphodiesterase, a key enzyme in phototransduction, is composed of P alpha beta and two P gamma S. P gamma has two functions in P alpha beta regulation: (I) an inhibitor of cGMP hydrolysis and (II) a stimulator of cGMP binding to their noncatalytic sites. Here we show for the first time that these functions can be discriminated. P gamma release by GTP-bound transducing from P alpha beta was stimulated by NaCl in a concentration-dependent manner. However, phosphodiesterase activity in membranes washed with NaCl-free buffer already reached the maximum level. [3H]-cGMP binding of P alpha beta in membranes washed with NaCl required more P gamma than that in membranes washed without NaCl. Other salts had a similar effect. Identical P gamma was released under different [NaCl]. These results indicate that P gamma for the function (I) is released in low [salt], but P gamma for the function (II) is released in high [salt]. These P gamma functions may be expressed separately in phototransduction.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1995

[Antihemolytic effect of chlorpromazine on erythrocytes in hyperosmotic and cold shock].

The combined effect of preliminary dehydration of erythrocytes and the cationic amphiphilic compound, chlorpromazine, on the stability of cells to subsequent hyperosmotic effect, has been studied. A protective effect of chlorpromazine depending on the degree of initial cell dehydration and temperature was found. The results obtained by an addition of the cationic amphiphile at preliminary stages of the experiment and directly in the process of development of hyperosmotic lysis have been compared. A conclusion was drawn that the observed effect is not due to preliminary treatment of cells with chlorpromazine and is therefore unrelated to its influence on the regulatory systems of the cell. It has been shown also that under cold shock conditions the earlier established protective effect of chlorpromazine is also directly realized at the moment of stress exposure, i.e., with a change in temperature.

Chlorpromazine↗
PubMed1995

[Variations of solar activity and radiation situation on board MIR station during the period 1986-1994].

This paper is dedicated to the analysis of the radiation situation onboard Mir station over a period of 1986-1994, there examined the main cosmophysics parameters and indices of the solar activity as well as the variations of the parameters of the earth's magnetic field and their association with the changes in the power of absorbed dose onboard the station. There noted the high levels of radiation exposure to the cosmonauts under terrestrial conditions when carrying out the roentgeno-radiologic examinations and procedures comparable or exceeding the absorbed doses during the flights. For revealing the regular associations of the radiation situation onboard the station with the parameters of solar activity there has been analyzed the time changes of average monthly values of dose power since the beginning of station functioning in 1986 until returning the fifteenth expedition to Earth. From the analyses of the results it might be assumed that the best statistical associations of average monthly power of the absorbed dose are found with the streams of protons of GCR. Wolff numbers and background stream of the radio emission of the Sun which reflects the existence of the radiation situation upon the phase of solar activity cycle. From this paper it transpires that calculating the dose loads during the period of the extreme phases of solar activity, it is possible to make between them the interpolations of time dependence by analogy with the dynamics in time of the background streams of GCR or Wolff numbers.

Data Interpretation, Statistical↗
PubMed1993

Site-directed mutagenesis of the cGMP phosphodiesterase gamma subunit from bovine rod outer segments: role of separate amino acid residues in the interaction with catalytic subunits and transducin alpha subunit.

The recombinant and 21 mutant phosphodiesterase (PDE) gamma subunit (PDE gamma) genes were expressed by sequential transcription and translation in vitro. Inhibitory properties of these mutants and their interactions with PDE catalytic and transducin alpha subunits were studied. The interaction of the PDE gamma subunit with the catalytic ones proceeds in two steps--primary binding and inhibition. The central region of the PDE gamma molecule enriched with the basic amino acid residues (particularly, Lys-29, Lys-31 and Arg-33), is involved in the primary binding, and the PDE gamma C-terminus plays the key role in inhibition. The spatial orientation of the C-terminus is of great importance here. The PDE gamma C-terminus also affects binding to catalytic moieties.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1991

[Modifying effect of concanavalin A and diamide on erythrocyte sensitivity to cold shock].

The temperature (0 degrees C and 37 degrees C) and the medium tonicity (0.15-1.20 M NaCl) were shown to affect erythrocyte agglutination by concanavalin A. Treatment of cells with lectin caused no significant decrease in the erythrocyte hemolysis upon cooling. Diamide, unlike concanavalin A used at concentrations above 2.0 M decreases the cell sensitivity to the cold shock. The changes in the erythrocyte susceptibility to cooling within the temperature range of 37-0 degrees C correlate with changes in the electrophoretic spectrum of membrane proteins. The progressive decrease in the spectrin bands intensity with a simultaneous formation of high molecular weight protein aggregates not included in the gel composition was observed after diamide treatment. The diamide effect depends on the medium tonicity, at which the treatment was performed, being especially well pronounced in hypertonic media with 0.8-1.2 M NaCl concentrations, the maximal spectrin aggregation being observed under these conditions. It is suggested that the main factor of the mechanism underlying the erythrocyte hypertonic cold shock is the increase in the association of peripheral cytoskeleton proteins with plasma membrane in osmotically dehydrated cells which limits the ability of lipids to adapt during cooling and results in the stabilization of defects in the membrane structure at low temperatures. Diamide eliminates these unfavourable changes eventually resulting in the dissociation of peripheral proteins from the cytoplasmic surface of the membrane on the protein aggregation.

Adaptation, Physiological↗
PubMed1991

[The effect of chlorpromazine on the temperature and osmotic sensitivity of erythrocytes].

The effect of chlorpromazine on the development of cold shock in erythrocytes exposed to sodium chloride was shown to depend on the tonicity of the medium in which the cells were cooled from 37 degrees C down to 0 degrees C as well as on the amphipate concentration. After cooling of erythrocytes in a NaCl (0.75-1.5 M)-containing medium with chlorpromazine (7 x 10(-5) M, 2.1 x 10(-4) M and 3.5 x 10(-4) M) the hypertonic cold shock was inhibited, the protective effect of the amphipate being less pronounced at its increasing concentrations. After cooling of cells under conditions of moderate hypertonicity (0.3-0.6 M NaCl) no modifying effect of chlorpromazine on the sensitivity of erythrocytes to the temperature decrease from 37 degrees C down to 0 degrees C was manifested. However, under iso- and hypertonic conditions chlorpromazine used at 2.1 x 10(-4) M and 3.5 x 10(-4) M stimulated the cold shock development in erythrocytes. A sharp increase in the medium tonicity (up to 1.8-3.0 M and higher) the cells underwent isothermal hemolysis which was more expressed at 0 degrees C than at 37 degrees C. These data suggest that chlorpromazine significantly activates the hemolytic process at low temperatures.

Chlorpromazine↗