Modular portable unit for thermal conductivity measurements in multiple cryogenic/magnetic field environments.

A modular design for a miniature thermal conductivity cell suitable for a dilution refrigerator and other sample-in-vacuum cryogenic environments with different magnet options is described. The sample is mounted once and the contacts remain in place when the unit is repositioned or transported between different magnets and cryostats. This mobility enables comprehensive measurements with access to specific options, such as vector magnet in one lab and ultra-high field in another. This design enables significant expansion of the temperature range by using not only dilution refrigerators but also 3He, 4He cryostats and even ubiquitous Quantum Design Physical Property Measurement System.

[Dynamics of structural change of ovarian tissue under ischemia].

In this work it was investigated the dependence of the correlation of folliculogenesis and endocrine function of ovarian tissue on the degree of structural damage and oocyte volume changes during ischemia. It was shown that after 2 hours of ischemia at 37 °C the morphological transformation of the structural components of the ovarian tissue were reversible. In case of restoration of blood flow conditions by heterotopic transplantation estradiol level of animals was 25,9 ± 5,18 pg/ml, progesterone--18,48 ± 3,69 ng/ml, significantly higher than the castrated animals-recipients. Supplement of the incubation medium by the fetal bovine serum lead to decreasing by 5-7% in the volume of oocytes of growing follicles during in ischemia and reduced steroidogenic function of ovarian tissue after perfusion. Increased time of the ischemia up to 4 hours was founded in irreversible morphological transformation, reduce the volume of oocytes by 40% and the formation of sclerosed tissue after transplantation of the ischemic fragments of ovarian tissue.

[Spatial distribution of local absorbed doses inside the Russian segment of the International Space Station].

The article discusses the procedure of operational radiation safety monitoring with the use of portable Pille-MKS dosimeters, presents the results of ISS dose measurements from September 22, 2003 (after Pille deployment on board the ISS) to March 16, 2011 (completion of the ISS-25 mission). The necessity of continuous dynamic tracking of the radiation environment in ISS compartments arises from the character and uniqueness of space ionizing radiation effects on crew. Radiation loading in the ISS compartments was analyzed and results of using different dosimeters were compared. Experimental radiation studies of the ISS piloted compartments are needed for reliable prediction of doses for the crew that still defy precise estimation.

[New approaches to diagnostics and identification of possible damages in normal humans under the conditions of microgravity and exposure to low doses of radiation].

The issues of radiation risk evaluation were addressed in the investigation of cytogenetic and molecular-biological changes in lymphocytes of cosmonauts and pilots of high-altitude airplanes. The goal was to determine individual sensitivity to the flight conditions and an additional factor (lymphocyte exposure to 1 Gy in situ), and adaptability as an index of induction of cell and organism resistance to extreme conditions.

[Estimation of radiation loads on cosmonauts of the International Space Station during 2000 - 2007].

Radiation loads on cosmonauts during the initial 15 increments of the International space station (ISS) were analyzed with the use of a human body geometry model. The total characteristic of absorbed and equivalent doses in compartments of the ISS Russian segment was determined with consideration of the actual cosmophysical situation and ISS ballistic parameters during every increment in the period from October 2000 to October 2007.

[Individual cytogenetic and molecular-biological characteristics of lymphocytes in blood of pilots and cosmonauts].

The purpose of this paper was the investigation of the pilots and of cosmonauts individual sensitivity to the fly conditions, to the additional irradiation (in the dose of 1 Gy), the adaptive response manifestation (in the doses 0.05 and 0.5 Gy). The DNA comet assay (the double strand DNA breaks was determined) and the method of unstable chromosome aberrations in metaphase was used. The human blood lymphocytes was the object of investigation. The significant individual differences were discovered in pilots and in cosmonauts in the initial DNA damage; in the sensitivity to the additional irradiation. The frequency of the adaptive response induction was decreased in the pilots in the comparison with the control group. The adaptive response was registered in cosmonauts (3 men). It is supposed that DNA damage, chromosome aberrations, sensitivity to the additional irradiation, the adaptive response manifestations can be used as biological markers of individual risk disease.

[The maturation of the spermatozoa: events, consequences and possible ways of control].

Maturation of spermatozoa represents a formation of competent cell with capacity to fertilize. For the purpose of this review the morphological and functional changes of nucleus, plasmatic and nuclear membranes, tail region as well as the change of localisation and activity of enzymes have been described to illustrate the epididymis stage of maturation. Particular attention has been paid to the role of active oxygen species in the cell transformation, pro- and antioxidant balance during the maturation and to the systems controlling gametes control. The possible ways of influence on the some events of spermatozoa maturation are shown.

[The model of geometrical human body phantom for calculating tissue doses in the service module of the International Space Station].

The model of a geometrical human body phantom developed for calculating the shielding functions of representative points of the body organs and systems is similar to the anthropomorphic phantom. This form of phantom can be integrated with the shielding model of the ISS Russian orbital segment to make analysis of radiation loading of crewmembers in different compartments of the vehicle. Calculation of doses absorbed by the body systems in terms of the representative points makes it clear that doses essentially depend on the phantom spatial orientation (eye direction). It also enables the absorbed dose evaluation from the shielding functions as the mean of the representative points and phantom orientation.

[Some results of radiaton monitoring of the ISS Russian Segment in 2000-2005].

Dynamics of the ISS RS radiation environment was studied using the data of daily (operative) monitoring and personal dose measurements during 11 increments in the period between August 1, 2000 and October 28, 2005 overlapping the maximum phase of the 23rd solar cycle. It was shown that personal absorbed doses varied within the range of measurements of dual wavelength radiometer R-16, a component of the ISS radiation monitoring system. Power of the absorbed doses fell in the range of 0.017 to 0.02 cGy/day and was solar-dependent.

[Analysis of protein concentration in the urine of children with metabolic nephropathy, crystal-calcium crystalluria].

We compared the efficacy of a routine sulphosalycilic acid method and a new technique of pirogallolic red staining for urine protein assay in children with metabolic nephropathy, crystal-calcium crystaluria. The former method was found ineffective in proteinuria diagnosis in children with metabolic nephropathy. The pirogallolic method appeared more effective as it detects microproteinuria and therefore allows accurate diagnosis, control of its treatment and finally to improve the results of therapeutic and prophylactic measures.

[The assessment of radiation hazards in the "MIR" and ISS orbits from the data of vehicle and personal dosimetric monitoring]. / Otsenka radiatsionnoi opasnosti dlia chlenov ékipazhei orbital'noi stantsii "Mir" i Mezhdunarodnoi kosmicheskoi stantsii na osnove dannykh bortovogo i individual'nogo dozimetricheskogo kontrolia.

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.

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.

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.

[Radiation environment of orbital complex "Mir" during minimum of the 22-nd solar cycle (1994-1996)]. / Radiatsionnaia obstanovka na orbital'nom komplekse "Mir" na faze minimuma 22-go tsikla solnechnoi aktivnosti (s 1994 po 1996 god).

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.

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.

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.

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.

[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]. / Funktsional'noe sostoianie podzheludochnoi zhelezy i opasnost' vozniknoveniia ostrogo pankreatita pri primenenii rannego énteral'nogo zondovogo pitaniia posle operatsii na zhelchevyvodiashchikh putiakh.

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.

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.