Coelenterazine-v ligated to Ca2+-triggered coelenterazine-binding protein is a stable and efficient substrate of the red-shifted mutant of Renilla muelleri luciferase.

It has been shown that the coelenterazine analog, coelenterazine-v, is an efficient substrate for a reaction catalyzed by Renilla luciferase. The resulting bioluminescence emission maximum is shifted to a longer wavelength up to 40 nm, which allows the use of some "yellow" Renilla luciferase mutants for in vivo imaging. However, the utility of coelenterazine-v in small-animal imaging has been hampered by its instability in solution and in biological tissues.

The intrinsic fluorescence of apo-obelin and apo-aequorin and use of its quenching to characterize coelenterazine binding.

The intrinsic fluorescence of two apo-photoproteins has been characterized and its concentration-dependent quenching by coelenterazine has been for the first time applied to determine the apparent dissociation constants for coelenterazine binding with apo-aequorin (1.2+/-0.12 microM) and apo-obelin (0.

Synthesis of biomass and utilization of plants wastes in a physical model of biological life-support system.

The paper considers problems of biosynthesis of higher plants' biomass and "biological incineration" of plant wastes in a working physical model of biological LSS. The plant wastes are "biologically incinerated" in a special heterotrophic block involving Californian worms, mushrooms and straw. The block processes plant wastes (straw, haulms) to produce soil-like substrate (SLS) on which plants (wheat, radish) are grown.

Increase of atmospheric CO2: response patterns of a simple terrestrial man-made ecosystem.

Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions. Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements. The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system.

Mass exchange in an experimental new-generation life support system model based on biological regeneration of environment.

An experimental model of a biological life support system was used to evaluate qualitative and quantitative parameters of the internal mass exchange. The photosynthesizing unit included the higher plant component (wheat and radish), and the heterotrophic unit consisted of a soil-like substrate, California worms, mushrooms and microbial microflora. The gas mass exchange involved evolution of oxygen by the photosynthesizing component and its uptake by the heterotroph component along with the formation and maintaining of the SLS structure, growth of mushrooms and California worms, human respiration, and some other processes.

High temperature effect on microflora of radish root-inhabited zone and nutrient solutions for radish growth.

The effect of high temperatures (35 and 45 degrees C) on microflora of the root zone of radish plants grown in phytotron was evaluated by the response of microorganisms from 9 indicator groups. Phytotron air temperature elevated to 35 degrees C for 20 hours caused no significant changes in qualitative and quantitative composition of the root microflora in experimental plants. By the end of the experiment, the species diversity of microflora had changed.

Tolerance of LSS plant component to elevated temperatures.

Stability of LSS based on biological regeneration of water, air and food subject to damaging factors is largely dependent on the behavior of the photosynthesizing component represented, mainly, by higher plants. The purpose of this study is to evaluate the tolerance of uneven-aged wheat and radish cenoses to temperature effects different in time and value. Estimation of thermal tolerance of plants demonstrated that exposure for 20 h to the temperature increasing to 45 degrees C brought about irreversible damage both in photosynthetic processes (up to 80% of initial value) and the processes of growth and development.

On monitoring the bacterial component as an indicator of the state of small man-made ecosystems.

High reproduction rates make the bacterial component of ecosystems a good indicator of the state of the system on the whole. This determines the necessity to develop rapid monitoring of the functional state of the bacterial component of small ecosystems. Information about substrate concentration in the population is indicative of the state of the bacterial culture.

Mathematical modeling of response of ecosystems with different structure to external impact.

A mathematical model was used to study the response of ecosystems of different structures to external impact. The response was measured as a sensitivity coefficient: the magnitude of the system's response vs. the change of the factor in the inflow.

Functional, regulatory and indicator features of microorganisms in man-made ecosystems.

Functional, regulatory and indicator features of microorganisms in development and functioning of the systems and sustaining stability of three man-made ecosystem types has been studied. 1) The functional (metabolic) feature was studied in aquatic ecosystems of biological treatment of sewage waters for the reducer component. 2) The regulatory feature of bacteria for plants (producer component) was studied in simple terrestrial systems "wheat plants-rhizospheric microorganisms-artificial soil" where the behavior of the system varied with activity of the microbial component.

Effect of volatile metabolites of dill, radish and garlic on growth of bacteria.

In a model experiment plants were grown in sealed chambers on expanded clay aggregate under the luminance of 150 W/m2 PAR and the temperature of 24 degrees C. Seven bacterial strains under investigation, replicated on nutrient medium surface in Petri dishes, were grown in the atmosphere of cultivated plants. Microbial response was evaluated by the difference between colony size in experiment and in control.

Structural and functional changes in photosynthetic apparatus of wheat under exposure to sulfur dioxide fumes.

Changes in photosynthesis of wheat (pigment content and chlorophyll fluorescence) have been measured to study effect of gaseous sulfur dioxide on wheat cenosis. Several biochemical and biophysical techniques have been estimated to evaluate their ability for early detection of destructive effect of SO2. A mathematical model of toxic gas absorption in a closed system with plants has been proposed.

Experimental microcosms as models of natural ecosystems for monitoring survival of genetically modified microorganism.

An experimental approach for investigation of genetically modified microorganisms (GMMO) introduced into model ecosystems to evaluate potential risk of propagation of recombinant plasmids in surrounding medium has been developed. The object of modeling was Escherichia coli Z905 strain with a recombinant plasmid with bacterial luminescence genes, which was introduced into water microcosms of different structure. The approach involves comprehensive investigation of GMMO at four hierarchical levels: molecular (retaining the structure of the plasmid and expression of cloned genes); cellular (variation of metabolic activity); population (competitive power and metabolic interactions of GMMO with indigenous microflora, migration of recombinant and natural plasmids); ecosystem (effect of GMMO and cloned genes on ecosystem parameters).

Experimental and mathematical models for small aqueous closed ecosystems with spatially separated components.

Experimental and theoretical models of closed "autotroph-heterotroph" (chlorella-yeast, chlorella-protozoa) ecosystems with spatially separated components have been created and studied. The chart of flows and interaction of components of gas-closed "chlorella-yeast" system have formed the basis describe mathematically the functioning of the given system, experimental results have been found to agree with computer solution of the problem in terms of quality. Investigation of the experimental model of the "producer-consumer" trophic chain demonstrated the role of protozoa in nitrogen turnover.

Modelling of genetically engineered microorganisms introduction in closed artificial microcosms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E.

Experimental models of small closed systems with spatially separated unicellular organism-based components.

Experimental models of small biotic cycles of different degree of closure and complexity with spatially separated components based on unicellular organisms have been studied. Gas closure of components looped into "autotroph-heterotroph" (chlorella-yeast) system doubled the lifetime of the system (as opposed to individually cultivated components). Higher complexity of the heterotroph component consisting of two yeast species also increased the lifetime of the system through more complete utilization of the substrate by competing yeast species.

A new enzymatic technique to estimate the efficiency of microbial degradation of pollutants.

Dynamics of active sludge microorganism activity in aerotanks under chemostat conditions has been studied. Dependence of microorganism catalase activity has been found to depend on residual substrate concentration in proportion to the biomass of microorganisms. Experimental data and field observations has formed the basis to develop a technique to evaluate in relative units the amount of the substrate consumed by biocenosis of the active sludge in the air tanks of purification facilities.

Element exchange in a water- and gas-closed biological Life Support System.

Liquid human wastes and household water used for nutrition of wheat made possible to realize 24% closure for the mineral exchange in an experiment with a 2-component version of "Bios-3" life support system (LSS) Input-output balances of revealed, that elements (primarily trace elements) within the system. The structural materials (steel, titanium), expanded clay aggregate, and catalytic furnace catalysts. By the end of experiment, the permanent nutrient solution, plants, and the human diet gradually built up Ni, Cr, Al, Fe, V, Zn, Cu, and Mo.

Interaction of physical-chemical and biological regeneration processes in ecological Life Support Systems.

Catalytic combustion of inedible biomass of plants in ecological Life Support Systems (LSS) gives rise to gaseous oxides (CO2, NO2, SO2, etc.). Some of them are toxic for plants suppressing their photosynthesis and productivity.

Plants--rhizospheric organisms interaction in a manmade system with and without biogenous element limitation.

The effect has been studied of inoculation of seeds of wheat with two species of rhizospheric microorganisms--Pseudomonas fluorescens and Pseudomonas putida--on young plant growth with complete and with nitrogen deficit mineral nutrition. With complete mineral medium, plants grown from seeds inoculated with bacteria of Pseudomonas genus (experiment plants) have been found to have better growth over plants not inoculated with these bacteria (control plants). The experiment plants had increased transpiration and their biomass had higher organic nitrogen content.

Impaired growth of plants cultivated in a closed system: possible reasons.

Plants in experiments on "man-higher plants" closed ecosystem (CES) have been demonstrated to have inhibited growth and reduced productivity due to three basic factors: prolonged usage of a permanent nutrient solution introduction into the nutrient medium of intra-system gray water, and closure of the system. Gray water was detrimental to plants the longer the nutrient solution was used. However, higher plant growth was mostly affected by the gaseous composition of the CES atmosphere, through accumulation of volatile substances.

Waste bioregeneration in life support CES: development of soil organic substrate.

An experimental model of matter turnover in the biotic cycle: plants (plant biomass) --> mushrooms (residual substrate + mushroom fruit bodies) --> worms (biohumus) --> microorganisms (soillike substrate) --> plants is presented. The initial mass of soillike substrate was produced from wheat plants grown in a hydroponic system. Three cycles of matter turnover in the biotic cycle were carried out.