Rational Design and Mutagenesis of Fungal Luciferase from Neonothopanus nambi.

The recently described bioluminescent system from fungi has great potential for developing highly efficient tools for biomedical research. Luciferase enzyme is one of the most crucial components of this system. The luciferase from Neonothopanus nambi fungus belongs to the novel still undescribed protein family. The structure data for this protein is almost absent. A detailed study of the N. nambi luciferase properties is necessary for the improvement of analytical methods based on the fungal bioluminescent system. Here we present the positions of key amino acid residues and their effect on enzyme function described using bioinformatic and experimental approaches. These results are useful for further fungal luciferase structure determination.

The Recombinant Luciferase of the Fungus Neonothopanus nambi: Obtaining and Properties.

A key component of the recently described bioluminescent system of higher fungi is luciferase, a new class of proteins. The properties of fungal luciferase and their relationship with its structure are interesting both for improving autoluminescent systems already created on its basis and for creating new ones. Therefore, it is extremely important to understand the spatial structure of this protein. We have performed heterologous expression and purification of Neonothopanus nambi luciferase, obtained a protein suitable for subsequent crystallization, and also determined some biochemical properties of the recombinant luciferase.

Luciferin-Luciferase System of Marine Polychaete Chaetopterus variopedatus.

This paper presents the preliminary results of the separation of the Chaetopterus variopedatus bioluminescent system into luciferin and luciferase and a brief description of some of their properties.

Comparison of Earthworm Bioluminescent Systems.

The results of a comparative study of the luciferin-luciferase systems of seven species of bioluminescing oligochaetes-Henlea petushkovi, Henlea rodionovae, Fridericia heliota (Enchytraeidae), Microscolex phosphoreus (Acanthodrilidae), Pontodrilus litoralis (Megascolecidae), Eisenia lucens, and Avelona ligra (Lumbricidae)-are presented.

Low-Molecular-Weight Components of Luminescent Reaction of the Siberian Enchytraeid Henlea sp.

The first results of the study of chromatographic and spectral properties of the detected lowmolecular-weight activators, putative emitters in the luminescent reaction of Siberian enchytraeid Henlea sp., are presented.

Isolation and Purification of Fungal Luciferase from Neonothopanus nimbi.

This is the first study to obtain a high-purity luciferase from the fungus Neonothopanus nambi biomass that is suitable for subsequent sequencing.

Why does the bioluminescent fungus Armillaria mellea have luminous mycelium but nonluminous fruiting body?

By determining the components involved in the bioluminescence process in luminous and nonluminous organs of the honey fungus Armillaria mellea, we have established causes of partial luminescence of this fungus. The complete set of enzymes and substrates required for bioluminescence is formed only in the mycelium and only under the conditions of free oxygen access. Since the synthesis of luciferin precursor (hispidin) and 3-hydroxyhispidin hydroxylase in the fruiting bodies is blocked, the formation of luciferin-the key component of fungal bioluminescent system-was not observed. That is why the fruiting body of Armillaria mellea is nonluminous despite the presence of luciferase, the enzyme that catalyzes the oxidation of luciferin with a photon emission.

Structure of fungal oxyluciferin, the product of the bioluminescence reaction.

The structure of fungal oxyluciferin was determined, the enzymatic bioluminescence reaction under substrate saturation conditions with discrete monitoring of formed products was conducted, and the structures of the end products of the reaction were established. On the basis of these studies, the scheme of oxyluciferin degradation to the end products was developed. The structure of fungal oxyluciferin was confirmed by counter synthesis.

[Investigation of the effect of oil and flour from wheat germ meal on lipid metabolism of students and teachers of the university].

The results of investigation of alimentary correction of lipid metabolism under the admin­istration of processed products from wheat germ - oil (with the content of policosanol at least 1.5-8.0 mg/100 g, vitamin E - 180-200 mg/100 g, PUFA - 60-65%) and cake flour (with the content of protein - 30-35%, oil with analogue composition -5-7%, digestible carbohydrates - 45-47%, fiber - 18-26%, vitamins B1, B3, B6, B9, E, PP, minerals and trace elements - Zn, Mn, K, Fe, Se, P) are presented. Volunteers among teachers and students of the university aged 16 to 65 years daily consumed wheat germ oil obtained by cold pressing in an amount of 3.5 g, regardless of the meal within 30 days. Then a part of them (30 persons) consumed daily 50 g of oil cake obtained after pressing oil, which provided the intake of the same amount of oil (3.5 g). Lipid metabolism param­eters were monitored in experiment participants before receiving the processed products of wheat germ, after germ meal intake and beyond 30 and 60 days after consumption of wheat germ. Data analysis was carried out on three age groups: 16-24, 25-44 and 45- 65 years. All participants of the experiment showed a reduction in total cholesterol level by 6-8%, increasing the concentration of HDL cholesterol by 3-24%, lowering LDL cholesterol concentrations by 4-21%, reduction of triglyceride concentration by 12-24%, a positive correction of atherogenic factor values by for 10-25%. Prolonged action of the investigated foods was established: lipid metabolism parameters in the tested group were better than in the control group after 30 days of intake discontinuation of oil or wheat germ flour, the positive adjustment effect disappeared 60 days after consuming the products. The findings demonstrate a positive effect on the normalization of lipid metabolism when cake flour of wheat germ was administered in daily food ration, similar to the effect of oil intake, which is important for the prevention of cardiovascular diseases and atherosclerosis. Given the significant production of cake flour of wheat germ (up to 90-95% of the raw material) and its not high cost as a secondary biological resource, this product can be recommended to the introduction in the diet of organized groups, including socially vulnerable groups.

[STUDY OF THE IMPACT OF WHEAT GERM CAKE OIL AND FLOUR ON INDICES OF ENERGY EXCHANGE IN STUDENTS AND TEACHERS OF HIGH SCHOOL].

There were obtained data which confirm the increase of efficiency of gas exchange processes in students and teachers of the engineering high school in the daily use of at least 3.5 g of wheat germ oil and 50 g of wheat germ cake flour without the correction of the basic ration. The study of the exhaled gas-air mixture showed in all age groups the gain in the concentration of carbon dioxide and the decline of concentration of oxygen by the 0.20-0.30%. There was established the increase of blood oxygenation level in all subjects. There was demonstrated the relationship between the depth of the alteration of these parameters with age, and regular physical activity in studied cases.

[Model experiments on sorption properties of beet-root fiber].

Model experiments provided results of determining sorbate properties of beet-root fiber in respect of copper, plumbum and zinc in diary foods. It was determined that this fiber makes possible the absorbing of the above mentioned heavy metal, which increases the hygienic safety of the studied diary foods.

[Correlations of sister chromatids cohesion complexes distribution with histones H3 and H4 modifications].

The formulation of "histone code" theory brings active investigations of the role of histone modifications and other supramolecular factors of DNA condensation in transcription regulation. In this work, we have analyzed the localization of methylated histones on 9, 36 and 79 lysines, hyperacetylated H4 histone, and subunits of cohesion complex DRAD21 relatively of Drosophila melanogaster polytene chromosomes chromatin condensation. We propose the hypotheses of a cascade regulation of transcription activity defined by histone modifications and the adaptive role of sister chromatids cohesion in the transcription of high active and extensive genes.

Purification and partial spectral characterization of a novel luciferin from the luminous enchytraeid Fridericia heliota.

A homogeneous luciferin preparation has been obtained from the luminous soil enchytraeid Fridericia heliota, which has an ATP-dependent luminescent system. A procedure for luciferin purification without losing fractions of active luciferase has been developed. The luciferin specific activity is 4000 times increased; its UV absorption spectrum maximum is 294 nm with a local minimum at 262 nm. The luciferin of the enchytraeid F. heliota is significantly different from firefly luciferin, whose luminescent reaction also requires ATP, and it also appears to have no similarities to other known luciferins.

Effect of different salts and detergents on luciferin-luciferase luminescence of the enchytraeid Fridericia heliota.

The study addresses the effect produced by different inorganic salts and detergents (SDS, Triton X-100, the Tween series) on the ATP-dependent bioluminescent reaction catalyzed by the luciferase of the new earthworm species Fridericia heliota (Annelida: Clitellata: Oligochaeta: Enchytraeidae). It has been shown that the effect of divalent metal salts on luminescence is determined by the action of cations. Three of them - Mg(2+), Mn(2+) and Ca(2+) - can stimulate luciferase activity at concentrations varying within a wide range, and Mn(2+) can act as a 100%-effective substitute for Mg(2+) in F. heliota luminescence reaction in vitro. The inhibitory effect of monovalent metal salts on luminescence is largely determined by the action of the anion part of the molecule. The effectiveness of the inhibitory effect of anions increases in the following order: Cl(-)

[A natural focus of icterohemorrhagic leptospirosis in the floodplain of the lower Don and means for its eradication]. / Prirodnyi ochag ikterogemorragicheskogo leptospiroza v poime Nizhnego Dona i puti ego ozdorovleniia.

An active natural focus of icterohemorrhagic leptospirosis has been detected in the area of fish-breeding ponds in Rostov Province, where the intensive epizootic among the population of Norway rats is observed the whole year round (574 animals have been examined, 56 cultures have been isolated). The epizootic process reaches its highest intensity in autumn (the proportion of infected animals exceeds 50%). This natural focus in the area of fish-breeding ponds is epidemiologically dangerous. The limitation of its infectious potential is possible by means of poisoned baits.