A Novel Express Method to Determine Activity of Antitumor Enzyme L-Lysine-α-Oxidase of Trichoderma harzianum Rifai F-180.

A novel express method is developed to determine activity of antitumor enzyme L-lysine-α-oxidase obtained by culturing Trichoderma harzianum Rifai F-180 fungus. The carcinogenic reagent ortho-dianisidine-hydrochloride was replaced in the reaction medium with environmentally friendly reagents of the chromogenic mixture that included tetramethylbenzidine. This method improved precision and sensitivity of ELISA by 10 and 40 times, respectively. In addition, it could detect activity of L-lysine-α-oxidase not only in the producer strains with a pronounced activity of this enzyme, but also in the strains where this activity has not been previously determined.

Method for Selection of New Primers for Identification of the Especially Dangerous Bacterium Acidovorax citrulli.

We propose a method for selection of new primers for identification of especially dangerous bacterium Acidovorax citrulli. A specific pair of new primers AC-1 F/R that can be used for detection and identification of A. citrulli was successfully tested. This opens up new possibilities for investigation of the role of this bacterium in the ecosystems.

L-Lysine-α-Oxidase: Acidovorax citrulli Bacterium Inhibitor.

Studies of the effects of Trichoderma harzianum Rifai F-180 culture fluid concentrate containing L-lysine-α-oxidase antitumor enzyme produced by the fungus and the homogenous enzyme, on ultrahazardous bacterium Acidovorax citrulli demonstrated the antibacterial activity of the concentrate. Trichoderma harzianum Rifai F-180 producing L-lysine-α-oxidase was cultured in a technological device at G. K. Skryabin Institute of Biochemistry and. Physiology of Microorganisms, Russian Academy of Sciences. Activity of L-lysine-α-oxidase in the resulted culture fluid concentrate was 0.54 U/ml, activity of the homogenous enzyme was 50 U/mg.

Antibacterial Activity of L-Lysine-α-Oxidase from the Trichoderma.

We studied the effects of a concentrate of metabolites of Trichoderma harzianum Rifai F-180, an active producer of L-lysine-α-oxidase, and homogenous enzyme on a highly virulent bacteria Erwinia amylovora. The producer of antitumor and antiviral Trichoderma enzyme L-lysine-α-oxidase was cultured on a processing system of G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences (Pushchino). Activity of L-lysine-α-oxidase in the prepared concentrate of metabolic products of the producer was 5.4 U/ml, and activity of the homogenous enzyme was 50 U/ml. Antibacterial activity of the enzyme was shown in our experiments.

Trichoderma L-Lysine-α-Oxidase Producer Strain Culture Fluid Inhibits Impatiens Necrotic Spot Virus.

A method for PCR diagnosis of impatiens necrotic spot virus is developed. Concentrated culture fluid with active L-lysine-α-oxidase (0.54 U/ml) from Trichoderma harzianum Rifai fungus producer strain F-180 inhibits vitally hazardous impatiens necrotic spot phytovirus.

Inhibition of Tobacco Ringspot Virus by the Culture Fluid of L-Lysine-α-Oxidase Producing Strain.

A producing strain of an anti-tumor and antiviral enzyme L-lysine-α-oxidase from Trichoderma was cultured using a technological device of G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences (Pushchino). L-lysine-α-oxidase activity in the obtained metabolite concentrate was 5.4 U/ml. We studied the effects of the concentrate of active L-lysine-α-oxidase producer on the highly infectious Tobacco ringspot virus and revealed anti-viral activity of it when enzyme concentration was at least 1.0 U/ml.

Plant Extracts in Development of Medicinal Products of Various Therapeutic Value.

Development, standardization and use of medicinal products of plant origin and various therapeutic value are described in the review. The literature data refer to the studies of some leading institutions of the Russian Federation and in particular to the Medical Institute of the Russian University of Peoples' Friendship, All-Russian Research Institute of Medicinal and Aromatic Plants, Pyatigorsk State Pharmaceutical Academy and some foreign laboratories. Possible use of the trichoderma extract and its prospects are shown.

[L-Lysine-α-Oxidase in vitro Activity in Experiments on Models of Viruses Sindbis, Forest-Spring Encephalitis, Western Nile, Tyaginya and Dhori].

The antitumor effect of L-lysine-α-oxidase from the culture fluid of Trichoderma harzianum Rifai F-180 was investigated for the first time. The in vitro studies revealed its high activity on a model of the forest-spring encephalitis virus and no activity against the Sindbis, Western Nile, Tyaginya and Dhori viruses.

[Antimycoplasmic Activity of Fermentation Broth of Trichoderma harzianum Rifai F-180, an Organism Producing L-Lysine-α-Oxidase, an Antitumor and Antiviral Enzyme].

A concentrate of the fermentation broth of Trichoderma harzianum Rifai F-180, an organism producing L-lysine-α-oxidase, an antitumor and antiviral enzyme, with the activity in the fermentation broth of 0.54-0.56 U/mI was recovered. The effect of the concentrate on the mycoplasmas growth was investigated for the first time. Two representatives of Mycoplasmafaceae, i.e. Mycoplasma hominis and Mycoplasma fermentans and one representative of Aholeplasmataceae. i. e. Aholeplasma laidlawii were used. It was shown that the fermentation broth inhibited the growth of Mycoplasma hominis after the preliminary exposure. The inhibition rate depended on the mycoplasma inoculation dose and the fermentation broth concentration.

[Persistence of bacteria and viruses in Ixodes].

Behaviour of viruses and salmonellas in ticks after their single or combined contamination was thoroughly studied on laboratory animals with bacteriemia or virusemia. When Ixodes ricinus was contaminated simultaneously with forest-spring encephalitis virus and salmonellas there were observed a decrease in the virus titer by the 30th-40th days and its death in 60 days. In case of the I. ricinus nymphs contamination, the virus titer after the combined contamination was by a factor of 10(2) lower in 60 days vs. the contamination with the virus alone and did not reach the contamination dose. The simultaneous contamination of the ticks with two pathogens (forest-spring encephalitis virus and salmonellas) resulted in inhibition of the growth and development of both the virus and the salmonellas.

[Antitumor enzyme L-lyzine-alpha-oxidase from Trichoderma harzianum Rifai F-180 and investigation of its action on L-lysine oxidation by capillar electrophoresis].

Trichoderma harzianum Rifai F-180, an organism producing the antitumor enzyme L-lysine-alpha-oxidase was cultivated and the enzyme was isolated and purified under the manufacturing conditions. The effect of L-lysine-alpha-oxidase on oxidation of L-lysine was investigated for the first time by capillary electrophoresis and the procedure conditions were developed. The reaction of L-lysine oxidative deamination is described and location of the reaction components picks on the elecrophoregrams was identified. The average rate of the catalytic reaction of L-lysine oxidation equal to 0.46 RU/min (7.7 x 10(-3) RU/sec) was determined. The use of the antitumor enzyme L-lysine-alpha-oxidase is recommended as a drug for the treatment of superficial tumors and tissue relative oxygen excess.

[Bactericidal effect of lysozyme].

Isolation of lysozyme from hemolymph of Alveonasus lahorensis (Acari: Parasitiformes, Argasidae) and Hyalomma marginatum (Acari: Parasitiformes, Ixodidae) with using ultrasound is described. It was shown that the bactericidal effect of the ultrasound-extracted lysozyme against Staphylococcus aureus and Micrococcus luteus significantly exceeded that of the chicken egg lysozyme and lysozyme from ticks without ultrasound exposure. Disintegration of the hemolymph cells increased lysozyme production.

[Investigation of antitumor substance from Trichoderma].

The effect of L-lysine-alpha-oxidase from Trichoderma harzianum Rifai on the functional activity of T-lymphocytes was investigated. It was shown that in a dose of 35 units/kg administered parentally the enzyme had no suppressive effect on the T-lymphocyte functional activity. An inhibitory effect of L-lysine-a-oxidase on some indices of the macrophages functional activity was observed. L-Lyzine-alpha-oxidase had a selective lymphotropic action and showed no mytostatic activity, which is in favour of the enzyme vs. other antitumor agents.

[Preclinical trials of L-lysine-alpha-oxidase, an antitumor enzyme].

The effect of L-lysine-alpha-oxidase from a representative of the Trichoderma genus on the humoral immune response to protein antigens was studied. It was shown that repeated fife-fold intravenous administrations of L-lysine-alpha-oxidase in a dose of 35 U/kg had no depressive action on the humoral immunity. The enzyme had no suppressive effect on the delayed type hypersensitivity to xenogenous erythrocytes. The use of L-lysine-alpha-oxidase in a therapeutic dose or in a twice as higher dose had no reliable effect on the leukocyte migration capacity vs. the control.

[Biosynthesis of L-lysine-alpha-oxidase, an antitumor enzyme, by Trichoderma spp].

Strains of Trichoderma spp., producing L-lysine-alpha-oxidase, prospective in chemotherapy of malignant tumors, were studied. The best results of the enzyme biosynthesis were observed on the wheat bran medium. When grown on the media with various pH levels, the strains showed different spectra of the L-aminooxidase activity. The highest activity of the strains was recorded with respect to destruction of L-lysine. The lysine-producing organism Brevibacterium sp. induced L-lysine-alpha-oxidase activity in Trichoderma spp.

[Characteristic effects of L-lysine-alpha-oxidase, an antitumor enzyme produced by Trichoderma sp].

Acute toxicity of L-lysine-alpha-oxidase, an antitumor enzyme of fungal origin was studied. After intravenous administration in a dose of 3000 U/kg the substance induced no signs of intoxication in the laboratory animals or their death. Direct dependence of the vessel permeability increase on the enzyme concentration was determined. In experiments on rabbits and guinea pigs L-lysine-alpha-oxidase showed no local irritating effect.

[Frequency of the Pro 12Ala polymorphism of PPARgamma2 gene in Ukrainian population and its possible role in development of metabolic syndrome].

Frequency of the Pro 12Ala gene polymorphism PPARg2 has been studied in 39 healthy men and 42 men with metabolic syndrome (MS) of the age of 40-65 years in Ukrainian population. The frequencies were 27.9% for 12Ala allele, 32.6% for Pro12Ala, and 4.4% for Ala1Ala that is close to the results of estimation of Czech population. The frequency of 12Ala allele was significantly less--18.4% (p < 0.05) in the persons with MS that corresponds to the pre-established information concerning the patients with diabetes of the 2nd type. A tendency has been detected to the increase of the body mass index in patients with 12Ala allele in the metabolic syndrome group.

[Biochemical characteristics of ticks of the families Argasidae and Ixodidae].

Polyacrylamide gel electrophoresis was used to study homogenate supernatants from the bloodsucking ticks (Argasidae and Ixodidae) belonging to 8 different genera and species. Differences were found in the electrophoretic mobility of protein fractions in adult ticks of these two different families and between several genera of Ixodes ticks (such as Haemaphysalis, Hyalomma, Dermacentor, Rhipicephalus and Ixodes). Argasidae ticks were found to have generic and even species-specific differences at the nymphal stage of development. A. lahorensis nymph-3 contained 5 protein fractions; O. papillipes had 2 fractions, and O. moubata had 3 fractions. Electrophoretic protein separation showed the equal number of protein fractions within one genus of ticks. At the larval stage, the tick homogenates were always found to have one protein fraction. The hemolymph of O. papillipes and O. moubata ticks displayed differences between the imago and nymph-3. A new biochemical marker was proposed to establish a species affinity.

[Metabolic syndrome in men of 35-65 years old from rural district of Poltava region and possibility of the prophylactic therapy with atorvastatine].

Observation of 250 men aged 35-65 years (middle age was 52 +/- 2 years) showed that prevalence of MS was 27%. Atorvastatine administration at dose of 10-20 mg per day for patients with ischemic heart disease and chronic obstructive lung disease associated with MS reduced during month administration the level of cholesterol, triglycerides, beta- and pre-beta-lipoproteids and increased the level of alpha-cholesterol in blood of these patients. Addition of atorvastatine to a common treatment patients with chronic obstructive lung disease improved as well the clinical picture of systemic inflammation process.

[Characteristics of the structure of plasma lipoprotein crystals]. / Osobennosti struktury kristallov lipoproteidov plazmy krovi.

Lipoproteids of human blood plasma in the norm and in pathology are studied by means of microscopy in polarized light. It is shown that lipoproteids in solid state have structure characteristics. Possible reasons of the observed structure characteristics of lipoproteid crystals are discussed.