Binding Potency of Heparin Immobilized on Activated Charcoal for DNA Antibodies.

In vitro experiments showed that heparin adsorbed on activated charcoal can bind antibodies raised against native and single-stranded DNA in a diluted sera pool with a high level of these DNA. Thus, heparin used as anticoagulant during hemosorption procedure can demonstrate supplementary therapeutic activity resulting from its interaction with various agents involved in acute and chronic inflammatory reactions such as DNA- and RNA-binding substances, proinflammatory cytokines, complement components, growth factors, etc. Research and development of heparin-containing carbonic adsorbents for the therapy of numerous inflammatory and autoimmune diseases seems to be a promising avenue in hematology.

Efficacy of dispersed fibrous carbon sorbent in treatment of malignant fungating wounds.

AIM: To evaluate the influence of dispersed fibrous carbon sorbent (DFCS) on malignant fungating wounds (MFWs) in Guerin's carcinoma-bearing rats with an assessment of wound bacterial microflora. MATERIALS AND METHODS: The study was performed on female Wistar rats inoculated subcutaneously with Guerin T8 carcinoma into the upper interscapular region. On day 20, the tumors were infected by injecting the suspension of S. aureus 8325-4. After 24 h, gauze dressings were applied daily to the wounds in the control group and DFCS in the experimental one. The state of the wounds was assessed by visual control with photo monitoring, odor control on a verbal rating scale and microbiological analysis of qualitative and quantitative characteristics of wound microbiota. RESULTS: On the 3rd day after the start of dressings, the number of S. aureus of 8325-4 strain was significantly higher in the wounds of rats in control group than experimental one (5.14 ± 0.27 vs 3.43 ± 0.33 lg CFU/ml). The total number of Staphylococci in the DFCS-treated wounds did not differ statistically from the number of S. aureus 8325-4 strain whereas in control ones were higher by an order of magnitude. On the 10th day the total number of Staphylococci and S. aureus 8325-4 in particular, remained consistently high in the wounds of the control rats, while was almost five orders of magnitude lower and represented mainly by S. aureus 8325-4 in the experimental ones. The total number of aerobic and optionally anaerobic microorganisms was significantly lower as well. A greater variety of microorganisms in the gauze-covered wounds, as well as an their increased number were accompanied by enhancement of the wound odor from "noticeable" to "strong". There was an appearance of a "barely noticeable" odor in only one animal from the experimental group. CONCLUSION: The study has demonstrated the ability of DFCS to control substantially the bacterial microflora as well as malodor of MWFs in vivo. The results obtained can contribute to solving the problem of improving the quality of palliative care for patients with malignant and other chronic wounds.

Comparative study of biochemical and morphological parameters in rats with Walker 256 and Walker 256/DOX carcinosarcoma.

AIM: To evaluate the changes of some biochemical blood plasma parameters and morphological structure of the internal organs of rats with transplanted doxorubicin (DOX)-sensitive (Walker 256) and doxorubicin-resistant (Walker 256/DOX) strains of Walker 256 carcinosarcoma. MATERIALS AND METHODS: The study was performed on female Wistar rats with transplanted Walker 256 or Walker 256/DOX and intact animals (control). On the 9th day after transplantation of tumor cells, a comparative analysis of some blood plasma biochemical parameters and morphological examination of the liver, kidneys, myocardium and spleen of rats was carried out. RESULTS: Walker 256 growth, in comparison with Walker 256/DOX, is accompanied by more pronounced systemic effect on tumor-bearing rats. Uric acid concentration in the blood plasma of Walker 256 bearing rats was significantly (by 15.5%) higher than in Walker 256/DOX bearing rats. Aspartate aminotransferase activity in the Walker 256 group was significantly (by 107.2%) higher than in Walker 256/DOX group, but alanine aminotransferase activity was 58.5% lower. 56.7% decrease of alkaline phosphatase in rats with Walker 256, and 21% increase of this index in rats with Walker 256/DOX were observed. The growth of Walker 256 carcinosarcoma led to greater structural damage of the liver, kidneys and spleen in experimental animals compared with Walker 256/DOX strain. CONCLUSION: Tumor growth in rats with Walker 256/DOX leads to less pronounced changes in the biochemical parameters of rat blood plasma and morphological structure of internal organs compared with wild-type carcinosarcoma.

[Fatty acid composition variability of rapeseed oil: classical selection and biotechnology].

The problems and achievements in the rapeseed Brassica napus L. var. oleifera breeding directed on the change of fatty acid composition in seed oil with the use of traditional and genetic engineering approaches are analyzed. It is noticed that the combination of biotechnological workings out and methods of classical breeding is the optimum for the further improvement of rapeseed oil composition.

Adsorptive therapy as a modificator for tumor-host interaction.

The potential of one of the adsorption methods, enterosorption (ES), using the new generation of carbon adsorbents to correct the negative manifestations of tumor-host interaction in the framework of paraneoplastic syndrome (PNS) as well as systemic toxicity of chemo- and radiation therapy, is discussed. The ES influence on the development of PNS was demonstrated in C57/BL6 mice with transplanted Lewis lung carcinoma. Two-week administration of carbon enterosorbents resulted in a significant suppression of metastasis and correction of tumor-related anemia, activation of granulocytic line in the bone marrow with nearly 3-fold enhancement of its mitotic activity. ES exerted a positive influence on the structural-morphologic indexes and regenerative potential of kidneys and liver, mitigated manifestations of oxidative stress, decreased the level of endogenous intoxication, increased resistance of erythrocyte membranes and decreased ligand loading of blood plasma transport proteins. The effect of ES on anticancer activity and toxic reactions of cisplatin (CP) was evaluated in Guerin carcinoma-bearing rats. ES reduced significantly creatinine and other kidney biochemical indexes elevated in the blood plasma of rats after CP treatment. ES attenuated dystrophic changes in the histological structure of internal organs (kidney, liver, spleen), caused by tumor growth and significantly aggravated under the influence of CP. Such changes were specially traced in the kidneys and well reflect the nephroprotective potential of ES. In rats irradiated with X-ray in sublethal dose, highly activated granulated carbonic enterosorbents facilitated the restoration of white blood cells and lymphocyte count. The results obtained confirm the insights of academician R.E. Kavetsky predicting the future of adsorptive detoxification with activated carbons in the treatment of cancer patients.

[Stable expression of promoterless bar gene in transgenic rape plants].

Phosphinothricin resistant plants of two rapeseed (Brassica napus L. var. oleifera DC.) spring industrial cultivars were obtained by Agrobacterium tumefaciens leaf disk transformation. Vector constructions contained the promoterless coding sequence of phosphinothricin acetyltransferase (bar) gene located between two inverted lox-sites (elements of Cre/lox recombination system of P1 phage) and selective neomycinphosphotransferase II gene (nptII). Integration of the alien genes was confirmed by the PCR analyses. Stable and linked inheritance of foreign genes in T1 and T2 progeny was shown.

[Immunological characteristics of a modifying effect of bacterial endotoxins on the growth of experimental tumors]. / Immunologicheskaia kharakteristika modifitsiruiushchego vliianiia bakterial'nykh éndotoksinov na rost éksperimental'nykh opukholei.

Two experimental models have been elaborated for E. coli 0111:B4 endotoxin effects on the growth of transplanted murine sarcomas induced by Meth A. A correlation is shown between stimulatory and inhibitory effects of endotoxin on the tumour growth and functional activity of T- and B-lymphocytes registered by in vivo tests.

[Antimetastatic action of bacterial endotoxins and changes in the activity of the enzymes of purine metabolism in macrophages]. / Antimetastaticheskoe deistvie bakterial'nykh éndotoksinov i izmeneniia aktivnosti fermentov purinovogo obmena v makrofagakh.

The antimetastatic action of bacterial endotoxins (BET), E. coli 0111:B4, B in particular, as well as their influence on the adenosine deaminase and 5'-nucleotidase activity were studied in peritoneal macrophages of mice bearing lung Lewis carcinoma. BET inhibition of lung metastasis growth was found to be due to such changes in macrophage adenosine metabolism, that testifies to the rise of their functional activity. The change in the level of macrophage purine metabolism can be considered as an important evidence of the effectiveness of the drugs capable to inhibit the metastasis growth.

[Obtaining and analysis of intergeneric somatic hybrids between Brassica napus and "albino" line of Orychophragmus violaceus]. / Poluchenie i analiz mezhrodovykh somaticheskikh gibridov mezhdu Brassica napus i liniei tipa "al'bino" Orychophragmus violaceus.

The Orychophragmus violaceus chlorophylldefective line of "albino" type has been obtained by spectinomycin treatment. Somatic hybridization between Orychophragmus violaceus and Brassica napus was performed by fusion of green mesophyll protoplasts of rape and callus protoplasts of the O. violaceus "albino" line. Near two hundred of regenerant plants were selected according to the regeneration type and ability to become green, and were determined as hybrids. Chloroplast DNA in selected hybrids was identical to rape chlDNA, which was confirmed by the PCR-RFLP analysis of plastid DNA fragments. Fragments of hybrid mitochondrial DNA analyzed by the PCR-RFLP analysis were identical to fragments of O. violaceus. The nuclear genome of the majority of hybrids was represented by the O. violaceus genome, which was demonstrated by analyses of isoenzymes, DNA telomeric sequences, ribosomal and satellite DNAs, and the RAPD analysis. The cytogenetic analysis of a number of lines has shown variability in the number of chromosomes in the obtained lines.

[Activity of the corn Spm transposon system in transgenic plants Orychophragmus violaceus (L.) O.E. Schulz obtained by both direct transfer of DNA to protoplasts and agrobacterial transformation of root explants]. / Aktivnost' sistemy Spm transpozonov kukuruzy u transgennykh rastenii Orychophragmus violaceus(L.) O.E.Schulz, poluchennykh putem kak priamogo perenosa DNK v protoplasty, tak i agrobakterial'noi transformatsiei kornevykh éksplantov.

Transposon mediated insertional mutagenesis is one of the approaches for the unique gene cloning. A wild species of Cruciferae family Orychophragmus violaceus (L.) O.E. Schulz, which is of interest for practical breeding as a donor of improved plant oil, was an object of the investigation. Plasmid construction used in the experiments included selective NPT II gene, reported GUS gene serving as an excision marker, structural BAR gene located within the dSpm element and Spm transposase. The GUS gene of this plasmid had not his own promoter and became functional only after Spm-transposition. Transformed Orychophragmus violaceus (L.) O.E. Schulz. plants were obtained by direct mesophyll protoplast transformation as well as Agrobacterium tumefaciens-mediated root explant transformation. Gene transfer and the transposition event were confirmed by the GUS activity and the PCR analysis. Relative transformation efficiency using protoplasts was 5.8%.

Enterosorption as a method to decrease the systemic toxicity of cisplatin.

UNLABELLED: A perspective adsorptive method to minimize systemic toxic effects of chemotherapy is enterosorption (ES). However, the capabilities of this method are far from being completely studied. The question remains opened - should ES be initiated in the first hours on completing cytostatic infusion without the risk of their anticancer activity to be decreased. AIM: to analyze ES influence on anticancer activity and toxic reactions of cisplatin (CP) upon the use of carbon enterosorbent in 1 h after intravenous administration of cytostatic. METHODS: CP at the dose of 1 mg/kg body weigh (BW) was administered to Guerin carcinoma-bearing rats each second day for two weeks. Enterosorbents on the basis of highly activated carbon fibers were administered by per os daily 1 h after CP injection. 3 days after the last CP administration the rats were weighted and blood under ether narcosis has been taken for biochemical examination. Tumors and innate organs were isolated, weighted, and fixed in 4% buffered formalin for morphologic examination. RESULTS: In rats administered with CP at the background of ES, BW loss was in 1.6 times lower than in animals after CP session. Relative kidney weight in CP-treated rats was 33.9% higher than in normal ones (p ≤ 0.05). No significant differences were detected between relative kidney weights in the CP + ES-treated and intact animals. Introduction of ES allowed prevent an 30% increase of creatinin content observed in blood plasma after CP treatment (р ≤ 0.05). Urea content was 1.7 times lower in blood plasma of CP + ES-treated rats than after CP treatment. CP caused significant toxic injuries in kidneys, liver, and spleen tissues. Morphologic structure of organs in rats treated with CP at the background of ES was affected at much lower degree. In tumors, large areas of newly formed connective tissue and blood vessels have been fixed after the CP+ES action instead of large necrotic area observed after CP treatment. ES caused insignificant suppression of Guerin carcinoma growth and had additional impact to inhibitory action of CP. CONCLUSION: Active carbon enterosorbents which are administrated just 1 h after CP administration possesses detoxicating potential sufficient for significant elimination of toxic effect of the cytostatic at the background of complete preservation of its antitumor activity.

Theoretical ground for adsorptive therapy of anthracyclines cardiotoxicity.

Anthracyclines play an important role in treatment of variety types of cancer due to their high effectiveness and broad spectrum of activity. However, a major limitation of their use is the dose-limiting cardiotoxicity. The inability to predict and prevent anthracycline cardiotoxicity is in part due to the fact that the molecular and cellular mechanisms remain controversial and incompletely understood. This review focuses on the biochemical basis of the anthracyclines toxic cardiac effects and pharmacological measures to their treatment and preventing. We describe the theoretical substantiation of the enterosorption abilities for diminishing of cardiac damage.

Carbon adsorbents in oncology: achievements and perspectives.

The results of own investigations and literature data are summarized to determine the place of the main methods of adsorption therapy in complex treatment of the patients with malignant tumors. New possibilities for the usage of new generation of carbon adsorbents and modern adsorptive technologies in cancer treatment are discussed.

New approaches to the removal of protein-bound toxins from blood plasma of uremic patients.

The article is devoted to the theoretical aspects of the development of the effective method for the removal of protein-bound uremic toxins. It is shown that the methods of flow and differential scanning microcalorimetry are sufficient enough for the evaluation of the degree of ligand loading of human serum albumin with protein-bound uremic toxins. The molecules of albumin isolated from blood plasma of the patients being kept on chronic dialysis are demonstrating significant alterations of conformation and complex-forming properties, the correction of which by conventional methods of extracorporeal detoxification (exhaustive dialysis, treatment on synthetic SCN carbons) are practically ineffective. Deliganding of uremic albumin may be successfully performed on conventional carbon haemosorbents upon preliminary separation of blood plasma and its dilution with acetate buffer 1:1 at pH = 5.08. Treatment of the whole blood of patients onto new mass-fractal deliganding carbon, i.e., hemosorbents of HSGD trademark. These HSGD haemosorbents quite effectively could be used for restoration of main parameters of uremic HAS molecules conformation and ligand-binding activity simultaneously with hemodialysis upon the protection by locally performed citrate anticoagulation as an easier and cheaper method for the removal of protein-bound uremic toxins.

The study of possibility to elevate antitumor activity and decrease of systemic toxic effects of cisplatin by its binding with deliganded albumin.

AIM: To evaluate antitumor and toxic action of cisplatin (CP) in non-bound form and in a complex with deliganded albumin. METHODS: To study complex-formation between CP and albumin, differential scanning and isothermic flow microcalorimetry were used. For quantitive evaluation of albumin-bound CP, the method of ultrafiltration was applied. Concentration of platinum in the samples was determined by atomic-absorption spectral analysis. Antitumor and toxic effect of CP and CP-albumin complex was studied in vivo using Guerin carcinoma (GC) model. RESULTS: It has been shown that the second drug-binding site, located in the III domain of albumin molecule is one of the main points of binding of CP. Purification of officinal human serum albumin (HSA) on highly active carbon hemosorbents of HSGD mark allows to obtain deliganded albumin (dHSA) with elevated complex-forming ability toward CP. Administration of CP-dHSA complex provides higher rate of GC growth inhibition, than that of CP, and the content of creatinine in blood plasma of GC-bearing rats increases by 15% versus 40% in the case of CP administration. CONCLUSION: The data obtained allow recommend application of CP-dHSA to complex for enhancement of antitumor action and decrease of toxic effects of cisplatin.

Application sorption: experience in clinical use and prospects of development.

The analysis of experimental - clinical results obtained during the last decade in the field of application of adsorptive carbon dressings in wound and burn therapy is given. Possible mechanisms of the therapeutic effect of this material are discussed, the feasibility of their application during different phases of wound process and burn disease treatment is demonstrated. The local absorption may be independent efficient method of treatment in the case of surface burns and wound with a small area of injury and provide a pronounced detoxifying effect in the case of suppurative-septic complication of a mechanical trauma and burns.

[Effect of applicative-sorptive therapy on the activity of various enzymes and inhibitors of the kallikrein-kinin system in traumatic disease]. / Vliianie applikatsionno-sorbtsionnoi terapii na aktivnos't nekotorykh fermentov i ingibitorov kallikrein-kininovoi sistemy v dinamike travmaticheskoi bolezni.

On the model of musculocutaneous wound in rabbits, the effect of applicative sorption with the use of activated carbonic fibrous materials on the course of traumatic disease was studied. The total esterase activity of the blood serum, activity of alpha 1-inhibitor of proteinases, kallikrein content in the blood serum were defined. It is shown, that changes in biochemical indices are in agreement with the findings of clinical observations and outcome of traumatic disease. The use of activated carbonic fibrous material at the early period of traumatic disease contributes to its favourable course, prevents the development of complications and death of the animals from sepsis.

[Experimental substantiation of the early use of applied sorption in treating wounds]. / Eksperimentalnoe obosnovanie rannego primeneniia applikatsionnoi sorbtsii v lechenii ran.

In the experiments on rabbits with bilateral musculocutaneous wound of a thigh, it is shown that the early use of the "Dnepr MN" activated carbon fibrous material (ACFM) intended for medical purposes provides timely haemostasis in the damaged tissues, reduces blood loss, prevents the development of traumatic edema, decreases the intensity of inflammatory process in a wound, facilitates rapid and uncomplicated healing of wounds under the artificial crust. The treatment-and-prophylactic effectiveness of ACFM is associated with its ability for rapid and irreversible sorption of biologically active substances forming in the wound tissues, and as well with protection of a wound against secondary infection.

Basic studies of local adsorption in burn treatment.

The model of a full-thickness thermal burn in rats was used to demonstrate that the application of adsorptive dressings made of a fibrous activated carbon material (ACFM) after early excision of a burn crust exerts a considerable influence on the development of both local and general symptoms of burn disease. Three days after the excision a water content in underlying tissues falls from 84.81 +/- 0.62% to 54.47 +/- 4.2%, the wound surface area decreases by 10%, while this surface area under gauze bandages increases by 6.3%. The local adsorption results in lower activity of proteolytic serum enzymes and toxicity of blood serum in burned animals as well as in the positive dynamics of changes in functional activity of the immune system. By the 14th day the T-cell activity in animals with adsorptive carbon dressings was by 3.2 times higher than that in the animals with gauze bandages. The level of B-cell response if ACFM is applied exceeds normal values to a great extent, starting from the first days after the excision of a burn crust.