[Polyelectrolyte complexes of lactoferrin and pH-sensitive microparticles on their basis].

Suspensions of insoluble polyelectrolyte complexes of dextran sulfate? (DS) of different molecular masses with lactoferrin (LF) have been fabricated and characterized. The encapsulation efficiency of LF and DS in a complex at pH 3.0 and 4.0 was assessed, and particles were characterized by their sizes and zeta-potential. The complexes formed at pH 3.0 differed by a higher stability level. The interaction with DS resulted in a twofold decrease in the antioxidant activity of LF, although the formation of complexes was not accompanied by conformational changes in LF molecules according to IR-spectrometry data. Microencapsulation was carried out by treating the suspensions with negatively charged LF-DS complexes with protamine and chitosane solutions with different molecular masses. The composition, size, and the zeta-potential of interaction products were assessed which allowed us to select the conditions for the preparation of pH-sensitive polyelectrolyte microparticles loaded with LF which would be able to gradually release glycoprotein under conditions that model the passage through the gastrointestinal tract of humans. These data indicate that this approach is promising for the creation of pH-sensitive biopolyelectrolytes suitable for oral administration of LF to target cells.

[Prophylaxis of hepatic insufficiency after major hepatic resections].

UNLABELLED: The article deals with a problem of prophylaxis of hepatic insufficiency in oncological patients after liver resections. MATERIALS AND METHODS: We analyzed data of effectiveness and safety of the use of Remaxol in oncological patients with hepatic metastasis of colorectal cancer--dynamics of indicators of cytolysis and cholestasis, hepatic protein synthesis, exchange of pigments, pro- and antioxidant system and the level of endogen intoxication in postoperative period. RESULTS: Use of Remaxol allows decreasing the duration of postoperative rehabilitation and intensive care unit staying.

[Experimental study of remaxol as a drug for supportive therapy in traditional and high-dose treatment of tumors].

We have experimentally investigated the detoxifying and modifying effects of remaxol in the framework of traditional and high-dose chemotherapy in mice with transplanted tumors. The influence of remaxol in comparison with heptral was studied on the toxic and therapeutic action of cytostatic drugs gemzar, lastet (etoposide), and methotrexate during their traditional and high-dose administration in mice with transplanted P388 lympholeukosis. Remaxol demonstrated a detoxifying action with respect to these cytostatic agents, which decreased in the following series: gemzar lastet methotrexate (according to the results of lethality evaluation). Remaxol significantly increased the therapeutic efficacy of gemzar used in both traditional and high-dose regimes. This was manifested by slowing down tumor growth and increasing animal lifetime during combined administration of the drugs. This may be due to the remaxol potentiation of the antitumor effect of gemzar. In combination with lastet and methotrexate, remaxol does not alter their therapeutic efficacy. The detoxifying action of remaxol with respect to these cytostatics is more pronounced than that of heptral.

[The test system to identify mucin MUC1 in human blood serum using the technique of immune-enzyme analysis based on monoclonal antibody ICO25].

On the basis of genuine mouse monoclonal antibody ICO25 the test system IEA ICO25 was developed and standardized to quantitative detect tumor-associated antigen, mucin1 in human blood serum in format of inhibitory immune-enzyme analysis. The analytic characteristics of test-system correspond to the standards applied to immune-enzyme diagnostic kits. The results of identification of MUC1 in blood serum of healthy donors and female patients with breast pathology using IEA ICO25 fully correlate with the data concerning the detection of antigen CA15-3 using certified commercial kits. The test system IEA ICO25 can be used to detect MUC1 in human blood serum for research purpose.

[Study of detoxifying efficacy of remaxol in experimental model of cisplatin-induced toxicosis].

The detoxifying efficacy of remaxol in the experimental model of cisplatin-induced toxicosis has been studied and the possibility of using this drug in cancer patients therapy is evaluated. Remaxol exhibited pronounced dose-dependent detoxifying effect in the model of toxicosis induced by cisplatin in a toxic dose (LD50). It reduced the death rate in test animals about three times when used at a 130 ml/kg dose, and prevented lethal outcome at a 500 ml/kg dose. Remaxol also normalized biochemical blood indices that characterized liver and kidney functions in survived animals. Remaxol did not stimulate growth of experimental carcinoma, sarcoma, melanoma, and lympholeukosis. The drug did not interfere with the anti-tumor efficacy of cisplatin in the schemes with combined treatment of animals bearing tumors of various histogenesis. Remaxol can be recommended for clinical study as a detoxifying preparation for cancer patients with various malignancies.

[Effect of polysaccharides on biological activity of human lactoferrin].

The influence of neutral and ionic polysaccharides on the antioxidant (AOA) and detoxifying activities of lactoferrin (LF) and the duration of its circulation in the body was studied. In addition to natural polymers, we studied artificial chitosan derivatives with different functional groups. On the basis ofAOA test, five polysaccharides were selected. The study of the detoxifying effect of LF in two models of induced toxicity revealed polysaccharides that maintained or increased the detoxifying activity of LF. We established that the formation of a complex of lactoferrin with two galactomannans and succinyl chitosan caused positive changes in LF properties: the detoxifying activity of the protein remained unchanged or increased, whereas its elimination from the body was decelerated.

[In vitro study of antimicrobial activity of lactoferrins from various sources].

Comparative antimicrobial activity of lactoferrins from various sources (native lactoferrin from Laprot, human hololactoferrin, recombinant human lactoferrin isolated from the cultural medium of permissive cell culture transfected using pseudoadenovirus nanostructure with the human lactoferrin gene, and native bovine lactoferrin) was studied to prove the possibility of their use for development of antimicrobial drugs. It was shown that all the substances were active against the Bacillus standard strains. The antibacterial activity was almost independent of the degree of saturation the lactoferrin molecules with Fe3+. The native human lactoferrin was more active than hololactoferrin against Candida when evaluated by the minimum inhibitory concentration (MIC). Fe(3+)-Non aturated recombinant human lactoferrin demonstrated the antimicrobial activity (by MIC) similar to that of the native human lactoferrin. The results showed that native and recombinant human lactoferrins might be used for the development of intravenous and intracavitary dosage forms, while the native bovine lactoferrin could be useful in development of oral drugs.

[An in vivo study of the biologic activity of the recombinant pseudoadenovirus nanostructure containing lactoferrin gene].

The Ad5-Lf recombinant pseudoadenovirus nanostructure (RPAN) based on adenovirus of the 5th serotype and containing lactoferrin (Lf) gene was constructed. The goal of this work was to develop a system for efficient production of human lactoferrin (Lf) in human body. It was shown using the model of cisplatin (DDP)-induced toxicosis that human Ad5-based RPAN with human Lf gene expressing cassette in its genome provides high rate of expression of Lf gene in animal body. In vivo recombinant human Lf demonstrates detoxification effect against acute DDP-induced toxic reactions similar to that of the native Lf. RPAN does not stimulate growth of primary and metastatic nodes of experimental tumors. Moreover, it inhibits the growth of Lewis lung carcinoma (LLC), Ehrlich carcinoma (ELD), and S37 sarcoma in early periods after tumor transplantation. The obtained experimental data are indicative of the good prospects of further biologic and medical study of RPAN and development of RPAN-based genetic engineering medicine of the new generation.

[Recombinant pseudoadenovirus nanostructure with human lactoferrin gene: production and study of lactoferrin expression and properties in vivo usage of the construction].

The Ad5-Lf pseudoadenovirus nanostructure (RPAN) was produced using homologous recombination in E. coli cells. This construction provided efficient expression of the Lf gene in permissive cell culture with high production rate of recombinant protein similar to native human Lf in some physical, chemical, and biological properties. Single intravenous injection of the construction into mice and rats was effective for prolonged production and circulation of recombinant human Lf in blood of experimental animals without toxic effects. The produced construction is promising for providing prolonged production of recombinant human Lf in the human body.

[Antitumor immunotherapy with the use of synthetic fragments of survivin].

The endogenous protein survivin is present in tumor cells and inhibits apoptosis. The influence of vaccination of mice by survivin fragments on growth of various types of tumors was studied to examine the possibility of creation of an antitumor vaccinating agent on its basis. Two peptides corresponding to the (118-144) and (80-88)-(153-165) sequences of survivin 2B were chosen and synthesized on the basis of literature data and theoretical calculations. Their ability to stimulate antibody production in mice of the C57BL/6J line (b haplotype) and in BDF1 hybrids (b x d haplotype) was investigated. Both peptides were shown to stimulate production of antibodies that bound the recombinant survivin in the BDF1 mice. Immunization of the BDF1 and C57BL/6J mice with the recombinant survivin resulted in the formation of antibodies that reacted with the (118-144) peptide. The effect of preventive vaccination with the peptides and the recombinant protein on the dynamics of growth of several species of tumors was studied. Vaccination with the (80-88)-(153-165) peptide was found to cause an antitumor effect in BDF1 mice suffering from sarcoma S-37. Thus, the creation of an antitumor agent on the basis of this peptide is a promising area of further studies.

[Antioxidants treatment of severe post-operative pyoinflammatory and septic complications].

New medical human lactoferrin product called Laprot possessing antioxidant, detoxicant, anti-inflammatory immunomodulating properties was developed and registered (serial number LS-002374) in the P.A. Hertsen Institute. System (intravenous) administration of Laprot is efficacious detoxicant and anti-inflammatory treatment in patients with severe postoperative pyoinflammatory and septic complications accompanied by polyorgan failure. Local administration contributes to clinically apparent cleansing of festering wounds and cavities, regress of local pyoinflammatory processes, reduction of local purulo-necrotic processes of trachea's mucosa. Laprot administrated intravenously as in the case of topical administration is well tolerated by the patients and doesn't cause any side affects.

[Role of the antioxidant ceruloplasmin in complex intensive therapy for severe posthemorrhagic complications in cancer surgery].

The paper summarizes the results of 10 years' experience in using the biocompatible antioxidant ceruloplasmin in cancer patients to prevent and treat life-threatening complications in critical states caused by various complications after extensive surgical interventions for malignant tumors or massive intraoperative blood loss. Hemorrhagic shock-complicated intraoperative massive blood loss is shown to exert a significant damaging effect on the redox system, which correlates with the objectified severity of a critical condition and with the degree of experienced hypoxia. The use of ceruloplasmin in cancer patients with postoperative complications or massive intraoperative blood loss contributes to the recovery of the potential of the antioxidative defense system, to the correction of oxidative stress, acute multiple organ deficiency, and endotoxemia, and to the reduction of the incidence of pyoseptic complications.

[Effect of substituents on photochemical and biological properties of 13,15-N-cycloimide derivatives of chlorin p6].

The effect of electron-accepting substituents in position 3 of the chlorine p6 macrocycle in neutral and carboxyl-containing negatively charged cycloimide derivatives of chlorin p6 (CIC) on the photochemical and biological properties of these photosensitizers was studied. A relationship between the structure and properties of CICs was analyzed on the basis of information on their photoinduced cytotoxicity, efficiency of the generation of reactive oxygen species, photostability, intracellular localization, quantitative parameters of accumulation in cells, and cellular pharmacokinetics. It was shown that these compounds can be used for the development of photosensitizers with intense light absorption at 740 nm, controlled intracellular localization, and a high photodynamic activity toward tumor cells.

[Photobiological properties of 13,15-N-(carboxymethyl)- and 13,15-N-(2-carboxyethyl)cycloimide derivatives of chlorin p6]. / Fotobiologicheskie svoistva 13,15-N-(karboksimetil)- i 13,15-N-(2-karboksiétil)tsikloimidnykh proizvodnykh khlorina p6.

Lipophilic derivatives of chlorin p6, 13,15-N-(carboxymethyl)cycloimide methyl ester (CIC1) and 13,15-N-(2-carboxyethyl)cycloimide methyl ester (CIC2), were shown to absorb light in 710 nm region and to be efficient IR photosensitizers. They exhibit similar phototoxicities on the cells of A549 human lung adenocarcinoma, which are 40- and 100-fold higher than those of chlorin p6 and the clinically used Photogem, respectively, and are not toxic in the absence of light irradiation. The confocal spectral imaging technique allowed us to demonstrate that the high phototoxicity of CIC1 and CIC2 is due to their ability to readily penetrate to cells and to be bound to the cell membranes and lipid-containing structures in the monomeric photoactive form. Under the irradiation, the membrane-bound CIC1 and CIC2 are characterized by high quantum yields of singlet oxygen generation (0.6 and 0.65, respectively) and the inability to produce hydroxyl radicals. A 1.5-microM content of CIC1 and CIC2 in the incubation medium provides for their average cytoplasmic concentrations of 21 and 16.5 microM, respectively. The incubation times to achieve 50% level of maximum accumulation for CIC1 and CIC2 in A549 cells are 30 +/- 6 and 24 +/- 12 min, and the times for 50% release of the dyes from the cells are 17 +/- 4 and 50 +/- 10 min, respectively. A diffuse distribution with the predominant accumulation in the membranes of the Golgi apparatus and mitochondria is characteristic of both CIC2 and CIC1, whereas, in addition, CIC1 is considerably accumulated in lipid droplets (cellular organelles responsible for the storage and metabolism of neutral lipids and steryl esters). Our results demonstrate that changes in the structure of the imide substituent could affect the intracellular localization and the rate of release of chlorin p6 cycloimide derivatives from cells while preserving their high photodynamic activity.

[DNA degradation and repair in human laryngeal carcinoma HEp-2 cells after combined exposure to vitamin B12b and ascorbic acid]. / Degradatsiia i reparatsiia DNK v kletkakh épidermoidnoi kartsinomy gortani cheloveka HEp-2 pri sovmestnom deistvii vitamina B12b i askorbinovoi kisloty.

The formation and accumulation of DNA fragments containing no more than 23,000 pairs of bases were observed under exposure of human larynx epidermoid carcinoma cells (Hep-2) to "chemical nuclease", oxycobalamin (vitamin B12b) and ascorbic acid (vitamin C). The obtained DNA damages were repaired more slowly than those induced by gamma-irradiation in the dose adequate to the level of DNA damages. DNA reparation was not revealed after washing the cells from vitamin B12b and ascorbic acid, and in the course of cell incubation with ascorbic acid. Vitamin B12b and ascorbic acid separately did not induce degradation of DNA. DNA damages induced by "chemical nuclease" action precede the cell death observed later.

[Efficacy of ceruloplasmin in the resuscitation oncology clinic]. / Effektivnost' tseruloplazmina v reanimatsionnoi onkologicheskoi klinike.

The use of ceruloplasmin in the intensive therapy of oncological patients has been shown to optimize the course of the rehabilitative period promoting dissipating of polyorgan failure, restoration of oxidant-antioxidant balance, reduction in the incidence of pyo-septic complications. Ceruloplasmin is found out to have an apparent detoxicating and antiinflammatory effect, it activities the bodily antioxidant defence, favors normalization of the condition of the immunity lymphocytic link.

[Antioxidants ceruloplasmin and lactoferrin in the prevention and treatment of postoperative complications in cancer patients]. / Antioksidanty tseruloplazmin i laktoferrin v profilaktke i lechenii posleoperatsionnykh oslozhnenii u onkologicheskikh bol'nykh.

Ceruloplasmin and laprote based on natural protein antioxidants were used in the treatment and prevention of postoperative complications in 174 patients after extensive interventions for cancer. Development of pyoseptic complications during the postoperative period is associated with activation of lipid peroxidation, decreased functional activity of the antioxidant component of detoxication, suppressed T-cellular immunity, and development of polyorgan failure in the presence of endogenous toxemia. Systemic treatment with laprote (50 pts) and ceruloplasmin (33 pts) after surgical cleansing and draining of purulent focus stimulated activation of antioxidant defense, decreased the intensity of oxidative processes, normalized the lymphocytic component of immunity, and promoted resolution of polyorgan, primarily hepatic failure. Local therapy with laprote (60 pts) promoted rapid regression of local pyoinflammatory processes. Intraoperative blood loss complicated by hemorrhagic shock had a deep impact on the oxidative/antioxidant system, which correlated with the severity of hypoxia. Addition of ceruloplasmin, a potent antioxidant, to therapy of these patients (n = 31) optimized the course of recovery by stimulating the resolution of posthypoxic polyorgan failure, recovery of the oxidant/antioxidant balance, and decreasing the incidence of postoperative pyoinflammatory complications.

[Study of localization and molecular interactions of biologically active compounds in living cells and tissue slices based on confocal microspectroscopy and reconstruction of the spectral images]. / Issledovaniia lokalizatsii i molekuliarnykh vzaimodeistvii biologicheski aktivnykh soedinenii v zhivykh kletkakh i srezakh tkanei na osnove metoda konfokal'noi mikrospektroskopii i rekonstruktsii spektral'nykh izobrazhenii.

The confocal spectral imaging (CSI) technique is described, its basic principles are considered, and a brief review of its applications to the study of biologically active compounds (BAC) within living cells and in tissue slices is presented. This technique is based on measurements and analysis of fluorescence or resonance Raman spectra in each point of the specimen under microscope with a three-dimensional resolution of about cubic micrometer. This technique is applicable to the study of stained fluorescent and nonfluorescent compounds. Unlike the conventional approaches based on the optical microscopy, the CSI technique opens the opportunity for the identification of complexes and microenvironment of BAC in intact cells and thin tissue slices (slices or sections), as well as for the analysis of localization and distribution of compounds of interest and their complexes in cellular organelles and tissue structures. The use of CSI technique in combination with the conventional biochemical and cytological methods makes it possible to significantly expand the informativeness of investigation of modes of action of new BAC.