Carcinogenesis--a new point of view.

Presented article suggests the novel hypothesis of carcinogenesis, where the key moment for all types (biological, physical, chemical) of carcinogenesis has been discussed. For confirmation of the hypothesis thorough theoretical analysis of the mechanisms of malignant transformation of cells after influence of any type of carcinogens and results of experiments have been presented. Hypothesis highlights are formulated as follows: 1) Covalent bond disorders between S+-methionine and Fe3+ atoms in cytochrome; 2) Electron transport chain blockade with certain ligand after its penetration in cytochrome pocket with further formation of 6th coordination bond between ligand and Fe atom (in one case increase in mitochondrial pH precede-, and in other, it follows electron transport chain blockade in cytochromes); 3) Fe3+ reduction up to Fe2+ leading to blockade of aerobic glycolysis; 4) Decrease in enzyme (Е1-TDP, oxidases etc.) activity due to mitochondrial pH alterations; 5) Production of S-adenosylmethionine owing to lipoic acid amide leading to accumulation of homocysteine in cytoplasm with further penetration in cell nucleus producing DNA mutations; 6) Fe2+ wash-out from cytochrome and its deposition in ferritin.

Anticancer efficacy of hydroxyethylthiamine diphosphate in vivo.

AIM: The aim of the presented article was investigation of anticancer efficacy of hydroxyethylthiamine diphosphate (HTD) in vivo. MATERIALS AND METHODS: The study was carried out on 30 C57BL/6J mice subcutaneously transplanted with Ehrlich carcinoma. Animals were treated with intraperitoneal injections of the solution composed from pyruvic acid and thiamine bromide every other day during 10 days and thereafter, every day during 2 weeks. Treatment efficacy was evaluated by tumor growth inhibition. RESULTS: In experimental animals treated with HTD, significant tumor growth inhibition has been registered: 73% at day 45(th) compared to the control group (p < 0.001). CONCLUSION: Treatment with HTD demonstrated high anticancer efficacy in vivo.

[Bacterial phagelysates and malignant tumor growth].

Anti-tumor preventive efficacy of E.coli phagelysate has been studied. Investigations were conducted on 2-3 months 48 male mice. Regimen of preventive vaccinations were: single - 0,25 ml phagelysate intraperitoneal injection, 3 days before Ehrlich carcinoma inoculation (1x10(6) tumor cells); 3 times vaccinations (0,25 ml, with 3 day intervals) 3, 6, and 9 days before inoculation of carcinoma; and 10 times (during 10 days, before inoculation of carcinoma). Treatment efficacy was evaluated according to the indices of cancer growth (development of cancer tissue, cancer growth inhibition percent, lifespan and survival percent). Experiments have shown that single and 3 times preventive vaccinations inhibited tumor development and delayed malignant growth, while, 10 times permanent vaccinations had no effects on cancer growth. Cancer growth inhibition percent in single and 3 times vaccinated animals were 58% on the average. Maximal lifespan in control group mice consisted 59 days. By the 125th day of cancer growth, at single vaccination 17% of mice were alive, while in 3 times vaccinated mice the survival percent was 25%. Anti-tumor potential of E.coli pagelysate supposedly could be explained by immunoregulatory properties of the preparation.

The novel hypothesis of carcinogenesis and anti-cancer treatment perspectives - Hydroxyethylthiamine diphosphate.

In presented article, the novel hypothesis of carcinogenesis, and thorough discussion of some essential biochemical mechanisms which might be responsible for the malignant transformation of cells (bond disorders between Fe(3+) and S(+)-methionine in cytochrome; blockage of the last 3-d orbital of Fe by the certain ligand and formation of 6th coordinated bond leading to Fe(3+) reduction up to Fe(2+) and cessation of tissue respiration; increase in mitochondrial pH and further in cell, leading to decreased activity of most of enzymes (thiamine diphosphate, oxidases); formation of S-adenosyl-methionine with its further dissociation and production of adenosine and homocysteine; effects of homocysteine on DNA structures, homocysteine-induced dimethylation of certain nitrogen basis and their extrusion from strands of DNA leading to mutations and cellular atypism) has been suggested. Along with theoretical discussions, article provides results of preliminary investigations carried out on C57Bl/6J mice with Ehrlich carcinoma aimed to capture lipoic acid amide with Hydroxyethylthiamine diphosphate, and study effects of Hydroxyethylthiamine diphosphate against malignant transformation of cells. Experiments have shown inhibition of cancer growth in treated animals. Morphological investigations of cancer tissue revealed necrotic zones, inflammatory infiltrations, central necrosis with adjacent inflammatory mono- and polymorph infiltrations, must cells, segmento-nuclear leukocytes, lymphadenoid follicular hyperplasia. According to the novel hypothesis of carcinogenesis and results of experiments the new approaches and perspectives of anti-cancer treatment with the use of Hydroxiethyl-thiamine diphosphate has been suggested.

Antitumor and adjuvant effects of phagelysates of E.coli in mice with Ehrlich carcinoma.

AIM: To augment anti-tumor host response and overcome the tumor-induced immunosuppression is of paramount importance especially when patient is subjected to radio-/chemotherapy and immune system suffers significantly. Various immunological methods have been employed as supplemental antitumor therapies. We were aimed to investigate the antitumor potential of phagelysates of gram-negative bacteria and their adjuvant effects for conventional chemotherapy in experiment. METHODS: Bacterial phagelysates of E.coli and purified suspensions of corresponding Un bacteriophage were obtained by standard methods of phage research. Experiments were carried out on BL57C/6J mice bearing transplanted Ehrlich carcinoma. Different regimens of phagelysate administration (0,5 ml E. coli phagelysate, 3/8 times with 5 day intervals) and conventional chemotherapy (combination of Doxorubicin 60 mg/m2, Cyclophosphan 800 mg/m(2), Ftoruracil 600 mg/m(2), 3 times with 21 day intervals) were tested. Treatment efficacy was evaluated by tumor growth inhibition percent, index of malignant growth, lifespan and survival percent. RESULTS: Experiments have shown that application of optimal doses of E. coli phagelysate can be well tolerated in mice. No stimulation or support of malignant growth was observed. E. coli phagelysate exhibited significant anticancer effect and adjuvant efficacy. Cancer development was delayed in 65% of inoculated animals in the test group. E. coli phagelysate inhibited tumor growth by 80-90% without apparent side effects. The mice survival was prolonged twice and more. On 65th-69th days of tumor growth in 13% animals complete regression of neoplasms was registered. Application of phagelysates in combination with chemotherapy significantly increased antitumor efficacy of conventional chemotherapeutic drugs. CONCLUSION: Application of bacterial phagelysates can be considered as promising novel strategy in cancer therapeutics.

[Application of bacterial thermo- and phagelysates for suppression of malignant tumor growth in experimental studies: 2 comparative analysis of anticancer efficacy of thermo- and phagelysates of Ps. aeruginosa and E. coli].

In the presented work antitumor effects of thermo- and phagelysates of Ps.aeruginosa and E.coli on Ehrlich carcinoma growth in mice have been studied. The treatment efficacy was evaluated according to the dynamic changes in volume of cancer tissue, cancer growth inhibition percent and calculations using Semi-empirical mathematical model describing cancer volume variations in relation to time passed after carcinoma inoculation. It was shown that at the early stage of cancer growth all tested bacterial preparations significantly inhibit cancer growth. Antitumor treatment effects were better expressed in animal studies using bacterial pagelysates in comparison to that of thermolysates. Comparative analysis of anticancer treatment efficacy of Ps.aeruginosa and E.coli phagelysates have shown that E.coli phagelysates reveal stronger and more prolonged anticancer treatment properties than phagelysates of Ps.aeruginosa.

[Application of bacterial thermo-and phagelysates for suppression of malignant tumor growth in experimental studies: 1. Anticancer efficacy of thermo- and phagelysates of E.coli].

Immunotherapy is considered as one of the promising treatment strategies for patients with malignant tumors. In the presented work antitumor effects of E.coli thermo- and phagelysates on Ehrlich carcinoma growth in mice have been studied. The treatment efficacy was evaluated by estimation of dynamic changes in volume of cancer tissue and by the percentage of cancer growth inhibition. Semi-empirical mathematical model, describing cancer volume variations in relation to time, passed after Ehrlich carcinoma inoculation was elaborated. It was shown that at the early stage of cancer growth both tested bacterial preparations significantly inhibit cancer growth. Antitumor treatment effects were better expressed in animal studies using bacterial phagelysates in comparison to that of thermolysates. After cessation of administration of bacterial preparations a prolonged antitumor activity in animals treated with E. coli phagelysates was demonstrated.

Addictive potential of Tianeptine - the threatening reality.

A total of 24 patients (male volunteers), consumers of opiates in the past and suffering from Tianeptine abuse, were under clinical observation. The age range of patients was from 21 to 33 years. Tianeptine consumption history was 5 months duration on the average. The daily dose of preparation was 40 tablets (500 mg intravenous injections on the average). Patients used Tianeptine in combination with antihistamines (Promethazine, Suprastin). Research was carried out with the use of clinical, psychological and laboratory methods. Has been used Ch. Spilberger's scale of anxiety and T. Balashov's scale of depression. Comparison of withdrawal syndrome developed after cessation of Tianeptine and opiates consumption has shown that in case of Tianeptine, in the dynamic of withdrawal syndrome predominates well expressed high-level of anxiety and depression, while at opiates consumption - withdrawal syndrome is characterized by algesic events and vegetodysfunctions. Supposedly, Tianeptine, in contrast to other anti-depressants, stimulates release of neurotransmitter dopamine in nucleus Accumbens, that probably determine addictive potential of this drug. High level of anxiety, excitability and vegetodysfunctions, presumably could be explained by activation of the NMDA (glutamate receptors) receptor system in Locus coeruleus, and in vegetative ganglion. In the present article potential threat that may develop at Tianeptine consumption, especially in former opiate consumers, without medical purposes has been emphasized.

[Influence of different type beta-adrenoblockers on functional state of liver at paracetamol induced toxic hepatitis in experiment].

The aim of the present work was comparative analyze of different type beta-adrenoblockers (nonselective beta-AB-Propranolol; cardio selective beta1-AB-Bisoprolol; nonselective beta-AB with additional vasodilatory effects - carvedilol) on functional state of liver at Paracetamol induced toxic hepatitis in experiment. Experiments were carried out on 42 laboratory white rats with the body mass 180-200 g. Model of acute toxic hepatitis was created using paracetamol (peroral administration with the dose of 1000 mg/kg, once a day). Beta-adrenoblockers (beta-AB) were used with following dozes: propranolol -1,4 mg/kg; bisoprolol - 0,7 mg/kg and carvedilol - 0,9 mg/kg; beta-AB were used during 9 days (perorally), immediately after paracetamol administration. Each group consisted of 7 rats. Functional state of the liver was estimated according to blood serum concentration of aspartat-transferase (AST) and alanin-transferase (ALT) using routine method of measurement on 5th and 9th days after administration of paracetamol. Results of experiments have shown that single administration of paracetamol with the dose of 1000 mg/kg impaired functional state of the liver that was confirmed by significantly increased activation of transaminases. It was stated that non-selective beta-AB Propranolol worsens-, cardio selective beta1-AB Bisoprolol - has no effect, while nonselective beta-AB carvedilol, with additional vasodilatory and antioxidant effects - slightly improves functional state of the liver at acute toxic hepatitis caused by paracetamol. Has been suggested that hepatotoxicity is not characteristic for all types of beta-AB. According to the obtained results, it has been recommended to estimate functional tests of the liver before propranolol administration, and carry out monitoring of the liver's functional state at treatment course, especially in individuals suffering with liver malfunction. It is important to investigate detailed mechanisms of hepatotoxicity of beta-AB also.

Effects of carvedilol, losartan and trimetazidin on functional parameters of isolated heart of rats at oxidative stress.

Effects of carvedilol, lozartan and trimetazidin on functional parameters of isolated heart (heart rate and volume speed of coronary blood flow) of rats at oxidative stress have been studied with the use of Langendorf's method. Experiments have been carried out on 32 pubertal white rats (with body mass 220-250 g). Oxidative stress was induced through perfusion of hydrogen peroxide (with concentration - 4.10 -5 M). Obtained results were analyzed statistically according to the Student's t criterion. Studies have shown that carvedilol and trimetazidin have well expressed antioxidant and cardio protective effects at oxidative stress provoked by hydrogen peroxide compared to lozartan. Moreover, effect of carvedilol on HR and VsCBf was especially obvious than effects of Trimetazidin. Antioxidant and positive treatment effects of carvedilol could be explained by its wide range of pharmacological ability: as nonselective beta-adrenoblocker (via inhibition of adenylatecyclase and decreasing cyclic adenosinemonophosphate), alpha 1-adrenoblocker (decreasing activation of phospholipase C and concentration of inositoltriphosphate, diacylglicerole and Ca(++)) and antioxidant. It is suggested that positive effects of carvedilol on HR and VsCBf at oxidative stress are related with wide range of pharmacological mechanisms of preparation. Cardio protective effects of carvedilol at oxidative stress (revealed in our experiments) along with other positive cardio - hemodynamic peculiarities perhaps is very important and could be used successfully at treatment of cardio-vascular system diseases.

[Alterations of tissue blood supply of experimental animals at malignant tumor growth].

The aim of the research was to study tissue blood flow in the dynamics of malignant tumor growth (Ehrlich carcinoma of 80 mice, sarcoma--37-80 rats, 30 rabbit--Brown-Pirs carcinoma) in functionally different organs of experimental animals (mice, rats, rabbits): skeletal muscles, liver, ear, mammary gland and ciliary body of eye. The study was conducted using H clearance method. Deformability of erythrocytes was studied using filtration-photometry method. Blood surface tension was studied by Rebinder bubble maximal pressure method. Obtained data were analyzed by Student's t criterion. The investigation showed that intensity of local hemocirculation and erythrocytes deformability in "intact" organs and tissues of experimental animals in the dynamics of malignant tumor growth progressively decreases while, blood surface tension increases. It was revealed that paraneoplastic disorders of local hemocirculation were stereotyped, universal and common phenomenon for malignant tumor growth. It was concluded that alterations of blood rheology decreased deformability of erythrocytes and increased blood surface tension play one of the leading roles in the mechanism of paraneoplastic disorders of local hemocirculation.

Anti-tumor preventative effect of mono-therapy with the use of proteus vaccine, Staphylococcus antitoxin and divaccine of Staphylococcus-proteus.

Anti-tumor preventative effect of mono-therapy with the use of Proteus vaccine, Staphylococcus antitoxin and divaccine of Staphylococcus-Proteus has been studied. Experiments were carried out on 80 non-purebred laboratory white mice (age--3-3,5 months, body mass--18-20 g) and 60 rats (body mass--100-120 g) using intraperitoneal inoculation of Ehrlich's adenocarcinoma (ascitic form--EAT, in mice, cancer cells--3 x 10(6)), and subcutaneous inoculation of Sarcoma S-45 (in rats). Anti-tumor preventative effect of bacterial vaccines and immunization was evaluated according to the following parameters: Frequency of cancer development, Inhibition of cancer growth, Body mass index of experimental animals, Volume of ascitic fluid. Results of experiments have shown that use of bacterial polysaccharides with preventative purposes has better effect at S-45 growth than at EAT growth; Vaccination with Proteus prolongs lifespan mach more than vaccination with antitoxin of Staphylococcus; Vaccination with complex divaccine of Staphylococcus-Proteus causes complete resorption of tumors from 32 to 60 days; Development of experimental malignant tumors depends on type of anti-microbial vaccines and starting date of inoculation after completion of vaccination.

[Correlated changes and value of redox-status, nitric oxide and blood supply in experimental malignant tumor growth].

With the use of methods of electronic paramagnetic resonance (EPR) and polarography EPR specter of tumor tissue, content of free nitric oxide and intensity of local hemo circulation in dynamics of sarcoma S-45 growth has been studied. It was stated that the intensity of local hemo circulation (especially in center of tumor tissue) progressively decreases. Sharp decrease of hemo circulation in the central part of the tumor tissue could be explained by drastically reduced concentration of NO, which in turn is related to disorders in electron transport in mitochondrial respiratory chain and probably to conversion of NO into toxic peroxinitrite. The relatively less expressed decrease of local hemo circulation in periphery, despite impaired reheological features of the blood, is explained by increased concentration of NO. It is suggest, that tumor tissue blood supply enhancement increases chemotherapeutic effects via affecting energetic metabolism in mitochondria, redox-status and concentration of free NO.

[The role of changes in tissue redox-status in mechanism of paraneoplasia].

To specify mechanisms of paraneoplastic alterations of redox-status of tissues in experimental malignant tumor growth, we investigated electronic paramagnetic centers of blood, skeletal muscle and liver with electronic paramagnetic resonance (EPR) method. We also studied the concentration and activity of antioxidant enzymes. Our experiments on adult white male rats of mixed population with sarcoma C-45 and mice with Ehrlich carcinoma have shown that malignant tumor growth leads to enhanced lipid peroxidation (LPO): production of potent LPO promoters - Fe2+, Mn+2+, NO, ubiquinone; depression of antioxidant defence - reduced production of total ceruloplasmin, elevated blood levels of oxidized ceruloplasmin and enhanced catalase activity. It is suggested that malignant tumor growth is associated with marked paraneoplastic shifts in tissue redox-potential. These alterations are involved in mechanisms of paraneoplastic changes of red cells, microhemocirculation and circulation intensity. All these interrelated processes result in generalized paraneoplastic hypoxia of the organs and tissues. Basing on our and literature data, we propose an original scheme of the mechanisms of paraneoplastic disorders of tissue redox-status, microcirculation and erythrocytes.

[Evaluation of anti-blastoma effects of tetravaccine in combination with chemotherapy].

The present work discusses methods of immune-correction and immune-rehabilitation of oncological patients using specific bacterial polyvalent vaccines. With this purpose the tetra-vaccine (Staphilococcus-Proteus-Clepsiela-Escherichia) has been used. Experiments have been carried out on white rats and mice with Sarcoma C-45. For chemotherapy of malignant tumors the following remedies were used: Cyclophosphan, Vincristin and Adriablastin. The vaccine and chemotherapy were applied in both ways, separately (control), and in combination. The results of investigations have shown that the best effect was achieved in case of combination of vaccine with chemotherapy--particularly, in the first experimental series malignant tumors completely underwent regression, while using only tetra-vaccine, the percent of malignant tumor growth inhibition constituted only 37.6%, and in the second experimental series percent of tumor growth inhibition constituted 56%-100% and the life span was prolonged by 22.4%-54.6%. Thus, the received results are quite encouraging and suggested method could be used for treatment of patients with cancer.

Changes in the prooxidant and antioxidant status of tissues during paraneoplastic processes.

Electron paramagnetic resonance was used to identify paramagnetic centers in the blood and liver of laboratory rats with S-45 sarcoma and mice with Ehrlich carcinoma. Paraneoplastic changes in prooxidant and antioxidant activity of the blood and liver tissue, mitochondrial respiration, and NO metabolism and inhibition of antioxidant processes contribute to impairment of cell membrane structures, erythrocyte hemolysis, and hypoxia. An important role of these processes in the pathogenesis of paraneoplastic anemia is confirmed by the positive effect of antioxidant and membrane-stabilizing therapy.

[Role of peroxidation processes and nitric oxide in mechanisms of paraneoplastic alterations of hemocirculation].

The present work was aimed to study mechanisms of paraneoplastic alterations of tissue redox-status, intensity of local blood flow in liver and their possible interrelations in case of malignant tumor growth. It has been investigated the electronic paramagnetic centres of blood and liver using the electronic paramagnetic resonance (EPR) method and intensity of local hemocirculation with the use of H(+) clearance polarography method. Experiments have been carried out on adult white rats of mixed population with carcinoma Walker and mice -- with carcinoma Ehrlich. It has been shown that malignant tumor growth displays conditions that lead to exaggerated lipid peroxidation (production of POL promoters -- Fe(2+), Mn(2+), NO, ubiquinone) and suppression of antioxidant protection of organism (reduction of total ceruloplasmin concentration in blood and increased concentration of oxidized ceruloplasmin). It has been suggested that in case of malignant tumor growth sharp paraneoplastic alterations of redox-status plays essential role in mechanisms of paraneoplastic disorders of tissues blood supply. All of these interrelated processes result in generalized paraneoplastic hypoxia in organs and tissues.