Experience of Using Tripeptide Leu-Ile-Lys for Experimental Therapy of Chronic 16-Week Oxalate Nephrolithiasis in Rats.

We studied the effect of tripeptide Leu-Ile-Lys on the course of chronic 16-week oxalate nephrolithiasis in rats modeled by administration of 1% ethylene glycol solution in drinking water for 16 weeks. The tripeptide Leu-Ile-Lys obtained by chemical synthesis (sample purity ≥98%) was administered intragastrically through a probe in a dose of 11.5 mg/kg in 1 ml saline. It was found that during tripeptide Leu-Ile-Lys significantly alleviated the course of experimental pathology, which was confirmed by characteristic biochemical and morphological indicators. We observed a decrease in the concentration of calcium ions by 4.4 times, weakening of oxidative stress in the renal tissue due to a decrease in the total prooxidant activity by 1.2 times, normalization of increased catalase activity, and reduction of superoxide dismutase activity by 2.4 times relative to disease control. Histological signs of nephrolithiasis were recorded in 9% cases (vs. 75% cases in disease control).

An Experimental Study of the Effect of Diindolylmethane on Alveolocyte and Hepatocyte Adhesion Strength in Mice.

We studied the effect of diindolylmethane in a dose of 600 mg/kg on the change in adhesion strength of alveolocytes and hepatocytes in CBA mice. Diindolylmethane was administered intragastrically to experimental animals for 10 days, controls intragastrically received an equivalent volume of saline. At the end of the therapeutic period, mice treated with diindolylmethane showed a significant increase in the adhesion strength of alveolocytes by 16% (p=0.003) and hepatocytes by 61% (p=0.0001) in comparison with the control group, which indicates the antipromotor activity of diindolylmethane.

Morphological Evaluation of the Influence of the Peptide Complex from Tissue of Porcine Kidneys on the Experimental Urolithiasis.

We performed morphological analysis of the effect of the peptide complex from porcine kidneys on the course of experimental urolithiasis modeled in rats by treatment with 1% ethylene glycol solution (in drinking water) for 6 weeks. The peptide complex obtained by acetic acid extraction was administered in a dose of 15 mg. Administration of the peptide complex to animals with experimental kidney stone disease leads to 100% destruction of large and medium stones to the "dust" granularity.

[Comparative estimation of antilithogenic activity of porcine kidney derived biomedical substance and sodium citrate in experimental urolithiasis].

AIM: to compare the anti-lithogenic activity of biomedical substance derived from freeze-dried porcine kidney and sodium citrate. MATERIALS AND METHODS: The experiments were conducted on Wistar rats divided into three groups of 15 animals each: control group (disease control), comparison group (sodium citrate treatment) and experimental group (treatment with biomedical substance from porcine kidneys). Experimental urolithiasis was modeled using the ethylene glycol model. On every 7th day of the 6 week experiment testing was done calcium and oxalate urine concentration and the activity of marker enzymes of renal epithelial damage: lactate dehydrogenase (LDH), -glutamyl transferase (GGT), and N-acetyl--D-glucosaminidase (NAG). At the end of the experiment, a part of the rats were decapitated and the renal tissue was tested for the oxidant status indicators of (renal thiobarbiturate reactive product content, TBRP, and total prooxidant activity, TPA) and antioxidant enzyme activities: glutathione peroxidase (GPO), superoxide dismutase (SOD) and catalase (CAT). To measure the number and size of calcium deposits formed in the renal papillary area, the Koss histochemical method was used. RESULTS: The experimental findings showed developing oxalate nephrolithiasis in the control group, as indicated by urinary supersaturation of oxalate ion, increased activity of marker enzymes, oxidative stress and the formation of numerous calcium deposits in the renal papillary area. In the comparison group, the 3-week use of sodium citrate contributed to a significant decrease in nephrolithiasis: a 3 to 4-fold decrease in the activity of marker enzymes in the urine, a 3.8-fold increase in the concentration of TBRP, normalization of GPO activity; the number and size of urinary calcium deposits decreased by 3.4 and 1.9 times, respectively. In the experimental group, using biomedical substance led to an even greater therapeutic effect. LDH activity and concentration of TPRP showed 1.9 times and by 26.2% greater decrease than in the comparison group, respectively, SOD and CAT activity almost doubled, there were 3.6 times fewer calcium deposits in the field of view and their mean size was 1.7 times smaller than in the comparison group. CONCLUSION: The study findings showed that the porcine kidney derived biomedical substance provide significantly greater antilithogenic effect than sodium citrate.

[Elimination of the activating influence of emotional stress on hemostasis mild hypothermia action].

The state of the hemostatic system in the psycho-emotional and hypothermia in rats. It is shown that the psycho-emotional impact is accompanied by a hypercoagulable. "Clean" effect of hypothermia is characterized by the activation of anticoagulation and fibrinolysis. Thus, we have shown that the general hypothermia in the aquatic environment has a more pronounced effect on the hemostatic system than the psycho-emotional impact.

Preclinical antitumor activity of the diindolylmethane formulation in xenograft mouse model of prostate cancer.

AIM: Preclinical study of the specific anticancer pharmacological activity of the formulation containing active substance 3,3ʹ-diindolylmethane (DIM), cod liver oil, polysorbate 80 and α-tocopherol acetate (vitamin E), in vivo in a xenograft animal model of LNCaP. MATERIALS AND METHODS: The DIM, cod liver oil, polysorbate 80 and α-tocopherol acetate (vitamin E) formulation was intragastrically administered to BALB/c-nude (nu/nu) mice during 33 days post inoculation at the dose of 133 mg/kg/day. Antitumor activity of the test drug was estimated by the rate of tumor growth inhibition (T/C% - treated versus control), dividing the tumor volumes from treatment groups with the control groups. RESULTS: Statistically significant tumor xenograft regressions have been shown in group which received the DIM, cod liver oil, polysorbate 80 and α-tocopherol acetate (vitamin E) on the 37(th) day of observation post inoculation. The highest antitumor activity was achieved on the 39(th) day (T/C = 16,8%). Therapeutic effect lasts for 6 days after the end of therapy period. CONCLUSION: Our findings demonstrate inhibitory effect of the formulation on tumor development in the xenograft animal model due to the tumor growth rate reduction.

[Peculiarities of hemostasis reactivity under stress load in persons with different degree of physical training].

Healthy young persons with different degrees of physical training have been impacted with exposure to a stress (a single physical exercise). It caused unidirectional hypercoagulative shifts and activation of anticoagulant and fibrinolytic blood systems. It was shown that changes of the untrained individuals' haemostatic parameters could be adjusted with adaptogen preliminary administration. The adaptogen administration in trained individuals resulted in disadaptive shifts in the haemostatic system. These contradictory changes indicate different levels of subject's adaptive potential.

[Hemostasis changes and their correction by adaptogen].

The paper presents an investigation into the changes in the hemostasis system of rats during extreme exercises. It has been observed that a single two-hour swimming exercise and an eight-hour imposed running in the treadbahn are accompanied by the expressed shifts in hypercoagulation with the signs of thrombinemia. On the background of the decrease in the anticoagulative and fibrinolytic activity it creates a serious threat of intravascular blood coagulation. The preliminary thirty-day course of extractum Eleutherococci eliminated the signs of intravascular blood coagulation.

[Heat-shock protein enhances immunogenicity of E7 oncoprotein of human papillomavirus included in chimeric construction (E7 HPV-HSP70)].

AIM: To study the effect of chimeric E7 protein of human papillomavirus type 18 on activation of adaptive immunity in absence of adjuvant. MATERIALS AND METHODS: Chimeric protein was genetically engineered and represents the protein molecule consisting of full-size E7 oncoprotein and heat-shock protein 70 (HSP70) of Mycobacterium tuberculosis in one polypeptide chain. Antibody titers as well as isotypes and subisotypes of immunoglobulins were measured by ELISA in sera of immunized animals. RESULTS: It was shown that studied construction E7 (HPV-18)-HSP70 significantly increases titers of antibodies to E7 protein of HPV type 18 and have cross-reactive antigenic activity with E7 protein of HPV type 16. Immunization with chimeric protein resulted in increase of IgG1 and IgG2b levels and decrease of IgG2a and IgM levels. CONCLUSION: . Oncoprotein E7 included in chimeric construction with HSP70 could be used for further studies on development of therapeutic vaccine for treatment of cervical cancer and precancerous lesions. Skew of immune response to Th2 type after intraperitoneal administration of the studied construction points to necessity for control of immunity during such studies.

[Clinical trials of the new skin test Diaskintest for the diagnosis of tuberculosis].

A new reagent for a skin test given the name Diaskintest has been designed for the screening diagnosis of tuberculosis and preclinical and clinical trials conducted. Preclinical trials were carried out on 315 laboratory animals (guinea-pigs, albino mice). The reagent Diaskintest was ascertained to be nontoxic, to have no sensitizing properties, to be safe and specific, and to induce no positive reactions in BCG-vaccinated animals and healthy guinea-pigs. Its specific activity was comparable with that of the national reference--purified tuberculin PPD-L-2. With progression of tuberculous lesions, the guinea-pigs showed higher responses to Diaskintest dilution and the BCG-vaccinated animals lacked responses to Diaskintest with increased delayed type hypersensitivity. The clinical trial was permitted by the Federal Service for Surveillance in Health Care and Social Development of the Russian Federation. Clinical trials were conducted in 150 persons. The safety, specificity, sensitivity of Diaskintest were first examined in the clinical studies and its action was compared with the results of tuberculin skin test (Mantoux test) with 2 TE of PPD L-2. Diaskintest was ascertained to be highly sensitive when given in a dose of 0.2 microg in 0.1 ml. In patients with active tuberculosis and new cases of Mycobacterium tuberculosis infection, the agent induced a positive skin reaction (a papule of more than 10 mm) in 98-100% of cases (p < 0.05). The agent caused no reaction associated with BCG vaccination. The specificity of the test was 93-100% with 95% significance. The rate of overexuberant reactions (vesicular necrotic changes, lymphangitis, and lymphadenitis) was 4-14% with 95% significance. Tuberculosis patients with significant immunopathological disorders might have no skin sensitivity to Diaskintest, as to PPD L-2 (a negative test). The findings substantiate the use of Diaskintest for mass epidemiological surveys for the differential diagnosis of tuberculosis and BCG vaccination-associated complications. The agent may be also used to evaluate the activity of the process in patients with tuberculosis and the efficiency of treatment in combination with other methods and to make a differential diagnosis of tuberculosis.

[Clinical and morphological parallels and molecular aspects of the morphogenesis of adenomyosis].

The purpose of the study was to reveal clinical and morphological parallels and to define molecular mechanisms, the regulation of proliferation, apoptosis, neoangiogenesis, and the extent of abnormal tissue in adenomyosis (AM). The surgical material obtained from 492 patients of late reproductive age was examined. The data of clinico-anamnestic and instrumental diagnostic studies and a morphological study with hematoxylin and eosin staining were analyzed. An immunohistochemical study was carried out on serial paraffin sections (n = 115), by applying antibodies to Apo-CAS, Ki67, PCNA, CD-34, MMP-1, MMP-2, MMP-7, MMP-9, TIMP-1, TIMP-3, TIMP-4, and E-cadherin. The specific features of their morphological structure and the clinical course of the disease allowed identification of its active and inactive forms. Immunohistochemically active AM is characterized by high proliferation, diminished apoptosis, and increased expression of MMPs along with lower expression of TIMPs by glandular and stromal cells as compared with inactive AM. At the same time, there was a high activity of stromal cells in the foci of active AM. The results of the study may be used to predict the course of the disease and to elaborate target therapy for AM.

[Role of heat shock proteins in developing of innate immunity reactions].

Increasing interest to heat shock proteins (HSP) from biologists and medics is connected to widespread distribution of HSP in live nature and reflects their key role in support of life functions which is based on the unique polyfunctionality of these biomolecules. Together with main function, which is defense of biologic systems from stress effects, some HSP in the process of evolution acquired the ability to incorporate in the reactions of the immune system. The in vestmen of this protein in practical reactions of innate immunity system are described. Analysis of mechanisms underlying the adjuvant effect of pro- and eukaryotic HSP in innate immunity system is presented. HSP receptor structures on target cells as well as triggered intracellular signaling pathways are described.

[Effect of eleutherococcus on hemostasis in immobilized rats].

The influence of a chronic (30 days) administration of an eleutherococcus extract on the haemostatic system state was studied in immobilized rats. A 3-h immobilization of untreated animals is accompanied by hypercoagulation and thrombinemia signs on the background of downregulation of the anticoagulant and fibrinolytic activity, which leads to a risk of thrombosis. Preliminary 30-day course of eleutherococcus uptake prevents the immobilization-induced thrombosis in rats.

[Functions of heat shock proteins in the system of adaptive immunity. Construction of vaccines].

One of the most important aspects of heat shock proteins (HSP) functioning, namely their role in reactions of innate and adaptive immunity, was reviewed in the article. Mechanisms of involvement of HSP in processing and presentation of antigens to T-lymphocytes were described. Principles of construction of prophylactic and therapeutic vaccines on the basis of HSP were set out. Assessment of range of indications for their use and possible risks related with inductive action of vaccines on the development of immunopathologic processes was a specially discussed topic.

[Efficacy of indinol in antirecurrence therapy of juvenile respiratory papillomatosis].

The study of indinol efficacy in antirecurrence therapy of respiratory papillomatosis was performed in 46 children aged 2-14 years. The treatment lasted at least 12 weeks. Duration of a recurrence-free period before and after indinol administration was analysed. The response was observed in 30 (65.2%) patients. In 5 (10.9%) patients the remission continued from 2 to 3 years 10 months. In 25 (54.3%) patients recurrence-free period increased 1,5-6-fold. Side effects were not registered. Simple in use, cheap, well tolerated, usable with other treatments, indinol is recommended as a method of choice in antirecurrence treatment of recurrent respiratory papillomatosis.

[Correlation between the expression of Rv3286c, Rv2626c, Rv2031c, AND Rv3133c and the tolerance of Mycobacterium bovis BCG to rifampicin and metronidazole in different physiological conditions].

The fact that antibiotics exert no effect on Mycobacterium tuberculosis may be attributable to the production of their persistent forms. A correlation was studied between the expression of the genes Rv3286c, Rv2626c, Rv2031, and Rv3133c and the phenotypical tolerance to antibiotics in relation to the physiological condition of M. bovis BCG. The anaerobic growth conditions ("Wayne dormancy model") were found to be attended by the increased expression of all the studied genes, by the higher tolerance to rifampicin, and by the decreased phenotypical resistance to metronidazole.

[Prevalence of human papilloma virus types and their influence on the course of the disease in children suffering from recurrent respiratory papillomatosis]. / Rasprostranennost' tipov virusa papillomy cheloveka i ikh vliianiia na techenie zabolevaniia u detei, stradaiushchikh retsidiviruiushchim respiratornym papillomatozom.

The type of human papilloma virus (HPV) was determined in 26 children aged between 1 year 10 months to 15 years 5 months suffering from recurrent respiratory papillomatosis (RRP). Polymerase chain reaction identified DNA of HPV type 6, 11, 16 and 18. HPV DNA was detected in all the patients including fifteen patients infected with HPV type 11; seven patients infected with HPV type 6; four children infected with HPV type 6 and 11. Types 16 and 18 of HPV were not detected. The analysis of RRP course has found that laryngeal papillomatosis runs a more aggressive course in cases with HPV type 11 infection than in those with HPV type 6.

[The structural and functional analysis of the biological activity of acridine derivatives]. / Strukturno-funktsional'nyi analiz biologicheskoi aktivnosti proizvodnykh akridina.

The paper contains an analysis of research on designing drugs based on acridine derivatives. The discussed series of compounds is of essential value since acridines belong to the group of natural compounds with the pronounced antibacterial and anti-tumor activity. Improved chemical-synthesis techniques made it possible to synthesize both simple and complex compounds of the acridine series; they displayed a clear pharmacological activity as anti-proliferative, anti-tumor and antiparasitic preparations. The ability to induce interferons (INF), type 1, is an expected property of simple acridine derivatives. A variety of INF inducers, now used clinically, have been designed recently on the basis of the above compounds. The most well-known acridine derivatives, their pharmacological properties, action mechanisms and outlooks for practical application are described in the paper. The unique qualities of acridines are primarily attractive due to the possibility of using them for the purpose-oriented designing of drugs. Thus, acridines were used as a basis to create the specific regulatory HIV-1 elements, proliferation inhibitors of leukemia cells and new anti-tumor drugs. The elaboration of complexes of acridines derivatives combined with peptides intercalating specifically into the DNA big or small grooves is the most outstanding trend of acridines' research--it opens up prospects for using them in the synthesis of compounds regulating the gene expression.

Developing additional resources.

We briefly outline existing information about several cohorts in the Southern Urals, Semipalatinsk and the Altai regions, in addition to those discussed in the companion papers in this issue of Radiation and Environmental Biophysics. These include: (a) the East-Urals Radiation Trace (EURT) cohort of individuals (exposed to fallout from the September 1957 explosion of a storage tank containing liquid radioactive waste from the Mayak Production Association) as well as their offspring, (b) the cohort of exposed parents (i.e. workers of the Mayak facility) and their children, having been established with the aim of examining reproductive health, and (c) several additional cohorts in the Altai region and in Semipalatinsk, where investment of additional resources would greatly facilitate the progress of ongoing studies. Furthermore, we include a brief description of the Russian Human Radiobiology Tissue Repository, which has been established in the city of Ozyorsk and is in the process of making an inventory of autopsied tissues from 700 deceased Mayak workers and of collecting and storing donations of blood and tumor tissues from the members of the Mayak workers cohort currently residing in the city.