Immunotropic Effect Research Articles

Experimental assessment of the immunotropic properties of the new biocomplund

The article concerns topical issues of studying the new biocompound within a consortium of biologically active substances (BAS). These compounds (metabiotics) are produced by probiotic strains 59T and 60T of saprophytic microorganisms which belong to Bacillus subtilis genus. These strains seem to be quite promising for the design of hepatopicts, the new drug candidates, for creation and development of a new class of hepatoprotector drugs. Previous studies have shown safety and hepatoprotective effects of abovementioned BAS. It is important to note that the combined use of these compounds provides a potentiated pharmacological effect. Moreover, there are significant reasons to believe that these BAS may influence different compartments of immune system. In this regard, it seemed appropriate to perform experimental assessment of the immunotropic effect of BAS in laboratory animals using a model of toxic liver damage with carbon tetrachloride, in order to confirm the effect on the humoral factors of immunity. The purpose of the present study is to provide experimental comparative assessment of humoral immunity indexes upon combined usage of biologically active substances in acute toxic liver damage. The experimental model of toxic damage was reproduced in white laboratory rats with repeated intragastric administration of carbon tetrachloride. The comparison drug was ursosan, a registered hepatoprotective drug. The following factors of humoral immunity were measured to assess the effects of biologically active substances: immunoglobulins M, G, A, quantitative indices of á-interferon and circulating immune complexes (CIC). As a result of the studies performed, we obtained reliable data showing increased amounts of all Ig classes as well as induction of endogenous á-interferon and antibody-forming cells under the conditions of acute toxic liver damage. Meanwhile, a significant restoration of the humoral immunity indices to initial values was registered after the 15-day treatment with biologically active substances. The data obtained allow us to recommend the studied BAS for creation of new drug candidates of microbial origin, hepatoprotective agents with immunotropic effects.

A novel immunopharmacological biocompound

The publication is devoted to topical issues of experimental study of new biocompounds, based on the creation of a consortium of biological composite compounds – biological active substances (metabiotics) produced by strains 59T and 60T of saprophytic compounds of the genus Bacillus subtilis. These strains are very promising for the creation of hepatopicts, new medicinal substances, in the creation and development of a new pharmacological class of hepatoprotectors. Previous studies have shown the safety and hepatoprotective effect of biologically active drugs. It is worth noting that the very effective compounds are the produced biologically active substances – metabolites of bacterial origin, on the basis of which it seems appropriate to create a new class of drugs – metabiotics. The absence of vegetative probiotic cells in such compounds will reduce the additional immune load on the body. The combined use of these compounds provides a potentiated pharmacological effect. In this regard, it seemed appropriate, under the conditions of modeling toxic liver damage by carbon tetrachloride, to conduct an experimental assessment of the immunotropic effect of biologically active substances (BAS) on laboratory animals in order to confirm the effect on cellular factors of immunity. The purpose of the study was to study the effect of the combined use of dietary supplements produced by Bacillus subtilis microorganisms on indicators of cellular immunity in laboratory animals with toxic liver damage. Acute toxic hepatitis was reproduced in white laboratory rats with repeated intragastric administration of carbon tetrachloride. The comparison drug was a registered hepatoprotective drug – ursosan. The immunotropic effect was assessed using factors such as: phagocytic activity of macrophages and neutrophils (FA), NBT test, quantitative assessment of antibody-forming cells, T and B lymphocytes. As a result of the studies performed, reliable data were obtained on an increase in the number of T and B lymphocytes, antibody-forming cells, as well as an increase in FA, which confirms the activation of all parts of cellular immunity in conditions of acute toxic liver damage. The data obtained allow us to recommend the studied biocompound for the creation of new drug candidates of microbiological origin, hepatoprotective agents with immunotropic effects.

Immunotropic effect of feed additives based on metaprobiotics and phytobiotics in providing specific immunity in broiler chickens

Immunotropic effect of feed additives based on metaprobiotics and phytobiotics in providing specific immunity in broiler chickens

Architectonics of parenchymal organs of white rats under conditions of induced immunosuppression and the use of Interferon lambda

Violation of the functioning of the organs of the immune system, both in humans and in animals, is due to the influence of unfavorable factors of natural and anthropogenic origin on the body, as well as infectious diseases of various etiologies. The article discusses changes in the architectonics of parenchymal organs of white laboratory rats after exposure to the drug interferon lambda on the background of cyclophosphamide (CF). It was found that modeling of immunosuppression through the use of CF led to a decrease in the number of lymphoid cells and the appearance of foci of accumulation of blast cells and megakaryocytes, which indicates a lesion of lymphoid and hematopoietic tissues. In addition, an immunosuppressive effect on the spleen was revealed, the mass coefficient of which in the group with the use of CF was increased 3.53 times, in the group with the simultaneous use of CF and the drug interferon lambda – 2.75 times relative to the control group. There was also a decrease in the mass coefficient of the thymus by 47.6-50.0% in the same groups, which can be regarded as damage to the organs of the immune system. The negative effect of CF on the spleen was confirmed by histological examination. Thus, when inducing immunosuppression, the loss of white pulp was 73.0-77.5% relative to the group of control animals, which is 20.0% higher than in rats treated with interferon lambda, which may indicate a positive immunotropic effect of the drug under study. In addition, no significant morphological changes were detected when the body of healthy rats was exposed to interferon lambda in the studied organs, the morphological picture is similar to that of intact animals, which may indicate the safety of its use to animals.

Immunopharmacological aspects of studying immunotropic properties of a novel biocompound

The article presents an experimental study of immunotropic activity of a new compound, a metabiotic, based on metabolites (biologically active substances) produced by a of saprophytic Bacillus subtilis B-9909 strain, being safe for health and standardized at Russian National Collection of Industrial Microorganisms (RNCIM). The aim of our study was to experimentally evaluate the immunotropic effects of metabolites produced by the probiotic Bacillus subtilis B-9909 upon the parameters of cellular immunity tested in the animal model of induced toxic liver damage. The metabolites were isolated from the cultural liquid of Bacillus subtilis culture (RNCIM strain B-9909) during its deep cultivation in a medium containing hydrochloric acid hydrolyzate of soy flour, or pancreatic hydrolyzate of casein. At 16-18 hours of cultivation, the cells were in the exponential growth phase. The indices of cellular status were studied in experimental groups of animals, when assessing therapeutic efficacy of an experimental metabiotic sample, as compared to a group of laboratory animals treated with the reference drug (ursosan). We assessed the quantitative indicators of blood as follows: determination of phagocytic activity (FA) of peripheral blood neutrophils; measurement of metabolic activity in peripheral blood neutrophils by means of NBT-test; quantitative determination of T and B lymphocytes; the number of antibody-forming cells (AFC). Liver damage was studied in a model of acute toxic hepatitis in albino rats. Experimental toxic hepatitis was induced by 40% solution of CCl4 in vaseline oil injected intragastrically for 2 weeks at the dose of 0.2 g/kg. The results of experimental studies showed activation of phagocytic activity by peripheral blood neutrophils at the early terms of experiment, being also confirmed by the results of NBT-testing. Moreover, at the early observation terms, a significantly increased representation of T and B lymphocyte populations as well as AFC numbers, were revealed thus suggesting activation of all components of cellular immunity in response to the carbon tetrachloride toxicity. Thus, the reported studies on the cellular status of laboratory animals treated with metabolites produced by probiotic microorganisms of the genus Bacillus subtilis (B-9909) on laboratory animals with toxic liver damage allow us to conclude that the test sample of this metabiotic preparation exerts a significant immunomodulatory effect, when compared with ursosan. This evidence allows us to consider this compound a promising candidate drug for a new hepatoprotector with immunotropic effect.

Relationship between vitamin D sufficiency and interferon-γ levels in children with chronic lung diseases

Introduction. The role and mechanism of the effect of vitamin D on the course of chronic lung diseases in children are not yet fully understood. In particular, there are not enough studies on the effect of vitamin D status on the production of interferon-γ (IFN-γ) in children with cystic fibrosis and bronchial asthma.Aim. To analyse the changes in IFN-γ levels in children with cystic fibrosis and bronchial asthma according to the serum 25(OH)D level before and during cholecalciferol supplementation.Materials and methods. A total of 114 children (aged from 3 months to 18 years old) were examined, including 51 (44.7%) children with bronchial asthma, 34 (29.8%) children with cystic fibrosis, and 29 (25.5%) children of the control group. If hypovitaminosis D was diagnosed, the children received a three-month cholecalciferol course at doses in accordance with the recommendations of the national consensus - Cystic Fibrosis: Definition, Diagnostic Criteria, Therapy and the national program - Vitamin D Deficiency in Children and Adolescents of the Russian Federation: Current Approaches to Management. The 25(OH)D and IFN-γ levels were twice determined in all children.Results. Median 25(OH)D levels in patients with cystic fibrosis and bronchial asthma did not reach the level of healthy children either before or after supplementation with cholecalciferol drugs at the recommended doses. Vitamin D intake resulted in a decrease in IFN-γ levels in healthy children from 3.07 [2.29; 4.81] pg/ml to 2.18 [1.74; 3.45] pg/ml (p < 0.05), whereas such changes were not detected in the paediatric patients with cystic fibrosis and bronchial asthma in the general population. However, after supplementation with cholecalciferol, the IFN-γ level was significantly higher in patients with cystic fibrosis and bronchial asthma - 2.86 [2.13; 3.86] pg/ml and 3.11 [0.89; 5.0] pg/ml, respectively, than in healthy children - 2.18 [1.74; 3.45] pg/ml (p < 0.05). A statistically significant decrease in IFN-γ level was observed in girls with cystic fibrosis and in healthy girls after supplementation with cholecalciferol, whereas no significant changes in IFN-γ levels were detected in the groups of boys.Conclusion. Modulation of IFN-γ levels appears to be one of the mechanisms of immunotropic effect of vitamin D on the course of chronic inflammation of infectious (cystic fibrosis) and allergic (bronchial asthma) etiology in children.

ISOLATION, CHEMICAL AND SPATIAL CHARACTERIZATION OF ACIDIC POLYSACCHARIDES FROM SOME PLANTS OF SIBERIAN FLORA WITH IMMUNOTROPIC ACTIVITY

Plant polysaccharides have a wide range of biological activity, including an immunotropic effect, the degree and magnitude of which depends on the composition and tertiary structure of the macromolecule. Acidic polysaccharides (APS) were isolated by extraction with an acidic aqueous solution (pH=3) and characterized From the rhizomes of Acorus calamus L., the aerial part of Centaurea pseudomaculosa Dobrocz., the herb Conium maculatum L., and the leaves of Betula pendula Roth. The content of total carbohydrates was determined for all samples (>90%), uronic acids (24.56–53.70%) and a small amount of impurity substances (protein and nucleic acids 0.81–1.02% and 0.016–0.035%, respectively); molecular weight distribution (Mn, Mw). Structural features were studied using physicochemical and instrumental methods, such as UV and IR spectroscopy, reaction with Congo red, and scanning electron microscopy. The polysaccharides of Acorus calamus, Conium maculatum, and Betula pendula have a triple-helical conformation and bulky ultrastructure; the polysaccharides of Centaurea pseudomaculosa are characterized by the absence of a triple-helical conformation and an amorphous ultrastructure.

Experimental study of immunotropic properties of metabolites of <i>Bacillus subtilis B-9909</i> strain, promising in the formulation of a new hepatoprotector

This brief report is devoted to the experimental study of the immunotropic activity of a new compound, a metabiotic, based on metabolites (biologically active substances, BAS) produced by the saprophytic and safe standardized strain of Bacillus subtilis B-9909. The aim of the study was to experimentally evaluate the immunotropic effect of metabolites produced by probiotic microorganisms of the genus Bacillus of the culture of probiotic microorganisms of the Bacillus subtilis B-9909 on laboratory animals when modeling their toxic lesion. Metabolites were isolated from the culture fluid of the bacterial culture Bacillus subtilis, strain RNCIM (Russian National Collection of Industrial Microorganisms) B-9909, during its deep cultivation in a medium consisting of hydrochloric acid hydrolysate of soy flour or pancreatic hydrolysate of casein. The study of humoral status indicators in experimental groups of animals in assessing the therapeutic efficacy of an experimental metabiotic sample, in relation to the group receiving the ursosan comparison drug, was carried out by determining such quantitative serum parameters as the titers of immunoglobulins M, G, A, E, the IFNa titer and the concentration of circulating immune complexes. Liver damage was studied by modeling acute toxic hepatitis in white rats. Thus, the studies conducted to study the humoral status of laboratory animals receiving metabolites produced by probiotic microorganisms of the genus Bacillus of the culture of probiotic microorganisms of the Bacillus subtilis B-9909 on laboratory animals when modeling toxic damage in them, give grounds to conclude that the test sample of BAS has a significant immunomodulatory effect, in comparison with ursosan. This circumstance allows us to recommend this compound as promising for the creation of a new hepatoprotector drug candidate with an immunotropic effect.

Combined Drug with Antibacterial Effect Supports the Normal Intestinal Microflora.

Widespread use of antibiotics leads to an imbalance of normal intestinal microflora and to the development of multidrug resistance. The problem can be solved by administration of the antibiotics in combination with the drugs that have an immunotropic effect. We studied the effect of the drug containing technologically processed affinity purified antibodies to IFNγ, CD4 receptor, β2-microglobulin of MHC class I, and β2-domain of MHC II combined with antibiotics on the composition of intestinal microflora of pigs and the total number of microbiome resistance genes. Using the methods of NGS sequencing and quantitative PCR, we found that the drug contributes to the maintenance of normal microflora and, consequently, to the symbiotic relationship of the host with microflora, and prevents the reproduction of pathogenic bacterial species. Analysis for the presence of the resistance genes of gastrointestinal microorganisms showed that the drug does not affect the qualitative and quantitative composition of these genes of the intestinal microbiome.

Effect of individual substances isolated from Silene jeniseensis Willd on the state of the main links of immunity at experimental immune deficiency

Introduction. The search, development and introduction of new drugs with an immunotropic effect are one of the priority tasks of modern immunopharmacology. Numerous studies have proven the immunotropic activity of individual substances isolated from medicinal plants (flavonoids, polysaccharides, ecdysteroids, terpenoids, etc.). In the present study, it is of interest to determine the immunomodulatory effect of individual substances isolated from Silene jeniseensis Willd.The aim of the study. Determination of the immunomodulatory activity of individual substances isolated from Silene jeniseensis: flavonoid isoorientin-2”-O-rhamnoside, polysaccharide arabino-3.6-galactan and ecdysteroid 20-hydroxyecdysone under conditions of cyclophosphamide induced experimental immunosuppression.Methods. Experiments were carried out on F1 (CBAxC57Bl/6) mice. Immunodeficiency was modeled by intraperitoneal administration of cyclophosphamide to control group animals in the dose 250 mg/kg once. Experimental groups of mice received the test substances intragastrically once a day for 14 days against the background of immunosuppression in the following doses: isoorientin-2”-O-rhamnoside – 10 mg/ kg, arabino-3.6-galactan – 3 mg/kg, 20-hydroxyecdysone – 3 mg/kg. The effect of substances on cellular immunity was determined in a delayed hypersensitivity reaction, humoral immunity was determined in an antibody formation reaction by local hemolysis according to A.J. Cunningham. The phagocytic activity of peritoneal macrophages was studied in relation to colloidal ink particles.Results. With the introduction of isoorientin-2”-O-rhamnoside, arabino-3.6-galactan and 20-hydroxyecdysone in experimental animals, there was an increase in the index of delayed-type hypersensitivity reaction by 1.3–1.4 times, the absolute and relative number of antibody-forming cells by 1.4–1.7 times, phagocytic index by 1.2–1.5 times compared with the data in the control group, which indicates the leveling of the suppressive effect of cyclophosphamide on cell-mediated immune response, antibody genesis and phagocytosis of macrophages.Conclusion. Isoorientin-2”-O-rhamnoside and arabino-3.6-galactan have the most pronounced immunomodulatory effect. The obtained data allow us to consider the studied substances as promising plant immunomodulators.

Разработка технологии выделения и исследования иммунотропного действия бурсальных пептидов на мышах с экспериментальным иммунодефицитом

Peptides are biologically active. This quality depends on the technological modes of protein hydrolysis. The research objective was to isolate peptides from the bursa fabricii and evaluate their immunotropic effect on mice of various lines with experimental immunodeficiency and the morphofunctional state of their immunopoiesis organs, i.e., thymus and spleen.
 The research featured bursa fabricii hydrolysate and peptides, as well as thymus and spleen of male mice. Amine nitrogen was determined according to State Standard R 55479-2013; molecular weight of peptides was determined by gel electrophoresis. The morphometry of the thymus and spleen was calculated using VideoTesT-Morphology 5.0. 
 The research revealed the rational modes of enzymatic hydrolysis of the bursa fabricii protein by papain with subsequent isolation of peptides by ultrafiltration. According to the content of amine nitrogen in the bursa fabricii hydrolysate, the rational concentration of the papain enzyme was 0.15%. The molecular weight of peptides after ultrafiltration of bursa fabricii hydrolysate through membranes with a 43 kDa permeability had different values and depended on the concentration of papain, hydrolysis time, and temperature. The maximal amount of 27–18 kDa peptides was isolated at 36°C (85%) when the concentration of papain was 0.15% (73%), the hydromodule was 1:3 (78%), and the hydrolysis time of the raw material was 6 h (82%). The rational technological parameters of the hydrolysis of the bursa fabricii protein were as follows: concentration of papain – 0.15%, temperature – 36°C, hydromodule – 1:3, and hydrolysis time – 6 h. In immunodeficient mice, bursal peptides prevented a decrease in total cellularity in the thymus, sustained the content of CD3+ cells, activated the maturation of T-lymphocytes and proliferation of B-lymphocytes, and reduced the immunocytotoxic effects of cyclophosphamide.
 The article offers a new technology for obtaining peptides with a pronounced immunotropic effect that can be used in functional food production.

Evaluation of clinical, microbiological and immunological effectiveness of a collagen sponge with antimicrobial and immunotropic properties

Relevance. The etiology of alveolitis distinguishes infectious and traumatic factors, which determine the relevant drugs for preventing the inflammatory process after tooth extraction and local treatment of the developed alveolitis. Purpose – the study aimed to evaluate the effectiveness of a collagen sponge containing lincomycin hydrochloride compared to the traditional method using iodoform packing strips based on clinical, microbiological and immunological research methods.Materials and methods. The study examined and surgically treated 75 patients with jaw alveolitis (K10.3), including those with eruption disorders, chronic periodontitis and chronic periodontitis in remission. Routine tooth extraction was recommended for them. We treated developed alveolitis in 37 patients and prevented complications during tooth extraction in 38 patients. According to the local exposure, the study allocated two subgroups: A –collagen sponge containing lincomycin hydrochloride and B –iodoform packing strips (traditional method). We comparatively analyzed microbiological parameters and cytokine content in the mixed saliva of patients using enzyme immunoassay (EIA).Results. In the studied alveolar socket material with a confirmed diagnosis of alveolitis, a spectrum of priority pathogens was established, including mainly microaerophilic streptococci, obligate anaerobes and staphylococci. The monitoring of the local treatment method effectiveness demonstrated that the sponge with lincomycin does not fall behind in the decontamination level regarding streptococci and obligate anaerobes but is less effective against the staphylococcal component of the microbial consortium. Analysis of cytokines confirmed the presence of the immunotropic effect of the collagen sponge with lincomycin.Conclusion. The possibility of using a collagen sponge with lincomycin hydrochloride has been substantiated as an alternative method of local treatment of alveolitis, which has a pronounced antibacterial and immunotropic effect at the level of regulation of the cytokine status.

Нейроендокринно-імунний супровід діуретичного ефекту бальнеотерапії на курорті Трускавець

Мета: виявити супутні зміни параметрів нейроендокринно-імунного комплексу і мікробіоценозу товстого кишечника, що супроводжують діуретичний ефект бальнеотерапії на курорті Трускавець. Результати. У спостереженні за 22 чоловіками, хворими на хронічний пієлонефрит в поєднанні з холециститом, виявлено, що 10–12-денний курс бальнеотерапії (пиття біоактивної води «Нафтуся», аплікації озокериту, мінеральні ванни) збільшує добовий діурез на 28 % (p < 0,001). Це поєднується із збільшенням екскреції фосфатів на 103 % (p < 0,001), кальцію — на 76 % (p < 0,01), креатиніну — на 32 % (p < 0,02), сечової кислоти — на 29 % (p < 0,001), сечовини — на 21 % (p < 0,01), магнію — на 19 % (p = 0,05) при відсутності суттєвих змін екскреції хлориду, калію і натрію. При цьому знижується рівень у плазмі креатиніну на 14 % (p < 0,001) і сечовини на 11 % (p < 0,01). Лейкоцитурія зменшується на 16 % (p < 0,02). Екскреторно-депураційний і протизапальний ефекти бальнеотерапії супроводжуються відчутною модуляцією параметрів нейроендокринно-імунного комплексу. Зокрема, зменшується як абсолютна, так і відносна щільність спектральної потужності (PSD) δ- і α-ритмів (p < 0,05–0,01), і підвищується відносна PSD β-ритму (p < 0,05–0,02) базальної електроенцефалограми. Разом з тим знижуються індекси варіабельності ритму серця: (VLF + LF)/HF — на 34 % (p < 0,05) і LF/HF — на 32 % (p > 0,05) за рахунок підвищення PSD HF більшою мірою, ніж LF. Базальний рівень у плазмі кортизолу знижується на 20 % (p < 0,01), тестостерону — на 15 % (p = 0,01), натомість підвищується рівень трийодтироніну — на 4 % (p < 0,05) і особливо кальцитонінової активності — на 92 % (p < 0,001), розрахованої за екскрецією з сечею фосфатів і кальцію. Лейкоцитарний індекс адаптації Поповича зростає на 46 % (p < 0,02). Стосовно параметрів фагоцитарної функції нейтрофілів констатовано підвищення зниженого індексу кілінгу ними Staphylococcus aureus на 19 % (p < 0,001) і Escherichia coli на 18 % (p < 0,01) за відсутності змін початково нормальних фагоцитарних індексів. Мікробне число стосовно Staphylococcus aureus залишається нормальним, а початково підвищена на 15 % інтенсивність фагоцитозу Escherichia coli знижується на 8 % (p < 0,05). Стосовно параметрів імунітету виявлено значуще підвищення рівня в крові лише CD16+ лімфоцитів (+17 %, p < 0,01) при відсутності змін рівнів CD3+CD4+ і CD3+CD8+ Т-лімфоцитів і CD19+ В-лімфоцитів. Не змінюються суттєво ні рівні в сироватці Ig G, M, A, ні циркулюючих імунних комплексів. Імунотропний ефект супроводжується редукцією проявів дисбактеріозу: вміст Bifidumbacter зростає на 19 % (p < 0,02), Lactobacter — на 20 % (p < 0,05), а Escherichia coli — на 48 % (p < 0,01), при цьому доля штамів з ослабленими ферментативними властивостями зменшується на 47 % (p < 0,001), з гемолітичними властивостями — на 77 % (p < 0,01). Висновок. Бальнеотерапія на курорті Трускавець чинить відчутний діуретичний ефект, що поєднується з активацією екскреторної і депураційної функцій нирок та бактерицидної функції нейтрофілів, редукцією дисбактеріозу товстого кишечника, лейкоцитурії і нейроендокринних проявів стресу.

To Study the Efficacy of Placental Extract Gel in Chronic Non Healing Foot Ulcer

Wound healing is a complex and regulated process that is critical in maintaining the skin barrier function. The numerous disease processes can affect the events involved in wound healing, resulting in chronic, non-healing wounds. These would subject them to significant discomfort and distress and drain the medical system resources enormously. A chronic wound can be defined as that does not heal in an orderly set of stages and in a predictable amount of time or within three months. They remain in the inflammatory phase for too longand never heal or may take years.3,4 Chronic leg and foot ulcers occur in many adults with diabetes, chronic venous insufficiency, arterial disease, prolonged pressure, or neuropathy.5 Chronic ulcers may last for 12 to 13 months and even recur in 60-70% of cases. They cause loss of function and decreased quality of life and significant morbidity.6,7 These chronic ulcers are predominantly seen in elderly, becoming more prevalent and more difficult to treat and are associated with high treatment costs. So wound care has become essential in these ulcers, and surgical debridement and dressings are the corners stones. Many wound dressings like gauge, films, hydrocolloids, gels, foams etc. were developed to both protect the healing wound from infection and to promote the healing process.1 Placental extract gel is a rich source of various peptides, amino acids, nucleotides, PDRNs and carbohydrates, which are believed to support the healing process.9 Application of placental extract preparation increases collagen synthesis, increases tissue protein, accelerates neoangiogenesis, and epithelialization.10 It has an immunotropic effect and causes the release of Epidermal growth factor, Fibroblast growth factor at the tissue level. It also reduces surrounding tissue inflammation and oedema and reduces microbial burden.11 Its application accelerates healing because of amino acids, nucleotide and vitamins that improve healing.

Immunotropic effect of medicinal plants with different microelemental composition

Increasing use of raw materials from medicinal plant is due to the fact that the natural compounds may be preferred to synthetic ones by, generally, low toxicity, absence of side effects and addiction. Over recent decades, much attention has been paid to the study of immunotropic and antioxidant effect of medicinal plants, as well as to analysis of their trace element composition, since the action of the main biologically active substances is often manifested in combination with the natural mineral composition of the plant. The aim of the present work was to assess immunotropic effect of aqueous extracts of officinal medicinal raw materials and to analyze their trace element composition. The objects of the study were water extracts (1:10) from industrial samples of 11 types of raw materials (black currant leaves (Ribes nigrum L.), yarrow herb (Achillea millefolium L.), licorice roots (Glycyrrhiza uralensis Fisch.), Flowers of the sand immortelle (Helichrysum arenarium (L.) Moench), St. John's wort (Nurericum perforatum), wild strawberry leaves (Fragaria vesca L.), wild bird cherry (Padus avium Mill.), blood-red hawthorn (Crataegus sangunea Mill.) (Tanacetum flowers) vulgare L.), common chicory root (Cich rium ntybus Fisch.) and oat grass (Avena sativa L.) supplied to the network of pharmacies in Orenburg. Immunotropic effects of raw plant materials were evaluated in a model of adaptive immunity cells, by their ability to induce production of cytokines: IL-1β, TNFα and IL-10. The level of cytokines (IL-1β, TNFα, and IL-10) were assessed using ELISA (Cytokin, St. Petersburg, Russia). Elemental composition of raw material samples was determined by mass spectrometry with inductively coupled plasma using the ELAN-DRC-e device (PerkinElmer SCIEX, USA). Analysis of immunotropic activity of aqueous extracts from officinal medicinal plants showed that the majority of plant raw materials are characterized by a suppressive effect both upon pro-inflammatory mediators (TNFα, IL-1β), and the main anti-inflammatory cytokine (IL-10). In contrast, aqueous extracts of yarrow, oats and chicory were characterized by a selective immunomodulatory effect aimed at suppressing only inflammatory mediators. A tendency has been established for a significant accumulation of such biologically important trace elements as Zn, Mn, Fe and Cu in medicinal plants, which can determine their biological activity, and allows them to be considered as promising components at the stage of developing drugs that both exert immunotropic effect, and are a source of microelements.

Effect of melatonin include into original dermal film upon adaptive immunity in experimental thermal trauma

Adaptive immunity changes in thermal trauma (TT) increase the risk of infectious complications and limit repair of the lesion. Hence, search and preclinical testing of effective and safe means to locally manage TT, containing bioregulators, is an urgent task of modern medicine. Dermal films (DF) are an innovative and popular variant of wound coatings for small-area burns, and pleiotropic properties of melatonin (MT) suggest its effectiveness in TT. The aim of the work is to investigate the effect of MT, as a component of original DF, upon the indexes of adaptive immunity in experimental TT. The experiment was performed on 115 male Wistar rats. Grade IIIA TT with area of 3.5% were produced by contact with boiling water for 12 s. DF with an area of 12 cm2 based on sodium carboxymethylcellulose contatning MT (5 mg/g) was applied daily for 5 days. Similar DF matrix, but without MT, was used in the control group, Amounts of CD3+ and CD45RA+ cells in blood, and lymphocyte subpolulations with early and late signs of apoptosis and partially necrotic cells were evaluated with flow cytofluorometer, as well as IgG and IgM concentrations were measured in blood serum using rat test systems. With TT, the amount of CD3+ in the blood decreases on days +5 and +10, CD45RA+, on days +5, +10 and +20, and the concentration of IgG in the serum, on days +5 and +10 of observation. On days +5 and +10 after TT, a relationship was established between CD3+ and the number of lymphocytes with signs of early apoptosis (R = -0.47; p 0.05; R = -0.51; p 0.05, respectively), and signs of late apoptosis and necrosis (R = -0.64; p 0.05; R = -0.42; p 0.05, respectively), between CD45RA+ and the number of lymphocytes with signs of early apoptosis (R = -0.47; p 0.05; R = -0.49; p 0.05, respectively), and signs of late apoptosis and necrosis (R = -0.57; p 0.05; R = -0.49; p 0.05, respectively). Usage of MT in DF composition leads to increase in blood CD3+ on the 5th and 20th days, CD45RA+, on the 5th day, and an increase in serum IgG concentration was observed on the 5th and 10th days following TT. Restriction of necrotic and apoptotic death of blood lymphocyte may be among the mechanisms of the immunotropic effect produced by MT which is, probably, due to its local antioxidant and anti-inflammatory action in the TT area.

STUDIES TO DETERMINE SHELF LIFE AND STORAGE CONDITIONS WOUND HEALING GEL WITH CYCVALONE

One of the urgent tasks in the process of drug development is to obtain stable pharmaceutical products in the process of storage and use. To establish the expiration date, it is necessary to determine the time during which the developed drug is able to maintain stability and meet all the requirements stated in the regulatory documentation. The aim of the study was to study the stability of laboratory samples of wound healing gel with cycvalone during long-term storage in natural conditions. The developed composition of the wound healing gel includes the antioxidant agent cycvalone, which has a pronounced manifestation of antioxidant and anti-radical activity, as well as has a wound healing, immunotropic and anti-inflammatory effect. Carbomer (Carbopol-940), potassium hydroxide solution and purified water were used as excipients. Gel samples were stored in the primary packaging at room temperature in a place protected from light. The stability of the dosage form was studied according to the following parameters: description, pH of water extraction, authenticity, colloidal and thermal stability, quantitative determination, rheological parameters in accordance with the developed methods. The measurement was performed on 6 series of dosage forms: after preparation, after 6, 12, 18 and 24 months of storage in vivo. The results of determining the quality indicators of the wound healing gel with an antioxidant agent (cycvalone) indicate a rational selection of active and auxiliary substances and their concentration, the choice of a rational technology and the absence of interaction between the components of the developed dosage form. Based on the conducted studies, it is proved that the wound healing gel with cycvalone remains stable during storage at a temperature of 15 to 25 ˚С for 24 months.

Immunotropic effects of nitrogenous metabolites (creatinine, urea, uric acid and bilirubin) in humans exposed to the factors of the accident at the Chоrnobyl nuclear power plant

Background. In previous studies, we found that nitrogenous metabolites exhibit significant immunotropic activity, both suppressor and enhancing, at healthy rats. The purpose of this and our subsequent research is to elucidate the links between nitrogenous metabolites and immune parameters in different categories of people, both healthy and sick. Material and methods. The object of observation in 1997 were 19 men and 3 women who were exposed to pathogenic factors of the accident at the Chоrnobyl nuclear power plant during the liquidation of its consequences in 1986-87. The survey was conducted twice - on admission and after two weeks of rehabilitation at the Truskavets’ Spa. The plasma and urinary concentration of the nitrogenous metabolites were determined. Immune status was assessed on tests of I and II levels according to the WHO memorandum. Results. Both negative and positive metabolic-immune correlations were revealed. Calculation of multiple correlation coefficients between individual metabolite parameters and constellations of immune parameters revealed the maximum immunotropic effect of Urea urine (R=0,756). This is followed by Uric acid plasma (R=0,727) and urine (R=0,691), Urea plasma (R=0,622), Creatinine plasma (R=0,588), Bilirubinemia (R=0,546) and Creatinine urine (R=0,510). The canonical correlation between the constellation of nitrogenous metabolites, on the one hand, and the parameters of immunity, on the other hand, was very strong: R=0,971; χ2(15)=239; p<10-5. Conclusion. Nitrogenous metabolites have immunomodulatory effects, both suppressor and enhancing, both in healthy rats and in humans exposed to pathogenic influences.

APPLICATION OF THE IMMUNOMODULATOR IN CHILDREN WITH OBSTRUCTIVE BRONCHITIS

Purpose. To assess the efficacy and safety of the use of the immunomodulator polyoxidonium in preschool children with obstructive bronchitis. Material and methods. The study included 35 children, who, based on anamnesis, clinical, laboratory and instrumental examination methods, were diagnosed with obstructive bronchitis. The age of the examined children ranged from 3 to 6 years. The control group consisted of 11 healthy children. The material for obtaining immunological data in children was peripheral venous blood. In order to analyze immunological changes, the content of B-lymphocytes and immunoglobulins in the blood was studied. The investigated immunological data included 6 indicators. The content of immunoglobulins was determined by enzyme immunoassay according to the attached instructions. Statistical analysis of the revealed data was carried out using the Wilcoxon test using the Statistica 6.0 software. All children underwent dynamic laboratory examination and immunological parameters. The primary examination was carried out when the children were admitted to the hospital. Re-examination was carried out after the end of therapy. Group I (n = 19) included patients who received treatment according to the classical scheme. Group II children (n = 16) underwent complex therapy with an immunomodulator. The drug polyoxidonium was selected as the study drug. Results. The data obtained reflect the effectiveness of the use of the immunomodulator polyoxidonium in the complex therapy of obstructive bronchitis in children. Conclusion. Immunological data obtained during the initial examination reflect the development of dysfunction of the immune system in children. It was revealed that polyoxidonium has an immunotropic effect, this is verified by the optimization of the parameters of the humoral link of the immune system.

Immunotropic effects of so-called slag metabolites (creatinine, urea, uric acid and bilirubin) at rats

Background. The study of the effect of end products of protein and nucleic acid metabolism on the immune system is of interest not only theoretical but also practical in line with the problems of urotoxins and hemodialysis. In previous studies, we found significant links between uricemia and uricosuria, on the one hand, and immune parameters, on the other hand. In this study, the spectrum of nitrogenous metabolites was expanded due to creatinine, urea and bilirubin. Material and methods. Experiment was performed on 60 healthy female Wistar rats. The plasma level and urinary excretion of the nitrogenous metabolites were determined. Immune status was assessed by thymocytogram, splenocytogram, blood leukocytogram and immunocytogram, as well as phagocytosis of blood neutrophils and monocytes. Results. Both negative and positive metabolic-immune correlations were revealed. Calculation of multiple correlation coefficients between individual metabolite parameters and constellations of immune parameters revealed the maximum immunotropic effect of uricosuria (R=0,637). This is followed by excretion of urea (R=0,617) and creatinine (R=0,606), bilirubinemia (R=0,589), creatinineemia (R=0,567), uricemia (R=0,566) and plasma urea level (R=0,500). The canonical correlation between the constellation of nitrogenous metabolites, on the one hand, and the parameters of immunity, on the other hand, was very strong: R=0,921; χ2(154)=282; p<10-6. Conclusion. Nitrogen metabolites exhibit significant immunotropic activity, both suppressor and enhancing.