FORMULATION AND TECHNOLOGY DEVELOPMENT OF HERBAL PHENOLIC BIOPOLYMER-CONTAINING FILMS FOR BURN TREATMENT.

Application of phytofilms based on biosolublepolymers is considered as a prospectivemethod for burn treatment . Herbal remedies contain biologically active substances, that are relatively less toxic, do not cause skin irritation or allergic reactions and, importantly, affectstrains of the microorganisms and viruses resistant to antibiotics and synthetic drugs. Nowadays, the advantages are given to such burn healing drugs, which along with high specific efficacy, have analgesic, anti-inflammatory and antimicrobial effects, and don't irritate the tissues. The mentioned peculiarities are characteristic for a new herbal phenolic biopolymer poly[3-(3,4-dihydroxyphenyl) glyceric acid](PDGA), isolated from the roots and stems of different comfrey species . The aim of the study was the development of the formulation and technology of biosoluble films for burn treatment on the basis of PDGA. The optimal content of phytofilm for burn healing was selected on the basis of the biopharmaceutical study results. The impact of the film-former on the quality, adhesion and moisture absorption of the phytofilmhas been studied. The optimal degree of the phytofilm moisture, determining its high adhesive properties,was established. The film prepared on the basis of sodium alginate, with 30.4% humidity, demonstrated the greatest adhesion strength. After investigation of the PDGA release it was found, that the hydrophilic bases such as: sodium carboxymethyl-cellulose (69.2%) andsodium alginate (78,65%) appeared to be optimal among the others. At the same time, taking into consideration the disadvantages of sodium carboxymethyl-cellulose (tautening effect on burnt surface, relatively low stability), a film based on sodium alginate has been chosen. The manufacturing technology for obtaining PDGA-containing phytofilm by casting is proposed. Theshelf-lifeofproposedPDGA-containingphytofilmis 2 years.

DEVELOPMENT OF FORMULATION AND TECHNOLOGY FOR THE POLY[3-(3,4-DIHYDROXYPHENYL)GLYCERIC ACID] GEL.

One of the most actual problems of pharmacy is the development of medication forms for external application with complex effects on (gel, emplastro, aerosol, etc.) skin wounds, burns and inflammatory factors. The centuries-old practice of using phyto-preparations (herbal remedies) proved that they have fewer side effects in comparison with synthetic drugs. Despite the wide application of herbal preparations, in the literature there is a little information about their application in development of wound and burn healing modern dosage forms. Among the medicinal plants with the mentioned pharmacological actions, comfrey (Symphytum L.) should be distinguished. Phenolic polymer poly[3-(3,4-dihydroxyphenyl)glyceric acid] (PDGA) or poly[oxy-1-carboxy-2-(3,4-dihydroxyphenyl)ethylene], amounting approximately 25% of polysaccharides and 1.5-2.5% of dry plant material, were isolated from the roots and stems of Caucasian comfrey species (S. asperum, S. caucasicum). Contrary to polysaccharides this phenolic polymer of Comfrey appeared to have a high immunomodulatory (anticomplement), antioxidative, antilipoperoxidantive, anti-inflammatory and wound-healing efficacy/activities. The aim of the study was development of the composition and technology of PDGA-containing gel. According to the results of complex biopharmaceutical studies PDGA gel optimal composition has been proved. The technological scheme for preparation of PDGA gel has been developed. PDGA gel stability under normal conditions of storage at +40С was studied. The gel has a shelf life (determined expiration date) of 2 year.

RAPID IDENTIFICATION OF THE ETIOLOGICAL FACTORS CAUSING DIARRHEAL DISEASES.

The main objective of this investigation was to develop and pilot a real-time Polymerase Chain Reaction (rt-PCR) diagnostic system for rapid and simultaneous identification of pathogens with a particular emphasis on diarrheal disease diagnostics. The diarrheal diseases were selected as a target for the pilot because they constitute a primary public health priority in Georgia and worldwide. The product developed by our research team "Neo_PCR_Diagnostics" represents an original system for the identification of pathogens associated with gastrointestinal tract infections and diarrhea. The advantages of the proposed technology over existing conventional methods include the ability of simultaneous identification of multiple pathogens and the detection of pathogenic agents directly from the fecal samples. For the evaluation of the new diagnostic system, stool samples were collected at collaborating hospitals and clinics and analyzed by real-time PCR using the Neo_PCR_Diagnostic system. The selection of the pathogens for detection was based on their epidemiological and clinical importance. The following bacterial pathogens were targets for detection: Salmonella spp., Campylobacter spp., Shigella spp., Clostridium difficile (Toxin A/B), Escherichia coli (ETEC, STEC and O157), Yersinia enterocolitica and Vibrio cholerae. The following viral pathogens were studied: adenoviruses, rotaviruses and noroviruses. Genetic material (DNA) of the following parasites were targets in our study: Giardia lamblia, Entamoeba histolitica and Cryptosporidium spp. We also compared the results obtained by our molecular technology with the conventional methods - bacterial culture (for bacterial growth) and ELISA (for bacterial toxins). For viral and parasitic pathogens, comparison tests were performed with immunochromatographic assays for direct detection of antigens in the stool samples or with the data obtained by use of home-made end-point PCR (where available). Advantages of the proposed technology over existing conventional technologies include the ability of simultaneous identification of diarrheal infections by multiple pathogens. The proposed test system allows the detection of pathogenic agents directly from the fecal samples and can be completed within one working day. In general, the spectrum of pathogens detected by our approach was wider than those detected by the conventional methods used in the clinical setting, taking into consideration the list of pathogenic agents requisitioned by physicians within the framework of the routine clinical visit. Given these promising results, Neo_PCR_Diagnostics test performance and accuracy may be sufficient for use in molecular microbiological diagnostics in clinical and/or research settings.

Ivy water extracts as gastric ulcer preventive agents.

In folk medicine the ivies (Hedera L. Fam.Araliaceae) are known as plants possessing diverse curative properties. A comparative phytochemical study of the biologically active water extracts of H. colchica and H. helix and evaluation of their ulcer preventive efficacy in ethanol-induced ulcer model in rats was carried out. Water extracts of H. colchica and H. helix (300 mg/kg, i.p.) significantly (p<0.01) decrease the ulcer index (0.50 and 1.38 vs 3.17 in control) and rise macroscopic curative ratio (84.2% and 56.6%, respectively). The results clearly indicate that pretreatment with water extract of H. colchica is preferable and further experiments are required to isolate the active principals responsible for itsantiulcerogenic activity.

Novel protective agent GF-6 for skin wounds.

The aim of present investigation was the determination of specific pharmacological (wound healing) properties and the evaluation of possible irritative, allergenic and toxic effects of GF-6 - novel protective agent for skin mechanical and burn wounds. In mouse excisional wound model GF-6 (topically 0.1 ml per wound) exhibited the ability to accelerate scab rejection and full reepithelization in test wounds with no signs of bacterial contamination. GF-6 caused no irritation responses such as erythema, oedema, dotted hemorrhage, hyperkeratosis, hyperplasia, alopecia and scaling in 14-day acute dermal irritation test. As well no signs of skin hypersensitivity (itch, erythema, oedema), and histological alteration (ulceration, dotted hemorrhage, changed capillary lumen, damaged hair follicle) were observed in skin application test at all time points during the 14-day observation period, evidencing that GF-6 is non-allergenic. In subchronic dermal toxicity 90-day study of five-fold concentrated GF-6 no statistically significant changes in respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behavior patterns were detected. The results of hematological study testified to no influence of the GF-6 90-day application on blood formed elements. Thus, GF-6 should be considered non-toxic, non-irritant, non-allergenic wound healing and wound protecting agent.