SUMMARY OF 1st STAGE / 2018

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In order to achieve the proposed objectives:

            In activity 1.1. "Selection and characterization of marine sources for the isolation of bioactive compounds (Pr 1)": two marine sources rich in protein and polysaccharide biologically active compounds were selected, namely Rapana venosa gastropods and green macrogens of Cladophora sp., After performing a scientific study on recoverable sources and their physicochemical and biochemical analysis (determination of protein, carbohydrate, lipid and glycosaminoglycans content).

             In activity 1.2. "Establishment of technologies for the production of protein and polysaccharide extracts from the selected sources (Pr 1)": the technological parameters of extraction have been established and optimized in order to elaborate the technologies for obtaining the protein extracts (collagen, gelatin) and polysaccharides (total polysaccharides, glycosaminoglycans) from the two marine sources. In the course of this activity, the technological stages of preparation (pretreatments, extraction and purification) were tested and the working parameters were optimized, such as: the type and concentration of the extraction agent, the tissue / solvent ratio, the tissue / enzyme, type and concentration of the substance used for purifications, temperatures and working times.

            In activity 1.3."Elaboration of Experimental Methods for Characterization of Protein and Polysaccharide Extracts from Selected Sources (Pr 1)": the optimal experimental methods were developed for the characterization of the protein and polysaccharide extracts obtained in the previous activity, which were grouped in two types of methods: 1 physicochemical and biochemical - protein dosing, determination of total hexoses, content in uronic acids, SDS-PAGE electrophoresis, denaturation temperature evaluation, FTIR analysis, respectively 2. Biological assessment of in vitro biocompatibility by quantitative methods (Neutral Red technique, tripartite blue exclusion, tetrazolium salt reduction test, lactate dehydrogenase dosing) and qualitative (optical and fluorescence microscopy) as well as antimicrobial activity assessment (diffusion method). Within this activity, 4 work protocols were established for quantitative collagen determination, glycosaminoglycan dosing, quantitative assessment of cytotoxicity and determination of antimicrobial activity.

            Within the activities 1.4. "Synthesis and Characterization of Functionalized Mesoporous Silica (Pr 2)" and 1.5. "Evaluation of biocompatibility of functionalized mesoporous silica supports (Pr 2)": four types of pure MCM-41 silica particles and 4 types of functionalized nanoparticles of silica were obtained with organic groups prepared by the co-condensation method by an original method. It has also been developed a procedure for the functionalization of MCM-41 type mesoporous silica particles with Zn and Mg heteroatoms, two metal ions involved in biological processes, which is expected to improve the adsorption capacity of bioactive compounds. The silica particles were characterized by various techniques: X-ray diffraction at low angles, thermal analysis, FTIR spectroscopy. The obtained particles have high porosity, an ordered structure of the pore mesh, various chemical compositions, and on the basis of tests of cell viability assays on normal and tumor cells it can be appreciated that some of them can be used as matrices for the encapsulation of some principles assets obtained from marine or wine bioresources depending on the intended purpose.

            Parallel to the carrying out of these activities, under Activity 1.6 "A) Harvesting, preparation and characterization of marine raw materials of plant origin and B) and harvesting, preparation and characterization of vegetal material (tescina) in the Murtfatlar area (Pr 2) ": The marine macroalgae of the species Ulva lactuca which was characterized in terms of the content of organochlorine pesticides, heavy metals and biochemical composition, was harvested and selected. Hydroalcoholic extracts of Ulva rigida (sin. Ulva lactuca) were prepared and antioxidant and antimicrobial activities determined. Also in this stage, viticultural material was harvested in the Murfatlar area, Constanta County. From this biological material, alcoholic extracts of different concentrations were prepared, by different methods, from various varieties of grapes. These were characterized in terms of the content of ascorbic acid and total polyphenols.

            The results obtained at this stage determined the achievement of all the objectives, being promising in the perspective of developing technologies for encapsulation of the active principles of the prepared extracts, which have the expected therapeutic effects.

            In conclusion, during the first stage of the complex project, all the result indicators were reached and the value of the results obtained is supported by the dissemination activity, by publishing 1 ISI Impact factor 2017/2018 article: 3,649; 8 papers communicated at international conferences, of which 3 papers published in volumes of ISI indexed conferences and 1 paper published in non-indexed volume volumes. Also, the implementation of the first phase of the project has improved the infrastructure of all partners and has diversified their service offerings. Six new young researchers with very good professional training (PhD students and master students) were hired, involved both now and in the future in the activities of the complex project.