Relevance of Bioassay of Biologically Active Substances (BAS) with Geroprotective Properties in the Model of the Nematode Caenorhabditis Elegans in In Vivo Experiments.

Aging is an inevitable process of nature. The age of living organisms contributes to the appearance of chronic diseases, which not only reduce the quality of life but also significantly damage it. Modern medicines can successfully fight multiple diseases and prolong life. At the same time, medications have a large number of side effects. New research indicates that bioactive phytochemicals have great potential for treating even the most severe diseases and can become an alternative to medicines. Despite many studies in this area, the effects of many plant ingredients on living organisms are poorly understood. Analysis of the mechanisms through which herbal preparations influence the aging process helps to select the right active substances and determine the optimal doses to obtain the maximum positive effect. It is preferable to check the effectiveness of plant extracts and biologically active components with geroprotective properties in vivo. For these purposes, live model systems, such as Rattusrattus, Musmusculus, Drosophila melanogaster, and Caenorhabditis elegans are used. These models help to comprehensively study the impact of the developed new drugs on the aging process. The model organism C. elegans is gaining increasing popularity in these studies because of its many advantages. This review article discusses the advantages of the nematode C. elegans as a model organism for studying the processes associated with aging. The influence of various BAS and plant extracts on the increase in the life span of the nematode, its stress resistance, and other markers of aging is also considered. The review shows that the nematode C.elegans has a number of advantages over other organisms and is a promising model system for studying the geroprotective properties of BAS.

Study of the l-Phenylalanine Ammonia-Lyase Penetration Kinetics and the Efficacy of Phenylalanine Catabolism Correction Using In Vitro Model Systems.

The kinetics of l-phenylalanine ammonia-lyase (PAL) penetration into the monolayer of liver cells after its release from capsules was studied. The studies showed the absence of the effect of the capsule shell based on plant hydrocolloids on the absorption of l-phenylalanine ammonia-lyase in systems simulating the liver surface. After 120 min of incubation, in all variants of the experiment, from 87.0 to 96.8% of the enzyme penetrates the monolayer of liver cells. The combined analysis of the results concludes that the developed encapsulated form of l-phenylalanine ammonia-lyase is characterized by high efficiency in correcting the disturbed catabolism of phenylalanine in phenylketonuria, which is confirmed by the results of experiments carried out on in vitro model systems. PAL is approved for the treatment of adult patients with phenylketonuria. The encapsulated l-phenylalanine ammonia-lyase form can find therapeutic application in the phenylketonuria treatment after additional in vitro and in vivo studies, in particular, the study of preparation safety indicators. Furthermore, it demonstrated high efficacy in tumor regression and the treatment of tyrosine-related metabolic disorders such as tyrosinemia. Several therapeutically valuable metabolites biosynthesized by PAL via its catalytic action are included in food supplements, antimicrobial peptides, drugs, amino acids, and their derivatives. PAL, with improved pharmacodynamic and pharmacokinetic properties, is a highly effective medical drug.

Evaluation of Biocompatibility and Antagonistic Properties of Microorganisms Isolated from Natural Sources for Obtaining Biofertilizers Using Microalgae Hydrolysate.

Determination of the biocompatibility of microorganisms isolated from natural sources (Kemerovo Oblast-Kuzbass) resulted in the creation of three microbial consortia based on the isolated strains: consortium I (Bacillus pumilus, Pediococcus damnosus, and Pediococcus pentosaceus), consortium II (Acetobacter aceti, Pseudomonas chlororaphis, and Streptomyces parvus), and consortium III (Amycolatopsis sacchari, Bacillus stearothermophilus; Streptomyces thermocarboxydus; and Streptomyces thermospinisporus). The nutrient media composition for the cultivation of each of the three studied microbial consortia, providing the maximum increase in biomass, was selected: consortium I, nutrient medium 11; consortium II, nutrient medium 13; for consortium III, nutrient medium 16. Consortia I and II microorganisms were cultured at 5-25 °C, and consortium III at 50-70 °C. Six types of psychrophilic microorganisms (P. pentosaceus, P. chlororaphis, P. damnosus, B. pumilus, A. aceti, and S. parvus) and four types of thermophilic microorganisms (B. stearothermophilus, S. thermocarboxydus, S. thermospinisporus, and A. sacchari) were found to have high antagonistic activity against the tested pathogenic strains (A. faecalis, B. cinerea, E. carotovora, P. aeruginosa, P. fluorescens, R. stolonifera, X. vesicatoria. pv. Vesicatoria, and E. aphidicola). The introduction of microalgae hydrolyzate increased the concentration of microorganisms by 5.23 times in consortium I, by 4.66 times in consortium II, by 6.6 times in consortium III. These data confirmed the efficiency (feasibility) of introducing microalgae hydrolyzate into the biofertilizer composition.

The effect of postharvest ultraviolet irradiation on the content of antioxidant compounds and the activity of antioxidant enzymes in tomato.

The effect of different doses of long-wavelength UV-A (320 nm-400nm) irradiation on physicochemical and antioxidant characteristics of tomatoes grown on the territory of the Russian Federation was studied. The obtained results show that this kind of processing does not cause deterioration of qualitative parameters of vegetables (texture, color, soluble solids content, titratable acidity). It was established that the total content of phenolic compounds, carotenoids and flavonoids increases (p-value<0.05) in tomatoes at all the investigated wavelengths (353 nm, 365 nm and 400 nm), while the content of chlorophylls reacts ambiguously: at some wavelengths, it increases, at other, it decreases. The maximum increase in antioxidant activity, as compared to untreated samples, is observed in tomatoe samples irradiated for 360 min within the range of 365 nm. For different types of tomatoes, the increment for common content of phenolic compounds is - 42.9-55.0 %, carotenoids - 24.0-56.0 %, flavonoids - 28.0-33.0 %, ß-carotene - 70.9-71.6 %, lycopene - 62.6-69.0 %, lutein - 64.8-72.0 % from original. The studies reveal some potential of post-harvest ultraviolet irradiation (A-range) of tomatoes to increase their antioxidant activity. However, more research is needed to confirm this fact and the possibility to develop some technology.

The effectiveness of plant hydrocolloids at maintaining the quality characteristics of the encapsulated form of L-phenylalanine-ammonia-lyase.

The effect of three types of polysaccharides (agar-agar, carrageenan, hydroxypropyl methylcellulose) on the activity and stability during storage at given temperature conditions of the enzyme preparation L-phenylalanine ammonia-lyase was studied. It was found that the most suitable storage temperature for encapsulated L-phenylalanine-ammonia-lyase is room temperature up to 25 °C for all samples of capsules from plant polysaccharides. Samples of capsules with agar-agar and hydroxypropyl methylcellulose under different temperature conditions inhibited the decrease in enzyme activity, which in other samples of capsules reached 90% in 6 months of storage. In samples of capsules with carrageenan at temperatures of 4 °C and 30 °C, there was a significant decrease in the activity of the enzyme preparation. Selection of capsule samples from plant polysaccharides suitable for L-phenylalanine-ammonia-lyase replacement therapy is done after studying the mechanisms of capsule destruction under conditions close to the conditions of the gastrointestinal tract, to which the next stage of our research will be devoted.

Sea Buckthorn and Rosehip Oils with Chokeberry Extract to Prevent Hypercholesterolemia in Mice Caused by a High-Fat Diet In Vivo.

Dietary supplementation based on sea buckthorn and rosehip oils with added chokeberry extract was studied. We added the dietary supplement to the feed mixtures for laboratory animals. The possible toxicological effects and hypocholesterolemic, hepatoprotective activity of the dietary supplement in vivo were studied. After the observation period (6 weeks), no significant changes were found in the mass of organs and blood serum of laboratory animals (p > 0.05). However, there was a decrease in hypercholesterolemic indicators. Regular consumption of sea buckthorn and rosehip oils with added chokeberry extract (dietary supplement "ESB-1") by laboratory animals inhibited the activity of liver enzymes and increased the antioxidant activity of blood serum (after the subcutaneous injection of sunflower oil/oil solution of carbon tetrachloride) but was not sufficient to bring them to physiological standards. The hypocholesterolemic and antioxidant properties of our dietary supplement already allow us to consider it a component of functional food products or a dietary supplement base. However, the full range of its biologically active properties, including the hepatoprotective function and regulation of metabolic disorders, has not been studied yet, which sets the direction of further research in vivo models and clinical practice to confirm its effectiveness in humans.

Study of the Potential of the Capsule Shell Based on Natural Polysaccharides in Targeted Delivery of the L-Phenylalanine Ammonia-Lyase Enzyme Preparation.

The treatment of classical phenylketonuria is currently represented by many new methods of disease management. A promising method is the use of the enzyme L-phenylalanine ammonia-lyase (PAL) in various forms. The widespread use of enzyme preparations in therapy is limited by a lack of understanding of the mechanisms and systems of the targeted transport of PAL into certain organs and tissues as a result of the incorporation of a drug into the carrier. To ensure the stability of enzymes during the delivery process, encapsulation is preferable, which, as a rule, ensures the preservation of the qualitative characteristics of the enzymes orally applied to the environmental effects of the gastrointestinal tract (acidity, temperature, oxidation, etc.). Capsule preparations showed sufficient stability in the model gastric fluids and sustained release of the drug in the simulated intestinal fluid. Currently, there is a wide range of polymers used for encapsulation. The use of natural sources in the production technology of capsule systems improves bioavailability, controls the release, and prolongs the half-life of active substances. The advantage of this method is that the used enzyme is completely protected by the cell membranes of the capsules, which preserve its stability in the aggressive environment of the gastrointestinal tract. Capsules were obtained on the basis of compositions of hydrocolloids of plant origin. The potential of the developed capsules for targeted delivery of the enzyme preparation was studied. The degradation of the encapsulated form of the PAL enzyme preparation was studied in vitro in model bio-relevant media simulating the gastric and intestinal environment. The dynamics of the breakdown of the capsule shell allow us to expect that the release of L-phenylalanine ammonia-lyase from capsules based on plant hydrocolloids will occur no earlier than reaching the upper intestines, where the interaction with the protein components of the consumed food products to neutralize phenylalanine should occur.

Identification and quantification of phenolic compounds of Western Siberia Astragalus danicus in different regions.

The potential of phenolic compounds of medicinal plants including Astragalus danicus L is determined by but not limited to their antioxidant activity. Their anti-inflammatory, antitumor, and other useful properties are known, which allows using these phytochemicals within preventive activities to reduce the risk of many serious diseases. Chromatographic analysis of the Astragalus danicus L. biomaterial from the plant samples collected in three regions of the Kemerovo region (Western Siberia, Russia) established the presence of compounds of flavonols (isorhamnetin glucoside, kaempferol glucoside), flavones (apigenin 7-glucoside), phenylpropanoids (chlorogenic acid) in the aerial part of plants. The total content of phenolic compounds in plant samples ranged from 100.75 ± 3.87 mg/g (Yashkinsky district) to 190.95 ± 7.34 mg/g (Belovsky district). The content of chlorogenic acid in the studied samples was from 0.14 ± 0.01 mg/g to 1.16 ± 0.04 mg/g. Isorhamnetin glucoside was found only in samples of plants from two districts - Prokopievsky (41.39 ± 1.58 mg/g) and Belovsky (95.0 ± 3.66 mg/g). The content of glucosides of kaempferol ranged from 0.38 ± 0.01 mg/g to 0.55 ± 0.02 mg/g. Its content is almost twice as high as the content in the well-known analogues of Astragalus. Apigenin-7-glucoside was isolated in Astragalus samples for the first time, in a small amount (3.34 ± 0.13 mg/g) in a sample of plants of one growing zone. Studies have confirmed that the content of flavonoids in plants significantly depends not only on the genetic characteristics of plants, but also on the hydrothermal regime, the climatic conditions of different botanical and geographical areas of the habitat. This work shows that Astragalus danicus L. growing in Kemerovo region is a promising raw material for pharmacological preparations.

Short Chain Fatty Acids (SCFA) Reprogram Gene Expression in Human Malignant Epithelial and Lymphoid Cells.

The effect of short chain fatty acids (SCFAs) on gene expression in human, malignant cell lines was investigated, with a focus on signaling pathways. The commensal microbial flora produce high levels of SCFAs with established physiologic effects in humans. The most abundant SCFA metabolite in the human microflora is n-butyric acid. It is well known to activate endogenous latent Epstein-Barr virus (EBV), that was used as a reference read out system and extended to EBV+ epithelial cancer cell lines. N-butyric acid and its salt induced inflammatory and apoptotic responses in tumor cells of epithelial and lymphoid origin. Epithelial cell migration was inhibited. The n-butyric gene activation was reduced by knock-down of the cell membrane transporters MCT-1 and -4 by siRNA. N-butyric acid show biologically significant effects on several important cellular functions, also with relevance for tumor cell phenotype.