[Influence of chronic immobilization stress on vitamin status in rats fed different diets].

The influence of a stress factor, widespread in modern conditions, on the vitamin status has not been studied enough. At the same time, the negative stress impact can be aggravated against the background of unhealthy nutrition, which in turn affects the vitamin status of the organism. In this regard, the goal of the research was to evaluate the effect of chronic restrict stress on the vitamin supply in rats fed a diet with adequate and increased content of fat, sugar and cholesterol. Material and methods. The experiment was carried out on 37 growing male Wistar rats (initial body weight of 45±5 g) divided into 4 groups. Animals of the 1st (control) and the 2nd groups received a complete semi-synthetic diet (CSSD) (20% protein, 10% fat, 58% carbohydrates in the form of starch, 384 kcal/100 g) for 92 days. The levels of all vitamins and mineral elements in the rats' diets were adequate for growing rats. Rats of the 3rd and the 4th groups were fed a high-calorie, high-fat high-carbohydrate diet (HFHCD) (20% protein, 28% fat, 2% cholesterol, 18% carbohydrates in the form of starch, 20% sucrose, 511 kcal/100 g). Animals of groups 2 and 4 were subjected to daily 90-minute immobilization. The concentration of vitamins A (retinol and retinol palmitate) and E (α-tocopherol) in the blood serum and liver were determined by high-performance liquid chromatography, vitamins B1 and B2 in the liver and urine, as well as riboflavin in the blood serum and 4-pyridoxic acid (4-PA) in urine were determined by fluorimetric methods. Biochemical parameters of blood serum were determined on a biochemical analyzer; the total content of fat, triglycerides (TG) and cholesterol (CH) was determined in the liver. Results. Replacing CSSD with HFHCD, both under restraint stress and without, was accompanied by an increase in liver weight by 1.8-2.0 fold, in its fat content by 2.6-3.3 fold, cholesterol by 32.6-35.3 fold and TG - by 33.0-57.6 fold (p=<0.001). An increase in alanine aminotransferase (ALT) activity by 1.7-2.0 fold (p=<0.01), in low-density lipoprotein (LDL) cholesterol level by 5.4 fold (p=<0.05) and the atherogenic coefficient by 2.5 fold (p<0.01) as well as a decrease in creatinine and urea level (p=<0.05) in blood serum were revealed. Immobilization was accompanied by a decrease in body weight, liver and liver fat in rats fed both CSSD and HFHCD (p<0.05), but didn't affect the blood serum biochemical parameters, with the exception of an increase in ALT activity. If the activity of alkaline phosphatase (ALP) did not change during immobilization of rats fed the CSSD, then in animals fed the high-calorie diet it decreased by 37.5% (p=<0.05 from the control) under its increase against the background of restrict stress by 78.7% (p=<0.01) compared to the indicator of rats of the 3rd group. Immobilization of rats treated with CSSD was accompanied by an increase in both absolute serum α-tocopherol level and concentration correlated with the level of cholesterol and triglycerides by 26.0-57.5% (p<0.05), with a simultaneous decrease in its content in the liver per 1 g of wet tissue by 22.1% (p=0.041) relative to the indicators of intact animals. Immobilization reduced the level of retinol palmitate in the liver by 2.3 times (p<0.01), but did not affect retinol level in the blood serum. At the same time, indicators of B vitamin status (the content of vitamins B1 and B2 in the liver per 1 g of wet tissue and per organ, blood serum riboflavin level, urinary excretion of riboflavin and 4-PA) did not change, with the exception of thiamine urinary excretion, which reduced compared to the control by 38.8%. In rats fed HFHCD, immobilization had no additional effect on the supply with vitamins A and E. The content of vitamins B1 and B2 in the liver in terms of the whole organ was reduced by 14.0-26.7% relative to the indicator in animals of the 3rd group, not subjected to chronic stress, only due to differences in liver weight in animals of these groups. Conclusion. The data obtained indicate that chronic stress has a negative effect on the vitamin status of the body, worsening the supply with vitamins A, E and B1, and substantiate the feasibility of studying the mechanisms of this effect in order to develop effective vitamin complexes for the treatment and prevention of diseases caused by long-term stress.

[Physiological and biochemical in vivo study of polyphenols and 20-hydroxyecdisone from quinoa grains effect on resistance to physical exercise in Wistar rats].

Increasing the ability of the human body to adapt to physical stress is relevant from the standpoint of using foods for special uses containing functional food ingredients (FFI) with effectiveness proven in vivo. The purpose of this study was to evaluate the effect of FFI from Chenopodium quinoa grains with a high content of polyphenols and phytoecdysteroids on the physical endurance of male Wistar rats. Material and methods. The experiment was carried out during 36 days using 50 weaned male Wistar rats. The animals were randomly divided into 3 groups (n=12): Control, Run and Run-FFI. Rats of the Control and Run groups received a standard semisynthetic diet during the experiment. Rats of the Run-FFI group received a semi-synthetic diet with the addition of FFI in an amount of 0.055±0.003%, containing phytoecdysteroids (50.4±0.6 mg/g) and polyphenols (212.0±2.0 mg/g). During the experiment, the rats were assessed for their neuromotor function (grip strength of front paws), memory, and behavioral reactions in the "Elevated Plus Maze" (EPM), "Conditioned Passive Avoidance Reflex" (CPAR) and "Open Field" (OF) tests. Once a week, animals from the Run and Run-FFI groups were subjected to moderate physical load on a "Treadmill". On the 36th day of the experiment, the animals of these groups were subjected to exhausting physical load. Immediately after running, the animals were placed in metabolic cages to collect daily urine. At the end of the experiment, the content of corticosterone, the activity of catalase, indicators of protein, lipid and mineral metabolism, indexes of the liver functional state and antioxidant defense system parameters were analyzed in the blood serum; the level of prostaglandin E2 and dopamine were determined in daily urine. Results. Physiological tests (CRAR, OF) showed that weekly exercise increased anxiety in laboratory animals. The FFI introduction into the diet led to normalization of the assessed parameters (EPM). As a result of 36-day consumption of FFI against the background of physical loads, a significant decrease by 22% in the main stress marker, corticosterone, was revealed in the blood of rats, as well as significant increase by 23% in the stress inhibitor - prostaglandin E2 urinary excretion, compared with animals of the Run group to the level not differed from the indicators of the control animals. There were no differences in endurance performance between the Run and Run-FFI groups on the results of the exhaustive exercise. Consumption of FFI prevented the formation of excess ammonia, significantly reducing the level of urea in the blood and normalizing its excretion to control levels in the urine, which was increased in the Run group by 19%. Conclusion. The results obtained demonstrated the adaptogenic properties of the developed FFI in response to stress caused by weekly moderate and acute exhaustive physical activity. The obtained data on the biological effect of the developed FPI on the adaptive potential of laboratory animals will serve as an experimental basis for its inclusion in the composition of specialized foods.

[In vivo study of the biological value of amaranth protein concentrate and its module with chicken egg protein].

Amaranth (Amaranthus L.), like other pseudocereals as quinoa (Chenopodium quinoa Willd.), chia (Salvia hispanica L.) and buckwheat (Fagopyrum sp.), is a promising source of dietary protein. Depending on the subspecies and breeds of amaranth, the protein content in its grain is estimated from 13.1 to 21.5%, and its amino acid score varies over a significant range and can be limited. The aim of this study was to obtain a protein concentrate from amaranth (Amaranthus L.) grain of the Voronezh breed, enrich it with chicken egg protein, determine the amino acid score of the obtained protein module, and experimentally evaluate in vivo its true digestibility and biological value. Material and methods. The amaranth protein concentrate was obtained from grain according to the technological scheme, including its enzymatic treatment, alkaline extraction, acid precipitation of proteins, microfiltration and lyophilization. The amino acid composition and amino acid score of the concentrate were determined. The protein module was obtained by mixing amaranth protein concentrate and chicken egg protein in a weight ratio of 58:42. The true digestibility and biological value of the protein module has been determined in vivo. The experiment was carried out on 32 Wistar male rats divided into 2 groups (n=16 rats): control group 1 with a body weight of 118.7±3.1 g and experimental group 2 with a body weight of 119.5±3.0 g. Animals of groups 1 and 2 received diets in which egg protein and a protein module were used as a protein source, respectively. Within 15 days of the experiment, individual indicators of food intake and body weight gain of each animal were determined. From the 14th to the 15th day food intake was determined and feces were collected. The amount of nitrogen in the food and feces was determined for each rat using the Kjeldahl method. The true digestibility of the protein was determined according to obtained data. Results. The resulting amaranth protein concentrate contained 70.4±0.6% of protein, 17.0±1.0% fat, 9.8±0.8% carbohydrates, 1.8±0.2% ash, its moisture content was 1.4±0.1%. There were no significant differences in food intake and body weight gain between animals of both groups. The calculated value of the true digestibility of chicken egg protein was 98.8±0.1% for the control group 1, of the protein module was 99.0±0.1% for the experimental group 2, the differences between the groups were not significant. Conclusion. The results of amino acid analysis and the in vivo study of the true digestibility of the protein module (composition amaranth protein/chicken egg protein) indicate the absence of limitation relative to the amino acid scale of the "ideal" protein (FAO/WHO, 2007) and high true digestibility. The biological value of the protein module, calculated according to PDCAAS, is 99.0±0.1%, which confirms the prospects for its inclusion in specialized foods.

The Effect of Semi-Synthetic and High-Fat High-Carbohydrate Diets Containing Cholesterol on Lipid and Cholesterol Metabolism in Male Wistar Rats.

The choice of an optimal biological model of exogenous hypercholesterolemia is necessary for correct assessment of the cholesterol-lowering properties of bioactive substances and specialized food products and for validity of the experimental results. We studied the effects of cholesterol consumption in the composition of the standard semi-synthetic diet and high-fat high-carbohydrate diet for more than 3 months on lipid and cholesterol metabolism in male Wistar rats. Our findings suggest that cholesterol consumption in both diets led to a significant impairment of lipid and cholesterol metabolism, as well as an increase in insulin resistance in rats.

[Innovative methods for extracting biolactive сompounds from plant materials].

Modern methods for extracting bioactive сompounds (BAC) from various raw materials are focused on efficacy and environmental awareness, involve the use of mathematical and statistical optimization methods, the choice of green solvents, and the use of additive extraction technology. The aim of this review was to present and briefly discuss up-to-date information on modern technological approaches to the production of plant BAС extracts for applying in food ingredients and foods for special dietary uses. Material and methods. For the main search of sources, the PubMed bibliographic database, Scopus and Web of Science databases, and the Google Scholar search engine were used. The search depth was 15 years. Results. The article presents a brief review of modern approaches to the extraction, concentration and purification of polyphenolic compounds from various plant materials. As an additive extraction technology aimed at destroying/increasing the permeability of the plant cell wall, a wide range of physical methods has been successfully used: ultrasound, microwave radiation, homogenization, application of a pulsed electric field, high hydrostatic pressure, cryo-crushing. A brief description of each method, its advantages and disadvantages are presented. Improving food safety and compliance with environmental regulations requires the choice of a safe, environmentally friendly and yet efficient extraction process. For these purposes enzymatic extraction, environmentally friendly solvents, such as deep eutectic solvents, supercritical fluid extraction and membrane technology are successfully used. The use of mathematical and statistical methods can reduce the total number of experimental trials and reduce the cost and time of experiments. The use of these methods together makes it possible to vary deliberately the process parameters in relation to a specific plant material and the final product, which provides the opportunity to isolate BAS concentrates from plant raw materials with a high yield. Conclusion. The introduction of innovative technological approaches for obtaining extracts of BAC of plant origin determines the prospects for the production of a wide range of foods for special dietary uses that meet high safety and efficiency requirements.

Study of mRNA of WT1, BAALC, EVI1, PRAME and HMGA2 genes in whole blood samples.

Simultaneous quantitative measurement of mRNA of the WT1, BAALC, EVI1, PRAME and HMGA2 genes in whole blood samples reflects the specific pathological proliferative activity in acute leukemia and their ratio is promising as a diagnostic marker. The transcriptome profile of acute leukemia cells is usually assessed using NGS or microarray techniques after a preliminary procedure for isolation of mononuclear cells. However, the results of using the multiplex PCR reaction for the simultaneous determination of all above mRNAs in whole blood samples have not been published so far. Determination of mRNA of WT1, BAALC, EVI1, PRAME and HMGA2 genes in venous blood level samples by multiplex RT-PCR. The study included 127 blood samples from patients who diagnosis of acute leukemia was subsequently confirmed. In the comparison group, 87 samples of patients without oncohematological diagnosis were selected, including 31 samples (K1) with a normal blood formula and 56 samples (K2) with a violation of the cellular composition - anemia, leukocytosis and thrombocytopenia. RNA isolation and reverse transcription were performed using the Ribozol-D and Reverta-L kits (TsNIIE, Russia). Determination of the mRNA expression level of the WT1, BAALC, EVI1, PRAME and HMGA2 genes by multiplex real-time PCR using a homemade multiplex PCR kit. The mRNA level was characterized by high interindividual variation and did not correlate with the rate of circulating leukocytes or blood blasts. Expression of WT1 mRNA was observed in whole blood only in one patient from the control group and in 112 (88%) patients with leukemia and was combined with a decrease in the level of HMGA2 mRNA expression and BAALC mRNA values. In contrast to the control groups, patients with leukemia had higher levels of BAALC mRNA in AML and ALL, increased PRAME mRNA in AML and APL, but lower levels of HMGA2 in APL.

[Arthrospira platensis: antioxidant, hypoglycemic and hypolipidemic effects in vitro and in vivo (brief review)].

The cyanobacterium Arthrospira platensis biomass is a promising food source of biologically active substances with pharmacological activity. The aim of this research was a brief review and analysis of experimental in vitro and in vivo studies of the antioxidant, hypoglycemic and hypolipidemic properties of A. platensis biomass, phycocyanins, and their chromophore - phycocyanobilin. Material and methods. For the main search of the literature, the PubMed Internet resource was used, the key component of which is the Medline article database, covering about 75% of the world's medical publications. In addition, Scopus and Web of Science databases were used. Search depth - 20 years. Search keywords: Arthrospira platensis, phycobiliprotein, C-phycocyanin, allophycocyanin, hypoglycemic effect, hypolipidemic effect, antioxidant activity, in vitro and in vivo studies. Results. A brief description of the composition of the cyanobacterium Arthrospira platensis biomass, methods of its cultivation, phycocyanins extraction methods is presented. The results of experimental studies indicate the presence of pronounced antioxidant properties of A. platensis biomass, mainly due to phycocyanins in its composition. The hypoglycemic and hypolipidemic effects of A. platensis biomass and extracted phycocyanins intake have been established in vivo when modeling carbohydrate and/or lipid metabolism disorders. The results of in vitro and in vivo studies indicate the presence of pronounced antioxidant properties of phycocyanins. Hypoglycemic effects are shown in particular in experiments on rats with hyperlipidemia and alloxan diabetes fed a diet enriched with A. platensis biomass and on KKAy mice, treated with C-phycocyanin extract. Conclusion. The analysis of the results of in vitro and in vivo studies of the antioxidant, hypoglycemic and hypolipidemic properties of A. platensis biomass and extracts with a high content of phycocyanins, presented in a brief review, suggests that their use in the diet of people with impaired carbohydrate and lipid metabolism is promising. Accordingly, from the standpoint of evidence-based medicine, clinical studies on the use of spirulina biomass and/or its extracts with a high content of phycocyanins as part of specialized foods intended for the prevention and/or dietary correction of carbohydrate and lipid metabolism disorders should be preceded by additional experimental physical-chemical, physiological and biochemical research.

[Amaranth grain proteins: prospects for use in specialized food products].

Amaranth is a widespread genus of predominantly annual herbaceous plants belonging to the Amaranthaceae family, which is one of the most widely used pseudocereals along with quinoa and buckwheat in nutrition. The aim of the research was to review and analyze the results of the studies on the characteristics of amaranth grain proteins, the effect of various food processing methods on their quality, and the prospects for using amaranth protein hydrolysates in therapeutic nutrition. Material and methods. For the main search for the literature, the PubMed bibliographic database was used, which covers about 75% of the world's medical publications. In addition, Scopus and Web of Science databases and non-commercial search engine Google Scholar were used. The depth of the search was 15 years. Results. The paper presents a brief review of modern approaches for obtaining amaranth protein isolates and concentrates, including the use of a complex of physicochemical methods: grinding, sifting, extraction at high pH values, defatting, ultrafiltration, centrifugation, isoelectric precipitation, and drying of the protein product. A comparative characteristic of amino acid content of protein fractions of pseudocereals is presented. Basically, leucine, isoleucine, and valine are limiting amino acids for the grain protein of various varieties of amaranth. When substantiating and developing modern effective food technologies for processing amaranth grain, the studies dedicated to the evaluation of their impact on the biological value of amaranth protein deserve special attention. Methods of grain fermentation, sprouting, steaming, malting, boiling can be used to increase the bioavailability and digestibility of its ingredients. The results of in vitro and in vivo studies indicate the presence of hypotensive, hypolipidemic and antioxidant activity of the amaranth protein and its hydrolysates what determines the prospects for their use as part of foods for special dietary uses and therapeutic nutrition. An analysis of the scientific publications presented in the review indicates an increase in demand for high-quality gluten-free products and an increase in the range of mass-consumption foods, such as bakery, pasta, flour confectionery, with pseudo-cereals in their composition, including amaranth. Conclusion. The high biological value and technological properties of amaranth protein concentrates/isolates determine the prospects for their use to create a wide range of specialized foods for various purposes.

[Amaranth, quinoa and buckwheat grain products: role in human nutrition and maintenance of the intestinal microbiome].

Pseudocereals such as amaranth, quinoa and buckwheat have been used as food since ancient times and in recent years there has been an increasing focus on their ability to have positive health effects. Moreover, some of the functional effects of pseudocereals could be mediated by effects on the gut microbiota. The review aims to assess the features of the chemical composition of amaranth, quinoa and buckwheat grain that determine their potential for maintaining the optimal composition of the intestinal microbiota, as well as to analyze the results of published studies evaluating the effects of pseudocereals on the intestinal microbiota. Material and methods. Scopus, Web of Science, PubMed, RSCI databases, and food composition databases were used for collection and analysis of scientific information. Results. The research presents an overview of the chemical composition of amaranth, quinoa and buckwheat grain regarding their influence on the intestinal microbiota. Compared to traditional cereals, the grain of these pseudocereals has high content of soluble dietary fiber, which could have a prebiotic effect in the gut stimulating the growth of protective microbiota populations and increasing production of short-chain fatty acids (SСFA), which play a crucial role in maintaining gut homeostasis and health in general. Amaranth, quinoa and buckwheat grain, as well as some grain fractions such as proteins and polysaccharides, may have positive effects on the gut microbiota, and the biologically active substances metabolized by them have a positive effect on the body's metabolism. The results of in vitro (by cultivation using model media) and in vivo experiments indicate that the introduction of various grain fractions of pseudocereals into the diet contributes to an increase in the content of SCFA, in alpha microbiota diversity indices, and also prevents the development of dysbiotic disorders caused by a high-fat diet. Conclusion. Pseudocereals' grain is promising raw material for the development of products that can have a positive effect on the intestinal microbiota.

[Investigation of the physiological and biochemical effectiveness of plasmalogens and astaxanthin in microencapsulated form].

Plasmalogens and astaxanthin have a wide range of biological effects, including pronounced antioxidant properties. One of the main disadvantages of using these biologically active lipids is their low stability, which leads to a decrease in biological activity in vivo. The aim of the work was the study of the physiological and biochemical effectiveness of plasmalogens and astaxanthin in microencapsulated form. Methods. The experiment was conducted using 70 male Wistar rats during 60 days. The first 28 days of the experiment animals received modified diet with lowered content of fat-soluble vitamins A, D, and E, via excluding fat-soluble vitamin mixture and sunflower oil from the diet. On the 29th day of the experiment, the animals were divided into groups. Standard fat-soluble vitamin mixture and sunflower oil were added into the diet of one group (K2 group), two other groups received emulsions, containing plasmalogens (0.80%), astaxanthin (0.04%) and fat-soluble vitamins in native (G3 group) or microcapsulated (G4 group) forms instead of sunflower oil (5.0% of the diet). During the next 32 days of the experiment, the animals' grip strength was measured; anxiety and motor activity were assessed in the elevated plus maze and open field tests; cognitive functions were assessed in the passive avoidance and Morris water maze tests. In the blood serum, the level of corticosterone, triglycerides, cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), malone dialdehyde, hydroperoxides, and total antioxidant activity were determined. Results. A significant increase in the grip strength in animals treated with an emulsion with encapsulated plasmalogens and astaxanthin indicates animal endurance growth. In the Morris water maze test, animals of the same group showed the best learning ability, which indicates an improvement in cognitive functions. A significant more than 3-fold decrease in blood corticosterone level in the animals treated with plasmalogens and astaxanthin, regardless of the form of administration, in comparison with the indicator of animals in the control groups, indicates an adaptogenic effect and requires further study. The consumption of the emulsions led to a significant improvement in lipid metabolism: a significant decrease in serum cholesterol by 20% was shown, against the background of a significant reduction in LDL cholesterol by 25%. Conclusion. The beneficial effect of including the experimental emulsion in the diet is expressed in improving memory and cognitive functions, increasing muscle tone and the static component of endurance in male Wistar rats.

Effect of Exhaustive Training or Forced Immobilization on Physiological Condition and Main Metabolic and Stress Markers of Wistar Male Rats.

For correct and reliable experimental in vivo assessment of antistress effect of various bioactive substances, appropriate biomodels reproducing stress and organism response to stress in laboratory animals should be chosen. We chose treadmill test for simulating exhaustive physical load and forced immobilization accompanied by disorders of physiological and psychological condition. Verification of the models used indicates their wide applicability for testing certain biological manifestations under reproduced stress exposure.

[Physiological and biochemical studies as a necessary component of the algorithm for assessing the effectiveness of food minor biologically active substances].

Preclinical physiological and biochemical studies of the effectiveness of the tested biologically active substances in the conditions of simulated pathology in laboratory animals are an important stage preceding the assessment of the clinical effectiveness of the created specialized foods for therapeutic or preventive nutrition. The aim of this work is a brief review of the developed algorithm for preclinical assessment of the effectiveness of specialized foods, which includes a set of sequential stages of testing the safety, stability, bioavailability and effectiveness of biologically active substances in experiments on laboratory animals. Results. The paper presents a brief review of methodological approaches for in vivo preclinical assessment of the effectiveness of minor plant biologically active substances - polyphenols from bilberry fruits and leaves and phytoecdysteroids extracted from spinach. A beneficial effect of bilberry leaves' polyphenols on carbohydrate and lipid metabolism of male Zucker and Wistar rats was shown, which determined the necessity to increase the effectiveness of the developed polyphenolic ingredient by sorption on a food carrier. When evaluating the in vivo effects of polyphenols from blueberries and leaves sorbed on buckwheat flour, a pronounced hypoglycemic effect was found. The experiments aimed to the assessment of the adaptogenic effects of phytoecdysteroids from spinach leaves showed a significant decrease in the content of the main biomarkers of the stress system, which indicates a «smoothing¼ of the response of the animal organism to a strong stress impact. Conclusion. The inclusion of minor biologically active substances into the composition of foods for preventive and therapeutic nutrition is the necessary condition to improve their effectiveness. The scientific and practical significance of the preclinical evaluation to a decisive extent depends on the presence of a correctly selected biomodel. Thus, the adequacy of the assessment of the effects of minor biologically active substances as functional food ingredients in the composition of the specialized food products depends on the reproducibility of pathological processes (clinical, biochemical and morphological disorders) characteristic for the studied disease on the selected in vivo model.

[Spinach and quinoa - prospective food sources of biologically active substances].

Formally, phytoadaptogens extracted mainly from medicinal plants are not food substances, and their use in preventive nutrition is significantly limited. However, screening studies can detect phytoadaptogens in food plants, however in much smaller quantities. For example, phytoecdysteroids are biologically active substances (BAS), used to relieve chronic fatigue syndrome, reduce nervous and muscle fatigue, improve memory and attention processes. Phytoecdysteroids can be isolated not only from various medicinal plants, but also from food plants like spinach (Spinacia oleracea) and quinoa (Chenopodium quinoa). Researchers show interest in possible synergies in the manifestation of a wide range of pharmacological effects, determined by a combination of phytoecdysteroids and other biologically active substances of these food plants. The aim of this research is a brief analytical review of publications characterizing the spectrum of minor biologically active substances of spinach and quinoa, as well as those devoted to the experimental assessment of their pharmacological activity in experiments on laboratory animals. Results. An analysis of the publications presented in the review indicates that spinach and its extracts are promising food sources of phytoadaptogens, carotenoids, phenolic compounds, in particular quercetin, kempferol, phenolic acids (p-coumaric, ferulic), as well as lignans metabolized into biologically active phytoestrogens (enterolactone and enterodiol). The possible prophylactic effect of spinach is being actively studied in vivo in laboratory animals when modeling diseases such as diabetes mellitus, cardiovascular and cancer diseases. Numerous adaptogenic effects and the neuroprotective effect of spinach and its extracts are shown, expressed in improving the memory and learning of animals. Publications of the last decade indicate an increased interest in such a promising food source of high-grade protein and a wide spectrum of biologically active substances, including phytoecdysteroids, as quinoa grains. More than 20 phenolic compounds in free or conjugated forms were found in quinoa grains; tannins, saponins, sterols, phytic acid, and ecdysteroids are present in small quantities. The consumptionof quinoa can have a beneficial effect on the organism, normalizing carbohydrate and lipid metabolism, reducing body weight and preventing excessive lipid peroxidation. The anti-inflammatory effects and antioxidant effects of quinoa are widely discussed. Conclusion. Spinach, quinoa and their extracts are promising food sources of biologically active substances, their pharmacological action is confirmed from the standpoint of evidence-based medicine by in vivo experimental studies. Accordingly, it is advisable to expand the use of these food plants for inclusion in biologically active dietary supplements, functional food ingredients and specialized food products for various purposes.

Effect of Vanadium Complex with Enzymatic Hydrolysate of Soy Protein on Carbohydrate and Lipid Metabolism Disorders in Male Wistar Rats.

The effects of vanadium complex with enzymatic hydrolysate of soy protein (V-EHSPI) were studied in male Wistar rats with induced disorders of carbohydrate and lipid metabolism. The content of vanadium (IV) in the studied complex was 15.8 mg/g dry product. High-lipid high-carbohydrate diet was used to induce disorders of lipid and carbohydrate metabolism. Addition of vanadium in three different doses to the diet over 100-day experiment reduced body weight gain and the levels of glucose, insulin, leptin, and triglycerides. V-EHSPI produced beneficial effects on carbohydrate and lipid metabolism even in a dose 5 µg/kg body weight/day (calculated from the mean food consumption over the entire treatment period). Significant inhibition of growth and changes in the weight of organs in animals treated with V-EHSPI attested to toxicity of vanadium in the studied dose range.

[Enzymatic hydrolysates of whey protein and chicken egg ppotein: production, physical-chemical and immunochemical characteristics].

Reducing the manifestations of food allergy by the inclusion of specialized foods in the nutrition of children and adults suffering from this disease is an important problem. The aim was to obtain and characterize in vitro food protein hydrolysates to evaluate their use in specialized foods with reduced potential allergenicity. MATERIAL AND METHODS: Whey protein concentrate (WPC) and chicken egg protein (CEP) and enzymes such as pancreatin and alkalase have been used. Proteolysis of proteins was carried out in an FA-10 fermenter for 3 hours at an enzyme : substrate ratio of 1:50 in dry matter, at optimal pH and temperature for pancreatin and alkalase. Enzymes were inactivated at +75 °C and fermentolizate was ultrafiltered. The solutions were concentrated by reverse osmosis and freeze-dried. The molecular weight distribution of the peptide fractions was evaluated by HPLC. Residual antigenicity was determined by the method of indirect enzyme-linked immunosorbent assay and expressed as the fold of antigenicity reduction relative to the original protein. RESULTS AND DISCUSSION: During WPC proteolysis with pancreatin the hydrolyzate was obtained with a fold reduction of antigenicity of 2.3×103 relative to the initial WPC. A decrease in antigenicity of 4.7×104 times was achieved with proteolysis of WPC by alkalase. The combination of WPC fermentolysis with pancreatin or alkalase followed by ultrafiltration reduced the content of high molecular weight peptides with a mass more than 8.7 kDa. The multiplicity of decrease in antigenicity with respect to the starting protein was 1.64×105 and 1.90×105, respectively. After repeated ultrafiltration the reduction in antigenicity of the obtained WPC alkalase or pancreatin hydrolysate was more than 1.0×106 and more than 5.0×105, respectively. The decrease in antigenicity of the CEP hydrolyzate obtained with proteolysis by alkalase and ultrafiltration compared to the initial CEP was 1.0×105 times, and 5.0×105 times when we used repeated ultrafiltration. CONCLUSION: A significant decrease in the content of high molecular weight peptides and a decrease in the antigenicity of peptide mixtures based on WPC and CEP to the values that permit their use in hypoallergenic products is achieved by combining proteolysis and double ultrafiltration through a UF10 membrane.

[Oxylipins - biologically active substances of food].

Oxylipins are biologically active molecules that are formed in all aerobic organisms enzymatically or as a result of the action of free radicals and reactive oxygen species. The value of oxylipins for plants is comparable to the value of eicosanoids for animals and humans. In the human organism, the oxylipins' formation occurs through enzymatic or non-enzymatic oxygenation of various ω-6 and ω-3 polyunsaturated fatty acids (PUFAs) obtained from food. Being "local hormones", oxylipins are involved in the regulation of inflammation, pain response, cell adhesion, migration and proliferation, apoptosis, angiogenesis, regulation of blood pressure, blood coagulation, and blood vessel permeability. There is a hypothesis that the molecular structure of oxylipins allows them to be positioned as adaptogens and justifies the use of plants as potential sources of oxylipins in traditional medicine. The aim of this research is a brief analytical review of publications characterizing the adaptogenic potential and promising sources of oxylipins (plant, cyanobacteria, and algae). Results. The publications of the last decade indicate an increased interest in the oxylipins of plants, cyanobacteria, and algae. In total, about 150 oxylipins and their derivatives are known in plants and fungi. Of the plant sources of oxylipins, Peruvian poppy root (Lepidium meyenii), white bryony (Bryonia alba L.), blackcurrant seed oil (Ribes nigrum), and licorice (Glycyrrhiza glabra) are of particular interest. Some macroalgae are capable of non-enzymatically or enzymatically synthesizing a variety of oxylipins, including antiinflammatory prostaglandins, resolvins, an d leukotrienes. In addition, to common oxidized derivatives of fatty acids, macroalgae also contain a number of complex and unique oxylipins. Other sour ces of oxylipin producers include macroscopic gelatin colonies of freshwater cyanobacteria Aphanothece sacrum. As the analysis of the presented in the review publications showed, most anti-inflammatory and pro-resolvent oxylipins have antiproliferative properties, have adaptogenic potential, and can protect the body at the system level, contribu ting to the formation of favorable bacterial clearance. Conclusion. The results of numerous studies indicate that plants, algae, and even bacteria can be a promising source of oxylipins, both for their use in their native form and for the targeted isolation of oxylipins from them in order to conduct further studies of their adaptogenic potential, cardio- and geroprotective properties. In the future , establishing the adequate daily intake of these substances and the development on their basis of dietary preventive and specialized products for various purposes will be relevant.

[The impact of bilberry fruits polyphenols, sorbed on buckwheat flour, on the induced disorders of carbohydrate metabolism of male C57BL/6c mice].

The hypoglycemic properties of polyphenolic compounds of plant origin are confirmed by the results of numerous preclinical and clinical studies. However, the biological effects of these compounds are limited by their low bioavailability. This makes it urgent to develop methods for its increasing due to new methods of entering polyphenols into the organism, for example, by extracting them from natural sources in the form of extracts and concentrating extracts on food polymer matrices for subsequent use as a functional food ingredient (FFI). The aim of this study was to evaluate in vivo the possible effect of consumption of the obtained FFI in the form of a food matrix - buckwheat flour enriched with bilberry polyphenols - on carbohydrate metabolism disorders induced by a high-fat diet with a high content of easily digestible carbohydrates (sucrose) and anxiety level of male C57Bl/6c mice. Material and methods. The food matrix was obtained by sorption of the bilberry fruits polyphenol extract on buckwheat flour. The total polyphenol content in the composition of food matrix was 8.9±0.7 mg-eq gallic acid/g flour. Total anthocyanin content in the composition of food matrix was 4.6±0.1 mg/g flour. The experiment was conducted for 150 days using 48 male C57Bl/6c mice (weaners). The animals were divided into 3 groups: the control group K1 (n=16, the mice received a standard semi-synthetic diet (22.5% protein, 10% fat, 58% carbohydrates as starch, 362 kcal/100 g), the control group K2 (n=14) and the experimental group G3 (n=18). Disorders of carbohydrate and lipid metabolism in animals of groups K2 and G3 were modeled by feeding an iso-nitrogenous high-fat diet with a high content of easily digestible carbohydrates (HFHC-diet: 22.5% protein, 30% fat, 18% carbohydrates in the form of starch, 20% sucrose, 493 kcal/100 g). FFI, a food matrix in the amount of 6.6 g/100 g of feed, was introduced into the diet of animals of the experimental group G3, which corresponded to the amount of polyphenols equal to 58.7 mg-eq gallic acid/100 g of feed and the content of anthocyanins 30.4 mg/100 g of feed. Once every three weeks, the level of glucose in the blood of animals was monitored. On days 60 and 114 of the experiment, animals were tested on an elevated plus maze. Animals were decapitated under light ether anesthesia at the end of experiment. The content of glycated hemoglobin was determined in the blood. Results and discussion. Animals of both groups treated with HFHC-diet consumed significantly less feed compared with animals of the control group K1 (2.91±0.05 g/day per mouse). Moreover, animals of the experimental group G3 consumed significantly more food (2.51±0.04 g/day per mouse) compared with animals of the control group K2 (2.36±0.04 g/day per mouse). In contrast, the energy consumption of animals of both groups fed HFHC-diet was significantly higher compared to the K1 group (10.5±0.2 kcal/ day per mouse). Energy consumption by animals of group G3 (12.3±0.2 kcal/day per mouse) was significantly higher compared to animals of the control group K2 (11.5±0.2 kcal/day per mouse). The data obtained indicate that the consumption of FFI in the form of polyphenols adsorbed on the food matrix can contribute to increased appetite in animals treated with the high-fat diet. The results of the Elevated Plus Maze test indicated the absence of the effect of polyphenols in the composition of the food matrix on the anxiety level of animals. Starting from day 42 until the end of the experiment, the glucose level in animals of group G3 was significantly lower than the corresponding indicator for animals of the control group K2. Conclusion. In accordance with the results obtained, further studies of the safety and clinical efficacy of including the developed FFI in the form of a food matrix with polyphenols into the composition of specialized foods for the prevention of carbohydrate metabolism disorders are advisable.

[The complex of bilberry polyphenols, sorbed on the buckwheat flour as a functional food ingredient].

Blueberries, which contain a wide range of polyphenolic compounds, are used in traditional medicine for prevention and treatment of carbohydrate metabolism disorders. However, the high content of mono- and disaccharides in the juice significantly reduces the possibility of using in preventive nutrition by persons with impaired carbohydrate metabolism. In this regard, it is necessary to search for technological approaches aimed at obtaining functional food ingredients with a higher content of polyphenols and a low content of mono- and disaccharides, for subsequent inclusion in corresponding therapeutic foods. The aim of this study was to develop a technological approach to obtain a food matrix by enriching buckwheat flour with polyphenolic compounds extracted from blueberries. Material and methods. The work presents a method for producing a food matrix by sorption of polyphenolic compounds from an aqueous-alcoholic extract of blueberries on buckwheat flour. The concentration of total polyphenols and anthocyanins in the extract and their content in the food matrix was determined by spectrophotometric methods. The content of mono- and disaccharides and profile of individual anthocyanins was determined by HPLC. Results and discussion. The sorption on buckwheat flour was 45% of the total polyphenol content and 48% of anthocyanin content in the initial blueberry fruit extract. The profile of anthocyanins sorbed on a food matrix did not differ significantly from the profile of blueberry extract. The results of determining the content of mono- and disaccharides indicate the absence of their sorption from the extract of blueberries on buckwheat flour. Conclusion. The results obtained are of significant interest, indicating that the sorption approach allows targeted concentration of polyphenols in the food matrix with a minimum amount of easily digestible carbohydrates, which significantly increases the efficiency of its use as a functional food ingredient in dietetic products for people with carbohydrate metabolism disorders.

[A comparative evaluation of the effect of soy protein and its enzymatic hydrolysate on lipid metabolism in male Wistar rats with induced obesity].

Among various food proteins, soybean proteins have the greatest traditions of application for the dietary correction and prevention of lipid metabolism disorders and related complications. Aim. In an in vivo experiment using male Wistar rats, the lipid-lowering properties of soy protein and its enzymatic hydrolysate were tested to evaluate their possible use as ingredients of specialized foods. Material and methods. Animals were randomly divided into 3 groups: control group G1 and 2 experimental groups G2 and G3. The total duration of the experiment was 70 days. The animals of the control group G1 were fed with high-lipid semi-synthetic diet. Animals of the experimental groups G2 and G3 received the same high-fat semi-synthetic diet, but with a 50% replacement of casein with soy protein isolate (SPI) and enzymatic hydrolyzate of SPI (EHSPI), respectively. The blood glucose was measured once per 2 weeks. At the end of the experiment on the 71st day the level of glycated hemoglobin was determined in the blood; the levels of triglycerides, cholesterol, high density lipoproteins (HDL), low density lipoproteins (LDL) and the concentration of malon dialdehyde were determined in the serum. Results and discussion. Starting from the 6th week of the experiment and prior to its completion, the average food intake of animals from the G3 group was significantly (р<0.05) lower compared to animals of the G1 control group. The food intake of animals of group G2 was significantly (р<0.05) reduced compared with this indicator for animals of group G1, starting from the week 8 of the experiment and prior to its completion. The monitoring of the body weight gain did not reveal significant differences between all groups of animals, despite differences in the food intake. Replacing casein in the diet by 50% with SPI had a pronounced antioxidant and cholesterol-lowering effect. The total cholesterol content (1.65±0.05 mmol/l) decreased significantly (р<0.05) due to a decrease in LDL (0.90±0.03 mmol/l), and malon dialdehyde level lowered (3.7±0.5 µmol/l, р<0.05) in the serum of group G2 rats compared with animals of the control group G1 (2.01±0.13 and 1.12±0.09 mmol/l; 5.1±0.4 µmol/l, respectively). Replacing casein by 50% with EHSPI in the diet of G3 rats was unfavorable, significantly (р<0.05) increasing the level of total cholesterol (2.76±0.16 mmol/l) and cholesterol in LDL (1.66±0.12 mmol/l) in blood of these animals compared with animals of both comparison groups G1 and G2. Conclusion. A preclinical comparative study of the cholesterol-lowering and antioxidant properties of the SPI substantiates the prospect of its following clinical trials with the aim of including into the composition of specialized foods for prevention and diet therapy of the disorders of endogenous cholesterol homeostasis.

[The impact of bilberry leaves' polyphenols on the anxiety level, spatial learning and memory of db/db mice].

Numerous experimental and clinical studies have shown high efficiency of plant polyphenolic compounds in restoring age-related memory and learning disorders. In the present study a functional food ingredient (FFI) was obtained by sorption of an aqueous solution of bilberry leaves extract on buckwheat flour, which allowed to concentrate polyphenols and increase their storage stability. The aim of this study was to evaluate the impact of a developed FFI, enriched with bilberry leaves` polyphenols, on the anxiety level, locomotor activity, memory and spatial learning of db/db mice with genetical type 2 diabetes. Material and methods. The experiment was conducted using 10 heterozygote male db/db mice and 10 homozygote male db/+ mice as the comparison control group (7 weeks of age). According to body weight, blood glucose level, the results of insulin resistance test and elevated plus-maze (EPM) test, animals were randomized into three groups: control group C1 - db/+ animals, control group C2 and experimental group G3 - obese db/db mice. Buckwheat flour was included into the diet of C2 group in a dose 22.5 g/100 g; FFI was included into the diet of G3 group in a dose 2.5 g/100 g (that was equal to 59.2± 1.4 mg-eq gallic acid per 100 g of the diet). The anxiety level and general locomotor activity were evaluated in the EPM test. The evaluation of behavior, memory and spatial learning was performed using passive avoidance test (PAT). Glycated hemoglobin level was determined in blood, insulin and leptin levels were determined in blood plasma, general antioxidant activity was determined in liver cytosolic fraction. Results and discussion. The obtained data on biochemical parameters and insulin resistance tests showed the absence of normalizing effects of developed FFI. However, the inclusion of polyphenol-containing FFI into the diet led to beneficial changes in physiological parameters. Animals of G3 group, provided with FFI, were significantly less anxious compared to both control groups. During PAT testing of short-term memory, no animals in G3 group entered to the dark compartment (0%), what demonstrated increased learning ability and well-established memory of these animals in comparison with C1 (50%) and C2 groups (80%). Conclusion. The results prove the effectiveness of bilberry leaves` polyphenols, sorbed on the brown buckwheat flour, in the correction of central nervous system disorders in db/ db mice with genetically altered type 2 diabetes, what points at possible prospect of FFI inclusion in therapeutic products for patients with type 2 diabetes mellitus.