Clinical metabolomics: current state and prospects in Russia.

Using analytical technologies it is possible now to measure the entire diversity of molecules even in a small amount of biological samples. Metabolomic technologies simultaneously analyze thousands of low-molecular substances in a single drop of blood. Such analytical performance opens new possibilities for clinical laboratory diagnostics, still relying on the measurement of only a limited number of clinically significant substances. However, there are objective difficulties hampering introduction of metabolomics into clinical practice. The Institute of Biomedical Chemistry (IBMC), consolidating the efforts of leading scientific and medical organizations, has achieved success in this area by developing a clinical blood metabogram (CBM). CBM opens opportunities to obtain overview on the state of the body with the detailed individual metabolic characteristics of the patient. A number of scientific studies have shown that the CBM is an effective tool for monitoring the state of the body, and based on the CBM patterns (signatures), it is possible to diagnose and monitor the treatment of many diseases. Today, the CBM creation determines the current state and prospects of clinical metabolomics in Russia. This article, dedicated to the 80th anniversary of IBMC, is a review of these achievements focused on a discussion of their implementation in clinical practice.

[Key challenges in the dietary intake structure and cutting edge technologies for optimizing nutrition to protect the health of the Russian рopulation].

This article presents an analysis of some of the results of the work of the Federal Research Center for Nutrition and Biotechnology (Center) in recent years, highlighting the most important, promising areas of Nutrition Science and Food Hygiene that need further development. The priority area of Center functioning is scientific support for the implementation of the Doctrine of Food Security of the Russian Federation (Decree of the President of the Russian Federation dated January 21, 2020 No. 20), Decree of the President of the Russian Federation dated July 21, 2020 No. 474 «On the national development goals of the Russian Federation for the period until 2030 «in terms of ensuring an increase in life expectancy and improving the life quality of the population, the Strategy for Improving the Quality of Food Products in the Russian Federation until 2030 (Order of the Government of the Russian Federation dated June 29, 2016 No. 1364-r). The Center coordinates all research on medical nutrition problems in the Russian Federation within the framework of the work of the Problem Commission on Nutrition Hygiene of the Scientific Council of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare, the Scientific Council of the Russian Academy of Sciences on Medical Nutrition Problems, the Scientific and Technical Committee of the Comprehensive Scientific Program «Priority Research in the Field of Nutrition of the Population¼, Profile Commission on Dietetics of the Expert Council in the Field of Health of the Ministry of Healthcare of Russian Federation, ensuring the implementation of their results with the participation of members of the Consortium "Healthcare, Nutrition, Demography". The most important area of the Center's work is scientific and expert support in the field of international and national technical regulation of the production and turnover of foods and raw materials, in particular, the work of the Russian national contact point of the Codex Alimentarius Commission (established by FAO and WHO), as well as the work of the Russian side in the Eurasian Economic Commission regarding the preparation of proposals for technical regulations of the Customs Union in the field of food safety, evaluation of draft technical regulations and amendments and additions to them.

[Alternaria toxins in tomato products marketed in the Russian Federation].

Tomatoes and tomato products are widely produced and consumed throughout the world. Alternaria spp. are the main cause of alternariosis (black mold disease) on fresh tomatoes, both in the field and after harvesting. Alternaria toxins are widespread contaminants of tomato products. The aim of the present study was to evaluate the contamination of tomato processing products from the domestic market with Alternaria toxins, as well as to assess their intake by humans through the consumption of tomato juices. Material and methods. The content of Alternaria toxins (alternatiol, alternariol monomethyl ether, altenuene, tentoxin, tenuazonic acid) was determined in 64 samples of tomato products (paste, ketchup, juice) by high-performance liquid chromatography coupled to tandem mass-spectrometric detection (HPLC-MS/MS). Results. The priority Alternaria toxins for tomato paste, ketchup and juice were tenuazonic acid (61% of 64 samples, in amounts from 20.0 to 1065.5 µg/kg), altenuene (52%, 8.9-200.1 µg/kg) and alternariol (27%, 12.2-561.6 µg/kg). Samples of tomato paste turned out to be the most contaminated with Alternaria toxins while tomato juice samples were the least contaminated. At the same time, several toxins were found in 91% of tomato paste samples, 35% of ketchups, and 23% of tomato juices. Conclusion. To the best of our knowledge, the present study is the first survey devoted to Alternaria toxins contamination of tomato paste, ketchup and tomato juice sold on the Russian market. The high frequency of their contamination with tenuazonic acid, altenuene and, to a lesser extent, alternariol has been established, which indicates a potential risk to human health when tomato processing products are consumed. This indicates the need for a hygienic assessment of contamination the above products with tenuazonic acid, altenuene and alternariol. When calculating the potential intake of Alternaria toxins for different age population groups, it was shown that high levels of alternariol (up to 56.77 ng/kg body weight per day) could be obtained under daily consumption of tomato juice by adults and children under three years of age, as well as tenuazonic acid when consuming tomato juice contaminated at the 95th percentile level as part of the diet in organized groups for orphans and children without parental care.

[International and Russian mechanisms for integrating innovations and experience to optimize the nutrition of the population].

The vigorous scientific and organizational activities of the Federal Research Center for Nutrition and Biotechnology, integration into the international scientific and technical space, the use of advanced innovative technologies, the best examples of world experience, cooperation with leading economic countries are the most important lever for optimizing the nutrition of the population of our country, which is essential for the preservation of the health of the nation and the implementation of the tasks of demographic policy in the Russian Federation.

[Mycotoxins in spices consumed in Russia].

Spices and herbs have been used since ancient times as flavor and aroma enhancers, colorants, preservatives and traditional medicines. As many other plant products, they can be exposed to contaminants, ones of which are mycotoxins, secondary metabolites of fungi. Such contamination can occur during harvesting, processing and storage, distribution, retailing and consumer use. Although they are used and consumed in small quantities, but added to a wide variety of products, especially ready-to-eat products. So the assessment of their contamination with mycotoxins is very important. The aim of the study was to investigate the contamination of spices and herbs with mycotoxins of fungi of the genera Aspergillus, Penicillium, Fusarium and Alternaria, as well as to assess the mycotoxins intake per person when consuming these food groups. Material and methods. Concentration of mycotoxins in 155 samples of spices and herbs was determined by ultra high-performance liquid chromatography coupled to tandem mass-spectrometric detection (UHPLC-MS/MS). The list of mycotoxins included deoxynivalenol, aflatoxins, ochratoxin A, zearalenone, T-2 toxin, fumonisins, sterigmatocistin, HT-2 toxin, diacetoxyscirpenol, enniatins, beauvericin, neosolaniol, citreoviridin, mycophenolic acid, citrinin, tentoxin, altenuene, alternariol and its monomethyl ether. Results. Among the regulated in plant products mycotoxins in the studied samples there were found aflatoxins (B1 - in 19% of samples, from 0.4 to 48.2 µg/kg, B2 - 8%, from < limit of quantitation (LOQ) to 3.2 µg/kg, G1 - 2%, 0.75-21 µg/kg, G2 - 5%, 0.5- 12.5 µg/kg), ochratoxin A (15% samples, 0.8-14 µg/kg), fumonisin B1 (8%, 16.1-722.6 µg/kg), and fumonisin B2 (14%, < LOQ - 79.6 µg/kg). T-2 toxin and deoxynivalenol were found in 10% of samples (< LOQ - 6.5 µg/kg and < LOQ - 65.5 µg/kg respectively), zearalenone - in 4 samples (1.7-106.2 µg/kg), HT-2 toxin - in 8 samples (5.4-19.8 µg/kg). Among little-studied (emergent) mycotoxins in the spices and herbs samples there were found tentoxin (in 36% of samples, in an amount from 0.7 to 10.9 µg/kg), altenuene (in 8%, 14.5-161.5 µg/kg). 10% of the samples were contaminated with alternariol and its methyl ether (from less than LOQ to 12.8 and < LOQ to 55.7 µg/kg, respectively), 4% - with sterigmatocystin (0.4-7.8 µg/kg), 5% - mycophenolic acid (13.1-297 µg/kg), 2% of the samples were contaminated with citrinin and enniatin B (< LOQ - 27.7 and 0.1-1 µg/kg), in 9 samples (6%) beauvericin was detected (< LOQ - 1.7 µg/kg). Over 60% of samples were contaminated with more than one mycotoxin. The content of aflatoxin B1 exceeded the maximum permissible level set in the EU (5 µg/kg) in nine samples. Conclusion. To the best of our knowledge, the present study is the first in the Russian Federation to report results indicating to the contamination of spices and herbs with mycotoxins. High occurrence of aflatoxins, tentoxin, ochratoxin A and fumonisin B2 has been observed. In calculating the potential exposure of mycotoxins, the possibility of high levels of aflatoxin B1 intake have been shown to be possible, which could lead to a public health risk when consuming contaminated spices, herbs and foods containing them.

[Bioactive compounds anthocyanins as a factor in the nutritional recovery of the body's adaptive potential after intense physical activity in the experiment: assessment of immunological and hematological indicators of adaptation].

Restoring the adaptive potential of an athlete is of paramount importance not only for the implementation of his training and competitive activities, but also for maintaining health. One of the leading place in complex recovery programs in sports is given to full-fledged optimal nutrition, which provides for meeting the body's requirements not only in energy, macro- and micronutrients, but also in minor bioactive compounds. The use of anthocyanin-containing products is a promising strategy for the normalization of metabolic and immune disorders that develop as a result of intense physical and neuro-emotional stress not only in athletes, but also in other groups of people exposed to these factors, including military personnel undergoing training in conditions close to combat. This determines the relevance of this study. The aim of the research was to study the effect of an anthocyanin-enriched diet on hematological profile and cellular immunity in rats after intense physical activity. Material and methods. The experiment was carried out for 4 weeks on 4 groups of male Wistar rats with an initial body weight of ~300 g. The motor activity of the animals of the 1st (control) and 2nd groups was limited by the standard keeping animals in the vivarium, while physically active rats of the 3rd and 4th groups received additional physical activity - training on a treadmill. Before the end of the experiment, the animals of 3rd and 4th groups were given debilitating physical activity on a treadmill (until the rats refused to continue the exercise). Rats of all 4 groups received a standard semi-synthetic diet, water ad libitum. Animals in 2nd and 4th groups were additionally fed blueberry and blackcurrant extract (30% anthocyanins) as part of the diet at a daily dose of 15 mg anthocyanins/kg body weight. Hematological parameters were determined on a Coulter ACT TM 5 diff OV hematological analyzer. Expression of CD45R, CD3, CD4, CD8a, CD161 receptors on rat peripheral blood lymphocytes was determined by direct immunofluorescent staining of whole blood cells using a panel of monoclonal antibodies conjugated with fluorescent dyes: APC, FITC, PE. The measurements were carried out on an FC-500 flow cytometer. Results. Intense physical activity in rats of the 3rd group did not lead to a significant change in erythrocyte parameters compared with the control group. Enrichment of the diet with blueberry and black currant extract (the 2nd and the 4th groups) provided a significant (p<0.05) increase in blood content of hemoglobin (Hb) (150.7±0.9 and 154.4±2.0 vs 145.4±0.9 g/l in control), hematocrit (44.95±0.21 and 46.18±0.64 vs 43.78±0.32%) and the average content of Hb in erythrocytes (18.00±0.20 and 18.03±0.24 vs 17.35±0.24 pg). The absolute content of leukocytes and other cellular elements of the leukocyte formula, as well as leukocyte indices in rats of the experimental groups didn't significantly differ from those of the control rats, which confirms the absence of an inflammatory process. Intense physical activity and anthocyanin enrichment of the diet didn't have a significant effect on rat platelet parameters. Enrichment of the diet of rats of the 4th group with blueberry and black currant extract led to the activation of cellular immunity, as evidenced by a significant (p<0.01) increase in the percentage (from the total content of T-lymphocytes) of T-helpers (70.13 ±1.34 vs 63.75±0.99%) and a decrease in the relative content of cytotoxic T-lymphocytes (28.65±1.38 vs 34.71±0.95%) in comparison with those in rats of the 3rd group and at the level of the trend (р<0.1) - from the 1st group indexes (66.87±1.20 and 31.87±1.26%, accordingly). Intense physical activity led to a decrease in immunoregulatory index in rats of the 3rd group (1.86±0.07) compared with the control (2.13±0.12) (p<0.1), and in animals of the 4th group this indicator was significantly higher (2.50±0.14, p<0.05). In animals of the 3rd group a statistically significant (p<0.05) decrease in the relative content of NK cells in peripheral blood was found compared to the control. Enrichment of the diet of physically active rats with blueberry and black currant extract led to a significant (p<0.05) increase in the percentage of NK cells compared to this indicator in rats of the 3rd group (4.87±0.75 vs 2.08±0.18%) and had no significant difference with the indicator in rats of the control group (4.32±0.98%). Conclusion. The enrichment of the rats' diet with blueberry and blackcurrant extract containing a daily dose of 15 mg of anthocyanins per kg of body weight provides an increase in blood Hb content, hematocrit and the average content Hb in erythrocytes. It has been established that intense physical activity induces the cellular immunity suppression. The activating effect of anthocyanins on adaptive cellular immunity and NK cells, which are lymphocytes of innate immunity, was revealed. The data obtained indicate the effectiveness of the use of bioactive compounds (anthocyanins) to increase the adaptive potential of the organism.

Systemic Biomarkers and Liver Morphology in Rats during Chronic Low-Dose Toxicant Administration against the Background of Vitamin Deficiency.

Liver morphology, intensity of apoptosis, and activity of xenobiotic metabolism enzymes were studied in a chronic model experiment in rats receiving a mixture of 6 pesticides against the background of life-long diets with adequate and insufficient supply of water-soluble vitamins. The dose of each pesticide in the mixture did not exceed the acceptable daily intake (1 ADI). It was found that chronic exposure to low doses of anthropogenic toxicants in combination with permanent vitamin deficiency provokes a number of liver changes, such as increased apoptosis activity, cytochrome P450 system depletion, steatosis, and inflammatory infiltration, which is a potential health risk factor.

[Effect of microand nanoplastics on the gastrointestinal mucosa and intestinal microbiome].

Worldwide production and use of the polymers has led to intensive environmental pollution with micro- and nanoplastics (MP and NP). Accumulating in ecosystems, MP are transmitted through food chains and enter the human body. The associated health risks are of grave concern and require assessment. The main entry gate for MP/NP supplied with food is the gastrointestinal tract (GIT). Despite the well-established concept of MP/NP toxicity, information about their actual effects on the GIT is contradictory. The aim of the research was to establish the nature and mechanisms of NPs and MPs action on the gastrointestinal mucosa and intestinal microbiome, basing on the literature data. Material and methods. The review was compiled after selecting of 90 documents from major databases including Web of Science, PubMed, Scopus, Elsevier, Springer and Google Scholar (up to March 2023). Results. In animal studies and in vitro models, it was shown that MP/NP affect mucus secretion, its rheological characteristics, and can cause an increase in the permeability of tight junctions of epithelial cells by reducing the expression of zonula occludens protein 1 (ZO-1), occludin and claudin-1, which promotes the penetration of MP through the intestinal wall. Various adsorption layers (coronas) formed on the surface of MPs both abiotically and during transit through the GIT can lead to both increased and reduced toxicity of MPs. Biofilms formed on MP/NP surface create favorable conditions for the activity of pathogenic bacteria and horizontal gene exchange between the components of the biofilm and the intestinal microbiome. Animal experiments have shown an unfavorable effect of MP/NP on the intestinal microbiota and its key metabolites, contributing to the development of dysbiosis. Conclusion. Most data on the effect of MP on the GIT have been obtained using a model object - polystyrene microspheres, which are rarely found in practice. A frequent limitation of the in vitro studies is the discrepancy between used doses (concentrations) of MP and those that may occur when MP are consumed with food. Data on the potential impact of MP/NP on the GIT protective barrier and intestinal microbiota obtained under various experimental conditions are contradictory. Thus, evidence of the impact of MP/NP on the GIT and intestinal microbiota of humans needs further confirmation, which will allow us to move on to the development of a set of measures that can reduce the risks of MP exposure to human health.

[Mycotoxins contamination of cocoa products and carob marketed in the Russian Federation].

Cocoa is a daily basic food for many people all over the world. Also, people engaged in healthy lifestyle often prefer carob. The aim of the present study was to evaluate occurrence of Aspergillus, Penicillium, Fusarium and Alternaria secondary metabolites in cocoa and carob available in the Russian Federation and assess mycotoxin's intake with these products. Material and methods. Concentration of 27 mycotoxins in 63 samples of cocoa and carob products was determined by ultra high-performance liquid chromatography coupled to tandem mass-spectrometric detection (UPLC-MS/MS). The list of mycotoxins included regulated ones (aflatoxins, ochratoxin A, deoxynivalenol, fumonisins, T-2 toxin, zearalenone), their derivatives and structural analogs (A and B trichothecenes), and emergent mycotoxins (alternariol, its monomethyl ether, altenuene, tentoxin, citrinin, sterigmatocistin, cyclopiazonic and mycophenolic acids, enniatins, beauvericin). Results. 29 of 41 cocoa samples were positive for beauvericin, contamination level was from 1.6 to 2184.8 µg/kg. Tentoxin, sterigmatocystin and alternatiol monomethyl ether were detected in 3 samples; their concentration varied in the ranges 0.7-1.2, 1.5-3.3 and 4.0-7.8 µg/kg correspondingly. Carob samples were positive for alternariol (14 of 22 samples, 1.5-43.1 µg/kg); tentoxin (13 samples, 0.5-8.7 µg/kg), mycophenolic acid (6 samples, 6.9-8.2 µg/kg) and for alternatiol monomethyl ether (3 samples, 1.0-1.2 µg/kg). Several samples of cocoa were contaminated with regulated mycotoxins: zearalenone (18 of 41 samples, 2.1-24.6 µg/kg), ochratoxin A (14 samples, 0.75-12.0 µg/kg) and aflatoxin B1 (2 samples, 0.59 and 0.86 µg/ kg). Several carob samples were contaminated with fumonisin B2 (7 of 22 samples, 4.2-5.2 µg/kg), ochratoxin A (5 samples, 0.5-1.4 µg/kg) and aflatoxin B1 (3 samples, 0.15-0.18 µg/kg). The ochratoxin A content in two instant cocoa beverages exceeded its maximum level (5 µg/kg) set for some plant products. Conclusion. To the best of our knowledge, the present study is the first survey devoted to emergent mycotoxin contamination of carob and cocoa alternative products marketed in the Russian Federation. The high occurrence of such mycotoxins as aflatoxins, beauvericin and ochratoxin A in these products indicates a potential health risk and the need for a hygienic assessment of cocoa and carob products' contamination not only with regulated in cocoa products aflatoxin B1, but with other mycotoxin including emergent ones. The risk of dietary intake of aflatoxins, beauvericin and ochratoxin A with cocoa products (especially with instant cocoa beverages) has been revealed for children over 7 years old in organized groups.

[The structure of blood gut microbiota markers in athletes and their relationship with the diet].

It is known that under conditions of ultra-high physical activity and a specific diet, the state of the microbiota plays a significant role in maintaining the health, metabolic and energy status of athletes. The purpose of the study was to evaluate the composition of blood microbial markers in professional football players and physically active people and their correlation with diets in order to substantiate recommendations for their optimization. Material and methods. In a cross-sectional study a group of football players (n=24, 28±3 years old, body mass index - 22.5±1.0 kg/m2) who received a diet according to the training regimen, and a comparison group of physically active individuals (n=25, 34±5 years old, body mass index - 21.8±2.8 kg/m2) have been examined. The method of gas chromatography-mass spectrometry was used to analyze microbial markers of microbiome, mycobiome, virome and blood metabolome populations. Data on actual dietary intake were collected using food diaries for 3 days, followed by data processing with the Nutrium 2.13.0 nutritional computer program. For analysis, individual daily requirements for energy and macronutrients have been calculated based on the basal metabolic rate (according to the Mifflin-San Geor formula, taking into account anthropometric data), the coefficient of physical activity (groups IV and II, respectively). Results. The analysis of the athletes' diet, compared with individual requirements and with the recommendations of the International Society for Sports Nutrition (ISSN), revealed a lack of complex carbohydrates (5±1 instead of 6.1±0.3 g/kg body weight day), an excess of sugars (23±4 instead of <10% of kcal). These figures are significantly higher than the intake of similar nutrients in physically active people in the comparison group. In football players, compared with the comparison group, significant changes in microbial markers were found for Alcaligenes spp., Clostridium ramosum, Coryneform CDC-group XX, Staphylococcus epidermidis (p<0.001), known for their pro-inflammatory activity in the intestine, as well as for Lactobacillus spp. (p<0.001) performing a protective function. In addition, mycobiome markers were increased in athletes: Candida spp. (p<0.001), Aspergillus spp. (p<0.001), among which there are potential pathogens of mycoses. This was not observed in the comparison group. At the same time, an increase in the microbial markers of Alcaligenes spp., Coryneform CDC-group XX, Lactobacillus spp., Streptomyces spp., Candida spp. Micromycetes spp., containing campesterol in the cell wall, in football players positively correlated with a high calorie diet (p<0.001). A similar correlation of mycobiome markers (Micromycetes spp., containing sitosterol in the cell wall, ρ=0.346, p=0.015) was observed with an excess of easily digestible carbohydrates. Taking into account the data obtained, a correction of the diet have been proposed: increasing the consumption of carbohydrates to 7.3-7.5 g/kg of body weight/day by including bakery products from whole grain flour and cereals in the diet (up to 300-370 g/day), limiting simple sugars (up to 90-95 g/day). Conclusion. High physical activity leads to changes in the structure of blood microbial markers, including a shift towards an increase in potentially pathogenic fungi. Wherein, a predictive role is played by an imbalance of macronutrients in terms of quantitative and qualitative composition, an excess of simple sugars, and a lack of slowly digestible carbohydrates. To correct the diet, an additional inclusion in the diet of their main sources - products from cereals (cereals and bakery products) is proposed.