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Vol 26, No 5 (2023)

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Plant protection and biotechnology

Regulation and metabolic engineering of the general phenylpropanoid pathway in response to stress in plants

Polivanova O.B., Cherednichenko M.Y.

Abstract

The diversity of plant secondary metabolites is based on the phenylpropanoid pathway. As part of its functioning, phenylalanine is exposed to enzymes that convert this amino acid into phenolic compounds. The initial steps are catalyzed by phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumaroyl-CoA ligase (4CL), which are part of the general phenylpropanoid pathway. The products of the general phenylpropanoid pathway are associated with the growth and development of plants, responses to the external stimuli, signaling and protective functions. The study of the biochemical and molecular bases of the biosynthesis of phenylpropanoids is important, since their functioning underlies the understanding of the molecular mechanisms of plant adaptation to external factors such as drought, salinity, lack of mineral nutrition components, and pathogens interaction. This review considers the relationship of the genes of the general phenylpropanoid pathway with the impact of pathogens and abiotic factors, as well as their genetic and metabolic engineering.

PAL, C4H, and 4CH genes of many plant species usually represent genes families encoding several enzymes isoforms. Increased levels of expression correlate with increased production of phenylpropanoids, and enzymes activity varies with developmental stage, cell differentiation, and environmental exposure. Thus, PAL, C4H, and 4CH are one of the key enzymes involved in plant responses to stress. For example, PAL is involved in signaling in response to pathogens. It is also directly related to lignin biosynthesis, which strengthens cell walls and has antimicrobial activity. Genes of the general phenylpropanoid pathway often become objects of genetic and metabolic engineering. These manipulations can be aimed to increase the biosynthesis of flavonoids and other secondary metabolites, as well as to obtain genotypes that are resistant to biotic and abiotic factors.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):3-9
pages 3-9 views

Medical chemistry

Characteristics of the amino acid profile of blood plasma in psoriasis

Kupriyanov S.V., Shatrova Y.M., Sysakov D.A., Fedorova K.S., Myakotina D.E., Kholina A.N., Sinitsky A.I.

Abstract

Introduction. Psoriasis is an autoimmune disease that affects the skin, has a number of systemic manifestations and is associated with the development of a number of somatic pathologies. It is known that one of the systemic features of psoriasis is changes in the blood plasma amino acid pool, but the pathogenetic significance of this fact has not yet been determined.

The purpose of this work was to study the amino acid profile of blood plasma in psoriasis and its relationship with biochemical markers of lipid and purine metabolism.

Materials and methods. The study group included 45 patients with psoriasis vulgaris. The comparison group included 46 donors of the Chelyabinsk Regional Blood Transfusion Station. The concentrations of 20 proteinogenic amino acids, as well as ornithine and citrulline, were determined.

Results. Recorded changes demonstrating the rearrangement of relationships between the concentrations of circulating amino acids in psoriasis; correlations of amino acid levels with a number of clinical diagnostic biochemical markers in the group of patients with psoriasis were determined. A significant increase in the levels of cysteine, phenylalanine and leucine was found in psoriasis compared with the corresponding indicators of the comparison group. Cysteine concentrations positively correlated with uric acid levels. Elevated levels of cysteine and leucine have been associated with disorders of purine and lipid metabolism.

Conclusions. The data obtained indicate changes in the amino acid profile in psoriasis, some of which may be associated with the development of comorbid metabolic pathologies.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):10-17
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Vitamin B1 and its status in vegetarians and vegans

Ranjit R., Galchenko A.V.

Abstract

Vitamin B1 is a water-soluble vitamin whose alternative name is thiamine or thiamin. The metabolically active form of vitamin B1 is thiamin diphosphate, also known as thiamin pyrophosphate. Thiamin diphosphate is a coenzyme in the pyruvate dehydrogenase complex and in the Krebs cycle. Thus, it is directly involved in catabolism. In addition, vitamin B1 also takes part in the pentose phosphate shunt, which contributes to the synthesis of nucleotides and their derivatives. Similarly, thiamin diphosphate plays an important role in myelin synthesis, amino acid metabolism, and the metabolism of neurotransmitters and hormones. There is also evidence for its direct regulation of ion channel activity.

Lack of vitamin B1 affects all organs and tissues, but the earliest manifestations of its deficiency are found in the nervous system. There are several reasons for this. Firstly, the nervous tissue is extremely and continuously dependent on glucose oxidation, which can be disrupted in case of insufficiency of the pyruvate dehydrogenase complex. Secondly, myelin production is interrupted. Thirdly, there is an imbalance of neurotransmitters. The fourth reason is the high activity of transmembrane ion-exchange processes in the nervous tissue. All these lead to damage to the central and peripheral nervous system and, in severe cases, the development of beriberi or Wernicke-Korsakoff syndrome.

B1 hypervitaminosis is an unlikely event. The hydrophilic thiamin diphosphate is rapidly excreted through the urine with an increase in its serum concentrations. In addition, the intake of large amounts of thiamin with food significantly reduces its intestinal absorption. As thiamin is relatively safe even in high quantities, the high vitamin B1 dose is used to treat symptoms of chronic cluster headaches and some neurodegenerative diseases.

Thiamin is quite evenly distributed in both animal- and plant-derived products. Its highest concentrations are found in the outer layer of cereals, legumes, seeds, or nuts. As a result, most studies show that thiamin intake is higher in vegans and vegetarians than in omnivores. But, with regard to the status of vitamin B1, not everything is so crystal clear. There are not many studies done on this topic to date. The lack of general consensus on a single accepted methodology for assessing the status of vitamin B1 seems to be the main hurdle. Although most of the data indicate a better supply of vitamin B1 in vegans and vegetarians, further research and standardization of methods are still needed.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):18-24
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Pharmaceutical chemistry

The study of metrological characteristics of the obtained substances of fat-soluble vitamins by interlaboratory comparison tests

Sysuev E.B.

Abstract

Relevance. This article discusses the possibility of using substances isolated from natural raw materials of fat-soluble vitamins retinol-palmitate (vitamin A), cholecalciferol (vitamin D3), tocopherol-acetet (vitamin E) in order to create domestic standard samples of approved type (SSS). The use of SSS in analytical practice, as well as in scientific research, makes it possible to ensure the accuracy and reliability of experimental results, to ensure the traceability of the result when performing analyses within the framework of repeatability and reproducibility (precision). The paper addresses issues related to the production and standardization of domestic pharmaceutical substances, as well as issues of standardization and conformity assessment of finished products of the pharmaceutical industry and other sectors of the national economy (food industry, agriculture, perfumery and cosmetics, the industry of production of goods for children's practice, etc.).

The purpose of the work. To establish the quantitative content of fat-soluble vitamins A, D3, E, in the obtained substances claiming the status of SS with the approved type, as well as to determine their stability and uniformity by means of interlaboratory comparison tests (hereinafter referred to as ICT).

Material and methods. The quantitative content of fat-soluble vitamins in the obtained substances was assessed by HPLC in 6 accredited analytical laboratories. The results were processed by evaluating the data provided for the presence of outliers and quasi-outliers using the Kohren, Grubbs and Mandel method.

Results. The conducted studies under the comparison program showed that the results of the quantitative determination of the obtained vitamins are within the norms of the certified characteristic "Mass fraction of vitamin, %" in relation to the content of the determined characteristic in reference substances of imported production.

Conclusions. As a result of the statistical data obtained, we have proved that the substances obtained from fat-soluble vitamins A, E, D3 comply with regulatory requirements based on the results of interlaboratory comparisons and can be recommended for the manufacture of domestic SS with the approval of their type, and it can also be assumed that the manufacturing processes of domestic and imported SS are harmonized. The quality of domestic SS is at an identical level with respect to foreign SS, but each is produced exclusively in the legal field of the manufacturer's state and is legitimate in its territory of application, which once again confirms the high need to create a domestic reference pharmaceutical base.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):25-31
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Phyto-stimulating effects of paracetamol biodestruction product on urtica dioica

Vikhareva E.V., Slabinskaya E.V., Mishenina I.I., Rubzov D.F., Rychkova M.I.

Abstract

Relevance. The search for environmentally friendly methods of processing pharmaceutical waste to obtain new biologically active compounds on their basis, including phytostimulating action, is especially relevant in recent years due to global pharmaceutical pollution of water bodies in many countries. The results of previous studies have shown that the product of bacterial degradation of paracetamol exhibits pronounced phytostimulating properties in relation to medicinal plants of Plantaginaceae, Asteraceae, Linaceae, Lamiaceae and other families.

The purpose of this study is to study the effect of PBP on morphometric parameters, dry biomass and the content of biologically active substances in the leaves of stinging nettle (Urtica dioica L.), a medicinal plant of the family. nettles (Urticaceae).

Material and methods. We used bacterial degradation of paracetamol obtained on the basis of the Laboratory of Alkanotrophic Microorganisms, PFRC, Ural Branch, Russian Academy of Sciences (Perm). Stinging nettle leaves collected from plants treated with bacterial degradation of paracetamol were used to determine the biomass by gravimetric method, morphometric parameters, as well as the content of the total free organic acids, hydroxycinnamic acids and ascorbic acid by titrimetric analysis and spectrophotometry. Water and alcohol extracts from medicinal plant materials were obtained according to the method of the Global Fund XIV ed.

Results. Under the influence of bacterial degradation of paracetamol, the morphometric parameters – the length and width of the nettle leaves increase by 115 % and 66 %, respectively, the total collection of leaf biomass increases by 40 % compared to the control (water). The increase in the content of the total free organic acids under the influence of bacterial degradation of paracetamol is 18.5%, hydroxycinnamic acids – 6.8%, ascorbic acid – 12.5%.

Conclusion. The product of bacterial degradation of paracetamol has a obvious phytostimulating effect on stinging nettle.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):32-37
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Ecological and hygienic assessment of quality flower of tanacetum vulgare agro- and urbanbiocenoses of the Voronezh region

Dyakova N.A.

Abstract

An important and little-researched aspect of the effect of human economic activity on medicinal plants is that in response to an increase in anthropogenic load, additional synthesis of secondary metabolites, especially flavonoids, is induced, which play an important role in adapting plants to changing conditions. The purpose of the study was to study the accumulation of flavonoids and phenol carboxylic acids in the flowers of Tanacetum vulgare, collected in various agricultural and urbobiogeocenoses of the Voronezh region from the point of view of anthropogenic impact. As part of the study, 51 samples of Tanacetum vulgare flowers determined the content of heavy metals and arsenic and the sum of flavonoids and phenol carboxylic acids in terms of luteolin. By comparing the data on the content of heavy metals in the upper soil layers of the region and the content of these elements in the flowers of Tanacetum vulgare, it can be argued that there are significant physiological barriers that prevent the accumulation of ecotoxicants in the generative organs of the plant, which is especially noticeable for elements such as lead, mercury, arsenic, cadmium, cobalt and chromium. It turned out that ordinary pyjma is able to selectively concentrate copper and zinc. It was revealed that in the flowers of Tanacetum vulgare collected in agrobiogeocenoses and urbocenoses, the content of flavonoids and phenolcarboxylic acids is higher than in samples from protected areas, which is explained by the fact that the key enzyme for the synthesis of flavonoids – phenylalaninammiacliase – has a pronounced stress inducibility. Under conditions of excessive toxic influence of pollutants, it is also possible to suppress the antioxidant system of plants.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):38-42
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Bioelementology

Evaluation of the content of essential microelements in the herb of Cichorium intybus L. growing in the orenburg region

Mikhailova I.V., Ivanova E.V., Pupykina K.A., Bondarenko A.I., Voronkova I.P., Vinokurova N.V.

Abstract

Relevance. It is of interest to search for new plant sources of essential trace elements. It is topical to study the content in plants of transitional microelements of the IV period, which are part of the enzyme systems of the human body – Cu, Zn, Fe, Mn, Ni, Co, Cr. A promising source of microelements is the herb of Cichorium intybus L. It should be noted that the accumulation of microelements in raw materials may depend on their content in the soil, on the nature of the interaction between microelements. For this reason, it was relevant to study the content of essential microelements in chicory herb growing in various territories of the Orenburg region, and in the soil of these territories.

The purpose of the work: to evaluate the content of essential microelements in the herb of Cichorium intybus L., growing in the Tyulgansky, Ponomarevsky, Buzuluksky, Sharlyksky and Sakmarsky districts of the Orenburg region, and in the soil of these areas.

Material and methods. Herb of Cichorium intybus L., collected in July 2021 in the Central (Tulgansky, Sakmarsky districts) and in the Western Orenburg region (Ponomarevsky, Buzuluksky, Sharlyksky districts). Determination of the content of trace elements was carried out by atomic absorption spectrometry. Statistica 10.0 software package (StatSoft, Inc.) was used for statistical processing.

Results. Analysis of the content of essential microelements in the herb of Cichorium intybus L. showed that the maximum content of Mn, Co and Cr was noted in the herb of chicory collected in the Buzuluksky district, Cu and Zn – in the raw material of the Sharlyksky district, Ni – in the raw material of the Tyulgansky district, Fe – in the raw material of Buzuluksky and Tulgansky districts. An analysis of the content of essential microelements in the soil found that the highest concentrations of Mn, Co, Fe, Zn and Cu were noted in the soil of the Buzuluksky district, Ni and Cr – in the soil of the Tyulgansky district.

Conclusion. Thus, the established differences in the content of essential microelements in the herb of Cichorium imtybus L., collected in the Orenburg region, may be due to differences in the content of these microelements in the soil, the phenomena of antagonism, synergism, and are taken into account when choosing raw materials containing the highest concentrations of microelements.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):43-47
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Problems of experimental biology and medicine

Stress-protective activity of the complex remedy in acute immobilization stress

Shantanova L.N., Lubsandorzhieva P.B., Baldandorzhieva M.V., Alekseeva E.A., Dargaeva T.D.

Abstract

Relevance. The range of medicinal remedies in the traditional Tibetan medicine contains several tens of formulae named bchud len. They are multicomponent remedies of natural origin having a stimulating non-specific effect on the human body. The analysis of such formula under the name ‘Rejuvenating-35’ has been presented in the paper. The complex remedy on the base of this formula was developed. Its composition includes 14 species of the plants and chitosan as a source of mineral substances. Previous phytochemical studies have revealed that the given remedy contains a wide spectrum of biologically active substances, namely triterpene saponins, flavonoids, ether oils, tannins, polysaccharides, etc.

The aim of the present work was to evaluate stress-protective activity of the complex remedy in acute restraint stress.

Material and methods. Experiments were carried out on white Wistar rats. The acute stress was simulated by immobilization of animals in supine position for 18 hours. The tested remedy was introduced in the form of decoction for 7 days before exposure to stress.

Results. It has been established that the course administration of the complex remedy decreases the manifestation of the Selye’s triad signs: 26% decrease of adrenal glands hypertrophy, 45% and 60% decrease in involution of thymus and spleen respectively as compared to the data in the control group of animals. Besides, its administration prevents the development of rough destructions in the stomach mucosa in the form of erosions and linear ulcers. Against the background of the complex remedy administration there was noted twofold decrease of TBA-active products concentration, fourfold increase of superoxide dismutase activity, 30% increase of catalase activity and the content of the reduced glutathione was 30% increased as compared to the data in the control.

Conclusions. The preventive administration of the complex remedy has stress-protective effect against the background of acute restraint stress. Molecular-cell mechanisms of its antistress effect are connected with the inhibition of free-radical oxidation processes and the increased activity of the endogenic antioxidant system.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):48-52
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Antioxidant activity and energy protective effect of Hedysarum alpinum L. extract in acute CCL4-induced hepatitis

Toropova A.A., Matypov B.D., Nikolaev S.M., Sambueva Z.G., Erentueva A.Y.

Abstract

Relevance. The course of physiological processes in the norm is significantly influenced by the pro- and antioxidant, as well as the energy status of the body. In the liver, biochemical disturbances in the functioning of these systems contribute to the development or aggravate the course of pathological conditions such as chronic hepatitis, cholelithiasis, chronic cholecystitis, etc. In this regard, the assessment of the influence of the studied herbal remedies that have a hepatoprotective effect on energy metabolism and the state of the antioxidant system of the body in experimental pathological processes is still relevant.

The purpose of the study was to evaluate the effect of Hedysarum alpinum L. dry extract on antioxidant status and energy metabolism in white rats with acute carbon tetrachloride-induced hepatitis.

Material and methods. The studies were carried out on white Wistar rats. Acute carbon tetrachloride hepatitis was induced by subcutaneous injection of a 50% oily solution of CCl4 for 4 days at a dose of 1.0 ml/kg. The dry extract H. alpinum was administered per os to animals at doses of 100 and 200 mg/kg. On the 7th and 14th days of the experiment, the energy metabolism and antioxidant status of the liver were assessed. The content of ATP, lactate, pyruvate, GSH the concentration of malondialdehyde (MDA), the activity of catalase, glutathione peroxidase, glutathione reductase, superoxide dismutase (SOD) in the liver homogenate were determined. The evaluation of the antiradical activity of the studied agent was carried out in vitro: binding of 2,2-diphenyl-1-picrylhydrazyl (DPPH˙), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS˙+).

Results. The dry extract H. alpinum was administered to animals at doses of 100 and 200 mg/kg, increases the activity of SOD, catalase, glutathione peroxidase, glutathione reductase and the content of GSH, reduces the content of MDA, increases the content of ATP, reduces the level of lactate; corrects the lactate/pyruvate ratio compared to the control. The dry extract of H. alpinum demonstrated the most pronounced antioxidant activity and energy-protective effect at a dose of 200 mg/kg. The extract of H. alpinum characterized by a pronounced radical-binding activity against DPPH˙ and ABTS˙+ (IC50DPPH˙=25.1 µg/ml; IC50ABTS˙+=47.1 µg/ml), which was due to the high content in its composition of phenolic compounds (2-C-β-D-glucopyranosyl-1,3,6,7-tetraoxyxanthone, rutin, catechins, xanthones), flavonols and polysaccharides.

Conclusions. The dry extract H. alpinum at doses of 100 and 200 mg/kg reduces the intensity of oxidative stress and corrects the state of energy metabolism in carbon tetrachloride-induced acute liver injury in white rats, showing a pronounced antioxidant, antiradical and energy-protective effects.

Problems of Biological Medical and Pharmaceutical Chemistry. 2023;26(5):53-59
pages 53-59 views

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