Use Of Biotechnological Methods Research Articles

Development of bread wheat lines resistant to warrior yellow rust (Puccinia striiformis f. sp. tritici) in a short time with the use of biotechnological methods

Development of bread wheat lines resistant to warrior yellow rust (Puccinia striiformis f. sp. tritici) in a short time with the use of biotechnological methods

Homeopathic medicine based on Carapichea Ipecacuanha for the treatment of alcoholism

Based on the results of the SWOT-analysis, the prospects and strategy for the development of a homeopathic medicinal product based on ipecacuanha are substantiated. It was found that there is only one domestic complex drug on the pharmaceutical market. It is shown that reduction of the cost and improvement of the quality of the starting material is possible with the use of biotechnological methods. Ultrasonic filtration is proposed to obtain the matrix tincture. It has been established that a homeopathic remedy with ipecacuanha can be a means of additional therapy at the stage of detoxification and the formation of patient's alcohol aversion. The necessity of additional research and improvement of quality control methods is proved.

Machine-hardware diagram of complex biotechnological processing of aquaculture objects

Currently, a number of fish farms operate in the Kaliningrad region, which specialize in breeding aquaculture objects, including common carp (Cyprinus carpio Linnaeus, 1758) and clariid catfish (Clarias gariepinus Linnaeus, 1758) and their further sale in live or chilled whole form. There is not complex processing, as a rule. The paper shows the need for biotechnological processing of aquaculture and ways to use the waste obtained during filleting for production of minced fish and, later, minced semi-finished products. The lack of integrated use of waste from fish processing industries is a common problem in the fishing industry, so the need to solve this problem is obvious. For the efficient processing of fish raw materials, it is necessary to create a technology that will make it possible to maximize the use of waste from fish production, including the use of biotechnological methods, but at the same time ensure high quality of finished products. The paper presents the mass composition of common carp (Cyprinus carpio Linnaeus, 1758) and clariid catfish (Clarias gariepinus Linnaeus, 1758), developed a technological diagram for the complex biotechnological processing of aquaculture objects in the conditions of small enterprises of fish processing farms, proposed a description of technological processes and a machine-hardware diagram. Integrated technology implies the presence of the main and additional production. An additional diagram is associated with the production of imitation fat, the basis for which can be ground fat-containing components pre-treated with enzymes, such as tioshka and milk, as well as fish broths obtained by cooking collagen-containing components (heads, fins, tails). This technological approach can be used in the conditions of “Belvodkhoz” enterprises engaged in the cultivation and subsequent processing of aquaculture objects, for example, Fish Farm “Volma”.

Bacteriophages: Clinical significance and application prospects

A review of the literature data covering topical issues of the use of bacteriophages in clinical practice is presented. Traditionally, phage therapy is based on the use of natural phages for the lysis of bacteria at the infection site. Despite some limitations, it has significant advantages over antibiotic therapy. The use of biotechnological methods currently makes it possible to eliminate the disadvantages of phage therapy by creating recombinant drugs and, in the future, expand its capabilities through the use of lytic phage proteins and their modified derivatives. Currently, bacteriophages are used not only for the treatment of infections but also for prevention and diagnosis (phagotyping to identify the source of infection). Bacteriophages are also used in genetic engineering as vectors for transferring DNA sections. Few side effects have been described about bacteriophages. Particularly, they do not negatively affect the intestinal microbiota, and compared with antibiotics, they are much less likely to be associated with allergic reactions. Moreover, existing randomized clinical trials, which are the gold standard of clinical trials, in phage therapy are extremely insufficient, which dictates the need to concentrate the efforts of all stakeholders in this direction.

INTRODUCTION OF DRACOCEPHALUM MOLDAVICA L. IN VITRO CULTURE

B a c k g r o u n d . Dracocephalum moldavica L. is widely used in folk medicine in many countries of the world due to its antispasmodic, antibacterial, analgesic, sedative and anti-inflammatory properties. D. moldavica essential oil is often used in aromatherapy and cosmetology. The use of biotechnological methods for obtaining secondary plant metabolites has become increasingly popular in recent years. On the one hand, they serve as a source of substances beneficial for pharmacology, while on the other hand, aseptic microclones of medicinal plants can be utilized as model organisms to explore methods to enhance the biosynthesis of specific target products. The in vitro use of plants for the production of biologically active substances is also a crucial and pertinent method for the conservation of natural biodiversity. The aim of this study was to determine the optimal conditions for introducing the medicinal plant Dracocephalum moldavica into in vitro culture. M e t h o d s. To introduce Dracocephalum moldavica plants into in vitro culture, the conditions for seed sterilization were carefully selected. The effectiveness of treatment with succinic acid in combination with various concentrations of sodium hypochlorite solution (20%, 50%, or 70%) was assessed to obtain aseptic material and ensure a high percentage of seed germination. The duration of seeds viability after storage for 1, 2, and 3 years was determined by testing seed germination in soil and in vitro. R e s u l t s . Seeds of D. moldavica have a relatively low germination rate when planted in soil and quickly lose their viability. In vitro cultivation can significantly enhance the germination of fresh seeds and allow for a more efficient use of two- and three-year-old seeds. The stimulating effect of succinic acid on the seed germination and seedling development of Dracocephalum moldavica has been observed, but it also activated the growth of fungal and bacterial infections. Therefore, it is advisable to use this stimulant only along with a more intensive sterilizing agent. C o n c l u s i o n s . The optimal method for introducing Dracocephalum moldavica from seeds into in vitro culture has been determined: pre-treatment of seeds with succinic acid followed by sterilization using 70% sodium hypochlorite.

Sweet Potato as a Key Crop for Food Security under the Conditions of Global Climate Change: A Review.

Sweet potato is one of the most economically important crops for addressing global food security and climate change issues, especially under conditions of extensive agriculture, such as those found in developing countries. However, osmotic stress negatively impacts the agronomic and economic productivity of sweet potato cultivation by inducing several morphological, physiological, and biochemical changes. Plants employ many signaling pathways to respond to water stress by modifying their growth patterns, activating antioxidants, accumulating suitable solutes and chaperones, and making stress proteins. These physiological, metabolic, and genetic modifications can be employed as the best indicators for choosing drought-tolerant genotypes. The main objective of sweet potato breeding in many regions of the world, especially those affected by drought, is to obtain varieties that combine drought tolerance with high yields. In this regard, the study of the physiological and biochemical features of certain varieties is important for the implementation of drought resistance measures. Adapted genotypes can be selected and improved for particular growing conditions by using suitable tools and drought tolerance-related selection criteria. By regulating genetics in this way, the creation of drought-resistant varieties may become cost-effective for smallholder farmers. This review focuses on the drought tolerance mechanisms of sweet potato, the effects of drought stress on its productivity, its crop management strategies for drought mitigation, traditional and molecular sweet potato breeding methods for drought tolerance, and the use of biotechnological methods to increase the tolerance of sweet potato to drought.

Electro-Biofabrication. Coupling Electrochemical and Biomolecular Methods to Create Functional Bio-Based Hydrogels.

Twenty years ago, this journal published a review entitled "Biofabrication with Chitosan" based on the observations that (i) chitosan could be electrodeposited using low voltage electrical inputs (typically less than 5 V) and (ii) the enzyme tyrosinase could be used to graft proteins (via accessible tyrosine residues) to chitosan. Here, we provide a progress report on the coupling of electronic inputs with advanced biological methods for the fabrication of biopolymer-based hydrogel films. In many cases, the initial observations of chitosan's electrodeposition have been extended and generalized: mechanisms have been established for the electrodeposition of various other biological polymers (proteins and polysaccharides), and electrodeposition has been shown to allow the precise control of the hydrogel's emergent microstructure. In addition, the use of biotechnological methods to confer function has been extended from tyrosinase conjugation to the use of protein engineering to create genetically fused assembly tags (short sequences of accessible amino acid residues) that facilitate the attachment of function-conferring proteins to electrodeposited films using alternative enzymes (e.g., transglutaminase), metal chelation, and electrochemically induced oxidative mechanisms. Over these 20 years, the contributions from numerous groups have also identified exciting opportunities. First, electrochemistry provides unique capabilities to impose chemical and electrical cues that can induce assembly while controlling the emergent microstructure. Second, it is clear that the detailed mechanisms of biopolymer self-assembly (i.e., chitosan gel formation) are far more complex than anticipated, and this provides a rich opportunity both for fundamental inquiry and for the creation of high performance and sustainable material systems. Third, the mild conditions used for electrodeposition allow cells to be co-deposited for the fabrication of living materials. Finally, the applications have been expanded from biosensing and lab-on-a-chip systems to bioelectronic and medical materials. We suggest that electro-biofabrication is poised to emerge as an enabling additive manufacturing method especially suited for life science applications and to bridge communication between our biological and technological worlds.

New approaches to registration and conservation of domestic cultivars of berry crops in the VIR Genebank on the example of red raspberry and black currant

A collection of nomenclatural standards is being created at VIR for domestic cultivars of various crops in accordance with the International Code of Nomenclature for Cultivated Plants (ICNCP). A new complex strategy was proposed for vegetatively propagated crops for registering domestic cultivars received from their authors in the VIR genebank. In addition to the creation of nomenclature standards, the strategy includes the development of a genetic passport of a cultivar and the use of biotechnological methods to preserve explants (buds, meristems) isolated from plant material transferred by breeders to the VIR Herbarium. This approach can be used for any vegetatively propagated crop applying a protocol developed specifically for an individual crop. For raspberry and black currant varieties, the collecting of plant material, its preparation for the registration of nomenclature standards and the preservation of viable samples under controlled in vitro conditions have specific features. This article provides detailed protocols for performing the mentioned work for raspberry and black currant varieties. In addition, the article summarizes the first results of the implementation of our proposed strategy on the example of domestic raspberry and black currant varieties created in various breeding centers of Russia. Three years of joint work of VIR researchers and breeders from four breeding centers in five regions of the country have resulted in creation of nomenclature standards for 20 raspberry varieties, as well as for five black currant varieties bred at VIR. Thirteen samples of raspberry varieties and four of black currant varieties, genetically identical to nomenclature standards, were introduced into in vitro culture; four raspberry cultivars have been placed in the VIR cryobank for the long-term cryopreservation.

Ruta chalepensis L. In Vitro Cultures as a Source of Bioactive Furanocoumarins and Furoquinoline Alkaloids

Recently, due to the decreasing areas of cultivation and climate change, the use of biotechnological methods to obtain biomass, which is a source of valuable bioactive metabolites, is becoming more and more interesting. In this study, Ruta chalepensis in vitro cultures were investigated in RITA® temporary immersion bioreactors. Biomass growth and the production of secondary metabolites in 4- and 5-week growth cycles on three variants of the Linsmaier and Skoog (LS) medium (naphthyl-1-acetic acid/6-benzylaminopurine (NAA/BAP): 0.5/1.0, 0.1/0.1, and 1.0/1.0 mg/L) were analyzed. Using high-performance liquid chromatography of methanolic extracts of biomass, the presence of linear furanocoumarins (bergapten, isoimperatorin, isopimpinellin, psoralen, and xanthotoxin) and furoquinoline alkaloids (γ-fagarine, 7-isopentenyloxy-γ-fagarine, and skimmianine) was confirmed. The highest content of linear furanocoumarins (1170 mg/100 g DW (dry weight)) was observed in the LS medium variant containing 0.5/1.0 mg/L NAA/BAP (4-week growth cycle). The highest content of furoquinoline alkaloids (449 mg/100 g DW) was observed in the LS medium variant containing 0.1/0.1 mg/L NAA/BAP (5-week growth cycle). Hence, R. chalepensis bioreactor cultures may be used as a biotechnological source of linear furanocoumarins (xanthotoxin and bergapten) and furoquinoline alkaloids (skimmianine and γ-fagarine).

Reguladores de crescimento vegetal na micropropagação de cultivares de Actinidia

ABSTRACT Actinidia Lindl., commonly known as kiwifruit, is a valuable berry crop. The area of commercial kiwifruit plantations is increasing; the global production of kiwifruit is about 0.62% of the total production of major fruit crops. The use of biotechnological methods, which can significantly accelerate the propagation of quality planting materials, is considered to be relevant for the propagation of this crop. In this study, we optimized the culture medium composition at the micropropagation stage for the effective cultivation of promising cultivars of A. arguta, A. kolomikta, and A. polygama. We investigated the features of Actinidia morphogenesis depending on the genotype, the concentration of 6-Benzylaminopurine (0.5, 0.8, and 1.0 mg L-1), and plant growth regulators (6-Benzylaminopurine, meta-topolin, and 2-isopentenyladenine at a concentration of 0.5 mg L-1) in the media Quoirin and Lepoivre. Actinidia arguta (multiplication rate of 8.0) and A. polygama (6.8) developed faster at the micropropagation stage compared to A. kolomikta (4.9). The studied Actinidia representatives were cultured most effectively on Quoirin and Lepoivre media supplemented with 0.5 mg L-1 meta-topolin, compared to the media containing 0.5 mg L-1 6-Benzylaminopurine and 0.5 mg L-1 2-isopentenyladenine. The use of meta-topolin in the medium contributed to the increase in various morphometric traits, such as the height of microshoots (up to 28% depending on the species), their number (up to 52%), and their multiplication rate (up to 42%). We also recorded a high morphogenic capacity of the investigated species.

Vegan alternative for fermented milk products: Preparation and some properties

Rice is relatively easy to digest by humans, and the presence of starch provides the organism with slow energy. In addition, the absence of lactose makes it possible to provide a complete diet for people suffering from allergies or individual intolerance to cow’s milk components. Therefore, the creation of analogues to traditional dairy and fermented milk products is a timely and in-demand task. The use of biotechnology methods allows not only to obtain a product with original organoleptic characteristics, but also to enrich it with metabolites of lactic acid bacteria, to give pre- and probiotic properties. The issues of obtaining fermented rice base (FRB) as a basis for alternative vegan products have been studied. It is shown that with the ratio of solid and liquid phases in the rice base – 1:2, respectively, favourable conditions were created for the development of lactic acid bacteria. It was found that additional treatment (partial enzymatic hydrolysis at a temperature of 40 ± 2 °C for 60 minutes) mixtures before fermentation improves the adaptation of the microbiota to this substrate. In addition, the taste of the rice base improves: a pronounced sweetness appears, the flour flavour disappears, the consistency becomes more viscous. The process of fermentation (for 12 hours at a temperature of 38-40 °C) of rice base was studied, the dynamics of acid accumulation depending on the introduced cultures of lactic acid bacteria was studied. The expediency of applying pectin (1% and 2% by weight) is shown to regulate the consistency of the FRB. The FRB samples received a high sensory evaluation.

Drought-tolerance of transgenic winter wheat with partial suppression of the proline dehydrogenase gene

The global climate changes and the consequent increase in the number of soil and air droughts during the vegetation period of grain crops require the development of new strategies to adapt plants to those yield-decreasing stressors. A relevant way of increasing drought-tolerance of cereals is the use of biotechnological methods, particularly RNA interference, which can down-regulate the activity of plants’ genes and increase concentration of stress metabolites that perform osmoprotective functions during drought. We studied the tolerance to soil moisture shortage in transgenic plants of winter wheat with partial suppression of the proline dehydrogenase gene, obtained using the technology of short interfering RNAs. We analyzed physiological and biochemical parameters and structural elements of yield productivity of 4 wild genotypes and their transgenic lines with reduced activity of proline dehydrogenase in the conditions of 7-day drought during the late booting–ear emergence. We determined that the presence of double-stranded RNA suppressor of the proline dehydrogenase gene in transgenic lines led to increase in the level of accumulation of free proline in flag leaves. At the same time, its concentration in transgenic lines was higher than in untransformed plants of the wild genotypes in both drought conditions and conditions of sufficient moisture. We found that against the background of water deficiency, the total chlorophyll content in leaves of plants of transgenic lines was significantly higher, and the ratio of carotenoids to chlorophyll was lower than in plants of the wild genotypes, suggesting mitigation of the negative impact of drought on the plants of transgenic lines. Lacking soil moisture, genetically altered lines of wheat had significantly higher parameters of the structure of grain yield compared with untransformed genotypes. At the same time, we observed genotypic difference according to grain productivity in biotechnological plants. Therefore, the results we obtained confirm the perspectives of using the technology of short interfering RNAs to increase tolerance of winter wheat to water deficiency.

BIOTECHNOLOGICAL ASPECTS OF PROPAGATION OF BLACK POPLAR HYBRIDS “SAN GIORGIO” AND “GHOY”

Energy independence of the country can be solved in several ways. One of them is the creation of energy plantations of woody plants. The area increase of such plantations is limited by the insufficient amount of high-quality planting material. Hybrid plants of the genus Populus are of considerable interest when creating plantations. Given that not all hybrids are successfully propagated by conventional vegetative methods, the use of biotechnological methods, namely microclonal propagation, solves the mentioned problem. This method allows obtaining a large amount of highquality, uniform planting material. The purpose of this study was to develop the technology of microclonal propagation of black poplar hybrids ‘San Giorgio’ and ‘Ghoy’. The study used generally accepted biotechnological methods in the author’s modification at all stages of obtaining planting material of regenerating plants of poplar hybrids. Studies have established a positive effect when obtaining an aseptic culture of sterilising substances 0.1% AgNO3, 2.5 and 1.25% NaClO, 0.05% sodium merthiolate and 0.1% HgCl2. The largest number of aseptic viable explants was obtained by cultures using a 0.05% solution of sodium merthiolate after exposure for 5 and 10 minutes. When rooting microshoots on a hormone-free nutrient medium ½ MS and with the addition of 0.1-1.0 mg/l IBA, it was established that the best parameters for rooting microshoots are on a hormone-free nutrient medium ½ MS. Adaptation was carried out for 4 weeks in a greenhouse on a substrate that included components of peat:sand:perlite in a ratio of 1:1:1. The viability of regenerating plants of black poplar hybrids ‘San Giorgio’ and ‘Ghoy’ was more than 91%. The conducted research and the obtained results will be useful for producers of planting material and the scientific environment, which develop renewable energy sources through the creation of energy plantations of fast-growing woody plants

In Vitro Regeneration Potential of Thin Cell Layer Explants of Lentisk (Pistacia lentiscus var. Chia) Plant

The problems encountered in the production of the lentisk trees, which are one of the important endemic plants of our country have led to the use of biotechnological methods. In this research for this purpose, the TCL (Thin Cell Layer) technique was consideredto investigate of in vitro regeneration potential of expants used for production of lentisk. Firstly, the leaf, node and stem parts of the plant were cut by TCL technique and these explants had been cultured in semi-solid MS media supplemented with 2,4-D and KIN at different concentrations (0.1, 0.2, 0.5 and 1.0 mg/L). The highest callus formation percentage was 100% in transverse stem layers and longitudinal node in MS media including 1 mg/L 2,4-D and 1 mg/L KIN. The lowest callus regeneration ratios were found as 26.67% for three explant types (transverse leaf, transverse stem, longitudinal node). Due to the high rate of darkening in regenerated calli, these were transferred primarily to semi-solid media containing different antioxidants (ascorbic acid, citric acid, PVP, active charcole) and after that culturedin liquid media containing different plant growth regulators (IAA, KIN and BAP) to induced somatic embryogenesis. Later, the calli were encapsulated to prevent darkening and the nurse technique was applied with Aloe vera L. and Gossypium hirsutum L. calli as a different application. As a result of all these trials, somatic embryogenesis didn’t occur, but darkening ratio was reduced to 6.67%.

Assessment of the quality of the southern Bug waters within the Khmelnytsky region and biotechnological ways to solve their purification

Water resources are the national wealth of each state, an important natural resource, and determine the development opportunities of most sectors of the economic complex of Ukraine. The Southern Bug River is the largest river, the basin of which is completely located in Ukraine. Due to the intensive growth of anthropogenic impact on the studied reservoir and the economic use of its resources, the quality of water in it has significantly decreased. To improve the quality of water in rivers, it is optimal to use biotechnological methods, which provide many opportunities for effective solutions to issues related to monitoring, assessment, and treatment of polluted watercourses. The aim of our study was to assess water safety, the current ecological status of the Southern Bug River within the Khmelnytsky region in three areas (Maryanivske Reservoir, Kopystyn, Shchedrivske Reservoir) and use in domestic needs, with analysis of potential risks to human health, development of new biotechnological and improvement of existing measures to improve water supply. The experimental part of the work was performed on the basis of the Khmelnytsky Regional Department of Water Resources in the laboratory at the address Khmelnytsky, st. Soborna, 29. We conducted a chemical assessment of the Southern Bug River on three indicators: salt composition (I1), tropho-saprobiological (I2), and specific toxic substances (I3). Research of river waters was carried out in order to control the MPC (maximum permissible concentrations) of substances for use in household and food needs of the population and to propose biotechnological methods of river water purification within the Khmelnytsky region. According to our research on hydrochemical characteristics, the water contained elevated levels of NH4, NO2, NO3, PO4, the average values of HSC (chemical oxygen demand) exceeded the MPC in all areas by 2 to 5 times. This may be due to exposure to organic compounds of natural origin. The content of nitrites, nitrates is within normal limits, but the content of ammonium salt exceeds the MPC up to 20 times in the village Kopystyn and from 2 to 7 times in the village of Letychiv. According to many indicators of the salt block, the maximum concentration limit was not exceeded, except for the village Kopystyn, the concentration of Magnesium (Mg) was 84.33 mg/dm3 at a rate of 40.00 mg/dm3. According to specific toxic substances, the water belonged to the III class 4 quality category, Chromium (Cr) was not detected, Manganese (Mn) and Copper (Cu) were within normal limits, although the Copper content was high. The concentration of Iron (Fe) varies in different areas and during the year, there is a case of excess Iron in the village. Kopistin up to 0.86 mg/dm3 at a rate of 0.30 mg/dm3. Iron affects the intensity of phytoplankton development and the qualitative composition of the microflora, so its concentration is subject to seasonal fluctuations. To improve the quality of water in the Southern Bug River, we have proposed the use of biotechnological methods, including those that provide many opportunities to effectively address issues related to monitoring, assessment, and treatment of polluted watercourses.

Use of Biotechnological Methods to Support the Production of New Peach Hybrids

Peach [Prunus persica (L.) Batsch] is among the most demanded fruit crops in the world. Biotechnological methods help to originate new hybrid forms in order to increase the cultivar diversity and create new valuable genotypes. Cross combinations between the cultivars Clyde Wilson, Jerseyglo, Loadel, Summerglo and the promising cultivar ‘Nikitskiy Podarok’ have been done. The embryos of these hybrids germinated and formed plantlets after stratification at 4 °C for 45–60 days. The best regeneration rates in the hybrids ‘Loadel’ × ‘Nikitskiy Podarok’ and ‘Summerglo’ × ‘Nikitskiy Podarok’ (96.30% and 92.59%, respectively) were noted on hormone-free Monnier culture medium supplemented with 400.0 mg L−1 casein hydrolyzate. When the newly formed plantlets had necrosis of the shoot apex or immature roots, nodal shoot segments were used. At the same time, a high regeneration capacity was noted in the hybrids ‘Summerglo’ × ‘Nikitskiy Podarok’ and ‘Loadel’ × ‘Nikitskiy Podarok’ on B5 culture medium with 0.75 mg L−1 6–benzyl–aminopurine (BAP) + 0.1 mg L−1 indole–3–butyric acid (IBA). After the second subculture, the number of new adventitious shoots was 5.18 ± 0.18 and 4.95 ± 0.18 shoots per explant, respectively. The plants obtained from the hybrid embryos in a soil mixture soil: peat: sand (3:1:1) were adapted. The main morphological and anatomical features of the leaf blades in newly originated peach hybrids have been studied: the thickness of their tissues and the distribution of stomatal apparatus, as well as the physiological parameters of the photosystem II activity in regenerants cultured in vitro and during their in vivo acclimatization. The high capacity to post aseptic adaptation in the obtained hybrids has been shown.

Influence of culture medium composition on the micropropagation of Thymus tauricus Klokov et Des.-Shost.

Thymus tauricus Klokov et Des.-Shost. is a perennial dwarf shrub of the Lamiaceae family. This plant has anti-inflammatory, antispasmodic, analgesic and antiseptic properties. The use of biotechnological methods makes it possible to increase the efficiency of traditional methods of breeding and seed production. Our investigation aimed to study the influence of the culture medium composition on the explants morphogenesis at 1-2th stages of Thymus tauricus clonal micropropagation. When comparing three cytokinins, the best explants development was revealed on culture media containing kinetin. Both high vitrification rate of microshoots (31.2-90.2%) and formation of small shoots were observed on media supplemented with BAP or TDZ. The most effective culture medium at the introduction stage is MS with 1.0 mg/l kinetin and 1.0 mg/l GA3 or 1.0 mg/l kinetin. On this medium, on average, 8.4-10.1 microshoots per explant and shoots length of 1.9 cm were obtained. It was found that the optimal culture medium at the actually propagation stage is MS with 1.0 mg/l Kin or 1.0 mg/l kinetin and 0.5 mg/l IAA, on which the multiplication index 26.9-28.5 were obtained. Induction of rhizogenesis with a frequency of 30.2-94.4% and a root length of 1.6-3.0 cm was noted on culture media for the second stage of clonal micropropagation. The results of the studies are the basis for development T. tauricus clonal micropropagation method.

СПЕЦИФИКА МОЛЕКУЛЫ МОРОКТОКОГА АЛЬФА: РЕЗУЛЬТАТЫ ПРОСПЕКТИВНОГО МНОГОЦЕНТРОВОГО НАБЛЮДАТЕЛЬНОГО ИССЛЕДОВАНИЯ ЭФФЕКТИВНОСТИ И БЕЗОПАСНОСТИ У ПОДРОСТКОВ С ГЕМОФИЛИЕЙ A

The main method of treatment for hemophilia A is replacement therapy with drugs of blood coagulation factors VIII (FVIII). The active use of biotechnological methods in the production of recombinant drugs contributes to the development and registration of new FVIII drugs. The results of clinical trials of drugs in children usually include a limited number of patients for a specific period of time. Post-marketing observational studies provide additional information on the results of using a new drug in patients in clinical practice. The objectives of the study were to collect and analyze data on the efficacy and safety of domestic recombinant FVIII with a deleted B-domain, moroctocog alfa (Octofactor, GENERIUM JSC) in the treatment of adolescents aged 12–18 years with hemophilia A in routine clinical practice. Materials and methods of research: a prospective multicenter open-label observational study of the results of using the Octofactor in adolescents with hemophilia A (№ OCF-HPA-N01) included 24 male patients with severe hemophilia A aged 12 to 18 years (mean age 14,8±1,7 years), who received moroctocog alfa in routine clinical practice. Enrollment in the study was carried out after the signing of the informed consent form by the parent of the minor patient and the minor patient aged 14 to 18 years, taking into account the results of the screening examination. The follow-up period was 52±4 weeks, which, presumably, was sufficient to achieve at least 100 days of moroctocog alfa administration. To assess the effectiveness, we analyzed the incidence of spontaneous bleeding that occurred within 48–72 h after drug administration; the severity of spontaneous bleeding against the background of prophylactic treatment with moroctocog alfa, the number of injections and the dose of the study drug for prophylactic treatment, as well as for treatment at the request of one episode of bleeding, taking into account its severity; the researcher's determination of the response to treatment of acute hemarthrosis according to the scale of the World Federation of Hemophilia (2020). To assess safety, the frequency and characteristics of adverse events (AEs) associated with drug administration were analyzed, including the frequency of formation of an inhibitor to FVIII. Results: during the study, 59 bleedings were recorded, of which 21 (36%) were spontaneous and 38 (64%) were post-traumatic. Among spontaneous bleeding episodes, 5 (24%) episodes occurred within 48–72 hours after administration of the study drug. Spontaneous bleeding within 48–72 hours after administration of Octofactor was absent against the background of prophylactic treatment in most patients (81%) and was observed rarely during the observation period (1–3 times) in the remaining 19% of patients. The median number of bleeding within 48–72 h after study drug administration was 1 episode over the follow-up period. The proportion of mild to moderate bleeding was 97 [88; 100]% among all types of bleeding and 100 [84; 100]% among spontaneous bleeding within 48–72 hours after administration of the Octofactor drug. The median of a single dose of Octofactor for preventive treatment was 2000 [1500; 2000] IU or 31,7 [25,6; 38,5] IU/kg, and with treatment on demand ‒ 2000 [2000; 3000] IU or 34,1 [28,8; 38,5] IU/kg per single injection. To stop the resulting bleeding required 1 [1; 2] introduction in a single dose of 3000 [2000; 4000] IU; the average dose was 4490 ÷ 4993 IU. When doctors subjectively assess the response to treatment of acute hemarthrosis on the scale of the World Federation of Hemophilia, an excellent response was noted in 9 (27%) episodes, good and moderate ‒ in 2 (6%) and 22 (67%) episodes, respectively. Lack of response to treatment of acute hemarthrosis with moroctocog alfa was not revealed in the study. During the study, 23 AEs were recorded in 13 (54%) patients not related to the study drug. Conclusion: thus, the obtained results of the study indicates the efficacy and safety of the Octofactor both the prophylactic treatment and treatment of on-demand bleedings in 12 to 18 year old patients with severe hemophilia A.

Development of a concept for the production of wheat snacks with the elimination of gluten by the biocatalysis

The increase in the number of cases of allergic reactions and celiac disease is an important problem. The solution to this problem is the search and development of relevant and effective ways to eliminate gluten. Specific amino acid sequences glutamine and proline determine the resistance to protease hydrolysis of the structural domains of gluten fractions. The analysis of the literature data showed that an alternative to the gluten-free diet is the use of biotechnological methods for modifying ingredients containing gluten. Such methods include the use of leavens on the base of lactic acid bacteria or enzyme preparations containing peptidases specific to gluten biocatalysis. In addition, the pretreatment of raw materials by extrusion cooking contributes to an increase in the degree of gluten hydrolysis. The effect of the thermoplastic extrusion and various enzyme systems containing proteases, amylolytic, cellulolytic and hemicellulolytic enzymes on the changes in the molecular weights of wheat protein fractions was studied. It was found that extrusion as a factor of protein modification significantly affects the proteolysis of wheat proteins using enzyme systems of different substrate specificity. The most effective hydrolysis was shown by the use of a complex enzyme preparation Amyloprotoorizin. including The effect was also noted after bioconversion of non-extruded wheat. An algorithm for the technology of wheat snacks based on the processes of extrusion and biocatalysis of proteins with specific proteases for the elimination of gluten is devepoped. The practical implementation of the technology will make it possible to obtain ready-to-eat snacks, which will be investigated for the preservation or elimination of antigenic properties during clinical trials.

Biotechnology and Drought Stress Tolerance in Plants

Drought stress in plants has become one of the major abiotic stress that limits the growth and development of plants which also contributes to low yields. Biotechnology which has new and emerging techniques can be use to solve the problem of drought stress in plants. This review aimed at identifying drought stress tolerance in plants at different stages, how plants respond to drought stress using different methods and the application of different biotechnology methods to improve drought tolerance in plants. Some important parameters about drought stress in plants such as drought tolerance mechanisms, plants responses to drought stress, gene regulation for drought stress tolerance in plants, effects of drought stress at different stages of plant growth and biotechnology methods in developing drought tolerance in plants was reviewed. The use of biotechnology methods such as classical breeding, use of genetic manipulation, genes from resurrection plants and Protoplast fusion was discussed. Drought stress affects our plants seriously and it leads to wilts, reduction of yields and death of plants at different developmental stages. Plants have developed different mechanisms to respond to drought stress but these mechanisms are not sufficient enough without the application of biotechnology to greatly improve the growth, development and increase yield in pants. The use of biotechnology greatly improves plants ability to tolerate drought stress depending on the plant species and period of exposure. The use of biotechnology methods has become very vital in improving plants drought stress so as to overcome the major problems of plants which includes increase in population and climatic change.