The genus Euonymus (L.) consists of shrubs and woody plants, distributed mainly in the Northern Hemisphere. Several hundred of secondary metabolites have been isolated from Euonymus spp. In addition, fatty oil was found in the fruits of some Euonymus spp., which accumulates not only in the seeds but also in the arils. This study presents the research of unique over ten-year-old suspension cell cultures of the endemic plant Euonymus maximoviczianus Prokh., obtained from the aril tissue of unripe capsules. The suspension cells retain the ability to form oil droplets containing neutral lipids. Both cells growing in the dark (Em-D culture) and cells growing in the light (Em-L culture) can synthesize very-long-chain fatty acids (VLCFAs) as well as cyanidin-3-O-hexoside, delphinidin-3-O-hexoside, and peonidin-3-O-hexoside. Here, we researched the VLCFA and anthocyanin accumulation dynamics during subcultivation, as well as the influence of methyl jasmonate (MeJA) and light on these processes. In the darkness, the formation of VLCFAs was more intense, while the biosynthesis of anthocyanins was significantly activated in the light. In Em-L cells, more than 76% of anthocyanins were represented by cyanidin-3-O-hexoside, and in Em-D cells delphinidin-3-O-hexoside was more actively synthesized (45%). MeJA substantially enhanced the accumulation of anthocyanins (especially in the light) and, surprisingly, the formation of VLCFAs in both Em-L and Em-D cell cultures. The possible competition between the biosynthetic pathways of VLCFAs and anthocyanins is discussed in connection with the commonality of the cytosolic pool of their precursor, malonyl-CoA.

An environmentally friendly, resource-efficient substitute for the production of vegetative biomass and the bioactive components of medicinal plants is provided by novel biotechnology-based suspension cells cultured in bioreactors. Despite the cultured biomass is produced in the controlled conditions, it requires detailed chemical analysis and toxicological evaluation before being approved for human consumption. This study reports the results of biochemical, toxicological, and elemental composition analysis of the suspension cell culture of a medicinal species Panax japonicus (T. Nees) C.A. Mey. cultivated in a large-scale bioreactor system. For the scaling up the process of cultivation, the suspension culture was grown sequentially in a series of laboratory (20-L), pilot (75-L), and industrial (630-L) bioreactors. The biomass productivity of cell culture was comparable or higher than of wild plants or in vitro cultured hairy roots reported in the literature. The resulting cell biomass was analyzed for ginsenoside contents using ultra-high-performance liquid chromatography-electrospray ioni-zation-mass spectrometry (UHPLC-ESI-MS). For the elemental composition of cells, inductively coupled argon plasma mass spectrometry (ICP-MS) was used. In addition, the biomass was evaluated for acute toxicity and skin sensibilization effects on in vivo animal models. The analyses confirmed that suspension cells cultured in bio-reactors accumulate a broad spectrum of ginsenosides and their malonylated derivatives at a total concentration of 7.54 % (w/w) based on dry weight and contain essential macro-(K, Ca, Mg, Na) and micro-(Zn, Mn, Fe, B, Al, Cu) elements in dietary-safe concentrations. Acute toxicity tests using the administered doses of 2000 and 5000 mg dry biomass per kg animal weight resulted in no changes in animal organ weights and no or minor changes in hematological and biochemical parameters of blood. No skin irritation or sensitizing effects were observed. Therefore, bioreactor cultivation of P. japonicus cell suspension is highly productive and provides a ginsenoside-rich and non-toxic cell biomass which is a potentially valuable component of functional foods, food additives, and natural health products.

Ex situ collections of algae, cyanobacteria, and plant materials (cell cultures, hairy and adventitious root cultures, shoots, etc.) maintained in vitro or in liquid nitrogen (−196 °C, LN) are valuable sources of strains with unique ecological and biotechnological traits. Such collections play a vital role in bioresource conservation, science, and industry development but are rarely covered in publications. Here, we provide an overview of five genetic collections maintained at the Institute of Plant Physiology of the Russian Academy of Sciences (IPPRAS) since the 1950–1970s using in vitro and cryopreservation approaches. These collections represent different levels of plant organization, from individual cells (cell culture collection) to organs (hairy and adventitious root cultures, shoot apices) to in vitro plants. The total collection holdings comprise more than 430 strains of algae and cyanobacteria, over 200 potato clones, 117 cell cultures, and 50 strains of hairy and adventitious root cultures of medicinal and model plant species. The IPPRAS plant cryobank preserves in LN over 1000 specimens of in vitro cultures and seeds of wild and cultivated plants belonging to 457 species and 74 families. Several algae and plant cell culture strains have been adapted for cultivation in bioreactors from laboratory (5–20-L) to pilot (75-L) to semi-industrial (150–630-L) scale for the production of biomass with high nutritive or pharmacological value. Some of the strains with proven biological activities are currently used to produce cosmetics and food supplements. Here, we provide an overview of the current collections’ composition and major activities, their use in research, biotechnology, and commercial application. We also highlight the most interesting studies performed with collection strains and discuss strategies for the collections’ future development and exploitation in view of current trends in biotechnology and genetic resources conservation.

This lecture presents classical information and new data on the molecular events of the "basic" (core) cell cycle (CC) of plants. The impact of water deficit, CO2, light, and temperature on CC is briefly examined. Data on the regulation of cell proliferation by auxins, cytokinins, abscisic acid, gibberellins, brassinosteroids, and ethylene are presented. Commonality and peculiarities of the effect of phytohormones on CC in various organs and tissues are discussed.

Callus and suspension cell cultures were successfully developed from Sutherlandia frutescens (Fabaceae), an endemic medicinal plant of South Africa. Two callus cell lines, originating from hypocotyl and cotyledon explants of in vitro seedlings under both dark and light conditions, showed intensive fresh weight accumulation with growth index ranging from 4.6 to 5.9. Suspension cell cultures induced from two callus lines had similar growth profiles and their growth index (15–18), specific growth rate (0.15–0.16 day−1), productivity (0.83–0.96 g/(l day)) and maximum biomass accumulation (16–18 g/l) remained relatively high for Fabaceae cell cultures during 27 sub-cultivations. Callus and suspension cell cultures showed similar profiles of secondary metabolites that were, however, different from leaves of greenhouse plants. Isoflavones were predominant in both callus and suspension cell cultures while flavonoids (sutherlandins) and triterpene glycosides of the cycloartane group (sutherlandiosides) were mostly found in leaves. Nineteen fatty acids (FA), both short- and very-long-chained (up to C25:0), were found in cell cultures. Linoleic and α-linolenic FA together comprised 60–64

The review examines the principles in formation of cell culture and tissue collections of higher plants in vitro as a variant of genetic collections, the modern state and perspective of using collections in studies of the peculiarities and patterns of synthesis of biologically active substances of higher plants, the important role of collections in biotechnological production of substances for medicinal preparations, and methods for maintaining the stability of cell cultures during long-term cultivation. Particular attention is paid to the retrospective and current state of the All-Russia Collection of Cell Cultures of Higher Plants, the founder of which was Russian Academy of Sciences Corresponding Member R.G. Butenko.

Suspension cell culture of camel’s thorn Alhagi persarum Boiss. et Buhse was produced for the first time and its characteristics were determined under different growing conditions (flasks and bubble bioreactors with useful volume of 15 L). The obtained culture was notable for an intense growth (accumulation of dry biomass of 15–20 g/L, growth index of 11–16, and specific growth rate of 0.15 ± 0.01 d–1), with growth characteristics in bioreactors close to parameters observed in flasks. Qualitative composition of secondary metabolites in biomass of this cell culture grown in flasks was preliminarily examined by means of ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-ESI-MS). Isoflavones from different structural groups were detected: free isoflavones (an isomer of afrormosin), glycosides of isoflavones (glycosides of calycosin and formononetin), and acylated glycosides of isoflavones (malonyl glycosides of calycosin, formononetin, genistein, and the isomer of afrormosin). Extracts from biomass of A. рersarum suspension cell culture grown in flasks were tested for antimicrobial activity. Staphylococcus aureus АТСС 25923 and Pseudomonas aeruginosa АТСС 27853 were used as test cultures. Extracts showed a specific antimicrobial activity in respect to S. aureus but were inactive to P. aeruginosa. Thus, the obtained cell culture of A. persarum is a promising object for further examination with a potential of using as an alternative raw material and a source of biologically active compounds characteristic of this species.

Recent evidence shows that small open reading frame (smORF; <100 codons)-encoded peptides (SEPs) containing transmembrane domains are preadapted to be progenitors of novel functional genes. A dozen of such SEPs translated from long non-coding RNAs (lncRNAs) are already functionally characterised in animals. However, functional plant lncRNA-smORF-coded peptides are not yet described. Here, we report detailed functional characterization of a 41-aa peptide encoded by lncRNA-smORFs in the moss Physcomitrium patens, which was named “FAst-growing MOSS” (FAMOSS). We found that the FAMOSS interacts with the Rab-type small GTPase proteins and its overexpression leads to faster moss growth rate and more intensive vesicular transport in apical cells, while its knockout results in the opposite effect. The FAMOSS contains a predicted transmembrane domain and possible orthologs from streptophyta algae to flowering plants have a very conserved structure. Thus, the FAMOSS peptide is a previously unknown conserved player of Rab-mediated processes in plants. Our findings are in line with functional studies of transmembrane SEPs in animals and prove the principles of SEPs evolution. This study provides new insights into functions of plant lncRNA-smORFs.

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

Background Bone fractures are a global public health issue; however, to date, no comprehensive study of their incidence and burden has been done. We aimed to measure the global, regional, and national incidence, prevalence, and years lived with disability (YLDs) of fractures from 1990 to 2019. Methods Using the framework of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we compared numbers and age-standardised rates of global incidence, prevalence, and YLDs of fractures across the 21 GBD regions and 204 countries and territories, by age, sex, and year, from 1990 to 2019. We report estimates with 95% uncertainty intervals (UIs). Findings Globally, in 2019, there were 178 million (95% UI 162-196) new fractures (an increase of 33.4% [30.1-37.0] since 1990), 455 million (428-484) prevalent cases of acute or long-term symptoms of a fracture (an increase of 70.1% [67.5-72.5] since 1990), and 25.8 million (17.8-35.8) YLDs (an increase of 65.3% [62.4-68.0] since 1990). The age-standardised rates of fractures in 2019 were 2296.2 incident cases (2091.1-2529.5) per 100 000 population (a decrease of 9.6% [8.1-11.1] since 1990), 5614.3 prevalent cases (5286.1-5977.5) per 100 000 population (a decrease of 6.7% [5.7-7.6] since 1990), and 319.0 YLDs (220.1-442.5) per 100 000 population (a decrease of 8.4% [7.2-9.5] since 1990). Lower leg fractures of the patella, tibia or fibula, or ankle were the most common and burdensome fracture in 2019, with an age-standardised incidence rate of 419.9 cases (345.8-512.0) per 100 000 population and an age-standardised rate of YLDs of 190.4 (125.0-276.9) per 100 000 population. In 2019, age-specific rates of fracture incidence were highest in the oldest age groups, with, for instance, 15 381.5 incident cases (11 245.3-20 651.9) per 100 000 population in those aged 95 years and older. Interpretation The global age-standardised rates of incidence, prevalence, and YLDs for fractures decreased slightly from 1990 to 2019, but the absolute counts increased substantially. Older people have a particularly high risk of fractures, and more widespread injury-prevention efforts and access to screening and treatment of osteoporosis for older individuals should help to reduce the overall burden. Copyright (C) 2021 The Author(s). Published by Elsevier Ltd.

Background While there is a long history of measuring death and disability from injuries, modern research methods must account for the wide spectrum of disability that can occur in an injury, and must provide estimates with sufficient demographic, geographical and temporal detail to be useful for policy makers. The Global Burden of Disease (GBD) 2017 study used methods to provide highly detailed estimates of global injury burden that meet these criteria. Methods In this study, we report and discuss the methods used in GBD 2017 for injury morbidity and mortality burden estimation. In summary, these methods included estimating cause-specific mortality for every cause of injury, and then estimating incidence for every cause of injury. Non-fatal disability for each cause is then calculated based on the probabilities of suffering from different types of bodily injury experienced. Results GBD 2017 produced morbidity and mortality estimates for 38 causes of injury. Estimates were produced in terms of incidence, prevalence, years lived with disability, cause-specific mortality, years of life lost and disability-adjusted life-years for a 28-year period for 22 age groups, 195 countries and both sexes. Conclusions GBD 2017 demonstrated a complex and sophisticated series of analytical steps using the largest known database of morbidity and mortality data on injuries. GBD 2017 results should be used to help inform injury prevention policy making and resource allocation. We also identify important avenues for improving injury burden estimation in the future.

Zinc is one of the most widespread transition metals in plants, and Zn deficiency has many adverse effects on plant productivity and human health. The zinc homeostasis system is actively studied; however, the mechanisms underlying regulation of Zn homeostasis have not been fully elucidated. Two potential ways to assess cellular Zn homeostasis are to sense changes in the free Zn2+ levels or disturbances in protein folding due to zinc deficiency. We found that changes in the cellular free Zn2+ level regulated the transcriptional activity of a set of primary Zn homeostasis genes in a dose-dependent manner and with fast kinetics. We propose that in addition to the well-known bZIP19/bZIP23 transcription factors, another Zn-sensing system exists in the cell and regulates free Zn2+ dependent changes in MTP2 gene expression. We first demonstrated that changes in the free Zn2+ levels in the cells likely influenced not only the transcriptional activation but also the mRNA degradation rates of Zn homeostasis genes, which were accelerated for several genes during recovery of the cellular Zn status. Zn deficiency clearly disturbed the protein folding processes in the cytosol of plant cells, but the cellular response system to these disturbances did not play a significant role in the regulation of Zn homeostasis genes.

The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019 provides a rules-based synthesis of the available evidence on levels and trends in health outcomes, a diverse set of risk factors, and health system responses. GBD 2019 covered 204 countries and territories, as well as first administrative level disaggregations for 22 countries, from 1990 to 2019. Because GBD is highly standardised and comprehensive, spanning both fatal and non-fatal outcomes, and uses a mutually exclusive and collectively exhaustive list of hierarchical disease and injury causes, the study provides a powerful basis for detailed and broad insights on global health trends and emerging challenges. GBD 2019 incorporates data from 281 586 sources and provides more than 3.5 billion estimates of health outcome and health system measures of interest for global, national, and subnational policy dialogue. All GBD estimates are publicly available and adhere to the Guidelines on Accurate and Transparent Health Estimate Reporting. From this vast amount of information, five key insights that are important for health, social, and economic development strategies have been distilled. These insights are subject to the many limitations outlined in each of the component GBD capstone papers.

Zinc is the most abundant and important transition metal in plants; however, the dynamic aspects of zinc homeostasis in plant cells are poorly understood. In this study we explored the pool of labile exchangeable zinc complexes in plant cells, and the potential influence of changes in intracellular zinc availability on cellular physiology. Work was performed on cultivated cell extracts of Arabidopsis thaliana (L.) Heynh. and Thellungiella salsuginea (Pall.) O.E. Schulz grown under control (3.48 µM Zn2+), 10-fold Zn excess or Zn starvation conditions. The free and labile Zn contents in the extracts were then determined by fluorimetric titration. We observed for the first time that plant cells contain micromolar concentrations of labile zinc complexes that account for a low percentage of the total zinc content. Labile zinc is mainly protein bound. Zn starvation inhibits cell proliferation and leads to the disappearance of the labile zinc pool, whereas Zn excess drastically increases the labile zinc pool. Free Zn2+ is buffered at picomolar concentrations in the intracellular milieu, and the increase in free Zn2+ concentrations to low nanomolar values clearly modulates enzyme activity by direct reversible binding. Such increases in free Zn2+ can be achieved by the substantial influx of additional zinc or by the oxidation of zinc-binding thiols. The observed features of the labile zinc pool in plant cells suggest it has a role in intracellular zinc trafficking and zinc signalling.

A set of transgenic Arabidopsis thaliana monoclonal cell lines has been created by selecting individual cells (cell aggregates) with random GFP gene integration events after the Agrobacterium-mediated transformation. The yield of the recombinant GFP ranged from 0.07 to 2.37% of the total soluble protein. Three lines with the highest, about 2% of the total soluble protein, accumulation of the target GFP protein were isolated. Areas of T-DNA insertions into the plant genome for 12 monoclonal cell lines were determined. The variation in this characteristic among 21 examined cell lines can serve as a reference for A. thaliana CRISPR/Cas9 genome editing aimed at the increase in the yield of recombinant proteins in plant expression systems. The lines with the highest level of the recombinant protein accumulation are of interest for the further identification and detailed characteristics of the sites for the transgene integration. These sites can be used as targets for the gene integration during the creation of lines for the production of recombinant proteins. Arabidopsis thaliana, cell suspension culture, Agrobacterium-mediated transformation, gene expression, green fluorescent protein. The work was performed under project no. 17-14-01099 of the Russian Science Foundation.