Prospects for mineral biofortification of wheat: classical breeding and agronomy.

Low intake of micro- and macroelements and vitamins in food negatively affects the health of more than two billion people around the world provoking chronic diseases. For the majority of the world's population, these are soft and durum wheats that provide beneficial nutrients, however their modern high-yielding varieties have a significantly depleted grain mineral composition that have reduced mineral intake through food. Biofortification is a new research trend, whose main goal is to improve the nutritional qualities of agricultural crops using a set of classical (hybridization and selection) methods as well and the modern ones employing gene/QTL mapping, bioinformatic analysis, transgenesis, mutagenesis and genome editing. Using the classical breeding methods, biofortified varieties have been bred as a part of various international programs funded by HarvestPlus, CIMMYT, ICARDA. Despite the promise of transgenesis and genome editing, these labor-intensive methods require significant investments, so these technologies, when applied to wheat, are still at the development stage and cannot be applied routinely. In recent years, the interest in wheat biofortification has increased due to the advances in mapping genes and QTLs for agronomically important traits. The new markers obtained from wheat genome sequencing and application of bioinformatic methods (GWAS, meta-QTL analysis) has expanded our knowledge on the traits that determine the grain mineral concentration and has identified the key gene candidates. This review describes the current research on genetic biofortification of wheat in the world and in Russia and provides information on the use of cultivated and wild-relative germplasms to expand the genetic diversity of modern wheat varieties.

The Influence of Adiponectin on Transport of Low-Density Lipoproteins through Human Endothelial Cell Monolayer In Vitro.

The influence of adiponectin, a protein secreted by adipocytes, on the activation of transendothelial LDL transport, the initial event of atherogenesis, was studied. The addition of adiponectin to the cultured endothelial hybridoma EA.hy926 cells did not affect both basal and TNF-stimulated transendothelial transport of LDL. In addition, adiponectin affects neither expression levels of CAV1, SCARB1, and ACVRL1 genes encoding proteins involved in transendothelial LDL transport, nor the MMP secretion by the EA.hy926cells. At the same time, adiponectin suppressed the TNF-stimulated IL-8 production and expression of the adhesion molecule gene ICAM1 in these cells. Thus, adiponectin reduces proinflammatory activation of EA.hy926 cells, which is not accompanied by changes in the transendothelial LDL transport. We speculate that anti-inflammatory action of adiponectin is the base for the influence of this adipokine on atherogenesis.

[Adiponectin in normal and atherosclerotic intima of human aorta]. / Adiponektin v normal'noi i ateroskleroticheski izmenennoi intime aorty cheloveka.

BACKGROUND: Adiponectin (AN) is a protein synthesized by adipocytes that has regulatory effects on lipid and lipoprotein metabolism, increases tissue sensitivity to insulin, and modulates endothelial functions and inflammatory response. However, its involvement in the processes of atherogenesis remains poorly understood. OBJECTIVE: To determine the localization and sources of AN in atherosclerotic and normal human aortic intima. MATERIAL AND METHODS: Immunohistochemical study was performed on sections of atherosclerotic and normal human aorta obtained during autopsy. Reverse transcription real-time PCR was performed using biopsies of para-aortic and abdominal adipose tissue, intima-media of the thoracic aorta, atherosclerotic plaques of the human carotid and femoral arteries, as well as on endothelial cells isolated from the human thoracic aorta. Transendothelial transport of AN was evaluated in a two-chamber model using a monolayer of human endothelial cell hybridoma EA.Hy926. RESULTS: It has been established that AN is present in atherosclerotic but not in normal human aortic intima. At the same time, AN ADIPOQ mRNA was not detected either in the intima media of the human aorta, nor in isolated endothelial cells of the aorta, nor in cells of atherosclerotic plaques of the carotid and femoral arteries. AN slowly penetrated the endothelial monolayer in vitro, but this transport was significantly enhanced by the action of tumor necrosis factor-alpha (TNFa). CONCLUSION: Obtained data indicate that AN is present in atherosclerotic but not in normal aortic intima. We assume that AN is not synthesized by the cells of normal and atherosclerotic arterial walls, but permeates from the plasma. Transendothelial transport of AN, like many other plasma proteins, is activated during the development of atherosclerotic lesions, apparently under the action of pro-inflammatory cytokines, in particular, TNFα.

Localization of the quantitative trait loci related to lodging resistance in spring bread wheat (Triticum aestivum L.).

The yield and grain quality of spring and winter wheat signif icantly depends on varieties' resistance to lodging, the genetic basis of this trait being quantitative and controlled by a large number of loci. Therefore, the study of the genetic architecture of the trait becomes necessary for the creation and improvement of modern wheat varieties. Here we present the results of localization of the genomic regions associated with resistance to lodging, plant height, and upper internode diameter in Russian bread wheat varieties. Phenotypic screening of 97 spring varieties and breeding lines was carried out in the f ield conditions of the West Siberian region during 2017-2019. It was found that 54 % of the varieties could be characterized as medium and highly resistant to lodging. At the same time, it was noted that the trait varied over the years. Twelve varieties showed a low level of resistance in all years of evaluation. Plant height-based grouping of the varieties showed that 19 samples belonged to semi-dwarfs (60-84 cm), and the rest were included in the group of standard-height plants (85-100 cm). Quantitative trait loci (QTL) mapping was performed by means of genome-wide association study (GWAS) using 9285 SNP markers. For lodging resistance, plant height, and upper internode diameter, 26 signif icant associations (-log p > 3) were found in chromosomes 1B, 2A, 3A, 3D, 4A, 5A, 5B, 5D, 6A, and 7B. The results obtained suggest that the regions of 700-711 and 597-618 Mb in chromosomes 3A and 6A, respectively, may contain clusters of genes that affect lodging resistance and plant height. No chromosome regions colocalized with the QTLs as sociated with lodging resistance or upper internode diameter were found. The present GWAS results may be important for the development of approaches for creating lodging-resistant varieties through marker-assisted and genomic selection.

[The Influence of Adiponectin on Production of Apolipoproteins A-l and E by Human Macrophages].

Adiponectin is an adipose tissue hormone affecting energy and lipoprotein metabolism and modulating inflammatory responses. However, the role of this adipokine in atherogenesis remains poorly understood. The aim of this study was to investigate the effect of adiponectin on the production of apolipoproteins (apo) A-l and E by human macrophages (MP). The study was conducted on macrophage-like cells of the THP-1 cell line of two differentiation terms, 3 and 5 days (3d and 5d). Adiponectin (10 µg/mL) stimulated the expression of apoA-1 gene at the mRNA level in 5d MP, but not in 3d MP. The level of apoE mRNA in MP under the action of adiponectin was not affected. Adiponectin suppressed macrophage TNF gene expression, while it induced the expression of IL-10 gene in 5d MP. The secreted levels of apoA-1 and apoE proteins under the action of adiponectin in macrophages of both periods of differentiation remained unchanged, while the level of the surface apoA-1 protein in 5d MP was decreasing. Incubation of 5d MP with the PPARα nuclear receptor antagonist MK-886 or with the nuclear receptor LXR agonist TO-901317 resulted in cancellation of the stimulating effect of adiponectin on apoA-1 gene expression. These data indicate that adiponectin, in addition to its anti-inflammatory action, has a modulating effect on production of apoA-1 by macrophages. The latter is probably one of the mechanisms of the influence of this adipokine on atherogenesis.

Neuroprotective and Antioxidant Activity of Arachidonoyl Choline, Its Bis-Quaternized Analogues and Other Acylcholines.

The antioxidant activity and protective effect in the toxicity model of H2O2 were studied for arachidonic (AA-CHOL), docosahexaenoic (DHA-CHOL), linoleic (Ln-CHOL), and oleic (Ol-CHOL) fatty acids, as well as arachidonoyl dicholine (AA-diCHOL) and O-arachidonoyl bistetramethylaminoisopropanol (ABTAP). AA-CHOL, DHA-CHOL and Ln-CHOL provided a 20% increase in cell survival. AA-CHOL, AA-diCHOL, Ol-CHOL, and ABTAP had a radical-scavenging effect in the ABTS test, approximately equal to the activity of a standard radical scavenger Trolox.

[Lodging in wheat: genetic and environmental factors and ways of overcoming]. / Полегание пшеницы: генетические и экологические факторы и способы преодоления.

Lodging is one of the main factors in reducing the yield and grain quality of winter and spring wheat varieties. The resistance of wheat cultivars to lodging largely depends on environmental factors, biological and morphological features of the stem and root systems. Selection of the varieties for resistance to lodging is relevant in many countries of the world and has a number of achievements. Plant height is one of the most important morphological characters associated with lodging resistance. Breeding of the varieties carrying the dwarfing genes (Rht) is the main direction to reduce the risk of lodging. The Rht-B1b, Rht-D1b, Rht8 and Rht11 genes are widely used throughout the world due to their significant influence on agronomically valuable traits, including lodging. It turned out to be important to study the anatomical and morphological features and chemical composition of stem tissues, which complement the assessment of resistance to lodging and allow the varietal material to be more fully characterized. The thickness of stem internodes and their anatomical structure play an important role in the stem strength. The diameter of the stem, its thickness and weight, a large number of vascular bundles and a wide ring of mechanical tissues correlate with resistance to lodging. The content of lignin, silicon and cellulose are important structural components and provide the stem strength of wheat plants. Molecular genetic analysis and mapping of genes and quantitative trait loci are of great importance in identifying the genetic basis of the relationship between the anatomical and morphophysiological characters of the stem and root system and lodging. Genetic factors reflecting correlations between the lodging and the thickness of the stem wall, the number of vascular bundles and other characters were mapped to chromosomes 1A, 1B, 2A, 2D, 3A, 4B, 4D, 5A, 5D, 6D and 7D. It has been found that loci with high phenotypic effects on lodging tolerance are colocalized with loci responsible for plant height, stem diameter and stem strength. To increase resistance to lodging, it is necessary to develop a set of agrotechnical methods that reduce the influence of soil and climatic factors and create wheat varieties tolerant to lodging.

[Effect of lipid-lowering activity of the natural original enzyme preparation in the experiment]. / Éksperimental'nye dannye o gipolipidemicheskoi aktivnosti otechestvennogo fermentnogo preparata prirodnogo proiskhozhdeniia kholesterinoksidazy.

The experimental study in vivo was aimed at evaluation of hypolipidemic action of the original natural microbial enzyme preparation of cholesterol oxidase (CHO). In preliminary chronic experiments in rats, rabbits, dogs, low toxicity, good tolerability, and anti-atherosclerotic activity of the CHO preparation were established. To assess the effect of CHO under conditions of moderate, nutritional, atherogenic dyslipoproteinemia, experiments were carried out in rats, guinea pigs, and rabbits. It was shown that administration of CHO had the pronounced lipid-lowering effect in models of atherogenic dyslipoproteinemia induced in these animals.

Neuroprotective Action of Amidic Neurolipins in Models of Neurotoxicity on the Culture of Human Neural-Like Cells SH-SY5Y.

It was established that in neurodegeneration models in the human neuron-like cell line SH-SY5Y, amide derivatives of arachidonic and docosahexaenoic acids were inactive in experiments with MPP+ and CoCl2 but protected from H2O2. The protective activity of neurolipins decreased in the series DHA-DA > AA-SER ≥ AA-GLY > AA-GABA ≥ AA-EA and was manifested starting from a concentration of 0.5 nM.

[The optimization of the nitric oxide quantitative analysis for its determination in the cultural medium of mammalian cell culture].

The protocol for the quantitative analysis of nitric oxide as nitrite-ion suitable for determination of its production by a mammalian cell culture was developed. The optimal results were obtained using microvolume-adjusted Griess method after the preliminary reduction of NO3- to NO2- with non-activated cadmium. The protocol was verified on a rat glioma C6 cell culture. The developed method may be used for the nitric oxide determination in 96-well and 48-well microplates; the detection limit is 2.1 ± 0.1 µM for NO2- and 2.9 ± 0.1 µM for NO3-.

[Successful anaesthesia for abdominal reconstructive surgery in a myasthenia gravis patient].

This article presents a clinical case of colon disease surgical treatment in a 34-year old patient with. generalized myasthenia. Perioperative management peculiarities in these patients are clarified. Different approaches to anaesthesia choice were discussed on a case study. The importance of tactics individualization, rational drugs selection, including neuromuscular block reversal agents as well as intraoperative neuromuscular transmission monitoring.

[Nanocomplexes of recombinant proteins and polysialic acid: preparation, characteristics, and biological activity].

A preparation of nanocomplexes containing recombinant proteins (interferons alpha2b and beta1b, insulin, and human granulocyte colony stimulating factor) and natural polysialic acid (PSA) has been described. The incorporation of protein into the complex changes its electrophoretic mobility. Atomic force microscopy reveals the average size of 23-kD insulin complexes with PSA of 10-20 nm and demonstrates that more than 60% of glycopolymer molecules carry a single protein molecule. Experiments with cultured cells show that cytokines bound to polysialic acid retain their ability to regulate cell proliferation. Insulin bound to PSA has a prolonged hypoglycemic effect in vivo.

[Modification of recombinant proteins by covalent polysialation illustrated with the example of human insulin].

Methods of selective and nonselective covalent immobilization of genetically engineered proteins on molecules of natural polysialic acid are described by the example of human insulin. Such modification increases insulin lifetime in vivo.

Hydrolysis of anandamide and eicosapentaenoic acid ethanolamide in mouse splenocytes.

The hydrolysis of anandamide has been studied in mouse splenocytes using tritiated anandamide analogs labeled in the acyl- or ethanolamide parts of the molecule. [3H]Anandamide undergoes rapid (t(1/2) = 2.5 min) uptake and hydrolysis, yielding ethanolamine and arachidonic acid. The anandamide hydrolysis in splenocytes is sensitive to inhibition by phenylmethylsulfonyl fluoride, and it is assumed that the observed activity is due to fatty acid amide hydrolase, which inactivates anandamide in central and peripheral tissues. Eicosapentaenoic acid ethanolamide and the 15-hydroxy-derivative of anandamide are shown to be amidohydrolase substrates as well. The fatty acids derived from hydrolytic cleavage of acylethanolamines undergo rapid oxidation by splenocyte lipoxygenase, yielding the corresponding 12-hydroxy-derivatives. Oxygenated ethanolamide derivatives were not found. The data suggest that polyenoic fatty acid ethanolamides are metabolic precursors of eicosanoids in splenocytes and that amide bond hydrolysis is the key point in switching of biological activity spectra between endocannabinoids and oxylipins.

[Lipoxygenase oxidation of arachidonic acid in murine splenocytes and its modulation by the lactone ganglioside GM3]. / Lipoksigenaznoe okislenie arakhidonovoi kisloty v splenotsitakh myshei i ego modulirovanie laktonom gangliozida GM3.

The main arachidonic acid metabolites released into the medium by mouse splenocytes have been identified on the basis of chromatographic and spectral studies as well as by mass spectrometry of the derivatives. In the absence or presence of exogenous arachidonic acid mouse splenocytes produce mainly 12-hydroxy-5,8,10,14-eicosatetraenoic and 12,20-dihydroxy-5,8,10,14-eicosatetraenoic acids. Both products are constantly released by intact cells into surrounding media without stimulation by exogenous substrate or other modulators. For the first time it is shown that exogenously added ganglioside GM3 lactone as well as ganglioside GM3 itself can influence the arachidonic acid metabolism in splenocytes.

[Gangliosides modulate lipoxygenase oxidation in human lymphocytes]. / Gangliozidy moduliruiut lipoxsigenaznoe okislenie v limfotsitakh cheloveka.

Using reverse phase high performance chromatography with UV-detection, the arachidonic acid cascade in human peripheral blood lymphocytes (PBL) was studied. It was found that PBL oxidized arachidonic acid via the lipoxygenase pathway, 12-hydroxyeicosatetraenoic acid (12-HETE) being the major metabolite of endogenous arachidonic acid. Exogenous arachidonic acid added to human PBL suspensions increased 12-HETE synthesis 5-7 times. In another experimental series the effects of gangliosides (GD3, GM1 and GM3) on lipoxygenase-catalyzed oxidation of arachidonic acid in human lymphocytes were investigated. All the gangliosides tested stimulated PBL to secrete 12-HETE both from endogenous and exogenous arachidonic acid. In most cases the stimulating effect of GD3 was much more apparent that those of GM1 and GM3.

[Glycosphingolipids from human T-lymphoma MOLT-4 cells]. / Glikosfingolipidy kletok T-limfomy MOLT-4 cheloveka.

The glycosphingolipids of human lymphoma MOLT-4 cells were studied, using biochemical methods and specific antisera to gangliosides. The major neutral glycosphingolipids were found to be glucosyl- and lactosyl ceramides. GM3, GM2, GM1 and GD1a were identified as ganglioside components.

[Neutral glycosphingolipids of murine lymphoma EL-4]. / Neitral'nye glikosfingolipidy limfomy EL-4 myshei.

Neutral glycosphingolipids of murine T-lymphoma EL-4 were studied. The major glycolipid components were identified as GlcCer, LacCer, GgOse3Cer and GgOse4Cer. It has been shown for the first time that not only gangliosides but also neutral glycolipids are shed from the cell surface into the outer medium.

[Comparative study of gangliosides from murine B- and T-lymphomas]. / Sravintel'noe izuchenie gangliozidov B- i T-limfom myshei.

Gangliosides from murine B-lymphomas (MOPC 21 and MOPC 406) and T-lymphoma EL-4 were studied by thin-layer chromatography, immunoprecipitation with specific antisera to gangliosides and by treatment with neuraminidase. It was found that the gangliosides of all three lymphomas differ in their interaction with antisera and neuraminidase although they are similar in their chromatographic behaviour.