Au-Based Nanoparticles Enhance Low Temperature Tolerance in Wheat by Regulating Some Physiological Parameters and Gene Expression.

One of the key problems of biology is how plants adapt to unfavorable conditions, such as low temperatures. A special focus is placed on finding ways to increase tolerance in important agricultural crops like wheat. Au-based nanoparticles (Au-NPs) have been employed extensively in this area in recent years. Au-NPs can be produced fast and easily using low-cost chemical reagents. When employed in microdoses, Au-NPs are often non-toxic to plants, animals, and people. In addition, Au-NPs mainly have favorable impacts on plants. In this study, we investigated the effect of Au-NP seed nanopriming (diameter 15.3 nm, Au concentration 5-50 µg mL-1) on cold tolerance, as well as some physiological, biochemical and molecular parameters, of cold-sustainable wheat (Triticum aestivum L.) genotype Zlata. The treatment with Au-NPs improved tolerance to low temperatures in control conditions and after cold hardening. Au-NPs treatment boosted the intensity of growth processes, the quantity of photosynthetic pigments, sucrose in leaves, and the expressions of encoded RuBisCo and Wcor15 genes. The potential mechanisms of Au-NPs' influence on the cold tolerance of wheat varieties were considered.

Effect of Low Temperature on Content of Primary Metabolites in Two Wheat Genotypes Differing in Cold Tolerance.

The study of cold-tolerance mechanisms of wheat as a leading cereal crop is very relevant to science. Primary metabolites play an important role in the formation of increased cold tolerance. The aim of this research is to define changes in the content of primary metabolites (soluble proteins and sugars), growth, and photosynthetic apparatus of freezing-tolerant and cold-sustainable wheat (Triticum aestivum L.) genotypes under optimal conditions and after prolonged (7 days) exposure to low temperature (4 °C). In order to gain a deeper comprehension of the mechanisms behind wheat genotypes' adaptation to cold, we determined the expression levels of photosynthetic genes (RbcS, RbcL) and genes encoding cold-regulated proteins (Wcor726, CBF14). The results indicated different cold-adaptation strategies of freezing-tolerant and cold-sustainable wheat genotypes, with soluble proteins and sugars playing a significant role in this process. In plants of freezing-tolerant genotypes, the strategy of adaptation to low temperature was aimed at increasing the content of soluble proteins and modification of carbohydrate metabolism. The accumulation of sugars was not observed in wheat of cold-sustainable genotypes during chilling, but a high content of soluble proteins was maintained both under optimal conditions and after cold exposure. The adaptation strategies of wheat genotypes differing in cold tolerance were related to the expression of photosynthetic genes and genes encoding cold-regulated proteins. The data improve our knowledge of physiological and biochemical mechanisms of wheat cold adaptation.

Human RPF1 and ESF1 in Pre-rRNA Processing and the Assembly of Pre-Ribosomal Particles: A Functional Study.

Ribosome biogenesis is essential for the functioning of living cells. In higher eukaryotes, this multistep process is tightly controlled and involves a variety of specialized proteins and RNAs. This pool of so-called ribosome biogenesis factors includes diverse proteins with enzymatic and structural functions. Some of them have homologs in yeast S. cerevisiae, and their function can be inferred from the structural and biochemical data obtained for the yeast counterparts. The functions of human proteins RPF1 and ESF1 remain largely unclear, although RPF1 has been recently shown to participate in 60S biogenesis. Both proteins have drawn our attention since they contribute to the early stages of ribosome biogenesis, which are far less studied than the later stages. In this study, we employed the loss-of-function shRNA/siRNA-based approach to the human cell line HEK293 to determine the role of RPF1 and ESF1 in ribosome biogenesis. Downregulating RPF1 and ESF1 significantly changed the pattern of RNA products derived from 47S pre-rRNA. Our findings demonstrate that RPF1 and ESF1 are associated with different pre-ribosomal particles, pre-60S, and pre-40S particles, respectively. Our results allow for speculation about the particular steps of pre-rRNA processing, which highly rely on the RPF1 and ESF1 functions. We suggest that both factors are not directly involved in pre-rRNA cleavage but rather help pre-rRNA to acquire the conformation favoring its cleavage.

Human nucleolar protein SURF6/RRP14 participates in early steps of pre-rRNA processing.

The biogenesis of ribosomes requires tightly controlled transcription and processing of pre-rRNA which comprises ribosomal RNAs forming the core of large and small ribosomal subunits. Early steps of the pre-rRNA processing and assembly of the ribosomal subunits require a large set of proteins that perform folding and nucleolytic cleavage of pre-rRNAs in the nucleoli. Structure and functions of proteins involved in the pre-rRNA processing have been extensively studied in the budding yeast S. cerevisiae. Functional characterization of their human homologues is complicated by the complexity of mammalian ribosomes and increased number of protein factors involved in the ribosomal biogenesis. Homologues of human nucleolar protein SURF6 from yeast and mouse, Rrp14 and Surf6, respectively, had been shown to be involved in the early steps of pre-rRNA processing. Rrp14 works as RNA chaperone in complex with proteins Ssf1 and Rrp15. Human SURF6 knockdown and overexpression were used to clarify a role of SURF6 in the early steps of pre-rRNA processing in human cell lines HeLa and HTC116. By analyzing the abundance of the rRNA precursors in cells with decreased level or overexpression of SURF6, we demonstrated that human SURF6 is involved in the maturation of rRNAs from both small and large ribosomal subunits. Changes in the SURF6 level caused by knockdown or overexpression of the protein do not result in the death of HeLa cells in contrast to murine embryonic fibroblasts, but significantly alter the distribution of cells among the phases of the cell cycle. SURF6 knockdown in both p53 sufficient and p53 deficient HCT116 human cancer cells results in elongation of G0/G1 and shortening of G2/M phase. This surprising result suggests p53 independence of SURF6 effects on the cell cycle and possible multiple functions of SURF6. Our data point to the shift from pathway 1 to pathway 2 of the rRNA biogenesis caused by the SURF6 knockdown and its likely association with p53 pathway.

Adaptive strategy of plant cells during chilling: Aspect of ultrastructural reorganization.

The review is focused on a comparative analysis of the literature data on the ultrastructural reorganization of leaf cells of higher plants, which differ in their response to low sub-damaging temperatures. The importance of adaptive structural reorganization of cells as a special feature contributing to the surviving strategy of plants existing under changed conditions is emphasized. The adaptive strategy of cold-tolerant plants combines the structural, functional, metabolic, physiological and biochemical reorganization of cells and tissues. These changes constitute a unified program directed to protecting against dehydration and oxidative stress, as well as maintaining basic physiological processes, and above all, photosynthesis. The ultrastructural markers of cold-tolerant plants adaptation to low sub-damaging temperatures include some particular changes in cell morphology. Namely: the following: an increase in the volume of the cytoplasm; the formation of new membrane elements in it; an increase in the size and number of chloroplasts and mitochondria; concentration of mitochondria and peroxisomes near chloroplasts; polymorphism of mitochondria; an increase in the number of cristae in them; the appearance of outgrowths and invaginations in chloroplasts; lumen expansion in the thylakoids; the formation in chloroplasts "sun type" membrane system with reduction in the number and size of grana and domination of non-appressed thylakoids membranes. Due to this adaptive structural reorganization cold-tolerant plants are able to function actively during chilling. On the contrary, structural reorganization of leaf cells of cold-sensitive plants under chilling is aimed at maintaining the basic functions at a minimum level. Cold-sensitive plants "wait out" low temperature stress, and with prolonged exposure to cold, they die from dehydration and intensification of oxidative stress.

Priming with gold nanoparticles leads to changes in the photosynthetic apparatus and improves the cold tolerance of wheat.

Nanotechnologies provide a great platform for researching nanoparticles effects on living organisms including plants. This work shows the stimulating effect of seed priming with gold nanoparticles (AuNPs) on photosynthetic apparatus of Triticum aestivum seedlings. It was found using inductively coupled plasma-atomic emission and mass spectrometry that AuNPs (the average diameter of 15.3 nm, concentration of 20 µg ml-1) penetrated into the seeds, but were not found in seedling leaves. Ultrastructural changes in chloroplasts were found using transmission electron microscopy in plants grown from treated seeds: increases in the size of plastids, starch grains, grana in chloroplasts, and the number of thylakoids in grana. The intensity of photosynthesis, the content of chlorophylls, and the portion of unsaturated fatty acids in the composition of total leaf lipids were increased in treated AuNPs plants. This study demonstrates that revealed changes determined the increased tolerance of wheat to low temperature. The adaptive significance of these changes, possible mechanisms of the AuNPs effects on plants and future perspectives of study are discussed. This is the first report showing nanopriming with AuNPs as a new method to study the mechanisms of stress tolerance.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)-Specific T Cells and Antibodies in Coronavirus Disease 2019 (COVID-19) Protection: A Prospective Study.

BACKGROUND: During the ongoing coronavirus disease 2019 (COVID-19) pandemic, many individuals were infected with and have cleared the virus, developing virus-specific antibodies and effector/memory T cells. An important unanswered question is what levels of T-cell and antibody responses are sufficient to protect from the infection. METHODS: In 5340 Moscow residents, we evaluated anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin M (IgM)/immunoglobulin G (IgG) titers and frequencies of the T cells specific to the membrane, nucleocapsid, and spike proteins of SARS-CoV-2, using interferon gamma (IFN-γ) enzyme-linked immunosorbent spot (ELISpot) assay. Additionally, we evaluated the fractions of virus-specific CD4+ and CD8+ T cells using intracellular staining of IFN-γ and interleukin 2 followed by flow cytometry. We analyzed the COVID-19 rates as a function of the assessed antibody and T-cell responses, using the Kaplan-Meier estimator method, for up to 300 days postinclusion. RESULTS: We showed that T-cell and antibody responses are closely interconnected and are commonly induced concurrently. Magnitudes of both responses inversely correlated with infection probability. Individuals positive for both responses demonstrated the highest levels of protectivity against the SARS-CoV-2 infection. A comparable level of protection was found in individuals with antibody response only, whereas the T-cell response by itself granted only intermediate protection. CONCLUSIONS: We found that the contribution of the virus-specific antibodies to protection against SARS-CoV-2 infection is more pronounced than that of the T cells. The data on the virus-specific IgG titers may be instructive for making decisions in personalized healthcare and public anti-COVID-19 policies. Clinical Trials Registration. NCT04898140.

Gold nanoparticles as adaptogens increazing the freezing tolerance of wheat seedlings.

The intensive development of nanotechnology led to the widespread application of various nanoparticles and nanomaterials. As a result, nanoparticles enter the environment and accumulate in ecosystems and living organisms. The consequences of possible impact of nanoparticles on living organisms are not obvious. Experimental data indicate that nanoparticles have both toxic and stimulating effects on organisms. In this study, we demonstrated for the first time that gold nanoparticles can act as adaptogens increasing plant freezing tolerance. Priming winter wheat (Triticum aestivum L., var. Moskovskaya 39, Poaceae) seeds for 1 day in solutions of gold nanoparticles (15-nm diameter, concentrations of 5, 10, 20, and 50 µg/ml) led to an increase in freezing tolerance of 7-day-old wheat seedlings. A relationship between an increase in wheat freezing tolerance and changes in some important indicators for its formation-growth intensity, the activity of the photosynthetic apparatus and oxidative processes, and the accumulation of soluble sugars in seedlings-was established. Assumptions on possible mechanisms of gold nanoparticles effects on plant freezing tolerance are discussed.