The influence of Au-based nanoparticles on some physiological, biochemical and molecular characteristics of wheat plants during low temperature hardening.

One of the most significant problems of the 21st century is the anthropogenic strain on the environment. The development of nanotechnology makes it possible to produce a variety of nanomaterials widely used in people's daily lives. However, nanomaterials can accumulate in ecosystems and spread through food chains. The environmental risks of nanoparticle proliferation are unclear. At the same time, certain nanoparticles act as adaptogens, improving plant tolerance to unfavorable stress factors. It is quite realistic to choose such experimental conditions, under which the effect on plant stress tolerance will be obvious and the accumulation of nanoparticles in tissues will be minimal. In this case, the main relevant factors are the type of nanoparticles, their concentration and their way of penetration into plants. We chose to study gold nanoparticles (Au-NPs), widely used in biomedical research. The concentration of Au-NPs was 20 µg/mL, which is considered safe for living organisms. The influence of Au-NPs on some physiological, biochemical and molecular characteristics of wheat plants during low temperature hardening was examined. The study of the photosynthetic apparatus and antioxidant system was the primary focus. The stimulating effect of Au-NPs on cold tolerance of wheat plants was shown. The results expand our knowledge of the processes by which nanoparticles impact plants and the potential applications of nanoparticles as adaptogens in science and agriculture.

Nanomaterials in plant physiology: Main effects in normal and under temperature stress.

Global climate change and high population growth rates lead to problems of food security and environmental pollution, which require new effective methods to increase yields and stress tolerance of important crops. Nowadays the question of using artificial chemicals is very relevant in theoretical and practical terms. It is important that such substances in low concentrations protect plants under stress conditions, but at the same time inflict minimal damage on the environment and human health. Nanotechnology, which allows the production of a wide range of nanomaterials (NM), provides novel techniques in this direction. NM include structures less than 100 nm. The review presents data on the methods of NM production, their properties, pathways for arrival in plants and their use in human life. It is shown that NM, due to their unique physical and chemical properties, can cross biological barriers and accumulate in cells of live organisms. The influence of NM on plant organism can be both positive and negative, depending on the NM chemical nature, their size and dose, the object of study, and the environmental conditions. This review provides a comparative analysis of the effect of artificial metal nanoparticles (NPm), the commonly employed NMs in plant physiology, on two important aspects of plant life: photosynthetic apparatus activity and antioxidant system function. According to studies, NM affect not only the functional activity of photosynthetic apparatus, but also structural organization of chloroplats. In addition, the literature analysis reflects the dual action of NM on oxidative processes, and antioxidant status of plants. These facts considerably complicate the ideas about possible mechanisms and further use of NPm in biology. In this regard, data on the effects of NM on plants under abiotic stressors are of great interest. Separate section is devoted to the use of NM as adaptogens that increase plant stress tolerance to unfavorable temperatures. Possible mechanisms of NM effects on plants are discussed, as well as the strategies for their further use in basic science and sustainable agriculture.

The immunostimulatory roles of gold nanoparticles in immunization and vaccination against Brucella abortus antigens.

One effective antigen carrier proposed for use in immunization and vaccination is gold nanoparticles. Prior work has shown that gold nanoparticles themselves have adjuvant properties. Currently, gold nanoparticles are used to design new diagnostic tests and vaccines against viral, bacterial, and parasitic infections. We investigated the use of gold nanoparticles as immunomodulators in immunization and vaccination with an antigen isolated from Brucella abortus. Gold nanoparticles with a diameter of 15 nm were synthesized for immunization of animals and were then conjugated to the isolated antigen. The conjugates were used to immunize white BALB/c mice. As a result, high-titer (1:10240) antibodies were produced. The respiratory and proliferative activities of immune cells were increased, as were the serum interleukin concentrations. The minimum antigen amount detected with the produced antibodies was âˆ¼ 0.5 pg. The mice immunized with gold nanoparticles complexed with the B. abortus antigen were more resistant to B. abortus strain 82 than were the mice immunized through other schemes. This fact indicates that animal immunization with this conjugate enhances the effectiveness of the immune response. The results of this study are expected to be used in further work to examine the protective effect of gold nanoparticles complexed with the B. abortus antigen on immunized animals and to develop test systems for diagnosing brucellosis in the laboratory and in the field.

Quantitative intra- and intergeneric taxonomic relationships among Micrococcaceae strains reveal contradictions in the historical assignments of the strains and indicate the need for species reclassification.

This article reports the results of quantitative intra- and intergeneric taxonomic relationships among Micrococcaceae strains and a novel endophytic bacterium (SG) isolated from a suspension culture of Arabidopsis thaliana (L.) Heynh in our laboratory. The known strain Rothia sp. ND6WE1A was used as a reference one for SG. Whole-genome sequencing and phylogenetic analysis were based on the 16S rRNA test. Quantitative analysis for the nucleotide identity (ANI) and calculation of evolutionary distances were based on the identified amino acids (AAI) test indicating the generic assignment of the reference strain within and between the identified monophyletic groups of Micrococcaceae. The amino acid data structure of Rothia sp. ND6WE1A was compared against the UniProt database (250 million records) of close lineage of Micrococcaceae, including other Rothia spp. These data presented unique and evolutionary amino acid alignments, eventually expected in the new SG isolate as well. The metagenomic entries of the respective genome and proteome, characterized at the genus and species levels, could be considered for evolutionary taxonomic reclassification of the isolated and the reference strain (SG + Rothia sp. ND6WE1A). Therefore, our results warrant further investigations on the isolated SG strain.

Antibody Phage Display Technology for Sensor-Based Virus Detection: Current Status and Future Prospects.

Viruses are widespread in the environment, and many of them are major pathogens of serious plant, animal, and human diseases. The risk of pathogenicity, together with the capacity for constant mutation, emphasizes the need for measures to rapidly detect viruses. The need for highly sensitive bioanalytical methods to diagnose and monitor socially significant viral diseases has increased in the past few years. This is due, on the one hand, to the increased incidence of viral diseases in general (including the unprecedented spread of a new coronavirus infection, SARS-CoV-2), and, on the other hand, to the need to overcome the limitations of modern biomedical diagnostic methods. Phage display technology antibodies as nano-bio-engineered macromolecules can be used for sensor-based virus detection. This review analyzes the commonly used virus detection methods and approaches and shows the prospects for the use of antibodies prepared by phage display technology as sensing elements for sensor-based virus detection.

Recombinant antibodies by phage display for bioanalytical applications.

Antibody phage display, aimed at preparing antibodies to defined antigens, is a useful replacement for hybridoma technology. The phage system replaces all work stages that follow animal immunization with simple procedures for manipulating DNA and bacteria. It enables the time needed to generate stable antibody-producing clones to be shortened considerably, making the process noticeably cheaper. Antibodies prepared by phage display undergo several affinity selection steps and can be used as selective receptors in biosensors. This article briefly describes the techniques used in the making of phage antibodies to various antigens. The possibilities and prospects are discussed of using phage antibodies as selective agents in analytical systems, including biosensors.

Immunization of Mice with Gold Nanoparticles Conjugated to Thermostable Cancer Antigens Prevents the Development of Xenografted Tumors.

Gold nanoparticles as part of vaccines greatly increase antigen stability, antigen accumulation in the lymph nodes, and antigen uptake by antigen-presenting cells. The use of such particles as part of anticancer vaccines based on heat shock proteins to increase vaccine effectiveness is timely. We prepared and characterized nanoconjugates based on 15-nm gold nanoparticles and thermostable tumor antigens isolated from MH22a murine hepatoma cells. The whole-cell lysate of MH22a cells contained the main heat shock proteins. BALB/c mice were injected with the conjugates and then received transplants of MH22a cells. The highest titer was produced in mice immunized with the complex of gold nanoparticles + antigen with complete Freund's adjuvant. The immunized mice showed no signs of tumor growth for 24 days. They also showed a decreased production of the INF-γ, IL-6, and IL-1 proinflammatory cytokines compared to the mice immunized through other schemes. This study is the first to show that it is possible in principle to use gold nanoparticles in combination with thermostable tumor antigens for antitumor vaccination. Antitumor vaccines based on thermostable tumor antigens can be largely improved by including gold nanoparticles as additional adjuvants.

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.

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.

Synthesis of Silymarin-Gold Nanoparticle Conjugate and Analysis of its Liver-Protecting Activity.

BACKGROUND: The liver disease problem prompts investigators to search for new methods of liver treatment. INTRODUCTION: Silymarin (Sil) protects the liver by reducing the concentration of free radicals and the extent of damage to the cell membranes. A particularly interesting method to increase the bioavailability of Sil is to use synthesized gold nanoparticles (AuNPs) as reagents. The study considered whether it was possible to use the silymarin-AuNP conjugate as a potential liver-protecting drug. METHODS: AuNPs were conjugated to Sil and the liver-protecting activity of the conjugate was examined. Experimental hepatitis and hepatocyte cytolysis after carbon tetrachloride action were used as a model system, and the experiments were conducted with laboratory animals. RESULTS: For the first time, silymarin was conjugated to colloidal gold nanoparticles (AuNPs). Electron microscopy showed that the resultant preparations were monodisperse and that the mean conjugate diameter was 18-30 nm ± 0.5 nm (mean diameter of the native nanoparticles, 15 ± 0.5 nm). In experimental hepatitis in mice, conjugate administration interfered with glutathione depletion in hepatocytes in response to carbon tetrachloride. It also was conducive to an increase in energy metabolism and stimulated the monocyte-macrophage function of the liver. The results were confirmed by the high respiratory activity of the hepatocytes in cell culture. CONCLUSION: We conclude that the silymarin-AuNP conjugate holds promise as a liver-protecting agent in acute liver disease caused by carbon tetrachloride poisoning.

Synthesis of silymarin-selenium nanoparticle conjugate and examination of its biological activity in vitro.

Silymarin (Sil) was conjugated to selenium nanoparticles (SeNPs) to increase Sil bioavailability. The conjugates were monodisperse; the average diameter of the native SeNPs was ~ 20-50 ± 1.5 nm, whereas that of the conjugates was 30-50 ± 0.5 nm. The use of SeNPs to increase the bioavailability of Sil was examined with the MH-22a, EPNT-5, HeLa, Hep-2, and SPEV-2 cell lines. The EPNT-5 (glioblastoma) cells were the most sensitive to the conjugates compared to the conjugate-free control. The conjugates increased the activity of cellular dehydrogenases and promoted the penetration of Sil into the intracellular space. Possibly, SeNPs play the main part in Sil penetration of cells and Sil penetration is not associated with phagocytosis. Thus, SeNPs are promising for use as a Sil carrier and as protective antigens.

Gold nanoparticles for preparation of antibodies and vaccines against infectious diseases.

INTRODUCTION: Vaccination remains very effective in stimulating protective immune responses against infections. An important task in antibody and vaccine preparation is to choose an optimal carrier that will ensure a high immune response. Particularly promising in this regard are nanoscale particle carriers. An antigen that is adsorbed or encapsulated by nanoparticles can be used as an adjuvant to optimize the immune response during vaccination. a very popular antigen carrier used for immunization and vaccination is gold nanoparticles, with are being used to make new vaccines against viral, bacterial, and parasitic infections. AREAS COVERED: This review summarizes what is currently known about the use of gold nanoparticles as an antigen carrier and adjuvant to prepare antibodies in vivo and design vaccines against viral, bacterial, and parasitic infections. The basic principles, recent advances, and current problems in the use of gold nanoparticles are discussed. EXPERT OPINION: Gold nanoparticles can be used as adjuvants to increase the effectiveness of vaccines by stimulating antigen-presenting cells and ensuring controlled antigen release. Studying the characteristics of the immune response obtained from the use of gold nanoparticles as a carrier and an adjuvant will permit the particles' potential for vaccine design to be increased.

Preparation and in vivo evaluation of glyco-gold nanoparticles carrying synthetic mycobacterial hexaarabinofuranoside.

A number of bacterial glycans are specific markers for the detection and the serological identification of microorganisms and are also widely used as antigenic components of vaccines. The use of gold nanoparticles as carriers for glyco-epitopes is becoming an important alternative to the traditional conjugation with proteins and synthetic polymers. In this study, we aimed to prepare and evaluate in vivo glyco-gold nanoparticles (glyco-GNPs) bearing the terminal-branched hexaarabinofuranoside fragment (Ara6) of arabinan domains of lipoarabinomannan and arabinogalactan, which are principal polysaccharides of the cell wall of Mycobacterium tuberculosis, the causative agent of tuberculosis. In particular, we were interested whether the antibodies generated against Ara6-GNPs would recognize the natural saccharides on the cell surface of different mycobacterial strains. Two synthetic Ara6 glycosides with amino-functionalized spacer aglycons differing in length and hydrophilicity were directly conjugated with spherical gold nanoparticles (d = 15 nm) to give two sets of glyco-GNPs, which were used for the immunization of rabbits. Dot assays revealed cross-reactions between the two obtained antisera with the hexaarabinofuranoside and the 2-aminoethyl aglycon used for the preparation of glyco-GNPs. Both antisera contained high titers of antibodies specific for Mycobacteria as shown by enzyme-linked immunosorbent assay using M. bovis and M. smegmatis cells as antigens while there was only a weak response to M. phlei cells and no interaction with E. coli cells. The results obtained suggest that glyco-GNPs are promising agents for the generation of anti-mycobacterial antibodies.

Amplification of speckle-microscope signal by using gold nanoparticle stable of contents.

Speckle-microscopy is used for the detection of Chlamydia trachomatis in aqueous suspensions or on slides. Monoclonal antibody tagged with gold nanoparticles that form biospeckles have also been used with a small number of scatterers. By devising a prototype of laser scanning speckle-microscope, we now demonstrate that the signal of speckle microscope can be significantly amplified in the presence of gold nanoparticles. This enhancement of signal emitted from gold nanoparticles offers a better approach to the detection of C. trachomatis.

Highly active extracellular α-class carbonic anhydrase of Cyanothece sp. ATCC 51142.

Here, for the first time, we report the presence of highly active extracellular carbonic anhydrase (CA) of α-class in cyanobacterial cells. The enzyme activity was confirmed both in vivo in intact cells and in vitro, using the recombinant protein. CA activity in intact cells of Cyanothece sp. ATCC 51142 reached ∼0.6 Wilbur-Anderson units (WAU) per 1 mg of total cell protein, and it was inhibited by a specific CAs inhibitor, ethoxyzolamide. The genes cce_4328 (ecaA) and cce_0871 (ecaB), encoding two potential extracellular CAs of Cyanothece have been cloned, and the corresponding proteins EcaA and EcaB, representing CAs of α- and ß-class, respectively, have been heterologously expressed in Escherichia coli. High specific activity (∼1.1 × 104 WAU per 1 mg of target protein) was detected for the recombinant EcaA only. The presence of EcaA in the outer cellular layers of Cyanothece was confirmed by immunological analysis with antibodies raised against the recombinant protein. The absence of redox regulation of EcaA activity indicates that this protein does not possess a disulfide bond essential for some α-class CAs. The content and activity of EcaA in a fraction of periplasmic proteins was higher in Cyanothece cells grown at ambient concentration of CO2 (0.04%) compared to those grown at an elevated CO2 concentration (1.7%). At the same time, the level of ecaA gene mRNA varied insignificantly in response to changes in CO2 supply. Our results indicate that EcaA is responsible for CA activity of intact Cyanothece cells and point to its possible physiological role under low-CO2 conditions.

Prospects for the use of spherical gold nanoparticles in immunization.

Recent years have seen extremely fast development of new viral nanovaccines and diagnostic agents using nanostructures prepared by biological and chemical synthesis. We used spherical gold nanoparticles (average diameter, 15 nm) as a platform for the antigen for swine transmissible gastroenteritis virus (TGEV). The literature data demonstrate that immunization of animals with the TGEV antigen coupled to gold nanoparticles (GNPs) not only activates antigen-presenting cells but also increases the proliferative activity of splenic lymphoid (antibody-forming) cells. The contents of γ-IFN, IL-1ß, and IL-6 in animals immunized with GNP-antigen conjugates were found to be higher than those in intact animals or in animals given the antigen alone. The increased concentration of IL-1ß in the immunized animals directly correlated with the activity of macrophages and stimulated B cells, which produce this cytokine when activated. The increased concentration of IL-6 indicates that the injected preparations are stimulatory to cellular immunity. Immunization with the TGEV antigen conjugated to GNPs as a carrier activates the respiratory activity of lymphoid cells and peritoneal macrophages, which is directly related to their transforming activity and to the activation of antibody generation. Furthermore, the use of this conjugate allows marked improvement of the structure of the animals' immune organs and restores the morphological-functional state of these organs. The microanatomical changes (increased number of follicles) indicate the activation of the B-dependent zone of the spleen and, consequently, the development of a humoral-type immunological reaction. The degradative processes observed in the animals immunized with TGEV antigen alone are evidence of weak resistance to pathogen attack. These results can be used to develop vaccines against this infection by employing TGEV antigen coupled to gold nanoparticles as a carrier.

Gold nanoparticles as an adjuvant: Influence of size, shape, and technique of combination with CpG on antibody production.

Gold nanoparticles (GNPs) are advantageous as an adjuvant in the design of effective vaccines and in the preparation of high-affinity antibodies to haptens and complete antigens. Another method of activating immunocompetent cells with colloidal gold is to conjugate GNPs with CpG oligodeoxynucleotides (ODNs). We examined how the size and shape of GNPs and various combinations of GNPs and CpG ODNs 1826 affect the immune response. When animals were injected with a model antigen (BSA) coupled to gold nanospheres (diameters, 15 and 50nm), nanorods, nanoshells, and nanostars, the titers of the resultant antibodies differed substantially. The antibody titers decreased in the sequence GNPs-50nm>GNPs-15nm>nanoshells>nanostars>nanorods>native BSA. We conclude that 50 and 15nm gold nanospheres are the optimal antigen carrier and adjuvant for immunization. The highest titer of anti-BSA antibodies was detected in the blood serum of mice immunized simultaneously with BSA-GNP and CpG-GNP conjugates.

Immunological properties of gold nanoparticles.

In the past decade, gold nanoparticles have attracted strong interest from the nanobiotechnological community owing to the significant progress made in robust and easy-to-make synthesis technologies, in surface functionalization, and in promising biomedical applications. These include bioimaging, gene diagnostics, analytical sensing, photothermal treatment of tumors, and targeted delivery of various biomolecular and chemical cargos. For the last-named application, gold nanoparticles should be properly fabricated to deliver the cargo into the targeted cells through effective endocytosis. In this review, we discuss recent progress in understanding the selective penetration of gold nanoparticles into immune cells. The interaction of gold nanoparticles with immune cell receptors is discussed. As distinct from other published reviews, we present a summary of the immunological properties of gold nanoparticles. This review also summarizes what is known about the application of gold nanoparticles as an antigen carrier and adjuvant in immunization for the preparation of antibodies in vivo. For each of the above topics, the basic principles, recent advances, and current challenges are discussed. Thus, this review presents a detailed analysis of data on interaction of gold nanoparticles with immune cells. Emphasis is placed on the systematization of data over production of antibodies by using gold nanoparticles and adjuvant properties of gold nanoparticles. Specifically, we start our discussion with current data on interaction of various gold nanoparticles with immune cells. The next section describes existing technologies to improve production of antibodies in vivo by using gold nanoparticles conjugated with specific ligands. Finally, we describe what is known about adjuvant properties of bare gold or functionalized nanoparticles. In the Conclusion section, we present a short summary of reported data and some challenges and perspectives.

Biosynthesis of nanoparticles of metals and metalloids by basidiomycetes. Preparation of gold nanoparticles by using purified fungal phenol oxidases.

The work shows the ability of cultured Basidiomycetes of different taxonomic groups-Lentinus edodes, Pleurotus ostreatus, Ganoderma lucidum, and Grifola frondosa-to recover gold, silver, selenium, and silicon, to elemental state with nanoparticles formation. It examines the effect of these metal and metalloid compounds on the parameters of growth and accumulation of biomass; the optimal cultivation conditions and concentrations of the studied ion-containing compounds for recovery of nanoparticles have been identified. Using the techniques of transmission electron microscopy, dynamic light scattering, X-ray fluorescence and X-ray phase analysis, the degrees of oxidation of the bioreduced elements, the ζ-potential of colloidal solutions uniformity, size, shape, and location of the nanoparticles in the culture fluid, as well as on the surface and the inside of filamentous hyphae have been determined. The study has found the part played by homogeneous chromatographically pure fungal phenol-oxidizing enzymes (laccases, tyrosinases, and Mn-peroxidases) in the recovery mechanism with formation of electrostatically stabilized colloidal solutions. A hypothetical mechanism of gold(III) reduction from HAuCl4 to gold(0) by phenol oxidases with gold nanoparticles formation of different shapes and sizes has been introduced.

Multifunctional gold-based nanocomposites for theranostics.

Although Au-particle potential in nanobiotechnology has been recognized for the last 15 years, new insights into the unique properties of multifunctional nanostructures have just recently started to emerge. Multifunctional gold-based nanocomposites combine multiple modalities to improve the efficacy of the therapeutic and diagnostic treatment of cancer and other socially significant diseases. This review is focused on multifunctional gold-based theranostic nanocomposites, which can be fabricated by three main routes. The first route is to create composite (or hybrid) nanoparticles, whose components enable diagnostic and therapeutic functions. The second route is based on smart bioconjugation techniques to functionalize gold nanoparticles with a set of different molecules, enabling them to perform targeting, diagnostic, and therapeutic functions in a single treatment procedure. Finally, the third route for multifunctionalized composite nanoparticles is a combination of the first two and involves additional functionalization of hybrid nanoparticles with several molecules possessing different theranostic modalities. This last class of multifunctionalized composites also includes fluorescent atomic clusters with multiple functionalities.