Ion Channels in Electrical Signaling in Higher Plants.

Electrical signals (ESs) appearing in plants under the action of various external factors play an important role in adaptation to changing environmental conditions. Generation of ES in higher plant cells is associated with activation of Ca2+, K+, and anion fluxes, as well as with changes in the activity of plasma membrane H+-ATPase. In the present review, molecular nature of the ion channels contributing to ESs transmission in higher plants is analyzed based on comparison of the data from molecular-genetic and electrophysiological studies. Based on such characteristics of ion channels as selectivity, activation mechanism, and intracellular and tissue localization, those ion channels that meet the requirements for potential participation in ES generation were selected from a wide variety of ion channels in higher plants. Analysis of the data of experimental studies performed on mutants with suppressed or enhanced expression of a certain channel gene revealed those channels whose activation contributes to ESs formation. The channels responsible for Ca2+ flux during generation of ESs include channels of the GLR family, for K+ flux - GORK, for anions - MSL. Consideration of the prospects of further studies suggests the need to combine electrophysiological and genetic approaches along with analysis of ion concentrations in intact plants within a single study.

The Morphological Parameters and Cytosolic pH of Cells of Root Zones in Tobacco Plants (Nicotiana tabacum L.): Nonlinear Effects of NaCl Concentrations.

Salinity impacts important processes in plants, reducing their yield. The effect of salinity on the cytosolic pH (pHcyt) has been little studied. In this research, we employed transgenic tobacco plants expressing the pH sensor Pt-GFP to investigate the alterations in pHcyt in cells across various root zones. Furthermore, we examined a wide spectrum of NaCl concentrations (ranging from 0 to 150 mM) and assessed morphological parameters and plant development. Our findings revealed a pattern of cytosolic acidification in cells across all root zones at lower NaCl concentrations (50, 100 mM). Interestingly, at 150 mM NaCl, pHcyt levels either increased or returned to normal, indicating a nonlinear effect of salinity on pHcyt. Most studied parameters related to development and morphology exhibited an inhibitory influence in response to NaCl. Notably, a nonlinear relationship was observed in the cell length within the elongation and differentiation zones. While cell elongation occurred at 50 and 100 mM NaCl, it was not evident at 150 mM NaCl. This suggests a complex interplay between stimulating and inhibitory effects of salinity, contributing to the nonlinear relationship observed between pHcyt, cell length, and NaCl concentration.

Controlled Formation of a Protein Corona Composed of Denatured BSA on Upconversion Nanoparticles Improves Their Colloidal Stability.

In the natural fluidic environment of a biological system, nanoparticles swiftly adsorb plasma proteins on their surface forming a "protein corona", which profoundly and often adversely affects their residence in the systemic circulation in vivo and their interaction with cells in vitro. It has been recognized that preformation of a protein corona under controlled conditions ameliorates the protein corona effects, including colloidal stability in serum solutions. We report on the investigation of the stabilizing effects of a denatured bovine serum albumin (dBSA) protein corona formed on the surface of upconversion nanoparticles (UCNPs). UCNPs were chosen as a nanoparticle model due to their unique photoluminescent properties suitable for background-free biological imaging and sensing. UCNP surface was modified with nitrosonium tetrafluoroborate (NOBF4) to render it hydrophilic. UCNP-NOBF4 nanoparticles were incubated in dBSA solution to form a dBSA corona followed up by lyophilization. As produced dBSA-UCNP-NOBF4 demonstrated high photoluminescence brightness, sustained colloidal stability after long-term storage and the reduced level of serum protein surface adsorption. These results show promise of dBSA-based nanoparticle pretreatment to improve the amiability to biological environments towards theranostic applications.

Development and application of photoconversion fluoropolymer films for greenhouses located at high or polar latitudes.

To convert and store energy in the process of photosynthesis, plants primarily use quanta of the red and blue parts of the spectrum. At high latitudes, the average daily intensity of red and blue parts of the spectrum is not very high; for many crops cultivated under greenhouse conditions, it reaches the sufficient level only on clear summer days. The problem of insufficient illumination in greenhouses is usually solved with artificial light sources. This article describes a technology for the manufacture of photoconversion fluoropolymer films for greenhouses. The fluoropolymer films described in the paper make use of original gold nanoparticles and nanoparticles with fluorescence in the blue or red region of the spectrum. In the polymer film, nanoparticles aggregate in the form of "beads", which enhances the field of the optical wave. The film photoconverts UV and violet light into blue and red light. Gold nanoparticles also partially convert energy in the green region of the spectrum (not used by plants) into heat, which is also important for agriculture at high latitudes. In addition, impregnation of gold nanoparticles into fluoropolymer significantly increases the lifetime of the film. The films described in the paper can significantly increase the productivity of greenhouses located at high latitudes. Plants cultivated under the films have more chlorophyll and a higher intensity of photosynthesis - although their system of distance stress signals is, to a certain degree, suppressed.

Production and Use of Selenium Nanoparticles as Fertilizers.

The synergy problem was discussed linking Se nanoparticles and different soil fertility agents. Se zero-valent-state nanoparticles were investigated as fertilizers and antioxidants. A technology was proposed for producing Se zero-valent-state nanoparticles. Se nanoparticles were obtained by laser ablation of Se in water using a fiber ytterbium laser, with a wavelength between 1060 and 1070 nm, a pulse repetition rate of 20 kHz, a pulse duration of 80 ns, and an average power of 20 W, and a copper vapor laser with wavelengths of 510.6 and 578.2 nm and an average power of 8 W. The main particle mass part shifted from 800 nm to a size less than 100 nm, corresponding to the increase in the laser fragmentation time. The resulting nanoparticles were monodisperse in size and mass. The Se nanoparticle water suspension was introduced into the soil. The soil Se nanoparticle concentrations were about 1, 5, 10, and 25 µg kg-1. An experiment was carried out in a climate chamber in two series: (1) growing plants in soil imitating the standard organogenesis environment conditions such as illumination of 16 h per day, temperature of 22 °C, soil humidity of 25% SDW, and an experiment duration of 30 days and (2) growing plants in soil under changing environmental conditions of organogenesis. The standard environmental conditions for the first 10 days are illumination of 16 h day-1, temperature of 22 °C, and soil humidity of 25% SDW. The plant stress for 5 days is hyperthermia of 40 °C. The standard environmental conditions for the next 15 days are illumination of 16 h day-1, temperature of 22 °C, and soil humidity of 25% SDW. At standard organogenesis, the plant leaf plate surface area was 30 ± 2 cm2 in the control option, and the Se nanoparticle doses were correspondingly 1 µg kg-1 for 32 ± 3 cm2, 5 µg kg-1 for 37 ± 2 cm2, 10 µg kg-1 for 38 ± 3 cm2, and 25 µg kg-1 for 28 ± 4 cm2. Hyperthermia stressed plant growth was studied. The highest plant growth rate was in Se nanoparticle concentrations of 5 and 10 µg kg-1. The eggplant growth on the soil with the Se nanoparticle addition at a concentration of 10 µg kg-1 of leaf plate surface area was twice compared to the eggplant growth in untreated soil. The same was for tomato plants. The leaf plate surface area of the cucumber plant grown using Se nanoparticles was 50% higher compared to the control option. The Biogeosystem technique methodology of 20-45 cm soil-layer intrasoil milling for soil multilevel aggregate system formation and intrasoil pulse continuous-discrete watering for soil water regime control was proposed for the Se nanoparticles for better function in the real soil, providing a synergy effect of soil mechanical processing, nanoparticles, humic substances, and polymicrobial biofilms on soil fertility.

Preclinical Study of Biofunctional Polymer-Coated Upconversion Nanoparticles.

Upconversion nanoparticles (UCNPs) are new-generation photoluminescent nanomaterials gaining considerable recognition in the life sciences due to their unique optical properties that allow high-contrast imaging in cells and tissues. Upconversion nanoparticle applications in optical diagnosis, bioassays, therapeutics, photodynamic therapy, drug delivery, and light-controlled release of drugs are promising, demanding a comprehensive systematic study of their pharmacological properties. We report on production of biofunctional UCNP-based nanocomplexes suitable for optical microscopy and imaging of HER2-positive cells and tumors, as well as on the comprehensive evaluation of their pharmacokinetics, pharmacodynamics, and toxicological properties using cells and laboratory animals. The nanocomplexes represent a UCNP core/shell structure of the NaYF4:Yb, Er, Tm/NaYF4 composition coated with an amphiphilic alternating copolymer of maleic anhydride with 1-octadecene (PMAO) and conjugated to the Designed Ankyrin Repeat Protein (DARPin 9_29) with high affinity to the HER2 receptor. We demonstrated the specific binding of UCNP-PMAO-DARPin to HER2-positive cancer cells in cultures and xenograft animal models allowing the tumor visualization for at least 24 h. An exhaustive study of the general and specific toxicity of UCNP-PMAO-DARPin including the evaluation of their allergenic, immunotoxic, and reprotoxic properties was carried out. The obtained experimental body of evidence leads to a conclusion that UCNP-PMAO and UCNP-PMAO-DARPin are functional, noncytotoxic, biocompatible, and safe for imaging applications in cells, small animals, and prospective clinical applications of image-guided surgery.

3D in vitro models of tumors expressing EGFR family receptors: a potent tool for studying receptor biology and targeted drug development.

Carcinomas overexpressing EGFR family receptors are of high clinical importance, because the receptors have prognostic value and are used as molecular targets for anticancer therapy. Insufficient drug efficacy necessitates further in-depth research of the receptor biology and improvement in preclinical stages of drug evaluation. Here, we review the currently used advanced 3D in vitro models of tumors, including tumor spheroids, models in natural and synthetic matrices, tumor organoids and microfluidic-based models, as a potent tool for studying EGFR biology and targeted drug development. We are especially focused on factors that affect the biology of tumor cells, causing modification in the expression and basic phosphorylation of the receptors, crosstalk with other signaling pathways and switch between downstream cascades, resulting ultimately in the resistance to antitumor agents.

Radioactive (90Y) upconversion nanoparticles conjugated with recombinant targeted toxin for synergistic nanotheranostics of cancer.

We report combined therapy using upconversion nanoparticles (UCNP) coupled to two therapeutic agents: beta-emitting radionuclide yttrium-90 (90Y) fractionally substituting yttrium in UCNP, and a fragment of the exotoxin A derived from Pseudomonas aeruginosa genetically fused with a targeting designed ankyrin repeat protein (DARPin) specific to HER2 receptors. The resultant hybrid complex UCNP-R-T was tested using human breast adenocarcinoma cells SK-BR-3 overexpressing HER2 receptors and immunodeficient mice, bearing HER2-positive xenograft tumors. The photophysical properties of UCNPs enabled background-free imaging of the UCNP-R-T distribution in cells and animals. Specific binding and uptake of UCNP complexes in SK-BR-3 cells was observed, with separate 90Y- and PE40-induced cytotoxic effects characterized by IC50 140 µg/mL (UCNP-R) and 5.2 µg/mL (UCNP-T), respectively. When both therapeutic agents were combined into UCNP-R-T, the synergetic effect increased markedly, ∼2200-fold, resulting in IC50 = 0.0024 µg/mL. The combined therapy with UCNP-R-T was demonstrated in vivo.

Liposomal Form of Tetra(Aryl)Tetracyanoporphyrazine: Physical Properties and Photodynamic Activity In Vitro.

Tetra(aryl)tetracyanoporphyrazines are the promising group of dyes for photodynamic therapy of tumors with unique combination of photosensitizer properties and sensitivity of fluorescence parameters to the environment viscosity. However, in vivo application of such hydrophobic photosensitizers requires using of drug carriers ensuring efficient delivery to the tumor site. The present study is focused on obtaining liposomes loaded with tetrakis(4-benzyloxyphenyl)tetracyanoporphyrazine and examining their properties depending on lipid composition. An efficient loading of the dye and a high long-term stability were proved for the liposomes composed of phosphatidylcholine with cholesterol and phosphatidylglycerol. This can be explained by the presence of negatively charged lipids in the bilayer and, as a consequence, a high value of the surface potential. A high rate of cellular uptake and a strong photoinduced toxicity give the prerequisites for the further use of the liposomal form of the photosensitizer for photodynamic therapy of tumors.

Effective delivery of porphyrazine photosensitizers to cancer cells by polymer brush nanocontainers.

Efficient drug delivery can be assigned to tasks that attract the most acute attention of researchers in the field of anticancer drug design. We have reported the first case of using amphiphilic polymer brushes as nanocontainers for photosensitizer delivery to cancer cells. Regular graft-copolymers of hydrophobic polyimides with hydrophilic polymethacrylic acid side chains were loaded with photosensitive dye tetra(4-fluorophenyl)tetracyanoporphyrazine (Pz) providing a sufficiently stable homogeneous fraction of fluorescent Pz-loaded nanoparticles with a size of 100-150 nm. Pz-loaded polymer brushes were substantially more efficient for Pz delivery into cells compared with other types of particles examined, Pz-polyethyleneglycol and Pz-methylcellulose. In vivo, an efficient Pz delivery to tumor can also be expected since the Pz-PB particle size is in the optimal range for passive targeting. Pz-PB showed pronounced photodynamic activity, while, that is important, in the absence of irradiation the PB carrier itself was significantly less toxic than the dye itself. Summing up, water-soluble polymer brushes with polyimide backbones and polymethacrylic acid side chains can be regarded as a novel type of nanocontainers providing efficient intracellular drug delivery for photodynamic therapy of cancers.

Targeted Radionuclide Therapy of Human Tumors.

Targeted radionuclide therapy is one of the most intensively developing directions of nuclear medicine. Unlike conventional external beam therapy, the targeted radionuclide therapy causes less collateral damage to normal tissues and allows targeted drug delivery to a clinically diagnosed neoplastic malformations, as well as metastasized cells and cellular clusters, thus providing systemic therapy of cancer. The methods of targeted radionuclide therapy are based on the use of molecular carriers of radionuclides with high affinity to antigens on the surface of tumor cells. The potential of targeted radionuclide therapy has markedly grown nowadays due to the expanded knowledge base in cancer biology, bioengineering, and radiochemistry. In this review, progress in the radionuclide therapy of hematological malignancies and approaches for treatment of solid tumors is addressed.

Reversible Change of Extracellular pH at the Generation of Mechano-Induced Electrical Reaction in a Stem of Cucurbita pepo.

As was shown recently the generation of cooling-induced action potential accompanies the reversible change of extracellular pH.1 The pH changes are linked to the Ca(2+)-induced transient inhibition of the plasma membrane H(+)-ATPase. Generation of mechano-induced local bioelectrical reaction (LBER) also accompanies the reversible change of extracellular pH. Shifts of pH medium during generation of mechano-induced electrical impulse are less than during generation of cooling-induced bioelectrical response. The exclusion of Ca(2+) influx from extracellular stores don't decrease the amplitude of mechano-induced LBER.