Project Narrative

The Project Narrative for the Citizen Curator Project at UCF

Budget Justification

This document contains the budget justification for the Citizen Curator Project at UCF

Grapevine Gene Systems for Resistance to Gray Mold <i>Botrytis cinerea</i> and Powdery Mildew <i>Erysiphe necator</i>

Grapevine is one of the world’s most economically important fruit crops. It is known that Vitis vinifera is a host for a large number of pathogenic agents, which significantly reduce the yield and berry quality. This forces the agronomists to use a huge amount of fungicides. Over the last few decades, alternative methods for solving this problem have been developed and continue to be developed. Such new technologies as marker-assisted selection, bioengineering of the rhizosphere, genetic engineering (transgenesis, cisgenesis and intragenesis) allow the production of pathogen-resistant cultivars. However, they are linked to a number of problems. One of the most promising methods is the creation of modified non-transgenic cultivars via CRISPR/Cas9-targeted mutagenesis. Therefore, researchers are actively looking for target genes associated with pathogen resistance and susceptibility. This review elucidates the main mechanisms of plant—pathogen interactions, the immune systems developed by plants, as well as the identified genes for resistance and susceptibility to the biotrophic pathogen Erysiphe necator and the necrotrophic pathogen Botrytis cinerea

Drug Transport System Based on Phospholipid Nanoparticles: Production Technology and Characteristics

One of the current trends in modern pharmaceuticals is the supply of drugs by transport systems. The use of delivery systems allows to increase the therapeutic efficacy, tolerability, and safety of drug therapy. Liposomes, polymer nanoparticles, carbon nanoparticles, blood cells, metal nanoparticles, oxides, etc., are used as transport systems. This work is aimed at obtaining a finished technological product based on soy phospholipids with particle size in the nanometer range and reproducible characteristics (size, charge). For this purpose, we carried out investigations to select the optimal conditions of technological process. The developed technology makes it possible to obtain phospholipid nanoparticles without the use of any solubilizers and/or surfactants, which increases its practical relevance for further work. The versatility of the technology is demonstrated by the example of incorporation of drugs of various chemical nature and pharmacotherapeutic groups

Study of Physico-Chemical Properties and Morphology of Phospholipid Composition of Indomethacin

Nonsteroidal anti-inflammatory drugs (NSAIDs), inhibitors of cyclooxygenase-2, an enzyme involved in the formation of anti-inflammatory prostaglandin PGE2, are the most common treatment for chronic inflammatory diseases, such as, for example, arthritis. One of the most commonly used drugs of this class is indomethacin, a derivative of indolylacetic acid. In this work, we studied the physicochemical properties of the phospholipid composition of indomethacin obtained earlier (codenamed “Indolip”) and the effect of freeze drying on its parameters. It was shown that the properties such as particle size, light transmission, phospholipid oxidation index did not change significantly, which indicated the stability of the drug after lyophilization. Measurement of the spectra of small-angle neutron scattering has shown that morphologically, Indolip is a vesicle whose radius is five times greater than the value of the bilayer thickness

Chlorin e6 Phospholipid Delivery System Featuring APN/CD13 Targeting Peptides: Cell Death Pathways, Cell Localization, In Vivo Biodistribution

We have previously designed a phospholipid delivery system for chlorin e6 to increase the efficacy of photodynamic therapy involving a second-generation photosensitizer. Further research into the matter led to double modification of the obtained nanoparticles with ligands exhibiting targeting and cell-penetrating effects: an NGR-containing peptide and heptaarginine (R7), respectively. This study investigated the cell death pathway on HT-1080 tumor cells after treatment with the proposed compositions: the chlorin e6 phospholipid composition and the two-peptide chlorin e6 phospholipid composition. It was demonstrated that most of the cells died by apoptosis. Colocalization analysis of chlorin e6 in the phospholipid composition with two peptides showed mitochondria are one of the targets of the photosensitizer. An HT-1080 tumor-bearing mouse model was used to evaluate the biodistribution of the drug in tumor, liver, and kidney tissues after administration of the study compositions in comparison with free chlorin e6. The photosensitizer mostly accumulated in the tumor tissue of mice administered the phospholipid compositions, and accumulation was increased 2-fold with the peptide-containing composition and approximately 1.5-fold with the unenhanced composition, as compared with free chlorin e6. The enhancement of the chlorin e6 phospholipid composition with targeting and cell-penetrating peptides was found to be effective both in vitro and in vivo

Bioceramics Based on &beta;-Calcium Pyrophosphate

Ceramic samples based on &beta;-calcium pyrophosphate &beta;-Ca2P2O7 were prepared from powders of &gamma;-calcium pyrophosphate &gamma;-Ca2P2O7 with preset molar ratios Ca/P = 1, 0.975 and 0.95 using firing at 900, 1000, and 1100 &deg;C. Calcium lactate pentahydrate Ca(C3H5O3)2&sdot;5H2O and monocalcium phosphate monohydrate Ca(H2PO4)2&sdot;H2O were treated in an aqua medium in mechanical activation conditions to prepare powder mixtures with preset molar ratios Ca/P containing calcium hydrophosphates with Ca/P = 1 (precursors of calcium pyrophosphate Ca2P2O7). These powder mixtures containing calcium hydrophosphates with Ca/P = 1 and non-reacted starting salts were heat-treated at 600 &deg;C after drying and disaggregation in acetone. Phase composition of all powder mixtures after heat treatment at 600 &deg;C was presented by &gamma;-calcium pyrophosphate &gamma;-Ca2P2O7 according to the XRD data. The addition of more excess of monocalcium phosphate monohydrate Ca(H2PO4)2&middot;H2O (with appropriate molar ratio of Ca/P = 1) to the mixture of starting components resulted in lower dimensions of &gamma;-calcium pyrophosphate (&gamma;-Ca2P2O7) individual particles. The grain size of ceramics increased both with the growth in firing temperature and with decreasing molar ratio Ca/P of powder mixtures. Calcium polyphosphate (t melt = 984 &deg;C), formed from monocalcium phosphate monohydrate Ca(H2PO4)2&sdot;H2O, acted similar to a liquid phase sintering additive. It was confirmed by tests in vitro that prepared ceramic materials with preset molar ratios Ca/P = 1, 0.975, and 0.95 and phase composition presented by &beta;-calcium pyrophosphate &beta;-Ca2P2O7 were biocompatible and could maintain bone cells proliferation

Microbial Biofilms at Meat-Processing Plant as Possible Places of Bacteria Survival

Biofilm contamination in food production threatens food quality and safety, and causes bacterial infections. Study of food biofilms (BF) is of great importance. The taxonomic composition and structural organization of five foods BF taken in different workshops of a meat-processing plant (Moscow, RF) were studied. Samples were taken from the surface of technological equipment and premises. Metagenomic analysis showed both similarities in the presented microorganisms dominating in different samples, and unique families prevailing on certain objects were noted. The bacteria found belonged to 11 phyla (no archaea). The dominant ones were Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. The greatest diversity was in BFs taken from the cutting table of raw material. Biofilms&rsquo; bacteria may be the cause of meat, fish and dairy products spoilage possible representatives include Pseudomonas, Flavobacterium, Arcobacter, Vagococcus, Chryseobacterium, Carnobacterium, etc.). Opportunistic human and animal pathogens (possible representatives include Arcobacter, Corynebacterium, Kocuria, etc.) were also found. Electron-microscopic studies of BF thin sections revealed the following: (1) the diversity of cell morphotypes specific to multispecies BFs; (2) morphological similarity of cells in BFs from different samples, micro-colonial growth; (3) age heterogeneity of cells within the same microcolony (vegetative and autolyzed cells, resting forms); (4) heterogeneity of the polymer matrix chemical nature according to ruthenium red staining

Chlorin e6 embedded in phospholipid nanoparticles equipped with specific peptides: Interaction with tumor cells with different aminopeptidase N expression

A promising direction in Biopharmaceuticals is the development of specific peptide-based systems to improve drug delivery. This approach may increase tumor specificity and drug penetration into the target cell. Similar systems have been designed for several antitumor drugs. However, for photodynamic therapy drugs, such studies are not yet enough. Previously, we have developed a method of inclusion of chlorin e6 (Ce6), a photosensitizer used in photodynamic therapy, in phospholipid nanoparticles with a diameter of up to 30 nm, and reported an increase in its effectiveness in the experiments in vivo. In this work, we propose to modify a previously developed delivery system for Ce6 by the addition of cell-penetrating (R7) and/or targeting NGR peptides. The interaction of the compositions developed with HepG2 and MCF-7 tumor cells is shown. The expression of CD13 protein with affinity to NGR on the surface of these cells has been studied using flow cytometry. The expression of this protein on the HepG2 cells and its absence on MCF-7 was demonstrated. After incubation of tumor cells with the resulting Ce6 compositions, we evaluated the cellular accumulation, photoinduced, and dark cytotoxicity of the drugs. After irradiation, the highest level of cytotoxicity was observed when R7 peptide was added to the system, either alone or in combination with NGR. In addition to R7, the NGR-motif peptide increased the internalization of Ce6 in HepG2 cells without affecting its photodynamic activity. In this work we also discuss possible mechanisms of action of the cell-penetrating peptide when attached to phospholipid nanoparticles

<i>Fusarium</i> Species Causing Pepper Wilt in Russia: Molecular Identification and Pathogenicity

Fusarium wilt pathogens represent an ongoing threat to pepper production worldwide. This is the first report providing data on the molecular identification of Fusarium fungi that cause wilt in pepper in the southern regions of Russia. Monitoring of the Fusarium infection on pepper was carried out in 2019–2022 in two economically important regions of this culture production: the Krasnodar Krai and Crimea. Based on a phylogenetic analysis of the translation elongation factor (EF1a) and the internal transcribed spacer (ITS), as well as the macro- and micromorphological characteristics of the fungi, the causative agents of Fusarium wilt have been identified. The causative agents identified as representatives of the Fusarium species composition included: F. clavus, F. solani, F. oxysporum, F. verticillioides, F. commune, F. torulosum, and F. sporotrichioides. Depending on the region, the specifics of biodiversity and the ratio of these species in pathocomplexes were noted. In Crimea, wilting could be attributed to all of the identified species; in the Krasnodar Krai, F. verticillioides and F. clavus were found to contribute to wilting. The pathogenicity test showed that the pathogens of pepper wilting in Russia, in addition to the already known F. oxysporum and F. solani, are the species F. clavus and F. verticillioides. This is the first report on the ability of these species to cause Fusarium wilt in pepper cultures. The obtained data will be of practical value for the development of biological control measures for fungi of the genus Fusarium, which cause pepper wilt in areas of industrial production and seed production. In addition, data on species composition and aggressive isolates will be used in a pepper breeding program for resistance to Fusarium wilt