Applied Biochemistry and Microbiology最新文献

Uterine leiomyoma (UL) is a benign tumor of monoclonal origin developing from smooth muscle cells and containing varying amounts of fibrous connective tissue. Leiomyomas and myometrium are similar in cellular composition but differ in the cellular transcriptome profiles. A rapid and reliable method for isolating and culturing primary UL cells is described in this paper. Conditions for growing and maintaining a monolayer culture of UL cells in vitro were optimized. The effect of sex steroid hormones, estradiol and progesterone, on UL genetic markers was evaluated. To prevent the loss of driver mutations in the MED12 and HMGA2 genes, the use of the UL cell model cultured in a specialized nutrient medium with the addition of progesterone at the first passage was proposed.

The growth-stimulating effect of biopreparations based on aqueous dispersions, or colloidal solutions, of fullerene C₆₀, including fullerene-containing substances obtained from intermediate product of the fullerene C₆₀ production process (fullerene extract and fullerene soot), on plants as a result of seed treatment for agricultural crops has been studied. In laboratory and field experiments, the growth-stimulating effect was demonstrated using examples of agricultural crops that are important in the food industry, such as the spring wheat (Triticum aestivum L.) varieties “Lada” and “Voevoda”, the winter wheat (Triticum aestivum L.) variety “Rubezhnaya”, the spring oats (Avena sativa L.) variety “Skakun”, the spring barley (Hordeum vulgare L.) variety “Veresk”, the winter rye (Secale cereale L.) variety “Orlovskaya”, the pumpkin (Cucurbita pepo L.) varieties “Ulybka” and “Muskatnaya”, and the pea (Pisum sativum L.) variety “Pharaoh”, due to the treatment (soaking) of seeds (both fresh and old (stale) or with low germination capacity) in the presence of fullerene-containing aqueous dispersions. In field experiments using the example of the winter wheat (Triticum aestivum L.) variety “Rubezhnaya”, an increase in the yield and vitality of plants and an increase in the weight and number of grains in an ear as result of pre-sowing treatment (soaking) of seeds in the presence of fullerene-containing aqueous dispersions were demonstrated. Experiments were also carried out on the example of the cultivated tobacco plant (Nicotiana tabacum L.) variety “Xhanti NN”, on the study of antiviral activity exerted by an aqueous dispersion of C₆₀ fullerene against the tobacco mosaic virus. In the current research, within the framework of a promising biotechnological approach proposed for agricultural production, the effectiveness, versatility, and reproducibility of the growth-stimulating effect have been proven on worthy examples of seed material when treated with environmentally friendly biopreparations, which are aqueous dispersions of fulle-rene-containing compounds.

Transposition of mobile elements is an important source of genetic diversity in plants. Rapid screening of plant populations for the presence of new insertions and identification of their localization sites in the genome are required when characterizing the collections of insertional mutants. We developed a new method onTEI-seq, which is based on specific amplification of mobile element insertion sites and subsequent nanopore sequencing of PCR products. The developed method was tested when sequencing endogenous EVD and ONSEN transposons of Arabidopsis thaliana. As a result, a high coverage of target transposons with nanopore reads (>1000×) was obtained in 2 h. The onTEI-seq technology was also used to search for ONSEN insertions in the population of A. thaliana insertional mutants containing the additional copies of LTR retrotransposons of this family. It was demonstrated that the sensitivity of the onTEI-seq method is sufficient to identify the insertions in plant population, where the portion of their carriers is only 2.5% (the ratio of insertion carriers to non-carriers was 1 : 39). Based on the results obtained, it can be concluded that the onTEI-seq is a sensitive and rapid method for the identification of new insertions and determination of their localization in the genome.

Antibiotics are intensively used when raising farm animals. As a result they accumulate in livestock waste, meat, and milk and can enter the soil and groundwater. Edible insects can effectively process agricultural waste reducing the concentration of drugs. In addition, such insects are one of the potential sources of alternative feed protein and have high nutritional value. Thus, studying the ability of insects to process waste containing antibacterial drugs while maintaining viability is a topical issue. In this study, the effect of the antibiotic ceftriaxone on the black soldier fly was studied. According to the results obtained, the antibiotic at a concentration of 0.1–100 mg/kg of feed has no effect on the weight, survival rate, productivity, ash content, or fatty acid and amino acid composition of the black soldier fly. At the highest concentration of this antibiotic, an increase in the moisture of the larvae is observed. Consequently, the black soldier fly can consume feed and process waste containing 0.1–100 mg/kg ceftriaxone, while maintaining viability and the basic characteristics necessary for its further use.

Cereals are the basis of human nutrition, accounting for more than 50% of the daily calorie intake in the world. A valuable feed crop is triticale, which is successfully used in bakery production in a number of countries. To date, this cereal is the host of more than 40 species of aphids, including the common grass aphid Schizaphis graminum R., one of the most common and most malicious. The main method of regulating the number of sucking pests is the use of pesticides, various fertilizers, and plant growth regulators (PGRs). In this study, the effect of the PGRs Verva and Ecopin on the reproductive ability of S. graminum when feeding insects on spring triticale was investigated. Common grass aphids were planted on 20 samples of spring triticale. Plants with planted aphids were treated once with the PGR Verva at a concentration of 1 mL/L and Ecopin at a concentration of 1 g/10 L. The pest population was assessed on the second and fourth days after planting. A sharp increase in the number of offspring in aphids was detected on 18 samples of treated PGR two days after planting, and by day 4 it had doubled. Only in two of the studied samples of experimental plants, had treatment with the PGR used had practically no effect on the number of pests. Future research should focus on studying the mechanisms of aphid-induced chlorosis and the role of aphid saliva proteins in inducing a physiological response in plants.

Polydimethylsiloxane (PDMS) remains one of the most popular materials for microfluidic chips, but its high gas permeability can lead to the formation of gas bubbles in microchannels, which hampers the reproducibility of experiments in organ-on-a-chip systems. The dependence of the gas permeability of PDMS on the ratio of base to hardener (2.5 : 1, 5 : 1, and 10 : 1) and polymerization temperature (25, 75, and 125°C) was investigated. According to the results obtained, the 5 : 1 ratio at 125°C provides an optimal combination of low gas permeability, fast polymerization, and economical consumption of components. Cytotoxicity testing on BeWo b30 and EA.hy926 cell lines, representing the trophoblast and endothelium in a placenta-on-a-chip model, demonstrated the absence of cytotoxic effects using different ratios of PDMS components, which confirms its safety for cellular experiments.

Genetic engineering approach has been used to create a collection of recombinant E. coli strains-producers of bacterial enzymes beta-lactamases related both to the existing natural analogues and to the mutant forms of natural enzymes. Currently, the collection includes beta-lactamases of the TEM (molecular class A), CTX-M (molecular class A), VIM (molecular class B), and NDM (molecular class B) types. A set of recombinant beta-lactamases can be used for screening new antibacterial drugs and inhibitors in vitro. The collection of beta-lactamase producers is included in the National Depository Bank of Live Systems “Noah’s Ark” of the Lomonosov Moscow State University.

Optimization of the protocols for cryopreservation and subsequent thawing of primary germ cells in Pushkin breed chicken was carried out. It was detected that with a controlled cooling rate of 1°C/min and combination of penetrating and non-penetrating cryoprotectants, a maximal viability of the cells was 62.9% after their slow thawing at 37°C, which is significantly higher than the values observed during thawing at 4°C. The results obtained emphasize a critical role of thawing temperature in the cell survival, demonstrating that rapid thawing increases their viability by minimizing sublethal damages. The study suggests a more efficient approach to the preservation of poultry genetic resources and creates a potential for biotechnology in the field of animal husbandry.