Production of axenic cultures of diatoms is a highly relevant task, as these widespread organisms are objects of research in different fields of biology, from ecology to whole‐genome studies, and promising as producers of various organic molecules, including biologically active substances. However, culturing of diatoms isolated from natural communities is accompanied by the development of associated microorganisms. We propose a procedure for establishing an axenic culture of the freshwater diatom Synedra asus subsp. radians (Kütz.) Skabitsch. from Lake Baikal. Such a culture has been produced and maintained by subculturing in our laboratory. The procedure involves filtration (to remove free bacteria), detergent treatment (to remove microorganisms associated with diatom frustules), selection of effective antibiotics and their concentrations nontoxic for diatoms (by the disk diffusion method), cell treatment with the selected antibiotic (ciprofloxacin), repeated filtration, and monoclonal culturing of diatom cells. Axenity of the culture was verified microscopically (after DAPI staining) and by molecular biological methods.

The design of cationic liposomes for efficient mRNA delivery can significantly improve mRNA-based therapies. Lipoplexes based on polycationic lipid 1,26-bis(cholest-5-en-3β-yloxycarbonylamino)-7,11,16,20-tetraazahexacosane tetrahydrochloride (2X3) and helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were formulated in different molar ratios (1:1, 1:2, 1:3) to efficiently deliver model mRNAs to BHK-21 and A549. The objective of this study was to examine the effect of 2X3-DOPE composition as well as lipid-to-mRNA ratio (amino-to-phosphate group ratio, N/P) on mRNA transfection. We found that lipoplex-mediated transfection efficiency depends on both liposome composition and the N/P ratio. Lipoplexes with an N/P ratio of 10/1 showed nanometric hydrodynamic size, positive ζ potential, maximum loading, and transfection efficiency. Liposomes 2X3-DOPE (1:3) provided the superior delivery of both mRNA coding firefly luciferase and mRNA-eGFP into BHK-21 cells and A549 cells, compared with commercial Lipofectamine MessengerMax.

Interest in studies concerning the effect of organic carbon sources on the growth of diatoms is largely aimed at subsequent physiological changes occurring in their cells. There are no data on structural changes in the cytoplasm and their relationship with changes in the composition of fatty acids in the course of mixotrophic culturing of freshwater diatoms. To elucidate the role of lipids in the growth of diatom cells in autotrophic and mixotrophic cultures, it is necessary to obtain information on the distribution of fatty acids among intracellular compartments and on possible ultrastructural changes in the cells. In this study, the results demonstrated that Synedra acus cells cultured in the presence of 80 mM glycerol contained lipid bodies of increased size, while cells from cultures supplemented with 40 mM glucose accumulated polysaccharide (chrysolaminarin) granules. An increase in the relative amounts of palmitic and stearic acids was revealed at the exponential growth phase of S. acus in the medium with 80 mM glycerol, which was indicative of the accumulation of fatty acids contained in triacylglycerols. Synedra acus subsp. radians have an ability to proliferate in the presence of high concentrations of organic substances and their transport into cells is evidence for its high adaptation potential, which, along with other factors, accounts for the dominance of this diatom in the spring-summer plankton of the oligotrophic Lake Baikal.

In this study, we present results on fatty acid analysis of phytoplankton of Lake Baikal, the world’s deepest lake, which differs from other lakes by its oceanic features. Since we used a large-mesh net, the net sample phytoplankton were primarily represented by the large elongated diatom Synedra acus. subsp. radians (Kützing) Skabichevskij. The similar algae composition of net samples of spring season phytoplankton collected at different sites of the lake allows us to compare results of the fatty acid analysis of these samples. The phytoplankton diversity of the sedimentation samples was contrary represented by 32 algae species. There are clear changes in the fatty acid composition of net phytoplankton exposed to anthropogenic impacts of varying intensity. The content of polyunsaturated fatty acids in phytoplankton collected from central stations (pelagic stations at a distance of ~10–30 km from the shoreline) without anthropogenic impact was higher by up to 15% than phytoplankton collected from nearshore stations (littoral stations at a distance of ~0.01–0.05 km from the shoreline) and offshore stations (pelagic stations at a distance of ~3 km from the shoreline). The interlaboratory precision of fatty acid determination of phytoplankton is estimated as ≤10%. We found high content of the lipid peroxidation marker (80–340 μg g⁻¹ of dry weight) in phytoplankton from nearshore and offshore stations with intensive anthropogenic impact. In phytoplankton from central stations, we did not find any lipid peroxidation. Determination of unsaturated fatty acids, coupled with analysis of fatty acid peroxidation products, can be used to evaluate the level of anthropogenic impact in terms of ecological health and biodiversity conservation.