Biodegradation of polyethylene in digestive gland homogenates of marine invertebrates.

Вiotic factors may be the driving force of plastic fragmentation along with abiotic factors. Since understanding the processes of biodegradation and biological depolymerization of plastic is important, a new methodological approach was proposed in this study to investigate the role of marine invertebrate digestive enzymes in plastic biodegradation. The aim of this study is to evaluate the possibility of enzymatic biodegradation of polyethylene fragments in the digestive gland homogenate of marine invertebrates differing in their feeding type (Strongylocentrotus nudus, Patiria pectinifera, Mizuhopecten yessoensis). Significant changes are found in the functional groups of the polymer after 3 days of incubation in the digestive gland homogenates of the studied marine invertebrates. A significant increase in the calculated CI (carbonyl index) and COI (сarbon-oxygen index) indices compared to the control sample was observed. The results suggest that digestive enzymes of studied organisms may play an important role in the biogeochemical cycling of plastic.

The Relationship between Lifespan of Marine Bivalves and Their Fatty Acids of Mitochondria Lipids.

Marine bivalves belonging to the Mytilidae and Pectinidae Families were used in this research. The specific objectives of this study were: to determine the Fatty Acids (FAs) of mitochondrial gill membranes in bivalves with different lifespans, belonging to the same family, and to calculate their peroxidation index; to compare the levels of ROS generation, malondialdehyde (MDA), and protein carbonyls in the mitochondria of gills, in vitro, during the initiation of free-radical oxation; to investigate whether the FAs of mitochondria gill membranes affect the degree of their oxidative damage and the maximum lifespan of species (MLS). The qualitative membrane lipid composition was uniform in the studied marine bivalves, regardless of their MLS. In terms of the quantitative content of individual FAs, the mitochondrial lipids differed significantly. It is shown that lipid matrix membranes of the mitochondria of long-lived species are less sensitive to in vitro-initiated peroxidation compared with the medium and short-lived species. The differences in MLS are related to the peculiarities of FAs of mitochondrial membrane lipids.

Dietary Exposure to Particles of Polytetrafluoroethylene (PTFE) and Polymethylmethacrylate (PMMA) Induces Different Responses in Periwinkles Littorina brevicula.

The marine and ocean water pollution with different-sized plastic waste poses a real threat to the lives of the next generations. Plastic, including microplastics, is found in all types of water bodies and in the organisms that live in them. However, given the chemical diversity of plastic particles, data on their toxicity are currently incomplete. Moreover, it is clear that different organisms, depending on their habitat and feeding habits, are at different risks from plastic particles. Therefore, we performed a series of experiments on feeding the gastropod scraping mollusk Littorina brevicula with two types of polymeric particles-polymethylmethacrylate (PMMA) and polytetrafluoroethylene (PTFE)-using a special feeding design. In the PMMA-exposed group, changes in gastrointestinal biochemical parameters such as increases in malondialdehyde (MDA) and protein carbonyls (PC) were detected, indicating the initiation of oxidative stress. Similarly, a comet assay showed an almost twofold increase in DNA damage in digestive gland cells compared to the control group. In mollusks fed with PTFE-containing food, no similar changes were recorded.

Degradation of micro-nano-sized polytetrafluoroethylene and acrylic fluorinated copolymer particles in the periwinkle digestive tract.

The aim of the work was to investigate the possible biodegradation of such stable plastics as fluorated acrylic copolymer Protacryl-M and polytetrafluoroethylene-teflon (PTFE)-that are widely applied in medicine for prosthetics in dentistry and orthopedics. Our tasks were as follows: 1. To select a suitable marine biological object (multicellular invertebrate) 2. To develop a method for delivering microplastic particles (MP) of selected plastics to the gastrointestinal tract (GIT) of an experimental animal 3. To develop a technique for separating MP particles from animal feces 4. To develop a technology for multiple passages of MP particles through the gastrointestinal tract of an animal 5. To select methods and techniques for determining the degradation of the surface of MP particles after passing through the gastrointestinal tract of the animal The effect of a biological agent on the types of plastic above is considered using marine gastropods of the genus Littorina (in particular, the periwinkle L. brevicula) as a model organism widely distributed in the marine intertidal zone.

A massive bloom of Karenia species (Dinophyceae) off the Kamchatka coast, Russia, in the fall of 2020.

In the fall of 2020, a long-lasting and massive harmful algal bloom (HAB) with extensive fields of yellow sea foam was observed in relatively cold waters (7-13 °C) off the coasts of the Kamchatka Peninsula, Russia. According to the estimates based on bio-optical parameters in satellite imagery, the Kamchatka bloom 2020 lasted for two months and covered a vast area of more than 300 × 100 km. An abundance of dead fish and invertebrates, including sea urchins, sea anemones, chitons, cephalopods, bivalves were found on shore during the bloom. Animals suffered almost 100% mortality within a depth range between 5 and 20 m. To identify the causative microalgal species, light and scanning electron microscopy, Raman spectroscopy, and molecular phylogenetic approaches were used. The HAB area was estimated by the spectral analysis of satellite-derived imagery. The causative organisms were unarmored dinoflagellates of Karenia species. Their density and biomass reached 100-620 cells·mL-1 and 1300-7700 mg·m-3, respectively, which accounted for 31-99% of the total cell density and 82-99% of the total phytoplankton biomass in late September to mid-October. The dominant species was Karenia selliformis, and the other co-occurring kareniacean species were K. cf. cristata, K. mikimotoi, K. papilionacea, K. longicanalis, and two unidentified morphotypes of Karenia spp. The molecular phylogeny inferred from LSU rDNA and ITS region showed that K. selliformis from Kamchatka in 2020 belonged to the cold-water group I and was identical to K. selliformis strains from Hokkaido, Japan, identified in 2021. This is the first HAB event caused by K. selliformis recorded from Russian coastal waters.

Micro-nano-sized polytetrafluoroethylene (teflon) particles as a model of plastic pollution detection in living organisms.

Micro- and nano-sized particles of polytetrafluoroethylene (PTFE) were used as model (reference) particles to study the biological effects of plastic pollution. Since the PTFE molecule contains fluorine, considered as an "atomic marker" sharply distinguishing it from other common plastics, micro- and nano-particles of PTFE have a specific crystalline structure and are, therefore, well identified by the methods of polarized light microscopy (POL), Raman microspectroscopy (micro-Raman), and energy-dispersive spectroscopy (EDS). Examples of PTFE particles detection in hemolimph of the cockroach Blatella germanica, in hemolimph of the larva and in faecal pellets of imago of a fly Lucilia sp., in the stomach and hingat of brine shrimp Artemia salina, and in association with cell wall of green unicellular alga Chlorococcus sp. are provided. The presented results strongly suggest that PTFE particles can be detected and identified in the biological medium using the method of "atomic markers", polarization microscopy and Raman spectroscopy.

Antioxidant activity of Far Eastern bivalves in their natural habitat.

The activities of the key antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GP) and glutathione reductase (GR) as well as levels of reduced glutathione (GSH) and integral antioxidant activity (IAA), were studied in the digestive glands and gills of 14 bivalve species. Species and tissue differences of the antioxidant (AO) systems of the investigated mollusks were discussed in connection with their physiological and biochemical peculiarities. This article describes the role of the AO system of mollusks in adaptation to natural habitat conditions and shows the relationship of AO activity with the maximum habitat depth (MHD) and maximum lifespan (MLS) of these species.

Synthesis, anticancer activity, and molecular modeling of 1,4-naphthoquinones that inhibit MKK7 and Cdc25.

Cell division cycle 25 (Cdc25) and mitogen-activated protein kinase kinase 7 (MKK7) are enzymes involved in intracellular signaling but can also contribute to tumorigenesis. We synthesized and characterized the biological activity of 1,4-naphthoquinones structurally similar to reported Cdc25 and(or) MKK7 inhibitors with anticancer activity. Compound 7 (3-[(1,4-dioxonaphthalen-2-yl)sulfanyl]propanoic acid) exhibited high binding affinity for MKK7 (Kd = 230 nM), which was greater than the affinity of NSC 95397 (Kd = 1.1 µM). Although plumbagin had a lower binding affinity for MKK7, this compound and sulfur-containing derivatives 4 and 6-8 were potent inhibitors of Cdc25A and Cdc25B. Derivative 22e containing a phenylamino side chain was selective for MKK7 versus MKK4 and Cdc25 A/B, and its isomer 22f was a selective inhibitor of Cdc25 A/B. Docking studies performed on several naphthoquinones highlighted interesting aspects concerning the molecule orientation and hydrogen bonding interactions, which could help to explain the activity of the compounds toward MKK7 and Cdc25B. The most potent naphthoquinone-based inhibitors of MKK7 and/or Cdc25 A/B were also screened for their cytotoxicity against nine cancer cell lines and primary human mononuclear cells, and a correlation was found between Cdc25 A/B inhibitory activity and cytotoxicity of the compounds. Quantum chemical calculations using BP86 and ωB97X-D3 functionals were performed on 20 naphthoquinone derivatives to obtain a set of molecular electronic properties and to correlate these properties with cytotoxic activities. Systematic theoretical DFT calculations with subsequent correlation analysis indicated that energy of the lowest unoccupied molecular orbital E(LUMO), vertical electron affinity (VEA), and reactivity index ω of these molecules were important characteristics related to their cytotoxicity. The reactivity index ω was also a key characteristic related to Cdc25 A/B phosphatase inhibitory activity. Thus, 1,4-naphthoquinones displaying sulfur-containing and phenylamino side chains with additional polar groups could be successfully utilized for further development of efficacious Cdc25 A/B and MKK7 inhibitors with anticancer activity.

Virtual screening, synthesis and biological evaluation of DNA intercalating antiviral agents.

This paper describes computer-aided design of new anti-viral agents against Vaccinia virus (VACV) potentially acting as nucleic acid intercalators. Earlier obtained experimental data for DNA intercalation affinities and activities against Vesicular stomatitis virus (VSV) have been used to build, respectively, pharmacophore and QSAR models. These models were used for virtual screening of a database of 245 molecules generated around typical scaffolds of known DNA intercalators. This resulted in 12 hits which then were synthesized and tested for antiviral activity against VaV together with 43 compounds earlier studied against VSV. Two compounds displaying high antiviral activity against VaV and low cytotoxicity were selected for further antiviral activity investigations.

Silver-Mediated Double Helix: Structural Parameters for a Robust DNA Building Block.

The DNA double helix is a versatile building block used in DNA nanotechnology. To potentiate the discovery of new DNA nanoscale assemblies, recently, silver cations have been introduced to pair DNA strands by base-Ag+-base bonding rather than by Watson-Crick pairing. In this work, we study the classical dynamics of a parallel silver-mediated homobase double helix and compare it to the dynamics of the antiparallel double helix. Our classical simulations show that only the parallel double helix is highly stable through the 100 ns simulation time. A new type of H-bond previously proposed by our collaboration and recently observed in crystal-determined helices drives the physicochemical stabilization. Compared to the natural B-DNA form, the metal-mediated helix has a contracted axial base pair rise and smaller numbers of base pairs per turn. These results open the path for the inclusion of this robust metal-mediated building block into new nanoscale DNA assemblies.

Synthesis of New 'Hybrid' Compounds Based on Benzofuroxans and Aminoalkylnaphthalimides.

Pathogenic bacteria and fungi eventually develop resistance to existing drugs, and therefore, we need constant development of new drugs. The research is aimed at addressing fundamental scientific problems-the search for new biologically active compounds among several benzofuroxan-containing 'hybrid' products. N-substituted naphthalimides were chosen as a second pharmacophore. Benzofuroxanes biological effects were studied by means of bacterial lux-biosensors. Compounds IIIa, IVa, IIIc, and IVc displayed more expressed bacteriotoxic action in comparison with the initial substances Ia-c and represent a certain interest for using as antibacterial substances.

The Role of Hydrogen Bonds in the Stabilization of Silver-Mediated Cytosine Tetramers.

DNA oligomers can form silver-mediated duplexes, stable in gas phase and solution, with potential for novel biomedical and technological applications. The nucleobase-metal bond primarily drives duplex formation, but hydrogen (H-) bonds may also be important for structure selection and stability. To elucidate the role of H-bonding, we conducted theoretical and experimental studies of a duplex formed by silver-mediated cytosine homopobase DNA strands, two bases long. This silver-mediated cytosine tetramer is small enough to permit accurate, realistic modeling by DFT-based quantum mechanics/molecular mechanics methods. In gas phase, our calculations found two energetically favorable configurations distinguished by H-bonding, one with a novel interplane H-bond, and the other with planar H-bonding of silver-bridged bases. Adding solvent favored silver-mediated tetramers with interplane H-bonding. Overall agreement of electronic circular dichroism spectra for the final calculated structure and experiment validates these findings. Our results can guide use of these stabilization mechanisms for devising novel metal-mediated DNA structures.

The regioselective synthesis of spirooxindolo pyrrolidines and pyrrolizidines via three-component reactions of acrylamides and aroylacrylic acids with isatins and α-amino acids.

The regioselective three-component condensation of azomethine ylides derived from isatins and α-amino acids with acrylamides or aroylacrylic acids as dipolarophiles has been realized through a one-pot 1,3-dipolar cycloaddition protocol. Decarboxylation of 2'-aroyl-2-oxo-1,1',2,2',5',6',7',7a'-octahydrospiro[indole-3,3'-pyrrolizine]-1'-carboxylic acids is accompanied by cyclative rearrangement with formation of dihydropyrrolizinyl indolones.

Valence photoelectron spectra of alkali bromides calculated within the propagator theory.

The valence ionization spectra covering the binding energy range 0-45 eV of alkali bromide XBr (X = Li, Na, K, Rb) vapors are studied within the framework of the propagator theory. Relativistic Algebraic Diagrammatic Construction calculations have been carried out in order to investigate photoionization processes and to describe molecular electronic structure. Theoretical results are compared with available experimental data.

Synthesis and biological activity of 7H-benzo[4,5]indolo[2,3-b]-quinoxaline derivatives.

New 7-(2-aminoethyl)-7H-benzo[4,5]indolo[2,3-b]quinoxalines (13-20) were synthesized with high yields starting from 3H-benzo[e]indole-1,2-dione. These compounds were screened for the cytotoxicity, anti-viral activity, interferon inducing ability and DNA affinity compared with the corresponding 6-(2-aminoethyl)-6H-indolo[2,3-b]quinoxaline derivatives (1-12). It was shown, that compounds 13-20 bind to DNA stronger (lg Кa=6.23-6.87) than compounds 1-12 (lg Кa=5.57-5.89). Anti-viral activity is significantly reduced with annulations of benzene ring in Indoloquinoxaline moiety 13-20.

Synthesis, cytotoxicity by bioluminescence inhibition, antibacterial and antifungal activity of ([1,2,4]Triazolo[1,5-c]quinazolin-2-ylthio)carboxylic acid amides.

We report in this work the synthesis, cytotoxicity, and antimicrobial activity of ([1,2,4]triazolo[1,5-c]quinazolin-2-ylthio)carboxylic acid amides 4-7 in connection with our previous research in the preparation of triazoloquinazoline derivatives. Due to simplicity, general availability of starting materials, and high yields, the most reliable method of synthesis appeared to be the one with N,N-carbonyldiimidazole activation stage. The chemical structures of all obtained substances were deduced from FT-IR, (1)H-NMR, EI-MS, and LC-MS spectral data. The results of cytotoxicity evaluated by bioluminescence inhibition of bacterium Photobacterium leiognathi, strain Sh1 showed that compounds 4.1, 4.6, and 6.1 were the most cytotoxic. Investigation of the antimicrobial and antifungal activity of amides 4-7 (concentration 5 mg/mL) was carried out by the stiff-plate agar-diffusion method. We found that the compounds possessed low (4.1, 4.7) antifungal activity against Candida tenuis and strong (4.21, 5.1, 5.9) or inefficient (4.7, 4.12, 4.16) activity against Aspergillus niger. Substances 5.1 and 5.9 slightly affected Mycobacterium luteum. Staphylococcus aureus was resistant to all obtained substances, and only the n-butyramide derivatives 7.1 and 7.5 inhibited the growth of Escherichia coli. Hence, there was no strong correlation between bioluminescence inhibition and antimicrobial activity of the investigated substances.

Synthesis of new 2-thio-[1,2,4]triazolo[1,5-c]quinazoline derivatives and its antimicrobial activity.

A series of novel ([1,2,4]triazolo[1,5-c]quinazolin-2-ylthio)carboxylic acids 2a-d and esters 3a-l were synthesized and evaluated for antimicrobial activity. Alkylation of potassium 2-thio-[1,2,4]triazolo[1,5-c]quinazoline 1 with halogenocarboxylic acids and its esters proceeded S-regioselectively. During acid catalyzed esterification of 2a-c, degradation of the pyrimidine ring was observed. The structures of the compounds were elucidated by FT-IR, (1)H- and (13)C-NMR, electron impact mass spectra (EI-MS) and LC-MS spectral data. Antimicrobial and antifungal activity of synthesized compounds was tested against Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, Mycobacterium luteum, Candida albicans and Candida tenuis. Acids 2a and 2c exhibited significant activity against C. albicans, which was additionally confirmed by the bioluminescence inhibition test and interrelated with their lipophilicity.

Comparative characteristic of Mytilus muscle cells developed in vitro and in vivo.

The mussel cells from premyogenic larval stages are capable of differentiation into smooth muscle cells in vitro. However, the behavior and protein composition of these cells are not completely identical to those of smooth muscle cells of adult mussels. In this study we compared some properties of mussel muscle cells forming from cells of trochophore (premyogenic larval stage) in vitro with those of muscle cells of veliger and adult mussel. We found a substantial difference between the contractile apparatus protein composition of veliger muscle and cultivated cells. Myorod, one of the molecular markers of the phenotype of mollusc smooth muscle cells (Shelud'ko et al., 1999, Comp Biochem Physiol 122:277-285), is not a constituent of the contractile apparatus of veliger muscle. At the same time the protein composition of contractile apparatus in cultivated cells was similar to that of adult Mytilus muscles. There were only few quantitative differences between them. The contractile activity of cultivated cells was changing in time. The kinetic parameters of first spontaneous contractions were similar to those of phasic contractions, while their period was close to that of tonic contractions. After 50-55 hrs cultivation the cells produced both phasic and tonic contractions, but the character of contractile activity of cultivated cells was regulated after six days of cultivation only. However, there were no muscle cells in vitro, whose contractile activity was similar to that of veliger muscle cells. So, we concluded that properties of muscle cells forming from premyogenic larval mussel cells in culture are similar to those of muscle cells of the adult mussel, but not of veliger.