Antibiotic Adjuvant 4-Hexylresorcinol Enhances the Efficiency of Antituberculosis Drugs.

We studied the possibility of using 4-hexylresorcinol to increase the efficiency of anti-mycobacterial chemotherapy. In an in vitro experiment, 4-hexylresorcinol increased the efficiency of rifampicin, kanamycin, and isoniazid against Mycobacterium smegmatis by 3-5 times. Experiments in sanitation of BALB/c mice infected with M. smegmatis showed the best efficacy of the isoniazid and 4-hexylresorcinol combination in comparison with isoniazid monotherapy. The growth-inhibiting activity of the combination of antibiotic rifabutin with 4-hexylresorcinol was shown on 6 strains of M. tuberculosis. A 2-fold decrease in the minimum inhibitory concentration of this antibiotic in the presence of half-minimum inhibitory concentration of 4-hexylresorcinol was demonstrated for monoresistant strain M. tuberculosis 5360/42Hr. On the mouse model of experimental tuberculosis caused by M. tuberculosis H37Rv, a 5-fold decrease in lung contamination and more rapid complete cure were achieved in animals treated with the combination of rifabutin and 4-hexylresorcinol in comparison with rifabutin monotherapy.

Sulfation of arabinogalactan with sulfamic acid under homogeneous conditions in dimethylsulfoxide medium.

Sulfation of larch wood arabinogalactan (AG) with sulfamic acid in dimethylsulfoxide (DMSO) medium in the presence of urea was studied for the first time. The use of DMSO as a solvent instead of more toxic 1,4-dioxane allows to sulfate AG under homogeneous conditions. The sulfated AG with a high sulfur content (12.0-12.5 wt %) was produced by sulfation at a temperature of 85-90 °C, the molar ratio of AG / sulfating agent equal to 1:0.85 during 2-3 h. The introduction of sulfate groups into the structure of arabinogalactan was confirmed by the appearance of new absorption bands in FTIR and FT Raman spectra, characteristic for the vibrations of the sulfate groups. It was proved by 13C NMR spectroscopy that the predominant substitution of the primary hydroxyl groups at C6 carbon atoms of the terminal galactose units of main and side chains of arabinogalactan takes place. Simultaneously, the hydroxyl groups associated with C2 and C4 carbon atoms of galactose unit of the main chain are only partially sulfated. According to results of GPC study, the sulfated AG is characterized by a narrow molecular weight distribution with an average molecular weight of 18.8 kDa and a polydispersity of 1.3. Supplementary Information: The online version contains supplementary material available at 10.1007/s00226-021-01341-2.

Study of the Blood Compatibility of Sulfated Organosolv Lignins Derived from Abies sibirica and Larix sibirica Wood Pulp.

The effects of sulfated organosolv lignins derived from fir (Abies sibirica) and larch (Larix sibirica) (SLf and SLl; 4-3-7.5% sulfur, median-weight molecular mass 2960-4888 Da), on human blood/plasma clotting, platelet aggregation, and erythrocyte hemolysis were studied in vitro. Antithrombin activities of the samples were below 2 U/mg. Specimens of SLf (sulfur content 6.5, 6.6, and 7.5%, molecular weights 3503, 3487, and 3580 Da, respectively) and SLl (4.3 and 6.3%, 2960 and 3497 Da) in a concentration of 0.01 mg/ml did not prolong the blood clotting time, did not provoke human platelet aggregation, did not destroy erythrocyte membranes, and could be used for construction of drug delivery systems. The SLf sample (6.5%, sulfur, 3503 Da) in concentrations from 0.09 to 1.82 mg/ml did not stimulate platelet aggregation, reduced ADP-induced platelet aggregation, and 2-fold prolonged the blood/plasma clotting time 2-fold in comparison with control and could be used for creation of biomaterial with clot-resistant surface.

Diversity of magnetotactic bacteria of the Moskva River.

Diversity of magnetotactic bacteria in the Moskva River at the Strogino area was studied using microscopy and phylogenetic analysis. Magnetotactic cocci were the predominant morphotype. Phylogenetic analysis of the 16S rRNA gene sequences revealed 13 OTUs of the orders Magnetococcales and Rhodospirillales, class Alphaproteobacteria. The shares of the relevant sequences were 90 and 10%, respectively. An axenic culture of magnetotactic spirilla was isolated from the studied community. According to the results of the 16S rRNA gene sequencing, the isolate was identified as a new Magnetospirillum species.

New composites of betulin esters with arabinogalactan as highly potent anti-cancer agents.

Betulin and its esters are the natural compounds with high in vitro cytotoxicity toward many cancer cells. However, the poor water solubility of these compounds has limited their applications. We prepared new composites of betulin esters using two methods, namely ball-milling of the mixtures of betulin esters with arabinogalactan and preparation of thin films of these mixtures by evaporating the aqueous solutions. These composites revealed higher water solubility as compared with the initial substances without losing the structural integrity and functionality. As a result, the new composites have shown much higher inhibitory effects against different cancer cell lines such as Ehrlich ascites carcinoma cells and lung carcinoma cells (A549) in comparison with the initial substances. The cell viability studies based on Annexin V and Propidium iodide probes have confirmed the high proapoptotic effect of betulin ester derivatives against cancer cells.

Succession Processes in the Anoxygenic Phototrophic Bacterial Community in Lake Kislo-Sladkoe (Kandalaksha Bay, White Sea).

The community of anoxygenic phototrophic bacteria (APB) in the water column of Lake Kislo- Sladkoe (Kandalaksha Bay, White Sea), which has recently become separated from the sea, was investigated in March-April 2012, March-April 2013, and in September 2013. The lake, which was previously considered meromictic, was in fact mixed and was strongly affected by the sea. In winter the lake is sometimes washed off with seawater, and this together with the seasonal cycles of succession processes determines the succession of the community. The consequences of the mixing in autumn 2011 could be observed in the APB community as late as autumn 2013. Green-colored green sulfur bacteria (GSB) usually predominated in the chemocline. In winter 2013 stagnation resulted in turbidity of water under the ice, which was responsible for both predom- inance of the brown GS B forms and the changes ratio of the species of purple sulfur bacteria (PS B) in anoxic water layers. Production of anoxygenic photosynthesis in the lake was at least 240 mg C m-2 day-- in September and 0-20 mg C m-2 day- in March-April, which corresponded to 40 and 69%, respectively, of oxygenic photosynthesis. Okenone-containing purple sulfur bacteria, strain TcakPS12 were isolated in 2012 from lake water. The ells of this strain form filaments of not separated cells. Strain TcakPS12 exhibited 98% similarity with the type strains of Thiocapsapendens DSM.236 and Thiocapsa bogorovii BBS, as well as with the strains AmPS10 and TcyrPS 10, which were isolated from Lake Kislo-Sladkoe in 2010.

[Candidatus "Jettenia moscovienalis" sp. nov., a New Species of Bacteria Carrying out Anaerobic Ammonium Oxidation].

A new species of bacteria oxidizing ammonium with nitrite under anoxic conditions was isolated from the activated sludge of a semi-industrial bioreactor treating digested sludge of the Kuryanovo wastewater treatment plant (Moscow, Russia). Physiological, morphological, and molecular genetic characterization of the isolate was carried out. The cells were ovoid (-0.5 x 0.8 µm), with the intracellular membrane structures characteristic of anammox bacteria (anammoxosome and paryphoplasm); unlike other anammox bacteria, it possessed extensive intracellular membrane structures located in layers parallel to the cytoplasmic membrane, but never close to the anammoxosome. The cells formed aggregates 5-28 µm in diameter and readily attached to solid surfaces. The cells were morphologically labile, easily plasmolyzed, and lost their content. Doubling time was 28 days, µ(max) = 0.025 day(-1); optimal temperature and pH for growth were 20-45 degrees C and 8.0, respectively. Phylogenetic analysis of the 16S rRNA gene sequences suggested its classification as a new species of the candidate genus Jettenia (order Planctomycetales). The name Candidatus "Jettenia moscovienalis" sp. nov. was proposed for the new bacterium.

[Anoxygenic phototrophic bacteria of the Kislo-Sladkoe Stratified Lake (White Sea, Kandalaksha Bay)].

The community of anoxygenic phototrophic bacteria (APB) in the water column of the Kislo-Sladkoe stratified lake recently isolated from the sea (White Sea, Kandalaksha Bay) was investigated in September 2010. The water of the sulfide-rich zone was greenish-brown due to intense development of green sulfur bacteria (GSB). Nine APB strains were isolated from the water samples: three belonging to GSB, five, to purple sulfur bacteria (PSB), and one, to purple nonsulfur bacteria (PNB). GSB predominated in the phototrophic community of the chemocline. Unexpectedly, two morphologically different green-colored GSB strains were found to be phylogenetically identical and related to the brown-colored @Chlorobium phaeovibrioides (99% similarity according to the 16S rRNA gene sequencing). Homology to the closest green-colored species (Chlorobium luteolum) was 98%. Two morphologically and physiologically similar PSB strains (TcrPS10 and AmPS10) had rounded cells containing okenonokenonee and gas vesicles. According to the 16S rRNA gene sequencing, these strains were most closely related (99%) to two different Thiocapsa species: Tca. marina (containing okenonokenonee and no gas vesicles) and Tca. rosea (containing spirilloxanthin and gas vesicles). The remaining isolates of purple bacteria were similar to the already described APB species.

[Characterization of a new strain of a purple nonsulfur bacterium from a thermal spring].

A new budding nonsulfur purple bacterium of the genus Rhodobacter (strain Ku-2) was isolated from a mat of a moderately thermal spring (Baikal rift zone, Buryat Republic, Russia). The bacterium had lamellar photosynthetic membranes, which are typica of only one Rhodobacter species, Rba. blasticus. The cells contined spheroidene carotenoids and bacteriochlorophyll a (Bchl a). In vivo absorption spectrum of the cells had the major maximum at 863 nm and an additional peak at 887 nm, which is characteristic of the pigment-protein complexes of Bchl a-containing membranes. The previously described Rba. blasticus strains did not exhibit a 887-nm maximum. The new isolate was photoheterotrophic, with optimal growth occurring at 35 degrees C, 3 g/L NaCl, and pH 7-8. The DNA G+C content was 64.4 mol %. The similarity between the 16S rRNA gene sequences of strain Ku-2 and the Rba. blasticus type strain was 98.7%. The similarity between the PufM amino acid sequences of strain Ku-2 and the previously studied Rba. blasticus strain was 89.0%. Thus, the bacterial strain Ku-2 belonged to the genus Rhodobacter and was phylogenetically related to Rba. blasticus.

[Activity and structure of the sulfate-reducing bacterial community in the sediments of the southern part of Lake Baikal].

The rates of sulfate reduction (SR) and the diversity of sulfate-reducing bacteria (SRB) were studied in the sediments of the Posol'skaya banka elevation in the southern part of Lake Baikal. SR rates varied from 1.2 to 1641 nmol/(dm3 day), with high rates (> 600 nmol/(dm3 day)) observed at both deep-water stations and in subsurface silts. Integral SR rates calculated for the uppermost 50 cm of the sediments were higher for gas-saturated and gas hydrate-bearing sediments than in those with low methane content. Enrichment SRB cultures were obtained in Widdel medium for freshwater SRB. Analysis of the 16S rRNA gene fragments from clone libraries obtained from the enrichments revealed the presence of SRB belonged to Desulfosporosinus genus, with D. lacus as the most closely related member (capable of sulfate, sulfite, and thiosulfate reduction), as well as members of the order Clostridiales.

[Anoxygenic phototrophic bacteria from microbial communities of Goryachinsk Thermal Spring (Baikal Area, Russia)].

Species composition of anoxygenic phototrophic bacteria in microbial mats of the Goryachinsk thermal spring was investigated along the temperature gradient. The spring belonging to nitrogenous alkaline hydrotherms is located at the shore of Lake Baikal 188 km north-east from Ulan-Ude. The water is of the sulfate-sodium type, contains trace amounts of sulfide, salinity does not exceed 0.64 g/L, pH 9.5. The temperature at the outlet of the spring may reach 54 degrees C. The cultures of filamentous anoxygenic phototrophic bacteria, nonsulfur and sulfur purple bacteria, and aerobic anoxygenic phototrophic bacteria were identified using the pufLM molecular marker. The fmoA marker was used for identification of green sulfur bacteria. Filamentous cyanobacteria predominated in the mats, with anoxygenic phototrophs comprising a minor component of the phototrophic communities. Thermophilic bacteria Chloroflexus aurantiacus were detected irn the samples from both the thermophilic and mesophilic mats. Cultures ofnonsulfur purple bacteria similar to Blastochloris sulfoviridis and Rhodomicrobium vannielii were isolatd from the mats developing at high (50.6-49.4 degrees C) and low temperatures (45-20 degrees C). Purple sulfur bacteria Allochromatium sp. and Thiocapsa sp., as well as green sulfur bacteria Chlorobium sp., were revealedin low-temperature mats. Truly thermophilic purple and gree sulfur bacteria were not found in the spring. Anoxygenic phototrophic bacteria found in the spring were typical of the sulfuret communities, for which the sulfur cycle is mandatory. The presence of aerobic bacteriochlorophylla-containing bacteria identified as Agrobacterium (Rhizobium) tumifaciens in the mesophilic (20 degrees C) mat is of interest.

[Production of modified magnetosome membrane proteins and analysis of their activity].

Genetic constructs coding for the fusion of the Mam12 membrane protein from the magnetotactic bacteria Magnetospirilium magnetotacticum with one B-domain (mb) or two B or Z domains (mbb and mzz, respectively) of Staphylococcus aureus protein A have been synthesized. Expression, fractionation, and purification of fusion proteins have been performed, and the activity of the modified proteins has been characterized. ELISA results showed that all modified proteins displayed immunoglobulin-binding activity.

[A novel bacterium carrying out anaerobic ammonium oxidation in a reactor for biological treatment of the filtrate of wastewater fermented residue].

A new genus and species of bacteria capable of ammonium oxidation under anaerobic conditions in the presence of nitrite is described. The enrichment culture was obtained from the Moscow River silt by sequential cultivation in reactors with selective conditions for anaerobic ammonium oxidation. Bacterial cells were coccoid, -0.4 x 0.7 mm, with the intracellular membrane structures typical of bacteria capable of anaerobic ammonium oxidation (anammoxosome and paryphoplasm). The cells formed aggregates 5-25 µm in diameter (10 µm on average). They were readily adhered to solid surfaces. The cells were morphologically labile, they easily lost their content and changed their morphology during fixation for electron microscopy. The organism was capable of ammonium oxidation with nitrite. The semisaturation constants Ks for nitrite and ammonium were 0.38 mg N-NO2/L and 0.41 mg N-NH4/L, respectively. The maximal nitrite concentrations for growth were 90 and 75 mg N-NO2/L for single and continuous application, respectively. The doubling time was 32 days, µ(max) = 0.022 day(-1), the optimal temperature and pH were 20 degrees C and 7.8-8.3, respectively. According to the 16S rRNA gene sequencing, the bacterium was assigned to a new genus and species within the phylum Planctomycetes. The proposed name for the new bacterium is Candidatus Anammoximicrobium moscowii gen. nov., sp. nov. (a microorganisms carrying out anaerobia ammonium oxidation, isolated in the Moscow region).