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Cancer stem cells (CSCs) are the most malignant subpopulation of tumor cells that possess a tumorigenic potential and resistantance to chemotherapy. These properties make CSCs a promising target for the development of targeted antitumor therapy which is especially in demand in highly aggressive cancers. However, the correct identification of cancer cells with stem properties remains a challenge. A newly developed lentivirus-based reporter SORE6 allows to directly identify CSCs by measuring gene expression of the embryonic stem cell factors SOX2 and OCT4. In the current study the reporter was modified to enable isolation of SOX2^(+)/OCT4^(+) cells by immunomagnetic separation and then was used to transduce HCC1806 and MDA-MB-453 triple-negative breast cancer (TNBC) cell lines. To validate the modified reporter, SOX2^(+)/OCT4^(+) populations were isolated and analyzed for the content of NANOG, a key transcription factor of pluropotency which expression is regulated by SOX2/OCT4. The percentage of SOX2^(+)/OCT4^(+) cells was assessed for each cell line. An increased content of NANOG protein was found in isolated SOX2^(+)/OCT4^(+) cell fractions indicating that the modified reporter is suitable for further studying the CSC subset.

The liquid phase of different units of an underground gas storage (UGS) in the period of gas injection was studied with respect to its hydrochemical composition and characterized microbiologically. The presence of viable aerobic and anaerobic bacteria was revealed in the UGS stratal and associated waters. An important source of microorganisms and biogenic elements in the ecosystem studied is water and various technogenic admixtures contained in trace amounts in the gas entering from the gas main in the period of gas injection into the storage. Owing to this fact, the bacterial functional diversity, number, and activity are maximal in the system of gas treatment and purification and considerably lower in the observation well zone. At the terminal stages, the anaerobic transformation of organic matter in the UGS aqueous media occurs via sulfate reduction and methanogenesis; exceptionally high rates of these processes (up to 4.9 x 10(5) ng S(2-)l(-1) day(-1) and 2.8 x 10(6) nl CH4 l(-1) day(-1), respectively) were recorded for above-ground technological equipment.

The numbers of microorganisms belonging to ecologically significant groups and the rates of terminal microbial processes of sulfate reduction and methanogenesis were determined in the liquid phase of an underground gas storage (UGS) in the period of gas extraction. The total number of microorganisms in water samples from the operation and injection wells reached 2.1 x 10(6) cells/ml. Aerobic organotrophs (including hydrocarbon- and oil-oxidizing ones) and various anaerobic microorganisms (fermenting bacteria, methanogens, acetogens, sulfate-, nitrate-, and iron-reducing bacteria) were constituent parts of the community. The radioisotopic method showed that, in all the UGS units, the terminal stages of organic matter decomposition included sulfate reduction and methanogenesis, with the maximal rate of these processes recorded in the aqueous phase of above-ground technological equipment which the gas enters from the operation wells. A comparative analysis by these parameters of different anaerobic ecotopes, including natural hydrocarbon fields, allows us to assess the rate of these processes in the UGS as high throughout the annual cycle of its operation. The data obtained indicate the existence in the UGS of a bacterial community that is unique in its diversity and metabolic capacities and able to make a certain contribution to the geochemistry of organic and inorganic compounds in the natural and technogenic ecosystem of the UGS and thus influence the industrial gas composition.

The effect of hydrogen peroxide ( 10(-9)-10(-1) M) on the mycelial growth of the fungi Alternaria alternata, Cladosporium cladosporioides, Mucor hiemalis, and Paecilomyces lilacinus has been studied. The growth of fungi isolated from habitats with a background level of radioactive contamination was stopped by H2O2 concentrations equal to 10(-3) and 10(-2) M, whereas the growth of fungi that were isolated from habitats with high levels of radioactive contamination was only arrested by 10(-1) M H2O2. The response of the different fungi to hydrogen peroxide was of three types: (1) a constant growth rate of fungal hyphae at H2O2 concentrations between 10(-9) and 10(-4) M and a decrease in this rate at 10(-3) M H2O2, (2) a gradual decrease in the growth rate as the H2O2 concentration was increased, and (3) an increase in the growth rate as the H2O2 concentration was increased from 10(-7) to 10(2)-5 M. The melanin-containing species A. alternata and C. cladosporioides exhibited all three types of growth response to hydrogen peroxide, whereas the light-pigmented species M. hiemalis and P. lilacinus showed only the first type of growth response. A concentration of hydrogen peroxide equal to 10(-1) M was found to be lethal to all of the fungi studied. The most resistant to hydrogen peroxide was found to be the strain A. alternata 56, isolated from the exclusion zone of the Chernobyl Nuclear Power Plant.

A detailed study of the processes of anaerobic methane oxidation and sulfate reduction in the bacterial mats occurring on coral-like carbonate structures in the region of methane seeps in the Black Sea, as well as of the phenotypic diversity of sulfate-reducing bacteria developing in this zone, has been performed. The use of the radioisotopic method shows the microbial mat structure to be heterogeneous. The peak activity of the two processes was revealed when a mixture of the upper (dark) and underlying (intensely pink) layers was introduced into an incubation flask, which confirms the suggestion that methanotrophic archaea and sulfate-reducing bacteria closely interact in the process of anaerobic methane oxidation. Direct correlation between the rate of anaerobic methane oxidation and the methane and electron acceptor concentrations in the medium has been experimentally demonstrated. Several enrichment and two pure cultures of sulfate-reducing bacteria have been obtained from the near-bottom water and bacterial mats. Both strains were found to completely oxidize the substrates to CO2 and H2S. The bacteria grow at temperatures ranging from -1 to 18 (24) degrees C, with an optimum in the 10-18 degrees C range, and require the presence of 1.5-2.5% NaCl and 0.07-0.2% MgCl2 x 6H2O. Regarding the aggregate of their phenotypic characteristics (cell morphology, spectrum of growth substrates, the capacity for complete oxidation), the microorganisms isolated have no analogues among the psychrophilic sulfate-reducing bacteria already described. The results obtained demonstrate the wide distribution of psychrophilic sulfate-reducing bacteria in the near-bottom water and bacterial mats covering the coral-like carbonate structures occurring in the region of methane seeps in the Black Sea, as well as the considerable catabolic potential of this physiological group of psychrophilic anaerobes in deep-sea habitats.

Spore germination and the viability of the mycelial fragments of the microscopic fungi Alternaria alternata, Penicillium spinulosum, and Mucor hiemalis were studied with respect to the action of some ecological factors: sucrose concentration (0, 0.2, 2, 10, and 100 g/l), temperature (4, 20, 25, and 30 degrees C), pH (3.5, 4.0, 5.0, 6.2, and 7.0), and cadmium concentration (0, 2, 10, and 100 mg/l). The spore germination rate and the viabilities of different mycelial fragments were found to reach their maxima at different values of the ecological factors studied. This finding suggests that the vegetative and asexual types of reproduction of microscopic fungi may have different ecological optima.

The distribution and population density of aerobic hydrocarbon-oxidizing bacteria in the high-temperature oil fields of Western Siberia, Kazakhstan, and China were studied. Seven strains of aerobic thermophilic spore-forming bacteria were isolated from the oil fields and studied by microbiological and molecular biological methods. Based on the 16S rRNA gene sequences, phenotypic characteristics, and the results of DNA-DNA hybridization, the taxonomic affiliation of the isolates was tentatively established. The strains were assigned to the first and fifth subgroups of the genus Bacillus on the phylogenetic branch of the gram-positive bacteria. Strains B and 421 were classified as B. licheniformis. Strains X and U, located between B. stearothermophilus and B. thermocatenulatus on the phylogenetic tree, and strains K, Sam, and 34, related but not identical to B. thermodenitrificans and B. thermoleovorans, undoubtedly represent two new species. Phylogenetically and metabolically related representatives of thermophilic bacilli were found to occur in geographically distant oil fields.