Silicon carbide fiber-mediated DNA delivery into cells of wheat (Triticum acstivum L.) mature embryos

We have demonstrated that foreign DNA can be delivered into cells of mature embryos of wheat (Triticum aestivum L.) using silicon carbide fibers (SCF). The highest transient expression of thegusA (GUS) gene was detected when dry embryos were vortexed for 10–30 min in a SCF-DNA solution containing 90–120 g/l of sucrose. Up to 100 (on average 20–40) blue expression units per embryo were observed. Scutellum side and epiblast of the intact wheat embryos are preferentially transformed. When embryos with the coleoptilar tip removed were treated and allowed to germinate, GUS staining was observed in emerging leaf tissues. The potential of this new approach for stable transformation of wheat is under investigation. It has been found that callus tissues induced from the SCF treated embryos contain GUS-expressing sectors one month after treatment.     

Versatility of the green microalga cell vacuole function as revealed by analytical transmission electron microscopy

Vacuole is a multifunctional compartment central to a large number of functions (storage, catabolism, maintenance of the cell homeostasis) in oxygenic phototrophs including microalgae. Still, microalgal cell vacuole is much less studied than that of higher plants although knowledge of the vacuolar structure and function is essential for understanding physiology of nutrition and stress tolerance of microalgae. Here, we combined the advanced analytical and conventional transmission electron microscopy methods to obtain semi-quantitative, spatially resolved at the subcellular level information on elemental composition of the cell vacuoles in several free-living and symbiotic chlorophytes. We obtained a detailed record of the changes in cell and vacuolar ultrastructure in response to environmental stimuli under diverse conditions. We suggested that the vacuolar inclusions could be divided into responsible for storage of phosphorus (mainly in form of polyphosphate) and those accommodating non-protein nitrogen (presumably polyamine) reserves, respectively.The ultrastructural findings, together with the data on elemental composition of different cell compartments, allowed us to speculate on the role of the vacuolar membrane in the biosynthesis and sequestration of polyphosphate. We also describe the ultrastructural evidence of possible involvement of the tonoplast in the membrane lipid turnover and exchange of energy and metabolites between chloroplasts and mitochondria. These processes might play a significant role in acclimation in different stresses including nitrogen starvation and extremely high level of CO₂ and might also be of importance for microalgal biotechnology. Advantages and limitations of application of analytical electron microscopy to biosamples such as microalgal cells are discussed.     

Cationic penetrating antioxidants switch off Mn cluster of photosystem II in situ

Photosynthesis Research - Photosystem I (PSI) generates the most negative redox potential found in nature, and the performance of solar energy conversion into alternative energy sources in...     

The effect of cytisine on mRNA transport in the cytoplasm of eukaryotic cells (plants and animals)

The action of alkaloid cytisine on protein biosynthesis in eukaryotic cells was studied. It was shown that the alkaloid had no effect on mRNA translation. Cytisine inhibited the release of mRNP particles from rat liver and wheat embryos nuclei. Sedimentation properties and distribution in CsCl gradient of the material extracted from alkaloid-treated nuclei did not differ from control one and were similar to informosomes from animal and plant cells described earlier. The main part of mRNA with sedimentation coefficients 14-18S, capable for translation in the cytoplasm is retained in alkaloid-treated nuclei.     

Application of the analytical transmission electron microscopy techniques for detection,identification and visualization of localization of nanoparticles of titanium and cerium oxides in mammalian cells

This work represents the results of the study on applicability of the modern methods of analytical transmission electron microscopy (TEM) for detection, identification and visualization of localization of nanoparticles of titanium and cerium oxides in A549, human lung adenocarcinoma cell line. Comparative analysis was performed for images of the nanoparticles in cells obtained in the bright-field mode of TEM, bright-field scanning TEM, and high-angle annular dark field scanning TEM. For identification of nanoparticles in the cells, the analytical techniques, energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy, were compared when used in the modes of obtaining energy spectra from different particles and of element mapping. It was shown that electron tomography is applicable to confirm that nanoparticles are localized in the sample rather than brought in by contamination. The possibilities and fields of using different techniques of analytical TEM for detection, visualization and identification of nanoparticles in biological samples are discussed.     

Characterization of lipodisc nanoparticles containing sensory rhodopsin II and its cognate transducer from Natronomonas pharaonis

We describe the preparation and properties of lipodisc nanoparticles–lipid membrane fragments with a diameter of about 10 nm, stabilized by amphiphilic synthetic polymer molecules. We used the lipodisc nanoparticles made of Escherichia coli polar lipids and compared lipodisc nanoparticles that contained the photosensitive protein complex of the sensory rhodopsin with its cognate transducer from the halobacterium Natronomonas pharaonis with empty lipodisc nanoparticles that contained no protein. The lipodisc nanoparticles were characterized by dynamic light scattering, transmission electron microscopy and atomic force microscopy. We found that the diameter of lipodisc nanoparticles was not affected by incorporation of the protein complexes, which makes them a prospective platform for single-molecule studies of membrane proteins.

     

A new subarctic strain of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis>—part I: identification and fatty acid profiling

We isolated a new subarctic strain Tetradesmus obliquus IPPAS S-2023 (Scenedesmaceae, Chlorophyceae) from rock baths in the White Sea. To verify its taxonomic assignment, internal transcribed spacer 2 (ITS2) of the strain was sequenced and its secondary structure was compared with predicted ITS2 secondary structures of Scenedesmaceae. The analysis of the ITS2 made it possible to assign the new strain IPPAS S-2023 to the species T. obliquus. The ultrastructural studies and energy-dispersive X-ray spectroscopy (EDX) analysis revealed a marked accumulation of vacuolar inclusions enriched in phosphorus and nitrogen (N) as well as cytoplasmiс oil bodies. Most of predicted properties of biodiesel derived from the fatty acids profile of the strain grown in the N-free medium complied with the requirements of European and American standards. The results suggest that the new subarctic strain T. obliquus IPPAS S-2023 is a promising candidate for nutrients biosequestration and for biodiesel production. In a companion paper, we assess its biomass production capability and suitability and demonstrated suitability of IPPAS S-2023 as a reference strain for studies on elevated CO₂ stress effects selection of carbon dioxide-tolerant microalgae by comparison with a CO₂-tolerant strain IPPAS S-2014.     

A new subarctic strain of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis>. Part II: comparative studies of CO<Subscript>2</Subscript>-stress tolerance

A huge interest in CO₂-tolerant microalgae is fueled by development of CO₂-biomitigation methods based on intensive cultivation of microalgae. Still, the mechanisms of CO₂-tolerance are scarcely investigated. Previously, we described a symbiotic Desmodesmus sp. IPPAS S-2014 from a White Sea hydroid tolerant to extremely high (20–100%) CO₂ levels. In the present work, we compared its ultrastructural and physiological responses to those of a novel free-living White Sea strain of Tetradesmus obliquus IPPAS S-2023 characterized in the companion paper. The strain S-2023 is closely related to Desmodesmus sp. IPPAS S-2014 but lacks its tolerance to extremely high CO₂ (it is unable to survive at 100% CO₂ and exhibits a reduced-growth phenotype when sparged with 20% CO₂: air mixture). We compared the responses of the cell organization and photosynthetic activity to 20% CO₂ in the tolerant and the intolerant White Sea chlorophytes using chlorophyll fluorescence measurements and ultrastructural analysis (transmission electron microscopy). The features peculiar to the CO₂-intolerant chlorophyte include (i) inability to maintain pH homeostasis, (ii) a steady decline in the photosynthetic activity of the cells, (iii) a reduction of the photosynthetic membranes, and (iv) delayed accumulation of starch (starch grains) and its subsequent conversion to reserve lipids (oil bodies). Nitrogen starvation enhances the effects of high-CO₂ stress in the CO₂-intolerant microalga. The results of this work are discussed in the context of selection of tolerant algal strains for CO₂ biomitigation applications.     

Lung volume, chest size, and hematological variation in low-, medium-, and high-altitude central Asian populations

To evaluate adaptive responses to high-altitude environment, we examined three groups of healthy adult males from Central Asia: 94 high-altitude (HA) Kirghiz subjects (3,200 m above sea level); 114 middle-altitude (MA) Kazakh subjects (2,100 m), and 90 low-altitude (LA) Kirghiz subjects (900 m). Data on chest size (chest perimeter and chest diameter), lung volume (forced expiratory volume (FEV) and forced expiratory volume in 1 sec (FEV1)), and hematological parameters (hemoglobin, erythrocytes, hematocrit, and SaO(2)) are discussed. The results show that 1) chest shape is less flat in the samples living at higher altitude. In the HA sample, chest perimeter is lower but chest excursion is high. 2) In the highlanders, forced vital capacity (FVC) and FEV1 are no higher than in the other samples, even when corrected for stature and body weight. The negative correlation between FVC-FEV1 and age decreases with increasing altitude. 3) The HA and MA samples have higher values of hemoglobin, erythrocytes, and hematocrit. The HA sample has lower SaO(2) and higher arterial oxygen content than the LA sample. No association between hematocrit and age was detected in the four samples. The results indicate that the high-altitude Kirghiz present features of developmental acclimatization to hypobaric hypoxia which are also strongly influenced by other major high-altitude environmental stresses.     

Use of ampicillin trihydrate in transplantation surgery]

Ampicillin trihydrate was used for the treatment of 29 patients with purulent inflammatory processes, such as peritonitis, suppurating operative wound, urinary tract infection after the kidney allotransplantation. The antibacterial activity of ampicillin was preliminarily tested on 517 microbial strains, i.e. staphylococci, streptococci, E. coli, Proteus and Ps. aeruginosa isolated from the surgical patients. The strains of penicillin sensitive staphylococci, streptococci and E. coli were most sensitive to the drug effect, the MIC ranging from 0.03 to 16 gamma/ml. It was shown that the blood retention time of the antibiotic was much more prolonged in the patients with a decreased excretion function of the kidneys. The treatment was performed under control of the clinical, bacteriological and biochemical parameters. The drug was used in a dose of 0.5 g 6--8 times a day for 5 to 15 days. A satisfactory therapeutic effect was registered in 73 per cent of the cases.