Lake Baikal: a unique place to study evolution of sponges and their stress response in an environment nearly unimpaired by anthropogenic perturbation.

Lake Baikal is considered as a unique place to study evolution. In this review, we report on recent data on the evolution of endemic freshwater sponges of this ancient lake. Nucleotide sequence data support the idea that these sponges are of monophyletic origin and evolved from Spongillidae. Baikalian sponges form the dominating biomass in the benthos of the lake. Data on the expression of the biomarker heat shock protein 70, revealed that the endemic sponge species of Lake Baikal are useful as bioindicators to assess the anthropogenic impact on the lake.     

The effects of synthetic protease inhibitors on human proinsulin production by recombinantBacillus subtilis strain

Summary Two inexpensive inhibitors non-toxic to B. subtilis cells, O,O-diethyl-1-(N--hydrohexafluoro-isobutyryl)amino-1-methyl-propylphosphonate and O,O-diísobutyl-1-[2-(ethoxycarbonyl)amino-perfluoroprop-2-yl]-1-methylpropylphosphonate, when added to the culture broth, are able to prevent the proteolytic degradation of recombinant human proinsulin, secreted byB. subtilis AJ73apr73 npr ^– cells.     

Biodiversity and Activity of the Microbial Community in the Kotelnikovsky Hot Springs (Lake Baikal)

Complex microbiological and chemical analyses of water and bacterial mats were performed in the Kotelnikovsky Hot Springs (Lake Baikal). Transmission electron microscopy demonstrated that short rods about 1.2–2 μm in diameter predominated in the natural microbial community. Scanning electron microscopy coupled with chemical analysis revealed a characteristic P peak in the bacteria-like mineral particles, which suggests their biogenic origin. Most strains of the thermophilic microorganisms were gram-positive spore-forming rods and can be assigned to the genus Bacillus. Assays for potential enzyme activity demonstrated that most of the strains tested were active at high temperature. The data obtained suggest high activity of the bacterial community in situ and its particular role in the functioning of the hydrothermal ecosystem. __________ Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 6, 2005, pp. 664–671. Original Russian Text Copyright ? 2005 by Bel'kova, Parfenova, Suslova, Ahn, Tazaki.     

Study of effectiveness of long-term per oral and parenteral cincacine administration at parenteral 241Am incorporation into the organism

Effect of long-term (during 4 weeks) cincacine administration following single parenteral 241Am intake has been studied on rats depending on method (per oral or parenteral), dosage and time of treatment initiation. Cincacine administration leads to limitation of radionuclide incorporation in the major organs of deposition for the both methods of introduction. At the parenteral 241Am intake in the organism parenteral cincacine administration was found to be more effective compared to per oral cincacine administration even in case of its dose increase by a factor of 6 and 12. At the parenteral introduction of the preparation, time of treatment beginning is more significant than at per oral administration.     

Role of malate dehydrogenase isoforms in the regulation of anabolic and catabolic processes in the colorless sulfur bacterium Beggiatoa leptomitiformis D-402

The functional role of tetrameric and dimeric isoforms of malate dehydrogenase in the carbon metabolism of the colorless sulfur bacterium Beggiatoa leptomitiformis, strain D-402, was studied. This strain can grow both lithotrophically and organotrophically. By using the inhibition analysis, the tetrameric isoenzyme was shown to operate in the glyoxylate cycle and the dimeric one was found to be involved in the TCA cycle. The dynamics of the dimeric isoenzyme conversion to the tetrameric isoform was found to be determined by the rate of thiosulfate oxidation. The regulation of the carbon metabolism in Beggiatoa leptomitiformis is supposed to be accomplished by means of structural and functional changes in the protein molecule of malate dehydrogenase.     

Carbon monoxide activates KCa channels in newborn arteriole smooth muscle cells by increasing apparent Ca2+ sensitivity of alpha-subunits

Carbon monoxide (CO) is a gaseous vasodilator produced by many cell types, including endothelial and smooth muscle cells. The goal of the present study was to investigate signaling mechanisms responsible for CO activation of large-conductance Ca(2+)-activated K(+) (K(Ca)) channels in newborn porcine cerebral arteriole smooth muscle cells. In intact cells at 0 mV, CO (3 microM) or CO released from dimanganese decacarbonyl (10 microM), a novel light-activated CO donor, increased K(Ca) channel activity 4.9- or 3.5-fold, respectively. K(Ca) channel activation by CO was not blocked by 1-H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (25 microM), a soluble guanylyl cyclase inhibitor. In inside-out patches at 0 mV, CO shifted the Ca(2+) concentration-response curve for K(Ca) channels leftward and decreased the apparent dissociation constant for Ca(2+) from 31 to 24 microM. Western blotting data suggested that the low Ca(2+) sensitivity of newborn K(Ca) channels may be due to a reduced beta-subunit-to-alpha-subunit ratio. CO activation of K(Ca) channels was Ca(2+) dependent. CO increased open probability 3.7-fold with 10 microM free Ca(2+) at the cytosolic membrane surface but only 1.1-fold with 300 nM Ca(2+). CO left shifted the current-voltage relationship of cslo-alpha currents expressed in HEK-293 cells, increasing currents 2.2-fold at +50 mV. In summary, data suggest that in newborn arteriole smooth muscle cells, CO activates low-affinity K(Ca) channels via a direct effect on the alpha-subunit that increases apparent Ca(2+) sensitivity. The optimal tuning by CO of the micromolar Ca(2+) sensitivity of K(Ca) channels will lead to preferential activation by signaling modalities, such as Ca(2+) sparks, which elevate the subsarcolemmal Ca(2+) concentration within this range.     

Diversity and physiological and biochemical properties of heterotrophic bacteria isolated from Lake Baikal neuston

For heterotrophic microorganisms (44 strains) isolated from the surface film of Lake Baikal, identification was carried out and their physiological and biochemical characteristics were determined. Compared to the water column, diversity of cultured heterotrophs was low, indicating formation of stable microbial communities at the air–water interface. Heterotrophic bacteria isolated from the surface microlayer exhibited the enzymatic activity comparable to that for strains from other biofilm associations. Deinococcus ficus strain NA202 was the most active component of the community, capable of utilization of the broadest spectrum of mono- and disaccharides, sugars, and amino acids. This strain possessed the highest diversity of extracellular enzymes and was the most resistant to UV radiation. The physiological and biochemical properties of this strain may be responsible for its adaptation to survival in extreme conditions of the surface microlayer. These results improve our understanding of occurrence of UV-resistant strains in freshwater ecosystems.     

Viability and angiogenic activity of mesenchymal stromal cells from adipose tissue and bone marrow under hypoxia and inflammation in vitro

Progenitor stromal cells derived from adipose tissue (ADSC) and bone marrow (BMDSC) hold great promise for use in the cell-based therapy of ischemic diseases. It was demonstrated that these cells secrete a number of angiogenic cytokines that stimulate vascularization. It was demonstrated that ADSC or BMDSC injected intramuscularly or intravenously into the animals with experimental hind-limb ischemia improve vascularization. However, low oxygen levels and inflammation may impair the viability and functional activity of transplanted cells. We have examined ADSC and BMDSC properties in vitro under hypoxic and inflammatory conditions. ADSC and BMDSC derived from Balb/c mice have been cultivated under hypoxia or in the presence of inflammatory cytokines. The viability of cells assessed by annexin V-PE binding and 7AAD storage, as well as by the quantitative TUNEL method, was not changed under hypoxic conditions Cell exposure to inflammatory cytokines induced apoptosis in 70% of cells. Inflammatory cytokines did not stimulate gene expression of angiogenic growth factors. Under hypoxia conditions up-regulation of genes for pro-angiogenic factors and down-regulation of anti-angiogenic genes were more apparent in ADSC. Using angiogenesis models in vitro and in vivo, we demonstrated that stromal cell maintenance under hypoxic conditions increased their ability to stimulate the growth of blood vessels.