Development of a PCR-based method for diagnostics of mycoses

Since the express-diagnostics of mycoses in immune-deficit patients still remains an acute problem, we developed an effective test system (Kan-Am) to detect DNA Candida albicans, which is a leader in the list of causative agents of candidosis. A comparison study of three PCR-systems used to detect a broad spectrum of fungoid pathogens was carried out, and a universal system (FungAm), which ensures the detection of DNAs of above 78 strains of 25 types of pathogenic fungi, was selected. The results of clinical testing of the species-specific and universal PCR-systems are well confirmed by the culture method, and they are indicative of the efficacy of applying them for the diagnostics of mycoses in neonatology. The use of the mentioned systems is a promising factor for the express-diagnostics of mycoses in immunodeficiency patients. The high sensitivity of the method makes it possible to detect 10 to 100 cells of a causative agent in 100 mcl of the examined biological material, which is compatible with the culture method. A kit of dry reagents (IonoMix) designed for an accelerated sample preparation and isolation, from them, of DNAs on the basis of Chelex-100 and of proteinase K was worked out; the kit is portable and meant for a long-term storing.     

Insertion of transposon P(lacW) with gradual expression of reporter genes

In Drosophila, insertions of the P?lacW? transposon led to uncommon expression patterns of the reporter genes white and lacZ: gene activity was gradually reduced in the neighboring cells along the dorsal-ventral or proximal-distal axes. Using in situ hybridization, the gradual gene expression was shown to be caused by P?lacW? insertions in regions 40A (lines G1 and G2) and 69C (line G3) of the Drosophila genome. The expression patterns of genes lacZ and white were similar in eye cells of each line. In salivary gland polytene cells of G3 flies, the lacZ expression was also gradually reduced, which indicates the existence of a mechanism maintaining the gene activity gradient during endomitosis.     

A Finding of the Red Squirrel (Rodentia,Sciuridae, <Emphasis Type="Italic">Sciurus vulgaris</Emphasis> Linnaeus, 1758) and Forest Dormouse (Rodentia,Gliridae, <Emphasis Type="Italic">Dryomys nitedula</Emphasis> Linnaeus, 1778) in the Pleistocene of the Southern Urals

The study deals with a new sample of the mammalian fossils from the Pleistocene deposits of the Ignatievskaya Cave in Southern Urals (54°53' N, 57°46' E). Among the rodent fossils, the teeth of red squirrel (Sciurus vulgaris) and forest dormouse (Dryomys nitedula) have been identified. The enamel differentiation quotient (SDQ) of the water vole teeth matches the Arvicola terrestris species from locations of the end of Middle and Late Pleistocene (oxygen isotope stages, OIS 6 and OIS 5–2, respectively). The high SDQ value for the water vole and the presence of the squirrel and dormouse in the fauna allow dating the finding near the end of the Middle Pleistocene and beginning of the Late Pleistocene (OIS 6 and OIS 5e, respectively). The widespread open and forest landscapes were inhabited by this fauna.     

Calcium-induced calcium release mediates all-or-nothing responses of mesenchymal stromal cells to noradrenaline

By using Ca²⁺ imaging and Fluo-4 dye, we examined the capability of certain agonists of G-protein coupled receptors to stimulate Ca²⁺ signaling in cultured mesenchymal stromal cells (MSC) derived from the human adipose tissue. In particular, a small subpopulation (～5%) MSC was found to respond to noradrenaline with Ca²⁺ transients. The all-or-nothing fashion was characteristic of adrenergic Ca²⁺ signaling in MSC, that is, while at low concentrations noradrenaline stimulated undetectable Ca²⁺ transients, virtually maximal responses were elicited by this agonist at any concentration above the threshold of 100–200 nM. In some experiments, MSC were loaded with the photosensitive Ca²⁺ chelator NP-EGTA to produce local or global jumps in cytosolic Ca²⁺ concentration by virtue of Ca²⁺ uncaging. Global uncaging eliciting a high enough Ca²⁺ jump triggered a Ca²⁺ transient in the MSC cytoplasm, which was similar to a noradrenaline response kinetically and by magnitude. When Ca²⁺ uncaging was produced locally, it initiated a Ca²⁺ signal that traveled along a cell with a speed that exceeded an expected one by two orders of magnitude, should Ca²⁺ signal transfer be mediated merely by passive Ca²⁺ diffusion in the presence of Ca²⁺ buffer. These findings implicated Ca²⁺-induced Ca²⁺ release (CICR) as a mechanism amplifying local Ca²⁺ signals in MSC. Of Ca²⁺ targets involved in CICR, the ryanodine receptor and IP₃ receptor are only known. The inhibitory analysis revealed IP₃ receptors to be principally responsible for CICR in MSC, whereas a contribution of ryanodine receptors was negligible. Altogether, our results suggest that an initial noradrenaline-dependent rise in cytosolic Ca²⁺ stimulates, should it reach the threshold level, IP₃ receptors, thereby triggering an avalanche-like Ca²⁺ release from Ca²⁺ stores and underlying the all-or-nothing dependence of cellular responses on the agonist concentration.     

Heavy Metal Contents in Tissues of Dominant Species of the Benthos and in Bottom Sediments of Zolotoi Rog Bay, Sea of Japan

Data on heavy metal contents in polychaetes and free-living nematodes inhabiting the bottom sediments of Zolotoi Rog Bay near the port of Vladivostok are reported. Chronically high contents of heavy metals (Fe, Cu, Zn, and Cd) were found in the bottom sediments and in the infauna. The levels of some toxic elements in tissues of the polychaete Dorvillea (Schistomeringos) japonica inhabiting polluted sediments of Zolotoi Rog Bay were higher than in worms of this species from relatively clean areas of the Sea of Japan. Similar high concentrations of Fe were found in the polychaete Capitella capitata and the nematode Oncholaimium ramosum. The highest concentrations of Fe, Zn, Cu, and Cd were found in tissues of the free-living nematode O. ramosum. Chronic heavy pollution of the bottom sediments leads to a decrease in the biodiversity of the infauna and to specific metal-tolerant communities.Original Russian Text Copyright © 2005 by Biologiya Morya, Davydkova, Fadeeva, Kovekovdova, Fadeev.     

Effects of a medium pH on the ability of Chinese hamster cells to repair radiation injuries]

In experiments with Chinese hamster cells at exponential and stationary growth phases, it has been shown that the postirradiation incubation of irradiated cells in a medium with low pH (up to 6.0) promotes the recovery of cells from potentially lethal damages; it has also been found that the recovery from sublethal radiation damages does not depend on the medium pH. The long-term incubation of nonirradiated cells with low pHc causes death of part of cells.     

Intensification of antibacterial and antiviral nonsusceptibility and of the immune response with an officinal RNA preparation]

Sodium nucleinate increased essentially the insusceptibility of mice to pathogenic escherichia, strain O26, Pr. vulgaris, Ps. aeruginosa, Ser. marcescens, and produced a total stimulating effect on the nonspecific bacterial resistance; analogous stimulating activity was found in the homologous low polymeric RNA from the liver. Sodium nucleinate intensified the insusceptibility of the animals to the tick-born encephalitis and encephalomyelitis viruses, and increased the antibody-forming cells count. The side-effect of heat-inactivated vaccine from pathogenic escherichia was reduced in animals inoculated with sodium nucleinate preliminarily.