Enzymes of SPZ7 phage: Isolation and properties

Bacteriophage enzyme preparations exolysin and endolysin were studied. Exolysin (a phage-associated enzyme) was obtained from tail fraction and endolysin from phage-free cytoplasmic fraction of disintegrated Salmonella enteritidis cells. A new method for purification of these enzymes was developed, and their molecular masses were determined. The main catalytic properties of the studied enzymes (pH optimum and specificity to bacterial substrates) were found to be similar. Both enzymes lyse Escherichia coli cells like chicken egg lysozyme, but more efficiently lyse S. enteritidis cells and cannot lyse Micrococcus luteus, a good substrate for chicken egg lysozyme. Similar properties of exolysin and endolysin suggest that these enzymes are structurally similar or even identical.     

Induction of permeability of the inner membrane of yeast mitochondria

The current view on apoptosis is given, with a special emphasis placed on apoptosis in yeasts. Induction of a non-specific permeability transition pore (mPTP) in mammalian and yeast mitochondria is described, particularly in mitochon-dria from Yarrowia lipolytica and Dipodascus (Endomyces) magnusii yeasts, which are aerobes possessing the fully competent respiratory chain with all three points of energy conservation and well-structured mitochondria. They were examined for their ability to induce an elevated permeability transition of the inner mitochondrial membrane, being subjected to virtually all conditions known to induce the mPTP in animal mitochondria. Yeast mitochondria do not form Ca²⁺-dependent pores, neither the classical Ca²⁺/P_i-dependent, cyclosporin A-sensitive pore even under deenergization of mitochondria or depletion of the intramitochondrial nucleotide pools, nor a pore induced in mammalian mitochondria upon concerted action of moderate Ca²⁺ concentrations (in the presence of the Ca²⁺ ionophore ETH129) and saturated fatty acids. No pore formation was found in yeast mitochondria in the presence of elevated phosphate concentrations at acidic pH values. It is concluded that the permeability transition in yeast mitochondria is not coupled with Ca²⁺ uptake and is differently regulated compared to the mPTP of animal mitochondria.     

Effects of chronic imipramine treatment on behavior in mice of congenic strains AKR and AKR.CBA-D13Mit76 differing in the distal fragment of chromosome 13

Congenic mouse strain AKR.CBA-D13Mit76 carries the 59-70 cM fragment of chromosome 13 transferred from genome of cataleptic CBA/Lac strain to genome of AKR/J none-cataleptic strain. This fragment contains the major gene of predisposition to pinch-induced catalepsy. We investigated contribution of the fragment to regulation of sensitivity of catalepsy, sexual motivation and social investigation to classical tricyclic antidepressant imipramine. The sexual motivation was higher in AKR.CBA-D13Mit76 than in AKR mice. Chronic imipramine treatment (25 mg/kg) reduced it in AKR.CBA-D13Mit76 mice and had no effect on weakly expressed sexual motivation of AKR males. No significant effects of genotype or chronic imipramine treatment on characteristics of social interest were observed. Imipramine failed to alter catalepsy expression in AKR.CBA-DI3Mit76 mice. Possible molecular genetic mechanisms underlying difference in behavioral responses to antidepressant administration are discussed.     

Serotonin binding by various populations of peritoneal cells and blood leukocytes from intact and thymectomized mice

Specific activity of 3H-serotonin binding with adhesive cells, as well as with T-lymphocyte enriched cells and B-lymphocytes, obtained from peritoneal excudate and blood leucocytes of CBA mice was determined. The highest affinity to amine was established in peritoneal T2 cells and adhesive blood cells. Results of studying the action of imipramin and blocators of some known serotonin receptors on amine binding with the investigated cells allow a conclusion on structural differences of serotonin-binding centres located on T-lymphocyte enriched cells and B-lymphocytes depending of their localization. The reduction of serotonin binding by immunocytes during ageing processes with animals (particularly tymectomized) was revealed, and this is supposed to be correlated with changes of immunological reactivity with ageing. It is supposed that serotonin participates in cooperative interactions of immunocytes in blood and peritoneal cavity.     

Assessing the anthropogenic impact on Lake Shira from antibiotic resistance of heterotrophic bacteria by neural networks methods

A general approach to assessing the anthropogenic impact on lake ecosystems is proposed and exemplified for the case of Lake Shira (Republic of Khakasia, Russia). The impact strength is estimated by applying neural network-based methods to samples of data on interdependent marking features of autochthonous and allochthonous bacteria isolated from the lake in 1997–2001. The proposed combination of analysis methods makes it possible to determine the state of an ecosystem from both small-and large-size samples of data having complex interrelations.     

Central mechanisms of fatigue during local static muscular activity

Changes in blood circulation, heart electrical activity, and the function of the brain cortex have been studied in healthy subjects of six age groups (from 7 to 65 years of age) during local static exercise. It has been shown that systemic responses of the body, such as a decrease in the contractility of muscles, changes in the heart function due to enhancement of central sympathetic regulation, and changes in the type of brain bioelectrical activity recorded with the use of encephalography, develop during fatigue.     

Cryopreservation of seeds and protocorms of rare temperate orchids

The efficiency of cryopreservation of seeds of five rare and endangered species of temperate orchids belonging to Platanthera and Dactylorhiza genera followed by their asymbiotic culture in vitro, as well as of in vitro cultured D. fuchsii protocorms (specific stage of orchid embryo development after release from the seed coat) was investigated. Germination rates of seeds after their exposure to liquid nitrogen were species-depended and could be either higher or lower than in the unfrozen control. There was no significant difference between growth rates of protocorms of the same species obtained from seeds collected in various Russia regions and cultured for 5 months. After vitrification, 9% of D. fuchsii protocorms with a larger diameter of 1200 μm survived cryopreservation; however, their growth was retarded for three months when compared to control protocorms.     

Chromosomal location of genes for vernalization requirement duration (<Emphasis Type="Italic">Vrd</Emphasis>) in winter bread wheat

In this study, we carried out monosomic analysis of two isogenic lines of cultivar Mironovskaya 808, monogenically dominant for genes Vrd1 and Vrd2 reducing the cultivar vernalization requirement duration from 50 to respectively 35 and 45 days, as well as analysis of substitution lines Cappelle Desperez/Bezostaya 1. Gene Vrd1 is localized on chromosome 4A; Vrd2, on chromosome 5D. The third gene, which is not allelic to the former two, is present in cultivar Cappelle Desperez on either the 1A, 6A, or 4B chromosome.     

G protein betagamma subunits interact with alphabeta- and gamma-tubulin and play a role in microtubule assembly in PC12 cells

The betagamma subunit of G proteins (Gbetagamma) is known to transfer signals from cell surface receptors to intracellular effector molecules. Recent results suggest that Gbetagamma also interacts with microtubules and is involved in the regulation of the mitotic spindle. In the current study, the anti-microtubular drug nocodazole was employed to investigate the mechanism by which Gbetagamma interacts with tubulin and its possible implications in microtubule assembly in cultured PC12 cells. Nocodazole-induced depolymerization of microtubules drastically inhibited the interaction between Gbetagamma and tubulin. Gbetagamma was preferentially bound to microtubules and treatment with nocodazole suggested that the dissociation of Gbetagamma from microtubules is an early step in the depolymerization process. When microtubules were allowed to recover after removal of nocodazole, the tubulin-Gbetagamma interaction was restored. Unlike Gbetagamma, however, the interaction between tubulin and the alpha subunit of the Gs protein (Gsalpha) was not inhibited by nocodazole, indicating that the inhibition of tubulin-Gbetagamma interactions during microtubule depolymerization is selective. We found that Gbetagamma also interacts with gamma-tubulin, colocalizes with gamma-tubulin in centrosomes, and co-sediments in centrosomal fractions. The interaction between Gbetagamma and gamma-tubulin was unaffected by nocodazole, suggesting that the Gbetagamma-gamma-tubulin interaction is not dependent on assembled microtubules. Taken together, our results suggest that Gbetagamma may play an important and definitive role in microtubule assembly and/or stability. We propose that betagamma-microtubule interaction is an important step for G protein-mediated cell activation. These results may also provide new insights into the mechanism of action of anti-microtubule drugs.