Effect of short-term hypoxia and re-oxygenation on mice peritoneal macrophages in vitro

Microfluorimetry of single cells could help to analyze their morphology and function state during changes of gas environment. It is very important to have a possibility of the cell visual control during hypoxia and collection of dynamic fluorimetric data in digital form. The effects of short-term pO2 decrease were studied. For estimating the effects of hypoxia and reoxygenation we used the mice peritoneal macrophages, which are very sensitive to physical, chemical and regulatory stimuli. A special small chamber for fluorimetric measurements during pO2 changes, was developed. The level of active oxygen forms, intracellular pH, and cell membrane instability were investigated during replacement of air by nitrogen or argon (of the basal level decreased to 20% of basic level) and in subsequent reoxygenation. The increase of active oxygen forms was shown during 30 min of hypoxia and their level continued to rise immediately after reoxygenation. A short-term decrease and subsequent increase of pO2 in the medium led to an increase of intracellular pH level. The shifts of measured cell indices were stabilized after 30-40 min of pO2 changes thus suggesting a fast comprehension of countermeasure cell mechanisms. No macrophages with membrane disorders were found despite the rise of the active oxygen forms level during hypoxia and reoxygenation in vitro. There were no significant differences between nitrogen and argon used for replacement of air in the medium. The data obtained suggest a high resistance of macrophages against pO2 changes and an involvement of the antioxidative mechanisms for cell protection especially during reoxygenation period.     

Visual afferentation deficit during diffuse photosensitivity period in nestlings affects morphogenesis in Wulst neurons

Golgi-stained preparations were processed with computerized morphometry to study the effect of the deprivation (eyes covered with nontransparent caps) on the development of neurons in the Wulst (the structure analogous to mammalian visual cortex) of pied flycatcher nestlings. Six-day-old nestlings that have only one form of the visual perception--diffuse photosensitivity--were studied. In the previous paper [Korneeva et al., 1994] in was shown that the Wulst of 6-day-old nestlings consisted of non-differentiated neurons and stellate-like cells at different stages of maturation; the latter group was subdivided into more- and less-mature cells. This work proved that even a 1.5-day-long deprivation (actual duration being counted off from the moment of appearance of the first retinal photoreceptors at the age of 4.5 days) resulted in significant changes in the geometry of stellate-like neurons. The changes in less-mature cells were predominantly destructive (decline of all quantitative indices of a cell, including the significant decrease in the total length of dendrites, maximal radius of the dendritic field and cell branching index), while the changes in more-mature cells were constructive (increase in all quantitative cell indices, including statistically significant increase in the soma section area, total length of dendrites, maximal radius of dendritic field and the number of foci of maximal branching). Different reactions of these cell types to the limitation of visual afferentation may be connected with differences in the afferent inflow to less- and more-mature cells or/and with different maturational stages of these neurons at the onset of deprivation.     

Evaluation of the usefulness of three collections of bacteriophages for typing methicillin-resistant Staphylococcus aureus,isolated in Moscow hospitals

The typing of S. aureus methicillin-resistant strains, isolated in different hospitals of Moscow; was carried out with the use of three collections of phages: the International Set of Phages; the set of phages of the International Center of S. aureus phage typing in London (L); and the experimental collection of phages of the Gamaleya Institute of Epidemiology and Microbiology in Moscow (M). In this study made with the use of both the phages of the International Diagnostic Set and phages L in the standard typing dose of 1 TP about 6% of the cultures under study proved to be sensitive. When the typing dose was increased to 100 TP the phages of the international diagnostic set lyzed 75.5% of the cultures. The typed strains were found to belong to phage types 77 (71.7%), 77/84/85 (19.6%) and 94/96 (6.5%). At a concentration of 100 TP phages L lyzed 83.7% of the cultures, but the dominating phage types could not be determined due to a great variety of phage markers. In contrast to the two preceding collections, the third phage collection M was composed in such a way that in the study of the investigated culture the specificity of its restriction modification was primarily evaluated and only then the presence of antiphage immunity was determined. This latter collection was used in the evaluation of 93.1% of the cultures. By the specificity of their restriction specification system the majority of them were classified with two new groups, heretofore not described. Only this collection M made it possible to differentiate epidemic and sporadic strains and to evaluate the epidemic situation in all 6 hospitals.     

Dynamics of local shifts in oscillations and energy metabolism of the rabbit cerebral cortex during formation of a conditioned defensive reflex

Brain energy metabolism in different functional states or activities of humans and animals is characterized by dynamic changes in the degree of coupling between glycolysis and tissue respiration in different cell compartments (Fox et al., 1988; Fox, 1989; Pellerin et Magistretti, 1994; Prichard et al., 1991; Schur et al., 1999). These processes determine variations in the brain redox state (Siesjo, 1978) that can be potentiometrically recorded with implanted platinum electrodes as the brain tissue redox state potential E (Puppi et Fely, 1983). This potential was recorded in rat brain cortex with four pairs of platinum electrodes implanted into different symmetrical cortical region (one electrode of a pair being located in the cortical layers, another being located epidurally). In the course of defensive conditioning (after 5-15 combination of a bulb light and a weak electrodermal stimulation of a ear), E oscillations (6-10 per minute) appeared. In this period, stimuli combinations produced the generalized E shifts. Later on (with accumulation of stimuli combinations), the episodes of E increase were replaced by the episodes of E decrease. To the 200-400th combinations, E oscillations disappeared, and E shifts became local and stable. The findings suggest that conditioning shifts the balance between the main energy-producing systems in the brain tissue: at the initial stages of conditioning brain functions are predominantly supported by the energy obtained from tissue respiration, while during the realization of defensive conditioning glycolysis is the main source of energy.     

Effect of of distamycin A on histone H1 methylation, extraction and formation of UV-inducible crosslinks with DNA in the interphase rat liver nucleus

Incubation in vitro of rat liver nuclei in the presence of S-adenosyl[methyl-(3)H]methionine ([(3)H] SAM) leads to incorporation of the radioactive label not only into core-histones H3 and H4, but also into linker histone H1. Addition of distamycine A to the incubation medium stimulates label incorporation into histone H1 ~ in 6 times and into histone H3 ~ in 2 times. The presence of distamycine facilitates histone H1 extraction by polyglutamic acid (poly(Glu)) and decreases of UV-induced DNA-histone cross-links formation. These effects give evidence of weakening of H1-chromatin interaction by distamycin to be results of histone H1 position change relative to nucleosome and(or) disturbance of histones H1-H3 interactions so as these histones are exposed to additional methylation.     

Analysis of the distribution of the nucleotide sequence and electrostatic potential of the Escherichia coli genome

The oligonucleotide composition of the E. coli genome and its sigma70-specific promoters has been analyzed. The promoter DNA was shown to contain mainly AT-rich hexanucleotides having functionally important physical properties such as the ability to form easily melting sites and induce the bending of the double helix. A comparative analysis of the electrostatic characteristics of hexanucleotides within the whole sequence of the E. coli genome and its promoter regions was made. Hexanucleotides possessing a more electronegative surrounding were found to predominate in the nucleotide sequence of the promoter DNA.     

Role of trophic and hormonal factors in exogenous regulation of the formation of reproductive organs in yellow lupine (Lupinus luteus L.)

During the formation and ripening of beans in yellow lupine, their intactness may be increased by treatment of the plants with the microelement molybdenum or the growth factor 24-epibrassinolide. Early stages of the effects of either factor involved activation of various metabolic pathways; eventually, the supply of fruit elements with free amino acids was improved and the seed productivity of the plants, increased.     

Physiological,biochemical, and cytological characteristics of a halotolerant and alkalitolerant methanotroph grown on methanol

The halotolerant alkaliphilic methanotroph Methylomicrobium buryatense 5B is capable of growth at high methanol concentrations (up to 1.75 M). At optimal values of pH and salinity (pH 9.5 and 0.75% NaCl), the maximum growth rate on 0.25 M methanol (0.2 h-1) was twice as high as on methane (0.1 h-1). The maximum growth rate increased with increasing medium salinity and was lower at neutral than at alkaline pH. The growth of the bacterium on methanol was accompanied by a reduction in the degree of development of intracytoplasmic membranes, the appearance of glycogen granules in cells, and the accumulation of formaldehyde, formate, and an extracellular glycoprotein at concentrations of 1.2 mM, 8 mM, and 2.63 g/l, respectively. The glycoprotein was found to contain 23% protein and 77% carbohydrates, the latter being dominated by glucose, mannose, and aminosugars. The major amino acids were glutamate, aspartate, glycine, valine, and isoleucine. The glycoprotein content rose to 5 g/l when the concentration of potassium nitrate in the medium was augmented tenfold. The activities of sucrose-6-phosphate synthase, glycogen synthase, and NADH dehydrogenase in methanol-grown cells were higher than in methane-grown cells. The data obtained suggest that the high methanol tolerance of M. buryatense 5B is due to the utilization of formaldehyde for the synthesis of sucrose, glycogen, and the glycoprotein and to the oxidation of excess reducing equivalents through the respiratory chain.