Determination of concentration and aggregate size in influenza virus preparations using the true UV-absorption spectra

It is well-known that influenza virus (IV) preparations are characterized by very large contribution of light-scattering to their UV absorption spectra. With the help of so called extrapolation method we managed to measure true absorption spectra of IV preparations and to determine absorption coefficients (E0.1(1) (cm, 280)) for the intact IV virions and for IV subviral particles. These coefficients turned out to equal 1.26 +/- 0.17 and 0.96 +/- 0.11 for the virions and subviral particles respectively. The knowledge of exact IV concentration is necessary for quantitative physico-chemical studies of IV virions and their components. It is also shown that UV absorption spectra measurements allow to register IV virion aggregation. Aggregation properties of IV subviral particles were also studied.     

Dynamics of Postural Indices in Case of Listening to Sounds of Steps Approaching from the Front and from Behind

Journal of Evolutionary Biochemistry and Physiology - In the study a hypothesis was tested on substantial difference between postural reactions to conspecific auditory motion information and...     

Postural Control Features of Field-Dependent and Field-Independent Subjects in the Absence of Visual and Audio Information

Human Physiology - Stabilometric parameters were recorded during maintaining the upright posture (European version of Rotberg’s posture: heels together, toes apart, hands down at the sides)...     

Oryzalin-Induced Changes in Water Status and Cytoskeleton Proteins of Winter Wheat Seedlings upon Cold Acclimation and ABA Treatment

The effect of oryzalin (a specific inhibitor of tubulin polymerization in plant cells) on water retention by the leaves and roots of winter wheat (Triticum aestivum L.) seedlings was studied. The cultivars differing in their frost resistance were compared after their acclimation to low temperature (3°C for 3 or 7 days) and after treatment with ABA. In control untreated plants, oryzalin reduced the water-retaining capacity (WRC) of leaves and roots. Both hardening and ABA lowered the effect of the inhibitor on WRC in leaves, whereas their effects on water retention by roots were opposite, i.e., hardening weakened and ABA intensified the effect of oryzalin. Oryzalin-induced reduction of WRC decreased in the following sequence of cultivars: weakly frost resistant moderately frost resistant highly frost resistant. It was more pronounced in the leaves than in the roots, the latter being characterized by the lower WRC and lower frost resistance. After three-day-long hardening of plants, an additive effect of hypothermia and ABA on oryzalin-induced decrease in WRC of leaves and the lack of such effect in the roots were observed. The immunochemical analysis of the composition and content of cytoskeletal proteins with Western blotting showed that in the leaves the actin/tubulin ratio was higher than in the roots. The treatment of nonacclimated plants with ABA lowered the content of - and -tubulins and actin in roots but did not affect the level of actin in leaves. Hardening negated the effects of ABA on cytoskeletal proteins. Oryzalin produced the greatest inhibitory effect on WRC and an increase in frost resistance in ABA-treated plants in the experiments with leaves of the weakly frost resistant cultivar before and after hardening. Organ- and cultivar-specific and ABA-mediated dependence of WRC on cytoskeletal proteins and microtubules and microfilaments formed by them is supposed to result from their effect on the state of intracellular water and water permeability of the plasma membrane. In the course of cold acclimation of plants and upon their treatment with ABA, this dependence was more distinctly expressed in leaves than in roots, and especially in the plants of the weakly frost resistant cultivar.     

Changes in the phenotypic properties of highly pathogenic influenza A virus of H5N1 subtype induced by N186I and N186T point mutations in hemagglutinin

The change in the phenotypic properties resulting from amino acid substitutions in the hemagglutinin (HA) molecule is an important link in the evolutionary process of influenza viruses. It is believed to be one of the mechanisms of the emergence of highly pathogenic strains of influenza A viruses, including subtype H5N1. Using the site-directed mutagenesis, we introduced mutations in the HA gene of the H5N1 subtype of influenza A virus. The obtained virus variants were analyzed and compared using the following parameters: optimal pH of conformational transition (according to the results of the hemolysis test), specificity of receptor binding (using a set of synthetic analogues of cell surface sialooligosaccharides), thermoresistance (heat-dependent reduction of hemagglutinin activity), virulence in mice, and the kinetics of replication in chicken embryos, and reproductive activity at different temperatures (RCT-based). N186I and N186T mutations in the HA protein increased the virulence of the original virus in mice. These mutations accelerated virus replication in the early stages of infection in chicken embryos and increased the level of replication at late stages. In addition, compared to the original virus, the mutant variants replicated more efficiently at lower temperatures. The obtained data clearly prove the effect of amino acid substitutions at the 186 position of HA on phenotypic properties of the H5N1 subtype of influenza A.     

Epitope mapping of the hemagglutinin molecule of a highly pathogenic H5N1 influenza virus by using monoclonal antibodies

We mapped the hemagglutinin (HA) antigenic epitopes of a highly pathogenic H5N1 influenza virus on the three-dimensional HA structure by characterizing escape mutants of a recombinant virus containing A/Vietnam/1203/04 (H5N1) ΔHA and neuraminidase genes in the genetic background of A/Puerto Rico/8/34 (H1N1) virus. The mutants were selected with a panel of eight anti-HA monoclonal antibodies (MAbs), seven to A/Vietnam/1203/04 (H5N1) virus and one to A/Chicken/Pennsylvania/8125/83 (H5N2) virus, and the mutants’ HA genes were sequenced. The amino acid changes suggested three MAb groups: four MAbs reacted with the complex epitope comprising parts of the antigenic site B of H3 HA and site Sa of H1 HA, two MAbs reacted with the epitope corresponding to the antigenic site A in H3 HA, and two MAbs displayed unusual behavior: each recognized amino acid changes at two widely separate antigenic sites. Five changes were detected in amino acid residues not previously reported as changed in H5 escape mutants, and four others had substitutions not previously described. The HA antigenic structure differs substantially between A/Vietnam/1203/04 (H5N1) virus and the low-pathogenic A/Mallard/Pennsylvania/10218/84 (H5N2) virus we previously characterized (N. V. Kaverin et al., J. Gen. Virol. 83:2497-2505, 2002). The hemagglutination inhibition reactions of the MAbs with recent highly pathogenic H5N1 viruses were consistent with the antigenic-site amino acid changes but not with clades and subclades based on H5 phylogenetic analysis. These results provide information on the recognition sites of the MAbs widely used to study H5N1 viruses and demonstrate the involvement of the HA antigenic sites in the evolution of highly pathogenic H5N1 viruses, findings that can be critical for characterizing pathogenesis and vaccine design.     

Activity of Lectins of Winter Wheat Seedlings Infected with Mycoplasma and Treated with Salicylic Acid

Changes in activity of soluble and cell wall lectins have been revealed in seedlings of winter wheat Triticum aestivum L. cultivar Mironovskaya 808 after infection with mycoplasma Acholeplasma laidlawii 118. The protective effect of salicylic acid was manifested as negating the bursts of lectin activity induced by mycoplasma infection.__________Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 4, 2005, pp. 423–427.Original Russian Text Copyright © 2005 by Trifonova, Maksyutova, Timofeeva, Chernov.     

Activities before and after stress of digestive enzymes in rats separated at different time from lactating females

The effect of separation of rat pups from lactating females at different time after birth (at the 18th, 21st, 30th, 36th or 43rd day) and stress exposed at the day of the experiment (at the 75th day of life) or twice (at the day of weaning and at the day of the experiment) on activities of membrane-bound (invertase, alkaline phosphatase, aminopeptidase M) and predominantly intracellular (glycyl-L-leucine dipeptidase) enzymes was studied in different parts of intestine of rats (males and females). A change of the enzyme activities has been shown for the first time in the animals separated from lactating females not only at early time, which has already been known, but also at later time, in comparison with their activities in the animals weaned at usual time, at the 30th day of life. The preliminary stress of rat pups at the day of separation from lactating females promotes in many cases a reduction of the response to the stress, to which the animal is exposed in the adult period of life. Deceased.     

The activity of hippocampal neurons during several types of behavior]

The activity of 87 hippocampal units and the EEG (field CAI) were studied in unrestrained rabbits during calm and active alertness and at different stages of sleep. Correlation has been established between the characteristics (mean frequency and the pattern) of unit firing, EEG and the animal's activity. For most of the neurones, fixed values of mean frequency and the discharge pattern corresponded to a definite functional state. With transition from sleep to alertness, 63.6% of the units became active, 29.1% were inhibited, and the rest of the units changed the firing pattern only. The cells which became activated during awakening, showed a reduced firing frequency during a more profound sleep and higher discharge frequency during the paradoxal phase of sleep, while the inhibitory cells revealed reverse dynamics of discharge frequency. In a state of alertness, the most pronounced shifts in firing activity were observed in 33.3% of the nerve cells at orienting investigating behaviour, in 27.1%, during attention reaction, in 22.9% at some kinds of movement, and in 16.7%, in the course of feeding and drinking. A conclusion has been drawn that the role of the hippocampus in achieving different behavioral reactions is probably to a great extent determined by its participation in setting up a level of the brain central tone, specific for each state.