Thermal stability of lactate dehydrogenase in the complex with the anion polyelectrolyte poly(styrene sulfonate)

The temperature stability of the cytoplasmic enzyme of the glycolysis of lactate dehydrogenase from a pig muscle (isoenzyme M4) in a complex with the anion polyelectrolyte poly(styrenesulfonate) has been investigated by the methods of adiabatic differential scanning microcalorimetry, the own protein fluorescence, and circular dichroism. Calorimetric investigations of complex of lactate dehydrogenase with poly(styrenesulphonate) in 50 mM phosphate buffer at pH 7.0 have shown that the temperature of the transition and enthalpy of lactate dehydrogenase thermal denaturation sharply decreases with growing weight ratio poly(styrenesulphonate)/lactate dehydrogenase, though at 20 degrees C the enzyme activity of lactate dehydrogenase remains unchanged for several hours irrespective of the addition of poly(styrenesulphonate). The addition of phosphate ions to the solution enhances the resistance of lactate dehydrogenase to both thermal denaturation and inactivation by polyelectrolyte. The data obtained are interpreted from the viewpoint of a special role of two anion-binding centers in intersubunits contacts of lactate dehydrogenase, which enhance its resistance to both thermal denaturation and destruction by polyelectrolyte.     

Monitoring phytoplasma-bearing leafhoppers/planthoppers in vineyards in the Golan Heights, Israel

In the course of identifying the vector(s) of the grapevine yellows (GY) and western-X (WX) phytoplasmas in the Golan Heights, leafhopper and planthopper species were examined. The planthopper, Hyalesthes obsoletus , was trapped on yellow sticky traps or collected on weeds; there was a relatively small peak during two weeks in June and during four weeks starting mid-September. The leafhoppers, Neoaliturus spp. and Circulifer sp. (of the haematoceps complex) were both trapped early or very late in summer, but could be collected on weeds throughout the summer. The two remaining leafhopper species, Macrosteles quadripunctulatus and Orosius orientalis (= albicinctus ) were only rarely caught on sticky traps, but were found on weeds throughout the summer. Phytoplasmas were found within the body of these five species; all are historically known to be efficient vectors of various phytoplasmas and were therefore chosen for further investigations.     

Effect of light of various spectral composition on kinetics of changes in a green leaf resistivity]

The kinetics of changes in the resistance of a green leaf by the action of exciting light was studied. It was shown that the illumination-induced changes in leaf conductance are mainly due to an elevation of temperature caused by the absorption of light energy. Based on the experimental results, the fraction of absorbed light energy converted into heat calculated under assumption that heating of the leaf across the thickness is uniform. It was found that the fraction of absorbed energy converted into heat does not depend on the intensity of radiation in the IR range and increases with decreasing intensity of photosynthetically active radiation. An explanation of the phenomenon is proposed, which is based on that in reality the heating of the leaf across the thickness is nonhomogeneous and this inhomogeneity of heating becomes more pronounced upon photoinduced changes in absorption resulting from a decrease in photosynthetically active radiation. It was shown that nonphotochemical fluorescence quenching does not substantially affect the leaf conductance at intensities of photosynthetically active radiation not higher than 200 Wt/m2.     

Application of the complex of DNA with the congo red anionic diazo dye for detection of nuclease-producing colonies of marine bacteria

This study was aimed at the development of a method for detection of colonies of nuclease-secreting marine bacteria. The BAL nuclease-producing marine bacterium Pseudoalteromonas espejiana BAL-31 was used as the test object. A new method was developed involving the congo red (CR) anionic dye. The P. espejiana culture was plated on nutrient agar with CR and denatured DNA. In such media. CR was found to form complexes with DNA. After two days of incubation at 30 degrees C, halos were found around the P. espejiana colonies. No halos appeared when DNA was not introduced, when BAL nuclease was inactivated, or when the medium was inoculated with Escherichia coli. It was concluded that the halos around the colonies indicated nuclease excretion. The halos were shown to result from the coagulation of CR released after digestion of the CR-DNA complex by the nuclease. This method for detection of nuclease-producing colonies can probably be used for all marine bacteria and possibly for halophilic bacteria as well.     

Streptozocin model of diabetes mellitus in the mollusc Anodonta cygnea: functional state of the adenylyl cyclase mechanisms of action of peptides of the insulin superfamily and their effect on carbohydrate metabolism enzymes

In terms of development of evolutionary biomedicine using invertebrate animals as models for study of molecular grounds of various human diseases, for the first time the streptozocin (ST) model of insulin-dependent diabetes in the mollusc Anodonta cygnea has been developed. This model is based on the following authors' data: (1) redetection of insulin-related peptides (IRP) in mollusk tissues: (2) discovery of the adenylyl cyclase signal mechanism (ACSM) of action of insulin and other peptides of the insulin superfamily in tissues of mammals, human, and mollusc. A. cygnea; (3) concept of molecular defects in hormonal signal systems as causes of endocrine diseases. Studies on the ST model have revealed in mollusc smooth muscle on the background of hyperglycemia at the 2nd, 4th, and 8th day after the ST administration a decrease of the ACSM response to activating action of insulin, IGF-1, and relaxin. These functional disturbances were the most pronounced at the 2nd day of development and rather less marked at the 4th and 8th day. Analysis of data on effect of hormonal and non-hormonal (NaF, GIDP, and forskolin) ACSM activators has shown that the causes of impair of signal-transducing function of this mechanism are (1) a hyperglycemia-induced increase of the basal AC activity and as a consequence--a decrease of the enzyme catalytic potentials in response to hormone; (2) a decrease of functions of Gs-protein and of its coupling with AC. Besides, administration of ST produced in the mollusc muscles an attenuation of regulation by insulin of carbohydrate metabolism enzyme (glucose-6-phosphate dehydrogenase, glycogensynthase). The pattern of disturbances in the studied parameters in the mollusc is very similar to that revealed by the authors in rat and human muscle tissues in type 1 diabetes.     

Phylogenetic reconstructions in the genus Capra (Bovidae, Artiodactyla) based on the mitochondrial DNA analysis

Mitochondrial genome fragments were examined in all species of the genus Capra (Bovidae, Artiodactyla). Phylogenetic analysis was carried out using 59 cytochrome b gene sequences (392 bp), and 22 sequences of the mtDNA variable fragment (402 bp). In the control region, two unique deletions were revealed. One of the deletions was found only in Capra cilindricornis (17 bp), while another one grouped C. caucasica with C. aegagrus (1 bp). The group of Caucasian wild goats splits into two clades, and furthermore, the sequences of C. caucasica demonstrate remarkable similarity to the sequences of C. aegagrus, while C. cylindricus seems to have evolved independently for a long period of time. It was demonstrated that C. pyrenaica and C. ibex were extremely close to one another. Capra sibirica formed an outer group relative to the other species, and according to our data, was the most ancient species of the genus. On the contrary, genetic distance separating C. falconeri (the most independent species of the genus related to its morphology) from the other species is small.     

Vasopressin and prostaglandin <Emphasis Type="Italic">E</Emphasis><Subscript>2</Subscript> as regulators of renal function during children’s development

The main physicochemical parameters of blood serum (the osmolality and concentrations of Na⁺, K⁺, Ca²⁺, and Mg²⁺) determined in 314 children (from newborn infants to 17-year-old adolescents) and 25 adults were found to be virtually constant throughout the postnatal period, which was due to the high effectiveness of the systems responsible for their stabilization. From the first postnatal days until puberty, prostaglandin E₂ (PGE₂) and arginine vasopressin (AVP) are involved in the regulation of renal water excretion. In infants, during their first postnatal months, the excretion of solute-free water is correlated with the excretion of PGE₂. Adult-type effects of AVP on the reabsorption of solute-free water were observed in children only after 12 postnatal months. A change in the reabsorption of ions in the thick ascending limb of Henle’s loop was shown to be involved in the regulation of the volume of urine excreted. AVP and PGE₂ are also involved in the regulation of the distal segment of the nephron and collecting tubules, but their influence on the volume and composition of urine is age-dependent.