Estimation of somatic,emotional, and behavioral reactions of monkeys in chronic experiments

Long-term chronic experiments with conditioning of monkeys and biophysical and pharmacological stimulation require monitoring somatic, emotional, and behavioral reactions. To form experimental groups, it is necessary to estimate baseline psychosomatic state of a monkey. We developed a method of monitoring the functional state of Macaca monkeys and scoring state parameters. With slight modification, this method may be applied to other monkey species.     

A technique for the operant conditioning to differentiate signals during reinforcement delay in primates

A new technique of training the primates to differentiate signals immediately during execution of the instrumental act (pushing the lever) was proposed. The technique makes it possible to study the processes of decision making during activation of the executive (motor) brain systems.     

The role of intracellular Ca(2+) pools in the regulation of protoplast volume. Effect of red light on the Ca(2+) mobilization in cytoplasm of Arabidopsis cells

The changes in cytosol Ca2+ concentration associated with the shrinkage of Arabidopsis cells induced by the inhibitor of Ca(2+)-ATPase, cyclopiazonic acid and the Ca2+ ionophore ionomycin were monitored using the fluorescence of Ca(2+)-sensitive probe chlortetracycline hydrochloride. It was found that these compounds elicited a substantial decrease in fluorescence intensity closely associated with Ca(2+)-release from the intracellular stores to the cytoplasm. The release of Ca2+ from the intracellular depots was accompanied by decrease of plant cell volume. Thapsigargin and 2,5'-ditert-butyl-1,4-benzohydroquinone (highly specific inhibitors of Ca(2+)-ATPase of endoplasmic reticulum) resulted in much weaker changes than cyclopiazonic acid did. It was also found with the help of the same technique that red light (lambda = 660 nm) illumination induced a similar Ca2+ release from the intracellular stores. Moreover, the amplitudes of light-induced fluorescence responses registered in mutant plants differing in the content of phytochrome A (phyAOX) and phytochrome B (phyBOX) were much higher than those registered in wild-type of Arabidopsis.     

The use of 16S and 16S-23S rDNA to easily detect and differentiate common Gram-negative orchard epiphytes

The identification of Gram-negative pathogenic and non-pathogenic bacteria commonly isolated from an orchard phylloplane may result in a time consuming and tedious process for the plant pathologist. The paper provides a simple "one-step" protocol that uses the polymerase chain reaction (PCR) to amplify intergenic spacer regions between 16S and 23S genes and a portion of 16S gene in the prokaryotic rRNA genetic loci. Amplified 16S rDNA, and restriction fragment length polymorphisms (RFLP) following EcoRI digestion produced band patterns that readily distinguished between the plant pathogen Erwinia amylovora (causal agent of fire blight in pear and apple) and the orchard epiphyte Pantoea agglomerans (formerly E. herbicola). The amplified DNA patterns of 16S-23S spacer regions may be used to differentiate E. amylovora at the intraspecies level. Isolates of E. amylovora obtained from raspberries exhibited two major fragments while those obtained from apples showed three distinct amplified DNA bands. In addition, the size of the 16S-23S spacer region differs between Pseudomonas syringae and Pseudomonas fluorescens. The RFLP pattern generated by HaeIII digestion may be used to provide a rapid and accurate identification of these two common orchard epiphytes.     

Peculiarities of functioning of liver mitochondria of river lamprey Lampetra fluviatilis and common frog Rana temporaria at periods of suppression and activation of energy metabolism

The work dealt with study of mitochondria in reversible metabolic suppression of hepatocytes of the river lamprey Lampetra fluviatilis in the course of prespawning starvation and of liver mitochondria of the common frog Rana temporaria during hibernation and activity. In winter the metabolic depression of lamprey hepatocytes, unlike that of frog hepatocytes, has been found to be due to deactivation of complex I of the electron transport mitochondrial chain, a low rate of NAD-dependent substrate oxidation, a low content of adenine nucleotide content, and a high degree of mitochondrial membrane permeability to H+ and other monovalent ions (KCl-, K+). The mitochondrial membrane permeability decreases in the presence of ethyleneglycoldiamineethyltetraacetic acid (EGTA), cyclosporine A (CsA), adenosine-5'-diphosphate (ADP), and Mg+. These facts indicate the presence in these mitochondria of the Ca2+ -dependent unspecific pore in the low-conductance state. Histological studies showed the lamprey and the frog to have principal differences in use of energy substrates at the period of metabolic depression. Lampreys utilize predominantly lipids, whereas frogs--glycogen. The clearly pronounces activation of lipid consumption is observed at the spring period before spawning and death of lamprey. Possible causes of metabolic depression are discussed as well as similarity and difference in behavior of mitochondria of cyclostomes and amphibians throughout metabolic depression and activity.     

The floral meristem undetermination mutation in Papaver somniferum L.: spontaneous phenotypic variation in ontogeny

A new morphogenetic mutation of the shoot, floral meristem undetermination, was found in Papaver somniferum L. with monocarpic shoot. The expression of the DFM (determination of floral meristem) gene, which limits the proliferative activity of stem cells in the floral meristem, was affected. The mutation displayed spontaneous phenotypic instability in ontogeny, variation in the mutant character expression on different flowers of the same plant in the same genotypic environment. The mutation phenotype varied from no expression or formation of individual phyllomes in the center of the primary ovary to formation of a new flower and a new capsule with viable seeds.     

Procedure for obtaining and catalytic properties of trypsin immobilized in cryogels of polyvinyl alcohol

Commercial preparations of trypsin, varying in activity, were immobilized in a cryogel of polyvinyl alcohol, activated by dialdehydes (terephthalic, succinic, or glutaric) or divinyl sulfone. All preparations of the immobilized enzyme exhibited hydrolytic activity and retained stability for 8 months. In an organic solvent environment, specimens of immobilized trypsin catalyzed the synthesis of N-carbobenzoxy-L-phenylalanyl-L-arginyl-L-leucine p-nitroanilide from N-carbobenzoxy-L-phenylalanyl-L-argininine methyl ester (or N-carbobenzoxy-L-phenylalanyl-L-arginine) and L-leucine p-nitroanilide, as well as the formation of N-carbobenzoxy-L-alanyl-L-alanyl-L-arginyl-L-phenylalanine p-nitroanilide from N-carbobenzoxy-L-alanyl-L-alanyl-L-arginine and L-phenylalanine p-nitroanilide. The presence of small amounts of water in organic solvents was prerequisite to the biocatalysts manifesting synthase activity in reactions of peptide bond formation.     

Regional medico-dosimetric register (RMDR) is a model for evaluation of long-term "low" dose ionizing radiation effects on Siberian group of chemical enterprises main production personnel

Medico-dosimetric register is an optimal model of epidemiological studies on evaluation of ionizing radiation effects. Regional medico-dosimetric register (RMDR) is a system of interrelating information blocks including data on Siberian Group of Chemical Enterprises (SGCE) personnel. At present SGCE personnel and Seversk residents RMDR database includes information on 138496 persons, 65538 of which are SGCE workers. SGCE personnel and Seversk residents RMDR is a scientific base for researches with the aim of evaluating long-term ionizing radiation effects in a "low" dose range. Information on mortality and morbidity rate as well as "thematic" registers of the main diseases potentiates in evaluating the spectrum of somatic stochastic effects and radiogenic risks in SGCE workers and Seversk residents as well as their offsprings. A practical significance of RMDR database is the formation of the main diseases "risk" groups depending on definite risk factors in certain groups that provides targeted diagnostic and preventive therapy both among high-dose establishments' workers and residents living near-by.     

Deletion map of the Escherichia coli K-12 dnaB gene

Summary Acid phosphatase isoenzymes of Chlamydomonas reinhardii were investigated by isoelectric focusing in polyacrylamide gel systems. In this paper we describe in detail an original method for isoelectric focusing of acid phosphatases extracted from wildtype and acid phosphatase-lacking mutant algae, obtained from Laboratoire de Génetique of University of Liège. Three isoenzymes can be separated from the buffer-soluble components of these cells. An additional isoenzyme type can be visualized using the nonionic detergent NP40 as solubilizer. We conclude that these four isoenzymes are releated to the structural gene of the soluble constitutive acid phosphatase, which was shown by their appearance in P ₂ and their total absence in mutant P _a. The pl values of soluble constitutive acid phosphatase isoenzymes range between pH 5.2 and 6.2. As a result of treatment with NP40 the extracts from both wild-type and mutant lines contain two additional active phosphatase forms which can be characterized by their high heat resistance and low pI values. These enzymes are fully active using either -naphthyl phosphate or different acetate esters as substrates.