Gas-liquid chromatography for the determination of acetylcholinesterase activity

Method has been modified and used for quantitative gas chromatographic determination of microtraces in a large volume of test mixture under study. The method can be applied for the differentiated determination of genuine and false blood cholinesterase by final products of specific substrate hydrolysis.     

Modeling experimental pollinosis

Experimental pollinosis was modelled on guinea pigs. The animals were placed in a cell in which an allergen-water-dialyzed extract of ambrosia pollen was dispersed by means of coaxial pulveriser. Homocytotropic antibodies serving as indices of animal sensitization to the specific allergen formed in guinea pigs as a result of aerosol sensitization. Sensitization of the animals to the ambrosia allergen was accompanied by an increase of sensitivity of the broncho-pulmonary apparatus and was characterized by the bronchospastic reaction in resolving inhalation or inrtavenous injection of a specific allergen.     

Relationship of Nitrogen Deficiency-Induced Leaf Senescence with ROS Generation and ABA Concentration in Rice Flag Leaves

Nitrogen (N) deficiency is one of the critical environmental factors that induce leaf senescence, and its occurrence may cause the shorten leaf photosynthetic period and markedly lowered grain yield. However, the physiological metabolism underlying N deficiency-induced leaf senescence and its relationship with the abscisic acid (ABA) concentration and reactive oxygen species (ROS) burst in leaf tissues are not well understood. In this paper, the effect of N supply on several senescence-related physiological parameters and its relation to the temporal patterns of ABA concentration and ROS accumulation during leaf senescence were investigated using the premature senescence of flag leaf mutant rice (psf) and its wild type under three N treatments. The results showed that N deficiency hastened the initiation and progression of leaf senescence, and this occurrence was closely associated with the upregulated expression of 9-cis-epoxycarotenoiddioxygenase genes (NCEDs) and with the downregulated expression of two ABA 8′-hydroxylase isoform genes (ABA8ox2 and ABA8ox3) under LN treatment. Contrarily, HN supply delayed the initiation and progression of leaf senescence, concurrently with the suppressed ABA biosynthesis and relatively lower level of ABA concentration in leaf tissues. Exogenous ABA incubation enhanced ROS generation and MDA accumulation in a dose-dependent manner, but it decreased the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in detached leaf. These results suggested that the participation of ABA in the regulation of ROS generation and N assimilating/remobilizing metabolism in rice leaves was strongly responsible for induction of leaf senescence by N deficiency.

     

Half a century of oligochaete research in Estonian running waters

About 51500 specimens from 1542 samples, collected over the years 1954–1975 and 1986–1999 in different running water bodies throughout Estonia, were identified. Tubificidae prevailed in the material, with Limnodrilus hoffmeisteri forming about 40%. This species was followed by the tubificids Tubifex tubifex, Potamothrix hammoniensis, Psammoryctides barbatus, L. udekemianus and Spirosperma ferox, the naidid Stylaria lacustris, and the lumbriculid Stylodrilus heringianus. Two main ecological assemblages were distinguished: the pelophilous assemblage, dominated by L. hoffmeisteri, and the psammophilous one, where usually P. barbatus was dominant. The relationships between different species and the chemical parameters of water were usually weak but in contrast, correlated well with sediment preferences. In organically enriched reaches, L. hoffmeisteri usually dominated. The fauna of the streams of the islands was poorer in species due to their small size rather than geographical isolation. Some recent antropochorous Ponto-Caspian invaders have only reached the lowermost reaches of the two largest rivers. Some brackish water species were found in the mouth of the Pärnu River. No essential differences were found between the comparable sets of oligochaete samples collected in 1954–1975 and 1987–1997 in the Estonian running waters.     

Investigations of �� Initiator Protein-mediated Interaction between Replication Origins �� and �� of the Plasmid R6K

A typical plasmid replicon of Escherichia coli, such as ori γ of R6K, contains tandem iterons (iterated initiator protein binding sites), an AT-rich region that melts upon initiator-iteron interaction, two binding sites for the bacterial initiator protein DnaA, and a binding site for the DNA-bending protein IHF. R6K also contains two structurally atypical origins called α and β that are located on either side of γ and contain a single and a half-iteron, respectively. Individually, these sites do not bind to initiator protein π but access it by DNA looping-mediated interaction with the seven π-bound γ iterons. The π protein exists in 2 interconvertible forms: inert dimers and active monomers. Initiator dimers generally function as negative regulators of replication by promoting iteron pairing (“handcuffing”) between pairs of replicons that turn off both origins. Contrary to this existing paradigm, here we show that both the dimeric and the monomeric π are necessary for ori α-driven plasmid maintenance. Furthermore, efficient looping interaction between α and γ or between 2 γ iterons in vitro also required both forms of π. Why does α-γ iteron pairing promote α activation rather than repression? We show that a weak, transitory α-γ interaction at the iteron pairs was essential for α-driven plasmid maintenance. Swapping the α iteron with one of γ without changing the original sequence context that caused enhanced looping in vitro caused a significant inhibition of α-mediated plasmid maintenance. Therefore, the affinity of α iteron for π-bound γ and not the sequence context determined whether the origin was activated or repressed.     

Recent development and future directions of enzyme technology in Japan

Fundamental research in enzymology have remarkably developed during the past thirty years. However, the actual application of enzymes has been rather limited for several technical and economical reasons. Such limiting factors are now being overcome year after year. Immobilized enzyme and cell technology might be a kind of break-through in this field. From the industrial view-point, enzyme technology is now coming into its real developmental stage. Enzyme technology has recently been finding several directions for its development. These directions may be classified as follows : (1) new industrial catalysts, (2) Tools for food production and processing, (3) pharmaceutical uses, (4) analytical and measurement tools, (5) aids for screening new physiologically active substances, and (6) aids for creation of new sources of energy and raw materials. The authors wish to demonstrate such directions of enzyme technology using actual examples studied and/or operated mainly in Japan.