Dimer interface of glutathione S-transferase from Arabidopsis thaliana: influence of the G-site architecture on the dimer interface and implications for classification

The three-dimensional structure of glutathione S-transferase from Arabidopsis thaliana has been solved at 2.2 A resolution (Reinemer et al., 1996). The enzyme forms a dimer of two identical subunits. The structure shows a new G-site architecture and a novel and unique dimer interface. Each monomer of the protein forms a separate G-site. Therefore, the requirements on the dimer interface are reduced. As a consequence, the interactions between the monomers are weaker and residues at the dimer interface are more variable. Thus, the dimer interface looses its relevance for a classification of plant glutathione S-transferases and the formation of heterodimers becomes even more difficult to predict.     

Automatic tuning of decentralized PID controllers for MIMO processes

An automatic tuning algorithm for decentralized PID control in multiple-input multiple-output (MIMO) plants is presented. This algorithm generalizes the authors' recent auto-tuner for two-input two-output systems to any number of inputs and outputs. The algorithm consists of two stages. In the first, the desired critical point, which consists of the critical gains of all the loops and a critical frequency, is identified. The auto-tuner identifies the desired critical point with almost no a priori information about the process. During the identification phase all controllers are replaced by relays, thus generating limit cycles with the same period in all loops. It is shown that each limit cycle corresponds to a single critical point of the process. By varying the relays parameters different points can be determined. The auto-tuner contains a procedure which converges rapidly to the desired critical point while maintaining the amplitudes of the process variables as well as of the manipulated variables within prespecified ranges. In the second stage, the data of the desired critical point is used to tune the PID controllers by the Ziegler-Nichols rules or their modifications. This paper focuses on the first stage. The steady-state process gains, which are required for the appropriate choice of the desired critical point, are determined by the auto-tuner in closed-loop fashion simultaneously with the identification of the critical point. The identification of the process gains is achieved at no extra plant time. Based upon a large number of simulated cases, the proposed auto-tuner seems to be efficient and robust. The paper discusses the underlying principles of the auto-tuner and its properties and capabilities are demonstrated via examples.     

In Situ ceramic particle-reinforced aluminum matrix composites fabricated by reaction pressing in the TiO2 (Ti)-Al-B (B2O3) systems

Particulate TiB₂ reinforced aluminum-based metal matrix composites (MMCs) were successfully fabricated by means of the reaction processing method. TiB₂ particulates were formed in situ through the reaction of Ti and B in Ti-Al-B, TiO₂ and B in TiO₂-Al-B, and TiO₂ and B₂O₃ in TiO₂-Al-B₂O₃ systems. The results showed that in situ TiB₂ particulates formed in the Ti-Al-B system had a size of 5 μm and they exhibited block and rodlike structures. Moreover, coarse Al₃Ti blocks several tens of micrometers in size were also formed simultaneously. On the other hand, equiaxed Al₂O₃ and TiB₂ particulates with a size of less than 2 μm were formed in situ in the TiO₂-Al-B and TiO₂-Al-B₂O₃ systems. The Al₃Ti phase was completely eliminated in the TiO₂-Al-B system with increasing B content. Tensile tests revealed that the Al₂O₃ · TiB₂/Al composite fabricated from the TiO₂-Al-B system exhibits excellent mechanical properties. The yield strength of the Al₂O₃ · TiB₂/Al composite appeared to increase with increasing TiB₂ content. The yield strength of the Al₂O₃ · TiB₂/Al composite could be further increased by introducing CuO into the TiO₂-Al-B system. Such an increment in mechanical strength arose from the strengthening effect caused by the Al₂Cu precipitates. The incorporation of CuO had no effect on the in situ reaction process of the TiO₂-Al-B system. Finally, the effect of SiC addition on the microstructure and mechanical properties of the composites fabricated from the TiO₂-Al-B and TiO₂-Al-B-CuO systems was also investigated.     

Neural mechanism of pupillary dilation elicited by electro-acupuncture stimulation in anesthetized rats

The neural mechanisms to reflex dilation elicited by electro-acupuncture stimulation were investigated in anesthetized rats. Two needles, with 160 microns diameter and about 5 mm apart, were inserted into the skin and underlying muscle of a hindpaw. Repetitive 20 Hz, 0.5 ms electrical pulses at various intensities were used for stimulation for 30s. The pupil size was magnified about 44 times via a microscope and was continuously recorded on a videotape. Electro-acupuncture stimulation at more than 0.5 up to 6 mA induced stimulus intensity-dependent pupil dilation. These responses were abolished by the severance of the sciatic and saphenous nerve of the stimulated hindlimb. Compound action potentials were recorded from the distal cut end of the tibial of a saphenous nerve following electro-acupuncture stimulation of the hindpaw. The mean threshold of the compound action potentials of the myelinated fibers in saphenous nerves was 0.18 mA, while that of unmyelinated fibers was 3.0 mA. The mean threshold of the compound action potentials of the myelinated fibers in the tibial nerve was 0.20 mA of unmyelinated fibers was 3.3 mA. Severance of bilateral trunks did not affect the response, while severance of the third cranial nerves abolished the responses. In conclusion, electro-acupuncture stimulation applied to the hindpaws of the anesthetized rats induced excitation of myelinated or of both myelinated and unmyelinated afferent fibers of the tibial and saphenous nerve, and involved a reflex response of pupil dilation through the third cranial parasympathetic efferent nerve.     

Detection of an anatoxin-a(s)-like anticholinesterase in natural blooms and cultures of cyanobacteria/blue-green algae from Danish lakes and in the stomach contents of poisoned birds

Ten natural bloom samples of cyanobacteria from the Danish lakes Knud s? (5), Ravn s? (4), and Salten Langs? (1) collected during 1993-1995 were assayed for toxicity by mouse bioassay, for acetylcholinesterase inhibiting activity by a colorimetric method, and for microcystins by enzyme-linked immunosorbent assay. In the mouse bioassay, seven samples were neurotoxic, two were non-toxic and one gave a protracted toxic response. One of the non-toxic and the single protracted toxic sample both contained anticholinesterase activity equivalent to 4 micrograms anatoxin-a(s) g-1. The neurotoxic samples contained equivalents to 20-3300 micrograms anatoxin-a(s) g-1. The highest anticholinesterase activities (equivalent to 2300 and 3300 micrograms anatoxin-a(s) g-1, respectively) were found in samples collected from Lake Knud s? in connection with bird-kills in 1993 and 1994. Small amounts of microcystins (0.1-0.9 microgram g-1) were detected in all samples but one. All Lake Knud s? and Lake Ravn s? samples were dominated by Anabaena lemmermannii, and the Lake Salten Langs? sample by several species of Anabaena. Gel filtration profiles indicated similarity between the toxic component from the Lake Knud s? 1994 bloom with registered bird-kills and anatoxin-a(s) isolated from Anabaena flos-aquae NRC-525-17. Anticholinesterase-producing cultures of A. lemmermannii were isolated from the Lake Knud s? 1993 bloom. These laboratory cultures produced anatoxin-a(s) equivalents of 29-743 micrograms g-1. Other cultures of A. lemmermannii isolated from Lake Knud s? and Lake Ravn s? were hepatotoxic or non-toxic. Dead birds collected from Lake Knud s? during the neurotoxic 1993 Anabaena bloom possibly died from cyanobacterial toxicosis. The stomach contents contained colonies and single trichomes of Anabaena, and anticholinesterase activities equivalent to 2.1-89.7 micrograms anatoxin-a(s) kg-1 body weight and microcystins (53-95 ng kg-1) were also detected.     

A magnetic resonance imaging study of planum temporale asymmetry in men with developmental dyslexia

The influence of ionic strength and composition on the binding and inhibition of human leukocyte elastase by glycosaminoglycans with variable degree and position of sulfation was investigated. The kinetic mechanism of inhibition had a hyperbolic, mixed-type character with a competitive component that was promoted by low ionic strength, reduced by phosphate ions, and which also depended on the substrate and glycosaminoglycan structure. Enzyme binding was a cooperative phenomenon that varied with ionic strength and composition. The inhibition patterns correlated with the cationic character of elastase and with the distribution of arginines on its molecular surface, most notably with residues located in the vicinity of the substrate binding region. The order of affinity for elastase binding was chondroitin 4-sulfate < chondroitin 6-sulfate < dermatan sulfate, iduronate-containing derivatives being superior with respect to the glucuronate-containing counterparts. Additional sulfation at both the 4- and 6- positions or at the N- and 4-positions of the N-acetylgalactosamine moiety decidedly improved the inhibitory efficiency. The results highlight a fundamental physiological role of enzyme-glycosaminoglycan interactions. In the azurophil granule of the human polymorphonuclear neutrophil, elastase and other enzymes are bound to a matrix of chondroitin 4-sulfate because this is the only glycosaminoglycan that simultaneously offers good binding for enzyme compartmentalization together with prompt release from the bound state at the onset of phagocytosis.     

Differences in the yields of dicentrics and reciprocal translocations observed in the chromosomes of irradiated human skin fibroblasts and blood lymphocytes from the same healthy individuals

The modulatory role of locally produced cyclooxygenase products and endothelium-derived nitric oxide in controlling vascular tone was investigated in bovine intra-mammary artery. Vascular reactivity initiated by vasoactive compounds, endothelin-1 (ET-1), bradykinin (BK), and substance P (SP) was measured isometrically in an isolated tissue bath. The effects of a cyclooxygenase inhibitor, indomethacin (10(-5) M) and an inhibitor of nitric oxide production, N omega-Nitro L-Arginine (L-NNA: 3 x 10(-4) M) were determined during agonist-mediated responses. Indomethacin alone markedly enhanced vascular contraction produced by ET-1, while L-NNA did not. Inhibition of endothelium-derived nitric oxide synthesis by L-NNA, however, significantly attenuated BK- and SP-induced vascular relaxations, whereas indomethacin had slight influence. The potentiation between indomethacin and L-NNA in regulating vasomotor tone was not observed in this vascular bed. Thus, it appeared that both the cyclooxygenase and endothelium-derived nitric oxide pathways participated in modifying vascular reactivity. Domination of one pathway over the other depended upon the agonist used to stimulate vascular tissue.