Determination of horseradish peroxidase concentration using the chemiluminescence of Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin-3-one

The chemiluminescence of the Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA) was observed at 462 nm in the presence of horseradish peroxidase (HRP) and the total spectrum of light emitted was found to depend linearly on HRP concentration. Methods for the determination of HRP concentration using the chemiluminescence was investigated. HRP could be detected in the range from 100 pmol/L to 100 nmol/L under the optimum condition, H2O2 (10 mmol/L) and MCLA (10 mumol/L) at pH 5.8.     

Dynamics and organization of MAP kinase signal pathways

In the budding yeast, Saccharomyces cerevisiae, four separate but structurally related mitogen-activated protein kinase (MAPK) activation pathways are known. The best understood of these regulates mating. Pheromone binding to receptor informs cells of the proximity of a mating partner and induces differentiation to a mating competent state. The MAPK activation cascade mediating this signal is made up of Ste11 (a MEK kinase [MEKK]), Ste7 (a MAPK/ERK kinase [MEK]), and the redundant MAPK-related Fus3 and Kss1 enzymes. Another MAPK activation pathway is important for cell integrity and regulates cell wall construction. This cascade consists of Bck1 (a MEKK), the redundant Mkk1 and Mkk2 enzymes (MEKs), and Mpk1 (a MAPK). We exploited these two pathways to learn about the coordination and signal transmission fidelity of MAPK activation cascades. Two lines of evidence suggest that the activities of the mating and cell integrity pathways are coordinated during mating differentiation. First, cells deficient in Mpk1 are susceptible to lysis when they make a mating projection in response to pheromone. Second, Mpk1 activation during pheromone induction coincides with projection formation. The mechanism underlying this coordination is still unknown to us. Our working model is that projection formation generates a mobile second messenger for activation of the cell integrity pathway. Analysis of a STE7 mutation gave us some unanticipated but important insights into parameters important for fidelity of signal transmission. The Ste7 variant has a serine to proline substitution at position 368. Ste7-P368 has higher basal activity than the wild-type enzyme but still requires Ste11 for its function. Additionally, the proline substitution enables the variant to transmit the signal from mammalian Raf expressed in yeast. This novel activity suggests that Ste7-P368 is inherently more permissive than Ste7 in its interactions with MEKKs. Yet, Ste7-P368 cross function in the cell integrity pathway occurs only when it is highly overproduced or when Ste5 is missing. This behavior suggests that Ste5, which has been proposed to be a tether for the kinases in the mating pathway, contributes to Ste7 specificity and fidelity of signal transmission.     

GBLT-VG for High User Densities by User Group Behavior and Hot Point in MMO Virtual Environment

Massively Multiplayer Online (MMO) games have large-scale virtual environments. Here, we propose a load distribution technique called the Group Based Load-distribution Technique with Virtual Group (GBLT-VG) that utilizes Peer-to-Peer (P2P) communication to maintain scalability under high user density conditions in MMO virtual environments. A common problem of P2P virtual environments is that user group behavior frequently generates high user density conditions. Existing techniques to manage the virtual environment cannot maintain performance in such high user density situations. The proposed technique, GBLT-VG, reduces area management loads of manager peers by introducing a new manager who manages a user neighbor group. GBLT-VG forms the user neighbor group efficiently by using party function under high user density conditions caused by users?? cooperative behaviors. In addition, GBLT-VG reduce the management load by using a virtual group when a number of parties is small, thereby decreasing the existing managers?? loads dynamically. This paper also presents simulation results demonstrating how GBLT-VG can reduce the average number of users managed by the server.     

Gas permeation property of polyaniline films

We examined the influence of polyaniline (PAn)'s unit sequence and doping with low molecular weight dopants or polymer dopants on permeation property. It was found that CO₂ permeability was increased by the formation of a quinonediimine unit in PAn with the oxidation. CO₂ sorption amount of PAn was decreased by oxidation. The increase of CO₂ permeability with oxidation, therefore, resulted from the increase of diffusivity, which was attributable to morphological variation by the increase of a quinonediimine unit. The permselectivity of PAn films was found to be remarkably improved by doping. In particular, the selectivity value of the PAn film doped with polyvinyl sulphonic acid as a polymer dopant went up to over 2,000. This remarkable increase of selectivity was found to result in the increase of selectivity, depending on diffusivity. It was also found that the permselectivity of the PAn film doped by polymer dopants was surpassed, as compared with that doped by low molecular dopants. © 1995 John Wiley & Sons, Inc.     

Study on modified phenolic resin. I. Modification with homopolymer prepared from p-hydroxyphenylmaleimide

We studied the improvement of heat resistance and mechanical properties of phenolic resin modified with polymers prepared from p-hydroxyphenylmaleimide (HPMI). Homopolymers of HPMI and copolymers of HPMI with some vinyl monomers were synthesized and M_n T_g and thermal decomposition temperature of them were measured by GPC, DSC, and TG, respectively. The miscibility of homopolymer with novolac was examined by DSC. It was found that the homopolymer had good heat resistance and good miscibility with novolac. Molding compounds were prepared by hot roll-kneading the mixtures of novolac, the homopolymer, hexamethylenetetramine, and glass fiber. It was found that molded test pieces had good heat resistance.     

Adsorption of trypsin onto poly(2-hydroxyethyl methacrylate)/polystyrene composite microspheres and its enzymatic activity

Poly(2-hydroxyethyl methacrylate)/polystyrene (PHEMA/PS) composite microspheres were produced by emulsifier-free seeded emulsion polymerization for styrene in the presence of PHEMA seed particles. Effects of the surface characteristics of the PHEMA/PS composite microspheres on the adsorption immobilization of trypsin and on its enzymatic activity were discussed. Above 5 mol% of HEMA content, trypsin molecules adsorbed had high activity, 65–100% of the activity of free trypsin. The excellence of the composite microspheres as a carrier for trypsin seems to be closely related with the surface heterogeneity consisting of both hydrophilic and hydrophobic parts.     

Estimation of corrosion conditions of a phosphoric acid fuel cell

The changes in the anode and cathode potentials in the horizontal plane of a phosphoric acid fuel cell (PAFC), under various conditions of reactant gas pressure and its utilization, were studied using a single cell with twelve reference electrodes located around the cathode. Pressure-utilization (P-U) potential maps were obtained from the data at various reactant gas partial pressures (PO₂, PH₂) and their utilization (UO₂, UH₂). These maps show the corrosion conditions clearly. A PO₂-UO₂ potential map of maximum cathode potential showed that the cathode is corroded at high oxygen partial pressures and at low oxygen utilization. Cathode corrosion can occur over the entire cell surface. A PH₂-UH₂ potential map of maximum cathode potential showed that the cathode is corroded at high hydrogen utilization and at any hydrogen partial pressure. However, in this case, cathode materials corrodes at the fuel outlet; the potential does not climb to high values at the fuel inlet area. Fuel gas flowing in series resulted in a lower possibility for corrosion than parallel gas flow.     

Catalysis of gasification of coal-derived cokes and chars

Calcium is the most important in-situ catalyst for gasification of US coal chars in O₂, CO₂ and H₂O. It is a poor catalyst for gasification of chars by H₂. Potassium and sodium added to low-rank coals by ion exchange and high-rank coals by impregnation are excellent catalysts for char gasification in O₂, CO₂ and H₂O. Carbon monoxide inhibits catalysis of the CH₂O reaction by calcium, potassium and sodium; H₂ inhibits catalysis by calcium. Thus injection of synthesis gas into the gasifier will inhibit the CH₂O reaction. Iron is not an important catalyst for the gasification of chars in O₂, CO₂ and H₂O, because it is invariably in the oxidized state. Carbon monoxide disproportionates to deposit carbon from a dry synthesis gas mixture (3 vol H₂ + 1 vol CO) over potassium-, sodium- and iron-loaded lignite char and a raw bituminous coal char, high in pyrite, at 1123 K and 0.1 MPa pressure. The carbon is highly reactive, with the injection of 2.7 kPa H₂O to the synthesis gas resulting in net carbon gasification. The effect of traces of sulphur in the gas stream on catalysis of gasification or carbon-forming reactions by calcium, potassium, or sodium is not well understood at present. Traces of sulphur do, however, inhibit catalysis by iron.