Lectin-like characteristics of recombinant human interleukin-1beta recognizing glycans of the glycosylphosphatidylinositol anchor

We found that 35S-labeled recombinant human interleukin-1beta (rhIL-1beta) binds phosphatidylinositol-specific phospholipase C-treated human placental alkaline phosphatase, phosphatidylinositol-specific phospholipase C-treated trypanosome surface variant glycoproteins, and urinary uromodulin immobilized on plates or immobilized on CNBr-activated Sepharose 4B. The interaction between rhIL-1beta and these glycoproteins was lectin-like, since it was inhibited in the presence of specific saccharides, i.e. mannose 6-phosphate or synthetic Ac-NH.CH2.CH2. PO4--->6Manalpha1-->(+/-2Manalpha1-->+/-6Manalpha1-->) propyl at about 1 microM. On the other hand, a wide variety of compounds including biantennary sugar chains derived from these glycoproteins as well as ethanolamine phosphate, inositol phosphate, mannose 6-sulfate, mannose 1-phosphate, glucose 6-phosphate, and mannitol 6-phosphate did not show any inhibitory effect at concentrations up to 1 mM. These results indicate that rhIL-1beta interacts with these glycoproteins via the mannose 6-phosphate diester of glycans on the glycosylphosphatidylinositol (GPI) anchor. Furthermore, when monolayers of polarized Madin-Darby canine kidney cells on polycarbonate filter membranes were incubated with 35S-rhIL-1beta in either the apical or basolateral chamber, 35S-interleukin-1beta was found to bind specifically to the apical membranes with a Ka value of 4.6 x 10(7) M-1, and the specific interaction was inhibited by 1 microM mannose 6-phosphate. Since the mannose 6-phosphate diester moiety exists only in the GPI glycans on plasma membranes, it was evident that interleukin-1beta can directly interact with the mannose 6-phosphate diester component of the intact glycan of GPI anchors on plasma membranes.     

DNA damage induced by m-phenylenediamine and its derivative in the presence of copper ion

We have previously reported that long-term priming of human polymorphonuclear neutrophilic granulocytes (PMN) with interferon-gamma (IFN-gamma) increased the fMLP-stimulated calcium influx. We now show that also after short-term incubation with IFN-gamma, PMN calcium metabolism is modulated. Single adherent cells in three different calcium-containing buffers (high, normal, and low [Ca2+]) were stimulated with the bacterial peptide fMLP or the Ca-ATPase inhibitor thapsigargin (Tg) after about 5 min preincubation with IFN-gamma. The results of this protocol indicated that IFN-gamma increases both calcium influx and calcium sequestration. Store dependent Ca2+ influx, directly measured on readdition of calcium to Tg-treated cells incubated in EGTA buffer, was significantly enhanced in IFN-gamma-treated cells. This effect of IFN-gamma was enhanced by the tyrosine kinase inhibitor herbimycin A. Strikingly, in low extracellular calcium concentrations, IFN-gamma induced calcium transients in 20%-60% of the cells. The proportion of PMN responding with Ca2+ transients increased with decreasing extracellular calcium concentration. Average lagtime from addition of IFN-gamma to a response that could be measured was 7.3 sec, and average increase in [Ca2+] above the basal level was 790 nM. These IFN-gamma-induced transients could not be depressed by herbimycin A. Thus, IFN-gamma can increase capacitative calcium influx, induce calcium transients, and possibly affect calcium sequestration in human PMN.     

Enhancement by ganglioside GT1b of annexin I phosphorylation in bovine mammary gland in the presence of phosphatidylserine and Ca2+

Ganglioside GT1b and, to a lesser extent, GD3, enhanced phosphorylation of a 36 kDa protein (the substrate of protein kinase C) in the particulate fraction from bovine mammary gland. Sialic acids, asialogangliosides, and GM3 were without effect, and GD1a conversely inhibited phosphorylation of the 36 kDa protein. The enhanced phosphorylation by GT1b required the simultaneous presence of phosphatidylserine (PS) and Ca²⁺. The 36 kDa protein reacted with anti-annexin I in Western blot analysis. Addition of purified annexin I to the reaction mixture containing the particulate fraction increased the extent of phosphorylated 36 kDa protein, and the phosphorylation was further enhanced by GT1b. The enhanced phosphorylation of annexin I by GT1b was also dependent on PS and Ca²⁺. When annexin I was phosphorylated by purified protein kinase C, GT1b inhibited the annexin I phosphorylation. Addition of epidermal growth factor or insulin to the particulate fraction had little effect on the enhancement. These results suggest that an enzyme or enzymes other than protein kinase C, epidermal growth factor receptor kinase, or insulin receptor kinase is responsible for the GT1b- and GD3-enhanced phosphorylation of annexin I in the presence of PS and Ca²⁺.     

Novel anion exchange membranes having fluorocarbon backbone: Preparation and stability

Anion exchange membranes with excellent durability were prepared by chemical modification of Nafion. The modification was achieved by transformation of the sulfonic acid group into quaternary ammonium group. Namely, Nafion membrane was first converted into an amide-type membrane. Reduction of the carbonxyl group to methylene followed by quaternarization with alkyl iodide resulted in the formation of an anion exchange membrane. The electric resistance of the resulting membranes depends on the equivalent weight of the starting membranes (4.4–6.0 Ω cm² in 0.5N NaCl). The characteristics of the membranes are the excellent stability toward chemical substances such as organic solvents, oxidizing agents, acids, etc. For example, the membranes are stable in aqueous saturated chlorine solution at 60°C for 1000 hr.     

Stabilization of bioethanol recovery with silicone rubber‐coated ethanol‐permselective silicalite membranes by controlling the pH of acidic feed solution

In order to produce highly concentrated bioethanol by pervaporation using an ethanol‐permselective silicalite membrane, techniques to suppress adsorption of succinic acid, which is a chief by‐product of ethanol fermentation and causes the deterioration in pervaporation performance, onto the silicalite crystals was investigated. The amount adsorbed increased as the pH of the aqueous succinic acid solution decreased. The pervaporation performance also decreased with decreasing pH when the ternary mixtures of ethanol/water/succinic acid were separated. Using silicalite membranes individually coated with two types of silicone rubber, pervaporation performance was significantly improved in the pH range of 5 to 7, when compared with that of non‐coated silicalite membranes in ternary mixtures of ethanol/water/succinic acid. Moreover, when using a silicalite membrane double‐coated with the two types of silicone rubber, pervaporation performance was stabilized at lower pH values. In the separation of bioethanol by pervaporation using the double‐coated silicalite membrane, removal of accumulated substances having an ultraviolet absorption maximum at approximately 260 nm from the fermentation broth proved to be vital for efficient pervaporation. Copyright © 2005 Society of Chemical Industry     

Pancyclicity of recursive circulant graphs

In this paper, we study the existence of cycles of all lengths in the recursive circulant graphs, and we show a necessary and sufficient condition for the graph being pancyclic and bipancyclic.     

Dispersion compensation with SBS-suppressed fibre phase conjugator using synchronised phase modulation

204 km, 10 Gbit/s standard optical fibre transmission is successfully demonstrated using the mid-span spectral inversion technique with a stimulated Brillouin scattering suppressed fibre phase conjugator, proposed recently by Tani and Yamashita. Transmission performance has been improved compared with a conventional phase conjugator.     

The principle and fundamental characteristics of brushless three-phase synchronous induction motor with two stators

Brushless three-phase synchronous motor involving a rotating ac exciter on the same shaft as the motor and with the windings linked through a shaft-mounted rectifier is widely used as medium and large capacity machines. However, the motor needs to fit with damping windings for self-starting outside of the field winding in the rotor and needs to equip discharge resistance with complex electronic device to dispose of electromagnetic force induced in the field winding at starting. Therefore, the rotor structure is very complex, and there are disadvantages, especially for the robust and highly reliable motor that is desired. To solve the problem found in traditional motors, the authors proposed a brushless three-phase synchronous induction motor with two stators and one wound rotor. This problem is solved because the proposed motor is of two-stators structure: the rotor winding operates as a secondary winding of induction motor at starting and as the field winding at synchronous operation. The motor can self-start as a wound-rotor induction motor without external secondary resistance. Therefore, discharge resistance is not necessary; the starting equipment is very simple and the rotor structure is robust. It is possible to start with high torque. In this paper, the basic constitution of the proposed motor and the principles of operation are described in detail. The experimental results at starting and pulling into synchronism are shown for confirming the principles of operation and the experimental and numerical results of the starting characteristics. The load ones and the generating ones are shown and then the practical use of the proposed motor is confirmed.     

The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2

Activation of the cyclin-dependent protein kinases p34cdc2 and p33cdk2 requires binding with a cyclin partner and phosphorylation on the first threonine residue in the sequence THEVVTLWYRAPE. We present evidence that this threonine residue, number 160 in p33cdk2, can be specifically phosphorylated by a cdc2-related protein kinase from Xenopus oocytes called p40MO15. Binding to cyclin A and phosphorylation of this threonine are both required to activate fully the histone H1 kinase activity of p33cdk2. In cell extracts, a portion of p40MO15 is found in a high molecular weight complex that is considerably more active than a lower molecular weight form. Wild-type MO15 protein expressed in bacteria does not possess kinase activity, but acquires p33cdk2-T160 kinase activity after incubation with cell extract and ATP. We conclude that p40MO15 corresponds to CAK (cdc2/cdk2 activating kinase) and speculate that, like p33cdk2 and p34cdc2, p40MO15 requires activation by phosphorylation and association with a companion subunit.     

Microscopic fatigue processes in a plain-weave glass-fibre composite

This paper focuses on the microscopic damage and progressive failure of a composite reinforced by plain-weave glass cloth under tensile fatigue loading. The fatigue process was divided into three stages like that of multi-directional laminates. It was found that the internal damage at each stage (matrix cracks, debonds in the weft, successive debonds in the warp and ‘metadelaminations’ between warps and wefts) occurred near the cross-over point of the fabric. The modulus decay mechanism was explained by considering the progression of this internal damage. From the end of the first stage to the beginning of the middle stage, a characteristic damage state (CDS) (called a ‘meta-CDS’) was observed. It was found that woven composites have a unit area of damage accumulation (called a ‘unit cell’) and the damage of each unit cell and its distribution control the total fatigue damage of the material.