Mechanical stress induces release of ATP from Ehrlich ascites tumor cells

The supernatant from a suspension of Ehrlich cells exposed to centrifugation at 700xg for 45 s induced a transient increase in the intracellular concentration of free, cytosolic Ca2+, [Ca2+]i, as well as activation of an outwardly rectifying whole-cell current when added to a suspension of non-stimulated cells. These effects were inhibited by suramin, a non-specific P2 receptor antagonist, and mimicked by ATP. Reversed phase HPLC analysis revealed that the supernatant from Ehrlich cells exposed to centrifugation contained 2. 6+/-0.2 microM ATP, and that the mechanical stress-induced release of ATP was inhibited by glibenclamide and verapamil, non-specific inhibitors of the cystic fibrosis transmembrane conductance regulator and P-glycoprotein, respectively. After trypan blue staining, less than 0.5% of the cells were unable to extrude the dye. Addition of extracellular ATP induced a suramin-sensitive, transient, concentration-dependent increase in [Ca2+]i, activation of an outwardly rectifying whole-cell current and a hyperpolarization of the plasma membrane. The ATP-induced hyperpolarization of the plasma membrane was strongly inhibited in the presence of charybdotoxin (ChTX), an inhibitor of several Ca2+-activated K+ channels, suggesting that stimulation of P2 receptors in Ehrlich cells evokes a Ca2+-activated K+ current. The relative potencies of several nucleotides (ATP, UTP, ADP, 2-MeSATP, alpha,beta-MeATP, bzATP) in eliciting an increase in [Ca2+]i, as well as the effect of repetitive addition of nucleotides were investigated. The results lead us to conclude that mechanical stimulation of Ehrlich cells leads to release of ATP, which in turn stimulates both P2Y1 and P2Y2 receptors, resulting in Ca2+ influx as well as release and activation of an outwardly rectifying whole-cell current.     

Energy storage system utilizing large‐capacity electric double‐layer capacitors for peak‐cut of power demand

Recently, due to a rapid increase of demand for air conditioning in summer, peak power demand is becoming increasingly acute. Therefore, the load factor has a tendency to drop every year. The drop of the load factor is leading to a drop in the utilization factor of the power facilities and an increase in the cost of installation. In this paper, we propose an energy storage system for peak‐cut of power demand, in which we use large‐capacity electric double‐layer capacitors. This energy storage system has some distinctive characteristics, including long life span, maintenance‐free operation, preservation of environment, high efficiency at charge/discharge, and so on. This paper deals with the circuit arrangement of the proposed energy storage system, the charge equalization method of the capacitors, and the control method of the converter at charge/discharge. Finally, the operating characteristics of this system are evaluated by simulation analysis. © 2000 Scripta Technica, Electr Eng Jpn, 133(3): 83‐92, 2000     

Fault current limiting system for 500‐kV power systems

Recently, expansion in the scale of power systems and development of localized power sources are leading to an increase in fault current of 500‐kV systems. In the future, it is quite likely that the fault current at the interconnection of such power systems may exceed the rated short‐time current of existing electric power facilities. As one of the solutions of this problem, a thyristor‐controlled series‐resonant‐type fault current limiter (FCL) is proposed to restrain the fault current. This paper deals with the FCL system configuration, the placement method of the FCL in power systems, the outline of the FCL's specification, and the operation method of the protective relay in the multimachine system. Finally, the effectiveness of the FCL is evaluated from the viewpoints of limiting the fault current by simulation analysis. The FCL is shown to be a useful protection device for large, high‐capacity power systems. © 1999 Scripta Technica, Electr Eng Jpn, 127(1): 11–22, 1999     

Improvement of inductive power collection in null-flux maglev system

In the superconducting maglev system it is important to develop a non-contact on-board power source without environmental pollution such as noise and exhaust gases; therefore, inductive power collection (IPC), which utilizes a harmonic magnetic field generated by ground coils in EDS, is being studied. However, alteration to a null-flux EDS that has a high drag ratio reduces the power collecting capacity in the IPC system. In addition, power collecting coils are located on the cryostat of the superconducting coil (SC), so eddy currents at the cryostat also reduce the power collecting capacity. Therefore, an exclusive SC type that locates the exclusive SCs and IPC and power collecting coils so as to face the upper and lower coils of ground coils, respectively, is examined; but we aim to improve the conventional type. After analyzing the influence of eddy currents at the cryostat in detail and improving the composition of the power collecting coil and cryostat, we found that the conventional type has the same capacity as the exclusive SC type. In order to prove the above-mentioned result, we measured the induced voltage of the new-type coils in a test run at Miyazaki test track and confirmed the output of this IPC system in a full-scale synthetic bench test with a PWM converter and magnetic field simulator. © 1998 Scripta Technica. Electr Eng Jpn, 122(2): 48–60, 1998     

Cytoplasmic tail regulates the intercellular adhesion function of the epithelial cell adhesion molecule

Ep-CAM, an epithelium-specific cell-cell adhesion molecule (CAM) not structurally related to the major families of CAMs, contains a cytoplasmic domain of 26 amino acids. The chemical disruption of the actin microfilaments, but not of the microtubuli or intermediate filaments, affected the localization of Ep-CAM at the cell-cell boundaries, suggesting that the molecule interacts with the actin-based cytoskeleton. Mutated forms of Ep-CAM were generated with the cytoplasmic domain truncated at various lengths. All of the mutants were transported to the cell surface in the transfectants; however, the mutant lacking the complete cytoplasmic domain was not able to localize to the cell-cell boundaries, in contrast to mutants with partial deletions. Both the disruption of the actin microfilaments and a complete truncation of the cytoplasmic tail strongly affected the ability of Ep-CAM to mediate aggregation of L cells. The capability of direct aggregation was reduced for the partially truncated mutants but remained cytochalasin D sensitive. The tail truncation did not affect the ability of the transfectants to adhere to solid-phase-adsorbed Ep-CAM, suggesting that the ability to form stable adhesions and not the ligand specificity of the molecule was affected by the truncation. The formation of intercellular adhesions mediated by Ep-CAM induced a redistribution to the cell-cell boundaries of alpha-actinin, but not of vinculin, talin, filamin, spectrin, or catenins. Coprecipitation demonstrated direct association of Ep-CAM with alpha-actinin. Binding of alpha-actinin to purified mutated and wild-type Ep-CAMs and to peptides representing different domains of the cytoplasmic tail of Ep-CAM demonstrates two binding sites for alpha-actinin at positions 289 to 296 and 304 to 314 of the amino acid sequence. The results demonstrate that the cytoplasmic domain of Ep-CAM regulates the adhesion function of the molecule through interaction with the actin cytoskeleton via alpha-actinin.     

Functional correlation between histamine metabolism and worm expulsion in Trichinella spiralis

Mucosal mast cell activity was quantified by measuring histamine forming capacity (HFC) of the gastric mucosa and histamine content in the intestinal tissues of mice infected with T. spiralis. The results wee correlated with the kinetics of worm expulsion. It was found that T. spiralis resulted in significant elevation of HFC by the day 6 post infection (p.i.) which reached a maximal value at day 9, a time when approximately 50% of the established worm burden had been expelled. Histamine content of the intestinal tissues followed the same pattern. No intestinal worms were present by day 28 of infection and there was a gradual reduction in HFC and histamine content which had returned almost to control values by that time. Significant inverse correlation between individual worm burdens and HFC was detected.     

Axin, a negative regulator of the Wnt signaling pathway, forms a complex with GSK-3beta and beta-catenin and promotes GSK-3beta-dependent phosphorylation of beta-catenin

Glycogen synthase kinase-3 (GSK-3) mediates epidermal growth factor, insulin and Wnt signals to various downstream events such as glycogen metabolism, gene expression, proliferation and differentiation. We have isolated here a GSK-3beta-interacting protein from a rat brain cDNA library using a yeast two-hybrid method. This protein consists of 832 amino acids and possesses Regulators of G protein Signaling (RGS) and dishevelled (Dsh) homologous domains in its N- and C-terminal regions, respectively. The predicted amino acid sequence of this GSK-3beta-interacting protein shows 94% identity with mouse Axin, which recently has been identified as a negative regulator of the Wnt signaling pathway; therefore, we termed this protein rAxin (rat Axin). rAxin interacted directly with, and was phosphorylated by, GSK-3beta. rAxin also interacted directly with the armadillo repeats of beta-catenin. The binding site of rAxin for GSK-3beta was distinct from the beta-catenin-binding site, and these three proteins formed a ternary complex. Furthermore, rAxin promoted GSK-3beta-dependent phosphorylation of beta-catenin. These results suggest that rAxin negatively regulates the Wnt signaling pathway by interacting with GSK-3beta and beta-catenin and mediating the signal from GSK-3beta to beta-catenin.