Tyrosine phosphorylation of Crk-associated substrates by focal adhesion kinase. A putative mechanism for the integrin-mediated tyrosine phosphorylation of Crk-associated substrates

Integrin-ligand binding induces the tyrosine phosphorylation of various proteins including focal adhesion kinase (pp125(FAK)) and Crk-associated substrate (Cas). FAK is activated and autophosphorylated by the ligation of integrins, although the substrate of FAK has not been revealed. We show here that p130(Cas) and Cas-L are FAK substrates. FAK directly phosphorylates Cas proteins primarily at the YDYVHL sequence that is conserved among all Cas proteins. Furthermore, the phosphorylated YDYVHL sequence is a binding site for Src family protein-tyrosine kinases, and the recruited Src family kinase phosphorylates the other tyrosine residues within Cas. The Cas-L YDYVHL sequence is phosphorylated upon integrin-ligand binding, and this integrin-mediated tyrosine phosphorylation is inhibited by the cotransfection of the FAK COOH-terminal domain that does not contain a kinase domain. These findings strongly suggest that FAK initiates integrin-mediated tyrosine phosphorylation of Cas proteins; then, Src family tyrosine kinases, which are recruited to phosphorylated Cas and FAK, further phosphorylate Cas proteins.     

Numerical study of radiation‐convection heat transfer

In this study, the authors attempted to introduce a simulation technique for radiation‐convection heat transfer in the high‐temperature fields of industrial furnaces, boilers, and gas turbine combustors. The convection effect was analyzed by a differential equation, but the radiation effect was analyzed by an integral equation. Thus, it was not easy to arrange both effects using the same type of equations. Then, the authors introduced the zone method and Monte Carlo method for the integral equation of the radiation effect and the finite difference method for the differential equation of the convection effect. A three‐dimensional analysis of the high‐temperature furnace was performed by this simulation technique to obtain its temperature distribution. Furthermore, another radiation‐convection heat transfer analysis in the low‐temperature living room was performed by the same technique. Finally, the authors tried to develop a computer software for radiation‐convection heat transfer and described their idea of software construction for the above. © 2002 Wiley Periodicals, Inc. Heat Trans Asian Res, 31(5): 391–407, 2002; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.10042     

Highly sensitive pyrosequencing based on the capture of free adenosine 5' phosphosulfate with adenosine triphosphate sulfurylase

In pyrosequencing chemistry, four cascade enzymatic reactions with the catalysis of polymerase, adenosine triphosphate (ATP) sulfurylase, luciferase, and apyrase are employed. The sensitivity of pyrosequencing mainly depends on the concentration of luciferase which catalyzes a photoemission reaction. However, the side-reaction of adenosine 5' phosphosulfate (APS, an analogue of ATP) with luciferase resulted in an unavoidable background signal; hence, the sensitivity cannot be much higher due to the simultaneous increase of the background signal when a larger amount of luciferase is used. In this study, we demonstrated a sensitive pyrosequencing using a large amount of ATP sulfurylase to lower the concentration of free APS in the pyrosequencing mixture. As the complex of ATP sulfurylase and APS does not react with luciferase, a large amount of luciferase can be used to achieve a sensitive pyrosequencing reaction. This sensitivity-improving pyrosequencing chemistry allows the use of an inexpensive light sensor photodiode array for constructing a portable pyrosequencer, a potential tool in a point-of-care test (POCT).     

Behavior of shock waves formed by unsteady supersonic jet injected into cell

The unsteady behavior of flow driven by a jet suddenly injected into a cell is numerically studied by solving the axisymmelric two-dimensional compressible Navier-Stokes equations. The system of the calculation is a model of the laser ablation of a certain duration followed by a discharging process through the exit hole at the down- stream end of the cell. In the calculations, the contour of the cell is changed while other parameters such as the Mach number of the jet, its duration, and the diameter of the cell exit are fixed. Monitoring the velocity at the exit hole is used to investigate the influence of the shape on the interaction between the shock wave and the jet. As the result, it was found that the velocity peak value and its arrival time at the downstream end of the cell exit are determined by the diameter of the cell.     

Suppression of cell growth by ectopic expression of N-cadherin

We found that ectopic expression of N-cadherin in 3Y1 caused tight association of cells and, thereby, substantially suppressed cell growth. N-cadherin expression inhibited neither tyrosine phosphorylation of cellular proteins, GTP uptake onto Ras, nor activation of MAP kinase, suggesting that it does not directly interfere the Ras-MAP kinase pathway. However, co-expression of N-cadherin with dominant negative Ras, S17N Ras, showed synergestic growth inhibitory effect, suggesting that N-cadherin signaling antagonizes the Ras-MAP kinase signaling. In addition, we found that N-cadherin yielded cell-cycle arrest at G0/G1 phase. These results strongly suggest that N-cadherin cell adhesion machinery works as a negative controller of cell cycle in 3Y1 and this growth suppressive function of cadherin is distinct from the epithelial morphogenetic function.     

The relationship between deterioration and reversal of optic disc cupping in monkeys with chronic experimental high-pressure glaucoma

PURPOSE: To investigate the correlation between the deterioration in optic disc cupping during the chronic elevation of intraocular pressure (IOP) and the reversal of cupping during a subsequent reduction in IOP in experimental glaucoma. METHODS: We examined changes in the vertical and horizontal cup to disc ratios, the rim area to disc area ratio, and the cup volume to disc area ratio in 11 monkey eyes with laser-induced glaucoma using computerized stereo-image analysis. Correlations between changes in disc parameters during a spontaneous IOP reduction and changes in disc parameters during a period of chronic IOP elevation from baseline before laser exposure (baseline) to before the IOP reduction (pre-IOP reduction) and during the period from baseline to after the reduction in IOP (post-IOP reduction) were determined by linear regression analysis. RESULTS: All disc parameters improved significantly during IOP reduction and deteriorated significantly during the periods from baseline to the pre-IOP reduction and from baseline to the post-IOP reduction. The degree of reversal in disc parameters was correlated with the deterioration from baseline to the pre-IOP reduction and from baseline to the post-IOP reduction in the vertical cup to disc ratio (r = 0.68, P = 0.0218 and r = 0.97, P < 0.0001, respectively), the horizontal cup to disc ratio (r = 0.57, P = 0.0649 and r = 0.98, P < 0.0001, respectively), the rim area to disc area ratio (r = 0.68, P = 0.0227 and r = 0.98, P < 0.0001, respectively), and the cup volume to disc area ratio (r = 0.67, P = 0.0256 and r = 0.88, P = 0.0004, respectively). CONCLUSION: The degree of deterioration in cupping from baseline before the induction of glaucoma may be an important determinant of the degree of cupping reversal during subsequent reductions in IOP in primate glaucoma.     

NiZnCu Ferrite Thick Film with Nano Scale Crystallites Formed by the Aerosol Deposition Method

This paper describes the magnetic properties of NiZnCu ferrite film deposited at room temperature by an aerosol deposition method (ADM). The thickness of the film was 6 μm and the deposition rate was estimated as 2 μm/min. The microstructure of as-deposited at room temperature films consists of randomly oriented nanocrystallites with a size of 20 nm. As-deposited and annealed films exhibited the following magnetic properties: intensity of magnetization M _s= 0.147 T (117 emu/cm³), coercivity H _c= 40.58 kA/m (510 Oe); and M _s= 0.3 T (250 emu/cm³), H _c= 14.95 kA/m (188 Oe), respectively.     

Interleukin-6 production by macrophages from BALB/c mice with a chronic infection of lactic dehydrogenase virus

The production of interleukin-6 (IL-6), which is known as a B cell differentiation factor, by peritoneal macrophages from mice with a chronic lactic dehydrogenase virus (LDV) infection was compared with that from uninfected mice. The same amounts of IL-6 were detected in the culture supernatant of macrophages from LDV-infected mice as those from uninfected mice. Furthermore IL-6 production of macrophages from LDV-infected and uninfected mice was not affected by the addition of indomethacin. These results suggested that many immunological alterations seen in LDV-infected mice may not be due to, at least in part, altered IL-6 production ability of macrophages and the IL-6 production may not be affected by cyclooxygenase-derived products.     

Influence of anti-LDL receptor antibody on the transfer of cholesterol from LDL to isolated lymphocytes in normal subjects

Results 2 1/2 years after an enuresis nocturna training are presented, including rate of success, percentage and duration of relapse for 113 children (mean age 11.6 year at the start of the training). The bibliotherapeutic treatment by parents did not require any intervention by a professional. Behaviour of parents in the event of a relapse differed between training conditions. Children in the Arousal condition recovered faster from a relapse, 90% of their parents used the Arousal training again at relapse or did not intervene at all and none of them consulted a professional. Clearly they had confidence in the method of Arousal training: combining the alarm device with reinforcement for correct behaviour at the time the alarm goes off. Parents in control conditions did not use the alarm device as often as the parents in the Arousal condition, but tried other means with less success, including consulting professionals.