Subcellular redistribution is involved in acute regulation of the brush border Na+/H+ exchanger isoform 3 in human colon adenocarcinoma cell line Caco-2. Protein kinase C-mediated inhibition of the exchanger

Na+/H+ exchanger isoform 3 (NHE3), an epithelial brush border isoform of the Na+/H+ exchanger gene family, plays an important role in reabsorption of Na+ in the small intestine, the colon, and the kidney. In several cell types, phorbol 12-myristate 13-acetate (PMA) acutely inhibits NHE3 activity by changes in Vmax, but the mechanism of this inhibition is unknown. We investigated the role of subcellular redistribution of NHE3 in the PMA-induced inhibition of endogenous brush border NHE3 in a model human colon adenocarcinoma cell line, Caco-2. Subcellular localization of NHE3 was examined by confocal morphometric analysis complemented with cell surface biotinylation and compared with NHE3 activity evaluated by fluorometric measurement of intracellular pH. PMA inhibited NHE3 activity by 28% (p < 0.01), which was associated with a decrease of the ratio of the brush border/subapical cytoplasmic compartment of NHE3 from approximately 4.3 to approximately 2.4. This translocation resulted in 10-15% of the total cell NHE3 being shifted from the brush border pool to the cytoplasmic pool. These effects were mediated by protein kinase C, since they were blocked by the protein kinase C inhibitor H7. We conclude that inhibition of NHE3 by protein kinase C in Caco-2 cells involves redistribution of the exchanger from brush border into a subapical cytoplasmic compartment, and that this mechanism contributes approximately 50% to the overall protein kinase C-induced inhibition of the exchanger.     

Refinement of conventional PSS design in multimachine system bymodal analysis

The design of the lead/lag network in a conventional power system stabilizer (PSS) is intended to provide the correct compensation in order to obtain an electrical torque component in phase with the speed variation. It is shown that, for a multimachine system, the conventional design analysis and synthesis tend to oversimplify the system representation and hence the interaction effects. A more rigorous approach that considers four torque (or power) components instead of the conventional single component is discussed. Using a modal analysis, the significance of these other components is revealed, the more comprehensive treatment using a generalized multimachine representation is justified and a reliable means for optimizing the PSS parameters is presented     

Regulation of angiogenesis and tumor growth

The data on tumour vascularization regulating substances, their structure and mechanism of action are reviewed. The systems for testing angiogenic reactions are analyzed. The role of mast cells, lymphocytes, macrophages and leucocytes in formation of new microvessels is under discussion. The results of the search for antitumor drugs inhibiting angiogenesis are promising. The data are analyzed on the role of prostaglandins, Cu2+ and urokinase in the processes of tumour vascularization. The possibility of the complex and multifactor regulation of microvessels is suggested.     

Synergetic effect of Re and Ru on γ/γ′ interface strengthening of Ni-base single crystal superalloys

The synergetic effect of Re and Ru on γ/γ′ interface strengthening of Ni-base SC superalloys has been investigated by performing DMol3 calculations. Results show that the synergetic effect of Re and Ru on the interface strengthening is better than that achieved by the individual Re or Ru due to Re-d/Ru-d, Re-d/Ni-d and Ru-d/Ni-d hybridizations. The electronic mechanism underlying the synergetic effect of Re and Ru on γ/γ′ interface strengthening is related to the charge transfer of electrons and the enhancement of d-bonding hybridization among Re---Ru, Re---Ni and Ru---Ni atoms.     

Effects of Temperature and Pressure on ZDDP

A recent theoretical study proposed that the anti-wear property of zinc dialkyl dithio phosphate (ZDDP) is due to the formation of chemically connected networks as a result of pressure-induced cross-linkage of phosphate groups of thermally decomposed ZDDP. To investigate the initial decomposition processes and the possibility of linking of phosphate groups in the decomposed product, in-situ high-pressure and high-temperature infrared (IR) spectroscopy using synchrotron radiation were performed on the original ZDDP. At room temperature no substantial structural change was observed up to 21.2 GPa, a pressure far exceeding the predicted onset of a structural transformation for the model zinc phosphate at 7 GPa. The observed Pressure induced broadening of the IR peaks is most likely associated with structural disorder or amorphization of ZDDP which is completely reversible upon decompression. When ZDDP is heated under pressure, an irreversible transformation was observed around 225 °C and 18.4 GPa. The experimental results show that ZDDP undergoes substantial decomposition at high pressures and high temperatures but no hint of cross-linkage of phosphate groups was found.     

Optimal asymptotic identification under bounded disturbances

The intrinsic limitation of worst-case identification of linear time-invariant systems using data corrupted by bounded disturbances, when the unknown plant is known to belong to a given model set, is studied. This is done by analyzing the optimal worst-case asymptotic error achievable by performing experiments using any bounded input and estimating the plant using any identification algorithm. It is shown that under some topological conditions on the model set, there is an identification algorithm which is asymptotically optimal for any input, and the optimal asymptotic error is characterized as a function of the inputs. These results, which hold for any error metric and disturbance norm, are applied to three specific identification problems: identification of stable systems in the l₁ norm, identification of stable rational systems in the H_∞ norm and identification of unstable rational systems in the gap metric. For each of these problems, the general characterization of optimal asymptotic error is used to find near-optimal inputs to minimize the error     

Amodal completion in the absence of image tangent discontinuities

The localization of the low-affinity adenosine binding protein adenotin-1 with respect to distribution in rat organs and subcellular compartments was investigated. Adenotin-1 was characterized by 5'-N-ethylcarboxamido[2,8-3H]adenosine ([3H]NECA) binding and Western blotting. Cytosolic as well as membrane fractions of all tissues contained adenotin-1. Highest levels of membrane-bound adenotin-1 were found in the liver (liver > kidney approximately spleen approximately lung > forebrain approximately cerebellum > fat heart - striated muscle), whereas highest levels of cytosolic adenotin-1 were detected in spleen, liver, lung and fat. Subcellular fractions from rat liver were prepared by differential and density gradient centrifugation. Like the homologous proteins endoplasmin or gp96, adenotin-1 is enriched in the endoplasmic reticulum. Cytosolic and membrane-bound adenotin-1 species are pharmacologically distinct, because in the liver particulate fraction adenotin-1 showed a more rapid binding kinetics, a twofold lower affinity for [3H]NECA (KD 227 nM vs. 105 nM) and a sevenfold higher affinity for 2-chloroadenosine than the cytosolic protein (Ki 1.48 microM vs. 9.25 microM). In rat liver cytosol, two different binding sites were found, which differed in [3H]NECA binding kinetics and displayed a hundredfold difference in their affinity for 2-chloro-5'-N-methylcarboxamidoadenosine (Ki 45.8 nM vs. 4.76 microM). The presence of adenotin-1 in subcellular fractions, as determined by radioligand binding, was confirmed by Western blotting. Adenotin-1 was detected as a 98-kDa band in all rat liver subcellular fractions, which agrees with the molecular mass determined for the purified protein. In the cytosol, a 65-kDa hand was labeled more intensely than the 98-kDa band. This additional band probably represents the pharmacologically distinct species of adenotin-1 found in the cytosol.     

Thermomagnetic fluctuations and hysteresis loops of magnetic cantilevers for magnetic resonance force microscopy

We have used frequency-shift cantilever magnetometry to study individual nickel magnets patterned at the end of ultra-sensitive silicon cantilevers for use in magnetic resonance force microscopy (MRFM). We present a procedure for inferring a magnet's full hysteresis curve from the response of cantilever resonance frequency versus magnetic field. Hysteresis loops and small-angle fluctuations were determined at 4.2 K with an applied magnetic field up to 6 T for magnets covering a range of dimensions and aspect ratios. Compared to magnetic materials with higher anisotropy, we find that nickel is preferable for MRFM experiments on nuclear spins at high magnetic fields.     

Gcn5p, a transcription-related histone acetyltransferase, acetylates nucleosomes and folded nucleosomal arrays in the absence of other protein subunits

Gcn5p is the catalytic subunit of several type A histone acetyltransferases (HATs). Previous studies performed under a limited range of solution conditions have found that nucleosome core particles and nucleosomal arrays can be acetylated by Gcn5p only when it is complexed with other proteins, e.g. Gcn5-Ada, HAT-A2, and SAGA. Here we demonstrate that when assayed in buffer containing optimum concentrations of either NaCl or MgCl2, purified yeast recombinant Gcn5p (rGcn5p) efficiently acetylates both nucleosome core particles and nucleosomal arrays. Furthermore, under conditions where nucleosomal arrays are extensively folded, rGcn5p acetylates folded arrays approximately 40% faster than nucleosome core particles. Finally, rGcn5p polyacetylates the N termini of free histone H3 but only monoacetylates H3 in nucleosomes and nucleosomal arrays. These results demonstrate both that rGcn5p in and of itself is catalytically active when assayed under optimal solution conditions and that this enzyme prefers folded nucleosomal arrays as a substrate. They further suggest that the structure of the histone H3 N terminus, and concomitantly the accessibility of the H3 acetylation sites, changes upon assembly into nucleosomes and nucleosomal arrays.