Spatial patterns of Ca2+ signals define intracellular distribution of a signaling by Ca2+/Calmodulin-dependent protein kinase II

Ca2+ plays a central role in cell signaling, and Ca2+/calmodulin-dependent protein kinase II (CaMKII) is a major mediator of Ca2+ actions. The spatial distribution of intracellular Ca2+ signaling is not homogenous, rather it is dynamically organized, and it has been speculated that spatial patterns of Ca2+ signals may function as a form of cellular information transmitted to downstream molecules. To address this issue, we studied the intracellular distributions of the signalings by CaMKII and Ca2+ in the same astrocytes. The former was visualized by monitoring site-specific phosphorylation of a cytoskeletal protein vimentin, using site- and phosphorylation-specific antibodies, while the latter was examined by fura-2-based Ca2+ microscopy. Local Ca2+ signals induced vimentin phosphorylation by CaMKII localized in the same area. On the other hand, Ca2+ waves in astrocytes induced global phosphorylation of vimentin by CaMKII. A small population of vimentin filaments highly phosphorylated by CaMKII underwent structural alteration into short filaments at electron microscopic level. These results indicate that CaMKII transmits spatial patterns of Ca2+ signals to vimentin as cellular information. The possibility is discussed that spatial patterns of vimentin phosphorylation may be important for intracellular organization of vimentin filament networks.     

Pervaporation separation of water/ethanol mixtures through polysaccharide membranes. III. The permselectivity of the neutralized chitosan membrane and the relationships between its permselectivity and solid state structure

The separation of water/alcohol mixtures through chitosan membrane neutralized by various acids was investigated. The permselectivity of the neutralized chitosan membrane was affected by the species of acids and, only when neutralized by polybasic acids, was a marked increase of the selectivity observed and a high selectivity maintained for ion free mixtures. The marked increase of the selectivity could be explained by the contraction of the “holes” produced by the thermal motion of polymer chains due to the conformation change of chitosan molecule caused by the neutralization. The conformation change was confirmed by the analysis of the solid state structure of the neutralized chitosan using CP/MAS ¹³C-NMR.     

Total Syntheses and Chemical Biology Studies of Hymeglusin and Fusarilactone A,Novel Circumventors of β-Lactam Drug Resistance in Methicillin-Resistant Staphylococcus aureus

Hymeglusin, a previously known eukaryotic hydroxymethylglutaryl-CoA (HMG−CoA) synthase inhibitor, was identified as circumventing the β-lactam drug resistance in methicillin-resistant Staphylococcus aureus (MRSA). We describe the concise total syntheses of a series of natural products, which enabled determination of the absolute configuration of fusarilactone A and provided structure-activity relationship information. Based on previous reports, we speculated that the target protein of this circumventing effect may be MRSA bacterial HMG−CoA synthase (mvaS). We found that this enzyme was dose-dependently inhibited by hymeglusin. Furthermore, overexpression of the MRSA mvaS gene and site-directed mutagenesis studies suggested its binding site and the mechanism of action.     

Characterization of ω-regular languages by first-order formulas

First-order formulas are used to specify various ways of acceptance of ω-languages by (deterministic) finite automata, and we study the relationship between the ‘arithmetic’ hierarchy of the formulas in prenex normal form and the topological hierarchy of the accepted ω-languages. Among other things it is proved that the ω-languages accepted by finite automata under the accepting conditions specified by Σ₁-type formulas are precisely open ω-regular languages, those accepted under the conditions specified by Σ₂-type formulas coincide with the ω-regular languages which are denumerable unions of closed sets, and that as long as the accepting conditions are specified by first-order formulas the accepted ω-languages remain to be ω-regular.     

KRAS and MAPK1 Gene Amplification in Type II Ovarian Carcinomas

In this study, we examined the clinical significance of KRAS and MAPK1 amplification and assessed whether these amplified genes were potential therapeutic targets in type II ovarian carcinoma. Using fluorescence in situ hybridization, immunohistochemistry, and retrospectively collected clinical data, KRAS and MAPK1 amplifications were identified in 9 (13.2%) and 5 (7.4%) of 68 type II ovarian carcinoma tissue samples, respectively. Interestingly, co-amplification of KRAS and MAPK1 seemed to be absent in the type II ovarian carcinomas tested, except one case. Active phospho-ERK1/2 was identified in 26 (38.2%) out of 68 type II ovarian carcinomas and did not correlate with KRAS or MAPK1 amplification. There was no significant relationship between KRAS amplification and overall or progression-free survival in patients with type II ovarian carcinoma. However, patients with MAPK1 amplification had significantly poorer progression-free survival than patients without MAPK1 amplification. Moreover, type II ovarian carcinoma cells with concomitant KRAS amplification and mutation exhibited dramatic growth reduction following treatment with the MEK inhibitor PD0325901. These findings indicate that KRAS/MAPK1 amplification is critical for the growth of a subset of type II ovarian carcinomas. Additionally, RAS/RAF/MEK/ERK pathway-targeted therapy may benefit selected patients with type II ovarian carcinoma harboring KRAS/MAPK1 amplifications.     

Visual perception of texture of textiles

The mechanism of visual texture perception was investigated by measuring visual evaluation values. Textile samples of different surface structures were examined, along with corresponding image samples rendered on paper. The relation between physical properties of textiles and light reflected from the textiles, and the relation between the reflected light stimulating the eyes of the subject and the psychological perception of texture are discussed. © 2001 John Wiley & Sons, Inc. Col Res Appl, 26, 469–477, 2001