Stroke Status Evoked Adhesion Molecule Genetic Alterations in Astrocytes Isolated from Stroke-Prone Spontaneously Hypertensive Rats and the Apigenin Inhibition of Their Expression

We examined the possibility that the expression of adhesion molecules is regulated differently in cultured astrocytes from stroke-prone spontaneously hypertensive rats (SHRSP/IZM) rats than in those from Wistar Kyoto rats (WKY/IZM) by tumor necrosis factor-alpha (TNF-α) or hypoxia and reoxygenation (H/R) and the inhibitory effects of apigenin. It was found that the expression of vascular cell adhesion molecule-1 (VCAM-1) by TNF-α in astrocytes isolated from SHRSP/IZM was increased compared with that in WKY/IZM. The expression of monocyte chemotactic protein-1 (MCP-1) mRNA induced by H/R in SHRSP/IZM astrocytes was increased compared with that in normal oxygen concentrations. Apigenin strongly attenuated TNF-α-induced VCAM-1 mRNA and protein expression and suppressed the adhesion of U937 cells and SHRSP/IZM astrocytes. These results suggest that the expression levels of adhesion molecules during H/R affect disease outcome and can drive SHRSP/IZM to stroke. It is suggested that apigenin regulates adhesion molecule expression in reactive astrocytes during ischemia

Cytochromec−Crown Ether Complexes as Supramolecular Catalysts: Cold-Active Synzymes for Asymmetric Sulfoxide Oxidation in Methanol

A series of supramolecular complexes of various cytochrome c proteins with 18-crown-6 derivatives behave as cold-active synzymes in the H_2O_2 oxidation of racemic sulfoxides. This interesting behavior contrasts with native functionality, where the employed proteins act as electron transfer carriers. ESI-MS, UV, CD, and Raman spectroscopic characterizations reveal that four or five 18-crown-6 molecules strongly bind to the surface of the cytochrome c and also that nonnatural low-spin hexacoordinate heme structures are induced in methanol. Significantly, crown ether complexation can convert catalytically inactive biological forms to catalytically active artificial forms. Horse heart, pigeon breast, and yeast cytochromes c all stereoselectively oxidize (S)-isomers of methyl tolyl sulfoxide and related sulfoxides upon crown ether complexation. These supramolecular catalysts show the highest efficiency and enantiomer selectivity at −40 °C in the H_2O_2-dependent sulfoxide oxidation, while oxidative decomposition of the heme moieties predominantly occurs at room temperature. The oxidation reactivity of the employed sulfoxides is apparently related to steric constraints and electrochemical oxidation potentials of their S O bonds. Among the cytochrome c complexes, yeast cytochrome c demonstrates the lowest catalytic activity and degradation reactivity. It has a significantly different protein sequence, suggesting that crown ether complexation effectively activates heme coordination but may additionally alter the native backbone structure. The proper combination of cytochrome c proteins, 18-crown-6 receptors, and external circumstances can be used to successfully generate “protein-based supramolecular catalysts” exhibiting nonbiological reactivities

Juvenile hormone acid O-methyltransferase in Drosophila melanogaster

Juvenile hormone (JH) acid O-methyltransferase (JHAMT) is the enzyme that transfers a methyl groupfrom S-adenosyl-L-methionine (SAM) to the carboxyl group of JH acids to produce active JHs in thecorpora allata. While the JHAMT gene was originally identified and characterized in the silkwormBombyx mori, no orthologs from other insects have been studied until now. Here we report on thefunctional characterization of the CG17330/DmJHAMT gene in the fruit fly Drosophila melanogaster.Recombinant DmJHAMT protein expressed in Escherichia coli catalyzes the conversion of farnesoic acidand JH III acid to their cognate methyl esters in the presence of SAM. DmJHAMT is predominantlyexpressed in corpora allata, and its developmental expression profile correlates with changes in the JHtiter. While a transgenic RNA interference against DmJHAMT has no visible effect, overexpression ofDmJHAMT results in a pharate adult lethal phenotype, similar to that obtained with application of JHanalogs, suggesting that the temporal regulation of DmJHAMT is critical for Drosophila development

Molecular Epidemiology of Trichophyton tonsurans Strains Isolated in Japan between 2006 and 2010 and Their Susceptibility to Oral Antimycotics

SUMMARY: Trichophyton tonsurans has been isolated among judo practitioners, wrestlers, and sumo wrestlers during an epidemic of tinea corporis and tinea capitis in Japan. A previous study using restriction fragment length polymorphism (RFLP) analysis of the non-transcribed spacer (NTS) region of the ribosomal RNA gene revealed different sources for the causative fungus in epidemics among judo practitioners and among wrestlers. Many different fungal strains have since been isolated from practitioners of these sports. The present study evaluated fungal characteristics of strains newly isolated between July 2006 and December 2010 using this molecular method. PCR-RFLP analysis using MvaI and AvaI was performed on 263 strains, composed of 186 isolates from judo practitioners, 32 from wrestlers, 30 from sumo wrestlers, 5 from other sports, 7 from family members or friends of the sports practitioner patients, and 3 from sporadic (non-epidemic) cases. Four molecular types, NTS I, II, III, and VII were detected. Of these, NTS I was the most predominant, occurring in 243 of 263 strains (92.4z). All of the 30 strains isolated from sumo wrestlers were classified as NTS I, suggesting that the epidemic among sumo wrestlers originated from an earlier epidemic among judo practitioners. Thirteen strains were classified as NTS II; all were related to wrestling and were isolated mainly from the Chubu and Kansai areas in the central part of Honshu island. NTS III was detected in 6 strains, and one strain classified as NTS VII was isolated from a sporadic case of tinea capitis in a Peruvian immigrant. The minimum inhibitory concentrations (MICs) of terbinafine, itraconazole, fluconazole, and griseofulvin on 10 strains of NTS I and NTS II and 4 strains of NTS III were examined; there were no differences in MIC between these molecular types

Amphotericin B assembles into seven-molecule ion channels: An NMR and molecular dynamics study

Amphotericin B, an antifungal drug with a long history of use, forms fungicidal ion-permeable channels across cell membranes. Using solid-state nuclear magnetic resonance spectroscopy and molecular dynamics simulations, we experimentally elucidated the three-dimensional structure of the molecular assemblies formed by this drug in membranes in the presence of the fungal sterol ergosterol. A stable assembly consisting of seven drug molecules was observed to form an ion conductive channel. The structure is somewhat similar to the upper half of the barrel-stave model proposed in the 1970s but substantially different in the number of molecules and in their arrangement. The present structure explains many previous findings, including structure-activity relationships of the drug, which will be useful for improving drug efficacy and reducing adverse effects