2-Methylsorbic acid,an antifungal metabolite of Penicillium vermiculatum

Summary 2-Methylsorbic acid (MA), (2Z,4E)-2-methyl-2,4-hexadienoic acid, is a new metabolite of Penicillium vermiculatum. Antifungal activity of this acid was higher than that of sorbic acid or the bromoderivatives bromomethylsorbic acid and bromosorbic acid. MA suppressed the growth of Talaromyces flavus and germination of its conidia. In P. vermiculatum this acid lowered production of vermiculin and inhibited proteosynthesis in Saccharomyces cerevisiae. Correspondence to: B. Proksa     

Inhibitory effect of stobadine on FMLP-induced chemiluminescence in human whole blood and isolated polymorphonuclear leukocytes.

The chemiluminescence (CL) technique with luminol and isoluminol was used to characterize the effect of stobadine on reactive oxygen metabolites (ROM) generation in human whole blood and in isolated polymorphonuclear leukocytes (PMNL) stimulated with N-formyl-methionyl-leucyl-phenyl-alanine (FMLP). In whole blood and in isolated PMNL, stobadine in the concentrations of 1, 10 and 100 micromol/L significantly inhibited the CL signal after FMLP, which activated predominantly extracellular generation of ROM. The same concentrations of stobadine were effective on CL in a cell-free system. On the other hand, myeloperoxidase (MPO) liberation was decreased by stobadine only in the concentration of 100 micromol/L. The results showed stobadine to act as a potent inhibitor/scavenger of extracellularly produced ROM in human PMNL and indicated interference of stobadine with ROM as well as with signalling events resulting in NADPH-oxidase activation and MPO liberation.     

The α-amylase inhibitor acarbose does not affect the parasitoid Venturia canescens when incorporated into the diet of its host Ephestia kuehniella

Amylase inhibitors (AIs) are suitable candidates for protecting plants and their products from attacks by herbivorous and granivorous insects. However, detailed studies of the suppressive effects of AIs on target and non‐target insects are necessary before their application in post‐harvest protection. To address this issue, laboratory bioassays were used to test the effect of the non‐proteinaceous inhibitor acarbose on a stored product pest, the flour moth Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae), and its parasitoid Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae). Two sublethal concentrations (0.001 and 0.0001%, wt/wt) of acarbose were incorporated into the diet of parasitized and unparasitized larvae of E. kuehniella. Development time and fresh body weight of the larvae, together with the size of the wasps, were compared for insects reared on acarbose‐treated and control diets. On the diet containing 0.001% acarbose, the developmental time was longer and relative weight gains of the E. kuehniella larvae were lower, but the weight of the larvae prior to pupation was similar to that of the control. The acarbose did not have a suppressive effect on the parasitoid V. canescens; in fact the wasps that emerged from the hosts reared on a diet containing 0.0001% acarbose were on average larger and heavier than the controls. These results demonstrate that it might be possible to enhance the control of stored product pests by using both biological control and AIs.     

Efficient synthesis of ruthenium complexes of the type (R-bpy)2RuCl2 and [(R-bpy)2Ru(L-L)]Cl2 by microwave-activated reactions (R: H, Me, tert-But) (L-L: substituted bibenzimidazoles, bipyrimidine, and phenanthroline)

Complexes of the type (R-bpy)₂RuCl₂ (R: H, Me, tert-but) were synthesised by microwave-activated reactions of [Ru(cod)Cl₂]_n with substituted 2,2′-bipyridines in dimethylformamide as the solvent. The complexes were isolated in high yields and high purity from the reaction mixture. Microwave-assisted or thermal reaction of the (R-bpy)₂RuCl₂ solutions with substituted bibenzimidazoles, 1,10 phenanthroline or bipyrimidine in dmf/water mixtures resulted in the formation of mixed ligand complexes of the type [(R-bpy)₂Ru(L-L)]Cl₂. The complexes were characterised by NMR spectroscopy and MS. Furthermore, their photochemical and electrochemical properties were investigated and the solid state structure of (4-tert-butyl-bpy)₂RuCl₂ (3), [(4-tert-butyl-bpy)₂Ru(tetramethylbibenzimidazole)](PF₆)₂ (4), and [(4-tert-butyl-bpy)₂Ru(bipyrimidine] (PF₆)₂ (5) was determined by X-ray diffraction analysis of single crystals.     

Calcium current in rat cardiomyocytes is modulated by the carboxyl-terminal ahnak domain

Ahnak, a protein of 5643 amino acids, interacts with the regulatory beta-subunit of cardiac calcium channels and with F-actin. Recently, we defined the binding sites among the protein partners in the carboxyl-terminal domain of ahnak. Here we further narrowed down the beta(2)-interaction sites to the carboxyl-terminal 188 amino acids of ahnak by the recombinant ahnak protein fragments P3 (amino acids 5456-5556) and P4 (amino acids 5556-5643). The effects of these P3 and P4 fragments on the calcium current were investigated under whole-cell patch clamp conditions on rat ventricular cardiomyocytes. P4 but not P3 increased significantly the current amplitude by 22.7 +/- 5% without affecting its voltage dependence. The slow component of calcium current inactivation was slowed down by both P3 and P4, whereas only P3 slowed significantly the fast one. The composite recombinant protein fragment P3-P4 induced similar modifications to the ones induced by each of the ahnak fragments. In the presence of carboxyl-terminal ahnak protein fragments, isoprenaline induced a similar relative increase in current amplitude and shift in current kinetics. The actin-stabilizing agents, phalloidin and jasplakinolide, did not modify the effects of these ahnak protein fragments on calcium current in control conditions nor in the presence of isoprenaline. Hence, our results suggest that the functional effects of P3, P4, and P3-P4 on calcium current are mediated by targeting the ahnak-beta(2)-subunit interaction rather than by targeting the ahnak-F-actin interaction. More specifically they suggest that binding of the beta(2)-subunit to the endogenous subsarcolemmal giant ahnak protein re-primes the alpha(1C)/beta(2)-subunit interaction and that the ahnak-derived proteins relieve the beta(2)-subunit from this inhibition.     

HCMV-encoded chemokine receptor US28 employs multiple routes for internalization

The human cytomegalovirus-encoded G protein-coupled receptor homologue US28 binds inflammatory chemokines and sequesters them from the environment of infected cells. Low surface deposition and endocytosis are dependent on constitutive C-terminal phosphorylation, suggesting a requirement for beta-arrestin binding in receptor internalization. In this report, a US28-dependent redistribution of beta-arrestin into vesicular structures occurred, although internalization of US28 was independent of beta-arrestin. Internalization of US28 was dynamin-dependent, and US28 partially partitioned into the detergent-resistant membrane fraction. Endocytosis was diminished by cholesterol depletion, yet sucrose inhibition was even stronger. The relevance of the clathrin-coated pit pathway was supported by colocalization of beta(2)-adaptin and US28 in endocytic compartments. Exchange of the C-terminal dileucine endocytosis motif inhibited rapid endocytosis, indicating a direct interaction of US28 with the AP-2 adaptor complex. We suggest that the arrestin-independent, dynamin-dependent internalization of US28 reveals a differential sorting of beta-arrestins and the virally encoded chemokine receptor homologue.     

The role of diversification in causing the correlates of dioecy

Dioecy is reported to be correlated with a number of ecological traits, including tropical distribution, woody growth form, plain flowers, and fleshy fruits. Previous analyses have concentrated on determining whether dioecy is more likely to evolve in lineages possessing these traits, rather than considering the speciation and extinction rates of dioecious lineages with certain combinations of traits. To address the association between species richness in dioecious lineages as a function of the ecological traits, we compared the evolutionary success (i.e., relative species richness) of dioecious focal lineages with that of their nondioecious sister groups. This test was repeated for the evolutionary success of randomly chosen nondioecious lineages (control lineages) compared with their nondioecious sister groups. If the possession of certain ecological traits enhances the evolutionary success of dioecious lineages, we predict an association between the presence of these traits and relative species richness in the former, but not latter, set of sister-group comparisons. Dioecious focal lineages with a higher number of these traits experienced higher evolutionary success in sister-group comparisons, whereas no trend was found for the control focal lineages. The increase in evolutionary success was especially true for dioecious focal lineages that had a tropical distribution or fleshy fruit. We discuss how these results provide strong support for differential evolutionary success theories for the correlations between dioecy and the ecological traits considered.     

Actin-binding proteins required for reliable chromosome segregation in mitosis

While studying mitosis in Dictyostelium mutants with deficiencies in actin-binding proteins, we found that two of these proteins, cortexillin and Aip1, are required for the precise segregation of chromosomes. Atypical spindles and nuclei with varying DNA content indicate that mutants lacking cortexillin or Aip1 are genetically unstable. These aberrations are caused by the detachment and irregular reattachment of centrosomes to the nuclear surface. Live imaging showed how coalescing mitotic complexes give rise to a multipolar spindle, and how excess centrosomes can be eliminated by mitotic cleavage between anucleate and nucleated portions of a cell. We hypothesize that mutations in regulatory proteins of the actin network might be one cause of genetic instability of malignant tumor cells.     

Pn-AMP1, a plant defense protein, induces actin depolarization in yeasts

Pn-AMP1, Pharbitis nil antimicrobial peptide 1, is a small cysteine-rich peptide implicated in host-plant defense. We show here that Pn-AMP1 causes depolarization of the actin cytoskeleton in Saccharomyces cerevisiae and Candida albicans. Pn-AMP1 induces rapid depolarization of actin cables and patches within 15 min. Increased osmolarity or temperature induces transient actin depolarization and results in increased sensitivity to Pn-AMP1, while cells conditioned to these stresses show less sensitivity. Mutations in components of a cell wall integrity pathway (Wsc1p, Rom2p, Bck1p and Mpk1p), which regulate actin repolarization, result in increased sensitivity to Pn-AMP1. A genetic screen reveals that mutations in components of the alpha-1,6-mannosyltransferase complex (Mnn10p, Mnn11p and Och1p), which regulate mannosylation of cell wall proteins, confer resistance to Pn-AMP1. FITC-conjugated Pn-AMP1 localizes to the outer surface of the cell with no significant staining observed in spheroplasts. Taken together, these results indicate that cell wall proteins are determinants of resistance to Pn-AMP1, and the ability of a plant defense protein to induce actin depolarization is important for its antifungal activity.