Toll-like receptor 4 deficiency: Smaller infarcts,but nogain in function

Backgound  

It has been reported that Toll-like receptor 4 (TLR4) deficiency reduces infarct size after myocardial ischemia/reperfusion (MI/R). However, measurement of MI/R injury was limited and did not include cardiac function. In a chronic closed-chest model we assessed whether cardiac function is preserved in TLR4-deficient mice (C3H/HeJ) following MI/R, and whether myocardial and systemic cytokine expression differed compared to wild type (WT).     

Second-harmonic generation from organic molecules incorporated in sol-gel matrices

Orientation of optically nonlinear organic molecules inside sol-gel matrices upon application of an external D.C. electrical field is demonstrated for the first time. The quadratic nonlinear response of silicon oxide or transition metal oxide based gels containing organic molecules has been determined from Electric Field Induced Second Harmonic (EFISH) measurements. Large concentrations of Optically Nonlinear Organic Molecules (ONOM) have been either incorporated inside the macromolecular network or chemically bonded to the oxide backbone of the gels. These results demonstrate the feasibility of permanently poled doped sol-gel matrices. Moreover, EFISH measurements performed on organic molecules appear to be a useful tool for monitoring the changes occurring during sol-gel transformations.     

Active site mutations in DNA topoisomerase I distinguish the cytotoxic activities of camptothecin and the indolocarbazole, rebeccamycin.

DNA topoisomerase I (Top1p) catalyzes topological changes in DNA and is the cellular target of the antitumor agent camptothecin (CPT). Non-CPT drugs that target Top1p, such as indolocarbazoles, are under clinical development. However, whether the cytotoxicity of indolocarbazoles derives from Top1p poisoning remains unclear. To further investigate indolocarbazole mechanism, rebeccamycin R-3 activity was examined in vitro and in yeast. Using a series of Top1p mutants, where substitution of residues around the active site tyrosine has well-defined effects on enzyme catalysis, we show that catalytically active, CPT-resistant enzymes remain sensitive to R-3. This indolocarbazole did not inhibit yeast Top1p activity, yet was effective in stabilizing Top1p-DNA complexes. Similar results were obtained with human Top1p, when Ser or His were substituted for Asn-722. The mutations altered enzyme function and sensitivity to CPT, yet R-3 poisoning of Top1p was unaffected. Moreover, top1delta, rad52delta yeast cells expressing human Top1p, but not catalytically inactive Top1Y723Fp, were sensitive to R-3. These data support hTop1p as the cellular target of R-3 and indicate that distinct drug-enzyme interactions at the active site are required for efficient poisoning by R-3 or CPT. Furthermore, resistance to one poison may potentiate cell sensitivity to structurally distinct compounds that also target Top1p.     

The cortical actomyosin system of cytochalasin D-treated lymphoblasts

Global cytoskeleton dynamics is likely to exist in animal cells and some experimental evidence for this has recently been obtained in cells from the human lymphoblastic cell line KE37. We have further investigated the dramatic and reversible microtubule-dependent cell elongation which occurs upon treatment of KE37 cells with cytochalasin D. This phenomenon results in a non-locomotory cell with definite polarity. It involves a sustained equatorial myosin II-dependent contraction of cortical, most of the myosin II being accumulated on segments of the main cellular extension. We report here that such a cell lengthening is energy-dependent and can be inhibited, or suppressed, by surface ligands such as wheat germ agglutinin but not by concanavalin A. Suppression of the cytochalasin D effect by wheat germ agglutinin is rapid and appears to be collapse of the cell extension and relocalization of the contracted actomyosin as a whole. It suggests that the binding of the wheat germ agglutinin to the cell surface results in the transient disassembly of microtubules, a possibility also raised by the potent antagonist effect of taxol on wheat germ agglutinin action. Taken together, the data are consistent with a specific role of microtubules in the control of the activity of the cortical actomyosin system.     

Extensive variation in synonymous substitution rates in mitochondrial genes of seed plants

Background  

It has long been known that rates of synonymous substitutions are unusually low in mitochondrial genes of flowering and other land plants. Although two dramatic exceptions to this pattern have recently been reported, it is unclear how often major increases in substitution rates occur during plant mitochondrial evolution and what the overall magnitude of substitution rate variation is across plants.     

Embryonic Ammonoid Shell Features: Intraspecific Variation Revisited

Two samples of ammonoids belonging to the Oppeliidae, Sublunuloceras virguloidesHecticoceras (Brightii) canaliculatum, are analyzed to estimate the intraspecific variability of embryonic shell features. The study of embryonic shell characters reveals two main shapes of protoconch, flattened and round. Prosiphons may be straight or slightly curved. New parameters for area are added to the linear parameters commonly found in the literature. Prosiphon length and caecum area vary greatly whereas protoconch and ammonitella diameter vary only slightly, and the ammonitella angle is almost constant. The protoconch-to-ammonitella size ratio behaves differently in each species, suggesting different patterns of embryonic growth. We compare our results with published data and discuss their significance for species determination and ontogenetic interpretation. The main finding is that intraspecific embryonic variation is greater than was previously believed.     

Behavior ofPseudomonas fluorescens within the hydrodynamic boundary layers of surface microenvironments

Phase, darkfield, and computer-enhanced microscopy were used to observe the surface microenvironment of flow cells during bacterial colonization. Microbial behavior was consistent with the assumptions used previously to derive surface colonization kinetics and to calculate surface growth and attachment rates from cell number and distribution. Surface microcolonies consisted of closely packed cells. Each colony contained 2ⁿ cells, where n is the number of cell divisions following attachment. Initially, cells were freely motile while attached, performing circular looping movements within the plane of the solid-liquid interface. Subsequently, cells attached apically, maintained a fixed position on the surface, and rotated. This type of attachment was reversible and did not necessarily lead to the formation of microcolonies. Cells became irreversibly attached by progressing from apical to longitudinal attachment. Longitudinally attached cells increased in length, then divided, separated, moved apart laterally, and slid next to one another. This resulted in tight cell packing and permitted simultaneous growth and adherence. After approximately 4 generations, individual cells emigrated from developing microcolonies to recolonize the surface at new locations. Surface colonization byPseudomonas fluorescens can thus be subdivided into the following sequential colonization phases: motile attachment phase, reversible attachment phase, irreversible attachment phase, growth phase, and recolonization phase.