On the mechanism of thrombin-induced angiogenesis: involvement of alphavbeta3-integrin

Thrombin has been reported to be a potent angiogenic factor both in vitro and in vivo, and many of the cellular effects of thrombin may contribute to activation of angiogenesis. In this report we show that thrombin-treatment of human endothelial cells increases mRNA and protein levels of alphavbeta3-integrin. This thrombin-mediated effect is specific, dose dependent, and requires the catalytic site of thrombin. In addition, thrombin interacts with alphavbeta3 as demonstrated by direct binding of alphavbeta3 protein to immobilized thrombin. This interaction of thrombin with alphavbeta3-integrin, which is an angiogenic marker in vascular tissue, is of functional significance. Immobilized thrombin promotes endothelial cells attachment, migration, and survival. Antibody to alphavbeta3 or a specific peptide antagonist to alphavbeta3 can abolish all these alphavbeta3-mediated effects. Furthermore, in the chick chorioallantoic membrane system, the antagonist peptide to alphavbeta3 diminishes both basal and the thrombin-induced angiogenesis. These results support the pivotal role of thrombin in activation of endothelial cells and angiogenesis and may be related to the clinical observation of neovascularization within thrombi.     

Ultra high-resolution fMRI in monkeys with implanted RF coils

Spatiotemporally resolved functional MRI (fMRI) in animals can reveal how wide-spread neural networks are organized and accompanying electrophysiological recordings can show how small neural assemblies contribute to this organization. Here we present a novel technique that yields high-resolution structural and functional images of the monkey brain with small, tissue-compatible, intraosteally implantable radiofrequency coils. Voxel sizes as small as 0.0113 microl with high signal-to-noise and contrast-to-noise ratios were obtained, revealing both structural and functional cortical architecture in great detail. Up to a certain point, contrast sensitivity increased with decreasing voxel size, probably because of the decreased partial volume effects. Spatial specificity was demonstrated by the lamina-specific activation in experiments comparing responses to moving and flickering stimuli. The implications of this technique for combined fMRI/electrophysiology experiments and its limitations in terms of spatial coverage are discussed.     

Visual competition

Binocular rivalry--the alternations in perception that occur when different images are presented to the two eyes--has been the subject of intensive investigation for more than 160 years. The psychophysical properties of binocular rivalry have been well described, but newer imaging and electrophysiological techniques have not resolved the issue of where in the brain rivalry occurs. The most recent evidence supports a view of rivalry as a series of processes, each of which is implemented by neural mechanisms at different levels of the visual hierarchy. Although unanswered questions remain, this view of rivalry might allow us to resolve some of the controversies and apparent contradictions that have emerged from its study.     

The 1.76 A resolution crystal structure of glycogen phosphorylase B complexed with glucose, and CP320626, a potential antidiabetic drug

CP320626, a potential antidiabetic drug, inhibits glycogen phosphorylase in synergism with glucose. To elucidate the structural basis of synergistic inhibition, we determined the structure of muscle glycogen phosphorylase b (MGPb) complexed with both glucose and CP320626 at 1.76 A resolution, and refined to a crystallographic R value of 0.211 (R(free)=0.235). CP320626 binds at a novel allosteric site, which is some 33 A from the catalytic site, where glucose binds. The high resolution structure allows unambiguous definition of the conformation of the 1-acetyl-4-hydroxy-piperidine ring supported by theoretical energy calculations. Both CP320626 and glucose promote the less active T-state, thereby explaining their synergistic inhibition. Structural comparison of MGPb--glucose--CP320626 complex with liver glycogen phosphorylase a (LGPa) complexed with a related compound (CP403700) show that the ligand binding site is conserved in LGPa.     

Field parasitism of Ceratitis capitata larvae by Aganaspis daci in Chios, Greece

High parasitism rates were recorded in the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in pupae derived from field infested figs, on the Greek island of Chios in 1999 and 2000. Adult parasitoids were identified as Aganaspis daci (Weld) (Hymenoptera:Eucoilidae), previously known as Trybliographa daci (Weld). Approximately 45%of C. capitata pupae yielded adultparasitoid in both years and the totalmortality of pupae due to the parasitoid was62–65%. Development of male A. daci at25°C, reared on 3rd instar larvaeof C. capitata, was shorter than that of the female (34 and 37 days respectively). Average adultmale longevity was 4–5 days longer than female(16–17 to 11–12 days, respectively) and almostidentical in wild and F₁ parasitoids ofboth sexes. We suggest that A. daci maybe an efficient form of biological control ofC. capitata in the Mediterranean regionand probably in other areas.     

Transfection-independent production of alphavirus replicon particles based on poxvirus expression vectors

This report describes a transfection-independent system for packaging alphavirus replicon vectors using modified vaccinia virus Ankara (MVA) vectors to express all of the RNA components necessary for the production of Venezuelan equine encephalitis (VEE) virus replicon particles (VRP). Infection of mammalian cells with these recombinant MVA vectors resulted in robust expression of VEE structural genes, replication of the alphavirus vector and high titers of VRP. In addition, VRP packaging was achieved in a cell type (fetal rhesus lung) that has been approved for the manufacturing of vaccines destined for human use.     

Beyond 100 genomes

By the end of 2002, we witnessed the landmark submission of the 100th complete genome sequence in the databases. An overview of these genomes reveals certain interesting trends and provides valuable insights into possible future developments.     

β-Adrenergic Receptors Mediate a Stress-Induced Decrease in IGF-II mRNA in the Rat Cerebellum

1. Exposure to a combined forced swimming–confinement stress resulted in a decrease in insulin-like growth factor II (IGF-II) mRNA levels in the whole brain (without the cerebellum) and in the isolated brain areas of the cerebral cortex, the hippocampus, and the cerebellum.2. In an effort to elucidate the neurotransmitter systems involved in this stress-induced decrease, animals were injected prior to exposure to the stress, with either propranolol, diazepam, or MK-801.3. Administration of diazepam or MK-801 did not affect the stress-induced decrease in IGF-II mRNA in any of the three brain areas examined.4. Administration of propranolol prior to the exposure to the stress inhibited the stress-induced decrease in IGF-II mRNA in the cerebellum. Propranolol had no such effect in the cerebral cortex or the hippocampus.5. Our results suggest that in the cerebellum, the stress-induced decrease in IGF-II mRNA is mediated by ₂-adrenergic receptors.