High Resolution Electron Beam Induced Current Measurements in an Scanning Tunneling Microscope on GaAs-MESFET

Recently, the first results of electron beam induced current (EBIC) measurements in a scanning tunneling microscope (STM) have been reported. Although the acquired results match with those obtained in conventional EBIC investigations, the interpretation of the obtained results is still restricted solely to a qualitative discussion. In this paper, a quantitative approach is used for two-dimensional numerical simulations of induced currents in GaAs-MESFET leading to a first starting point for a sophisticated interpretation of the dependence of induced currents on experimental and device parameters

Development of an Scanning Tunneling Microscopy-Based Electron Beam Induced Current (EBIC) Microscope

Measurements of electron beam induced currents (EBIC) can either be performed in a scanning electron microscope (SEM) or in a scanning tunneling microscope (STM), since both microscopes are very similar in their basic assembly. However, a straightforward application of an STM in EBIC-measurements, i.e. the use of a microscope tip as a fine source for low energetic electrons is not possible due to the specific demands on the instrument in an EBIC application. The present paper gives a compilation of these demands and describes their conversion into an optimized STM-EBIC microscope

Phosphorylation of ezrin on Thr567 is required for the synergistic activation of cell spreading by EPAC1 and protein kinase A in HEK293T cells

Recent studies have demonstrated that the actin binding protein, ezrin, and the cAMP-sensor, EPAC1, cooperate to induce cell spreading in response to elevations in intracellular cAMP. To investigate the mechanisms underlying these effects we generated a model of EPAC1-dependent cell spreading based on the stable transfection of EPAC1 into HEK293T (HEK293T–EPAC1) cells. We found that direct activation of EPAC1 with the EPAC-selective analogue, 8-pCPT-2′-O-Me-cAMP (007), promoted cell spreading in these cells. In addition, co-activation of EPAC1 and PKA, with a combination of the adenylate cyclase activator, forskolin, and the cAMP phosphodiesterase inhibitor, rolipram, was found to synergistically enhance cell spreading, in association with cortical actin bundling and mobilisation of ezrin to the plasma membrane. PKA activation was also associated with phosphorylation of ezrin on Thr567, as detected by an electrophoretic band mobility shift during SDS-PAGE. Inhibition of PKA activity blocked ezrin phosphorylation and reduced the cell spreading response to cAMP elevation to levels induced by EPAC1-activation alone. Transfection of HEK293T–EPAC1 cells with inhibitory ezrin mutants lacking the key PKA phosphorylation site, ezrin-Thr567Ala, or the ability to associate with actin, ezrin-Arg579Ala, promoted cell arborisation and blocked the ability of EPAC1 and PKA to further promote cell spreading. The PKA phospho-mimetic mutants of ezrin, ezrin-Thr567Asp had no effect on EPAC1-driven cell spreading. Our results indicate that association of ezrin with the actin cytoskeleton and phosphorylation on Thr567 are required, but not sufficient, for PKA and EPAC1 to synergistically promote cell spreading following elevations in intracellular cAMP

Leading infrared logarithms for sigma-model with fields on arbitrary Riemann manifold

We derive non-linear recursion equation for the leading infrared logarithms (LL) in four dimensional sigma-model with fields on an arbitrary Riemann manifold. The derived equation allows one to compute leading infrared logarithms to essentially unlimited loop order in terms of geometric characteristics of the Riemann manifold. We reduce the solution of the SU(oo) principal chiral field in arbitrary number of dimensions in the LL approximation to the solution of very simple recursive equation. This result paves a way to the solution of the model in arbitrary number of dimensions at N-->ooComment: Talk given by MVP at the conference devoted to memory of A.N. Vasilie

Categorizing click trains to increase taxonomic precision in echolocation click loggers

L.R. and K.J.P. were supported by Marine Scotland Science and the Marine Alliance for Science and Technology for Scotland (MASTS) pooling initiative and their support is gratefully acknowledged. MASTS is funded by the Scottish Funding Council (grant reference HR09011) and contributing institutions.Passive acoustic monitoring is an efficient way to study acoustically active animals but species identification remains a major challenge. C-PODs are popular logging devices that automatically detect odontocete echolocation clicks. However, the accompanying analysis software does not distinguish between delphinid species. Click train features logged by C-PODs were compared to frequency spectra from adjacently deployed continuous recorders. A generalized additive model was then used to categorize C-POD click trains into three groups: broadband click trains, produced by bottlenose dolphin (Tursiops truncatus) or common dolphin (Delphinus delphis), frequency-banded click trains, produced by Risso's (Grampus griseus) or white beaked dolphins (Lagenorhynchus albirostris), and unknown click trains. Incorrect categorization rates for broadband and frequency banded clicks were 0.02 (SD 0.01), but only 30% of the click trains met the categorization threshold. To increase the proportion of categorized click trains, model predictions were pooled within acoustic encounters and a likelihood ratio threshold was used to categorize encounters. This increased the proportion of the click trains meeting either the broadband or frequency banded categorization threshold to 98%. Predicted species distribution at the 30 study sites matched well to visual sighting records from the region.PostprintPeer reviewe

RNase-induced apoptosis: Fate of calcium-activated potassium channels

The connection between the action of microbial RNases and Ca2+-activated K+ (KCa) channels was investigated in human embryo kidney cells HEKhSK4 artificially expressing the channels. These channels protected HEKhSK4 cells from apoptosis induced by binase and 5K charge reversal mutant of RNase Sa. After the first 24 h, potassium current increased without increase in intracellular Ca2+, and mitochondrial potential remained high. After 72 h, the concentration of calcium increased and mitochondria lost their potential. Whole-cell recordings of membrane currents through KCa channels in RNase-treated cells demonstrated a biphasic pattern: initially their activity in cell population increased, peaked at 24 h, and then gradually decreased. In each individual cell we observed either an increase of the amplitude of KCa current, or a complete shutdown of the channels. The activity of KCa channels could be restored by removing RNases from the media. Based on this pattern and especially its timing, we hypothesize that toxic RNases downregulate KCa channels at the level of transcription or translation. Our results indicate that new anticancer agents could be created on the basis of microbial RNases targeting KCa channels. © 2008 Elsevier Masson SAS. All rights reserved

Low-frequency components in harbor porpoise (Phocoena phocoena) clicks : communication signal, by-products, or artifacts?

Author Posting. © Acoustical Society of America, 2008. This article is posted here by permission of Acoustical Society of America for personal use, not for redistribution. The definitive version was published in Journal of the Acoustical Society of America 124 (2008): 4059-4068, doi:10.1121/1.2945154.Underwater sound signals for biosonar and communication normally have different source properties to serve the purposes of generating efficient acoustic backscatter from small objects or conveying information to conspecifics. Harbor porpoises (Phocoena phocoena) are nonwhistling toothed whales that produce directional, narrowband, high-frequency (HF) echolocation clicks. This study tests the hypothesis that their 130 kHz HF clicks also contain a low-frequency (LF) component more suited for communication. Clicks from three captive porpoises were analyzed to quantify the LF and HF source properties. The LF component is 59 (S.E.M=1.45 dB) dB lower than the HF component recorded on axis, and even at extreme off-axis angles of up to 135°, the HF component is 9 dB higher than the LF component. Consequently, the active space of the HF component will always be larger than that of the LF component. It is concluded that the LF component is a by-product of the sound generator rather than a dedicated pulse produced to serve communication purposes. It is demonstrated that distortion and clipping in analog tape recorders can explain some of the prominent LF components reported in earlier studies, emphasizing the risk of erroneous classification of sound types based on recording artifacts.This work was supported by the Carlsberg Foundation and Oticon, and via a Steno Scholarship from the Danish Natural Science Research Council to PTM

Phosphodiesterase 2A2 regulates mitochondria clearance through Parkin-dependent mitophagy

Programmed degradation of mitochondria by mitophagy, an essential process to maintain mitochondrial homeostasis, is not completely understood. Here we uncover a regulatory process that controls mitophagy and involves the cAMP-degrading enzyme phosphodiesterase 2A2 (PDE2A2). We find that PDE2A2 is part of a mitochondrial signalosome at the mitochondrial inner membrane where it interacts with the mitochondrial contact site and organizing system (MICOS). As part of this compartmentalised signalling system PDE2A2 regulates PKA-mediated phosphorylation of the MICOS component MIC60, resulting in modulation of Parkin recruitment to the mitochondria and mitophagy. Inhibition of PDE2A2 is sufficient to regulate mitophagy in the absence of other triggers, highlighting the physiological relevance of PDE2A2 in this process. Pharmacological inhibition of PDE2 promotes a ‘fat-burning’ phenotype to retain thermogenic beige adipocytes, indicating that PDE2A2 may serve as a novel target with potential for developing therapies for metabolic disorders

Adenylate Cyclase Toxin Promotes Internalisation of Integrins and Raft Components and Decreases Macrophage Adhesion Capacity

Bordetella pertussis, the bacterium that causes whooping cough, secretes an adenylate cyclase toxin (ACT) that must be post-translationally palmitoylated in the bacterium cytosol to be active. The toxin targets phagocytes expressing the CD11b/CD18 integrin receptor. It delivers a catalytic adenylate cyclase domain into the target cell cytosol producing a rapid increase of intracellular cAMP concentration that suppresses bactericidal functions of the phagocyte. ACT also induces calcium fluxes into target cells. Biochemical, biophysical and cell biology approaches have been applied here to show evidence that ACT and integrin molecules, along with other raft components, are rapidly internalized by the macrophages in a toxin-induced calcium rise-dependent process. The toxin-triggered internalisation events occur through two different routes of entry, chlorpromazine-sensitive receptor-mediated endocytosis and clathrin-independent internalisation, maybe acting in parallel. ACT locates into raft-like domains, and is internalised, also in cells devoid of receptor. Altogether our results suggest that adenylate cyclase toxin, and maybe other homologous pathogenic toxins from the RTX (Repeats in Toxin) family to which ACT belongs, may be endowed with an intrinsic capacity to, directly and efficiently, insert into raft-like domains, promoting there its multiple activities. One direct consequence of the integrin removal from the cell surface of the macrophages is the hampering of their adhesion ability, a fundamental property in the immune response of the leukocytes that could be instrumental in the pathogenesis of Bordetella pertussis