Introduction of Macromolecules into Bovine Adrenal Medullary Chromaffin Cells and Rat Pheochromocytoma Cells (PC12) by Permeabilization with Streptolysin O: Inhibitory Effect of Tetanus Toxin on Catecholamine Secretion

Conditions are described for controlled plasma membrane permeabilization of rat pheochromocytoma cells (PC12) and cultured bovine adrenal chromaffin cells by Streptolysin O (SLO). The transmembrane pores created by SLO invoke rapid efflux of intracellular 86Rb+ and ATP, and also permit passive diffusion of proteins, including immunoglobulins, into the cells. SLO-permeabilized PC12 cells release [3H]dopamine in response to micromolar concentrations of free Ca2+. Permeabilized adrenal chromaffin cells present a similar exocytotic response to Ca2+ in the presence of Mg2+/ ATP. Permeabilized PC12 cells accumulate antibodies against synaptophysin and calmodulin, but neither antibody reduces the Ca2+-dependent secretory response. Reduced tetanus toxin, although ineffective when applied to intact chromaffin cells, inhibits Ca2+-induced exocytosis by both types of permeabilized cells studied. Omission of dithiothreitol, toxin inactivation by boiling, or preincubation with neutralizing antibodies abolishes the inhibitory effect. The data indicate that plasma membrane permeabilization by Streptolysin O is a useful tool to probe and define cellular components that are involved in the final steps of exocytosis

Applied neutron tomography in modern archaeology

The use of neutron tomography for archaeometric purposes is quite a new technique. The property of neutron to transmit easily large, dense samples is of great importance in modern archaeology. The three-dimensional visualisation of the inner structure of samples of archaeological interest helps to make suggestions about the technological process of manufacturing or reveals information about the origins of delivering of noble materials used in ancient masterpieces. Another application field in modern archaeology is the non-destructive inspection of the quality of specimen conservation where the neutron tomography allows visualization of impregnation solutions in wood or metal matrices. The high sensibility of neutrons to hydrogen makes it possible to detect organic remains in fossils. All of these advantages make the neutron tomography a unique technique for non-destructive investigation in archaeological sciences

Further Characterization of Dopamine Release by Permeabilized PC 12 Cells

Rat pheochromocytoma cells (PC 12) permeabilized with staphylococcal α-toxin release [3H]dopamine after addition of micromolar Ca2+. This does not require additional Mg2+-ATP (in contrast to bovine adrenal medullary chromaffin cells). We also observed Ca2+-dependent [3H]-dopamine release from digitonin-permeabilized PC 12 cells. Permeabilization with α-toxin or digitonin and stimulation of the cells were done consecutively to wash out endogenous Mg2+-ATP. During permeabilization, ATP was removed effectively from the cytoplasm by both agents but the cells released [3H]dopamine in response to micromolar Ca2+ alone. Replacement by chloride of glutamate, which could sustain mitochondrial ATP production in permeabilized cells, does not significantly alter catecholamine release induced by Ca2+. However, Mg2+ without ATP augments the Ca2+-induced release. The release was unaltered by thiol-, hydroxyl-, or calmodulin-interfering substances. Thus Mg2+-ATP, calmodulin, or proteins containing -SH or -OH groups are not necessary for exocytosis in permeabilized PC 12 cells

Reduzierung von Missing Wedge Artefakten mit DIRECTT

Wir stellen eine Prozedur vor, mit der typische Missing Wedge MW Artefakte in CT Rekonstruktionen effektiv unterdrückt werden können. Dazu wird der iterative DIRECTT Algorithmus Direkte Iterative Rekonstruktion Computertomographischer Trajektorien mit einigen Zyklen diskreter Rekonstruk tion eingesetzt. Die Kernfunktionen des Algorithmus , die wiederholte Auswahl und Gewichtung von Elementen einer Zwischenrekonstruktion, werden beibehalten. Abgesehen von Sonderfällen der Röntgen und Neutronentomographie tritt die MW Restriktion neben einer Vielzahl anderer in Standardkonfigurationen der Elektronentomographie auf. Um den MW Effekt isoliert zu untersuchen und beurteilen zu können, werden hier vollständige experimentelle Datensätze einer bekannten Porenstruktur um einen Sektor beschnitten sowie MW Rekonstruktionen von Porenmodellen erzeugt. Die Ergebnisse werden mit zurzeit verbreiteten Algorithmen Gefilterte Rückprojektion FBP und ART Varianten verglichen. Die Bewertung im Fourierraum zeigt, dass DIRECTT die fehlende Information im MW erfolgreich ergänzt. Zur quantitativen Bewertung der lokalen Rekonstruktions qualität werden Methoden der räumlichen Statistik eingesetz

Pseudospherical surfaces on time scales: a geometric definition and the spectral approach

We define and discuss the notion of pseudospherical surfaces in asymptotic coordinates on time scales. Thus we extend well known notions of discrete pseudospherical surfaces and smooth pseudosperical surfaces on more exotic domains (e.g, the Cantor set). In particular, we present a new expression for the discrete Gaussian curvature which turns out to be valid for asymptotic nets on any time scale. We show that asymptotic Chebyshev nets on an arbitrary time scale have constant negative Gaussian curvature. We present also the quaternion-valued spectral problem (the Lax pair) and the Darboux-Backlund transformation for pseudospherical surfaces (in asymptotic coordinates) on arbitrary time scales.Comment: 20 page

Neutron tomography in modern archaeology

The search for non invasive and non destructive techniques is fundamental when dealing with samples of great historical, cultural and artistic value as well as with samples strongly degraded. Among different techniques, Neutron Tomography NT allows a close analysis of samples of Archaeological interest without damaging them. In what follows, a few cases in which the Neutron Tomography instrument of the BENSC at HMI Berlin has been successfully applied will be show

Molecular Aspects of Secretory Granule Exocytosis by Neurons and Endocrine Cells

Neuronal communication and endocrine signaling are fundamental for integrating the function of tissues and cells in the body. Hormones released by endocrine cells are transported to the target cells through the circulation. By contrast, transmitter release from neurons occurs at specialized intercellular junctions, the synapses. Nevertheless, the mechanisms by which signal molecules are synthesized, stored, and eventually secreted by neurons and endocrine cells are very similar. Neurons and endocrine cells have in common two different types of secretory organelles, indicating the presence of two distinct secretory pathways. The synaptic vesicles of neurons contain excitatory or inhibitory neurotransmitters, whereas the secretory granules (also referred to as dense core vesicles, because of their electron dense content) are filled with neuropeptides and amines. In endocrine cells, peptide hormones and amines predominate in secretory granules. The function and content of vesicles, which share antigens with synaptic vesicles, are unknown for most endocrine cells. However, in B cells of the pancreatic islet, these vesicles contain GABA, which may be involved in intrainsular signaling.' Exocytosis of both synaptic vesicles and secretory granules is controlled by cytoplasmic calcium. However, the precise mechanisms of the subsequent steps, such as docking of vesicles and fusion of their membranes with the plasma membrane, are still incompletely understood. This contribution summarizes recent observations that elucidate components in neurons and endocrine cells involved in exocytosis. Emphasis is put on the intracellular aspects of the release of secretory granules that recently have been analyzed in detail

Morphological evolution of electrochemically plated stripped lithium microstructures by synchrotron X ray phase contrast tomography

Due to its low redox potential and high theoretical specific capacity, Li metal has drawn worldwide research attention because of its potential use in next generation battery technologies such as Li S and Li O2. Unfortunately, uncontrollable growth of Li microstructures LmSs, e.g., dendrites, fibers during electrochemical Li stripping plating has prevented their practical commercialization. Despite various strategies proposed to mitigate LmS nucleation and or block its growth, a fundamental understanding of the underlying evolution mechanisms remains elusive. Herein, synchrotron in line phase contrast X ray tomography was employed to investigate the morphological evolution of electrochemically deposited dissolved LmSs nondestructively. We present a 3D characterization of electrochemically stripped Li electrodes with regard to electrochemically plated LmSs. We clarify fundamentally the origin of the porous lithium interface growing into Li electrodes. Moreover, cleavage of the separator caused by growing LmS was experimentally observed and visualized in 3D. Our systematic investigation provides fundamental insights into LmS evolution and enables us to understand the evolution mechanisms in Li electrodes more profoundl

Opioid receptor activation triggering downregulation of cAMP improves effectiveness of anti-cancer drugs in treatment of glioblastoma

Glioblastoma are the most frequent and malignant human brain tumors, having a very poor prognosis. The enhanced radio- and chemoresistance of glioblastoma and the glioblastoma stem cells might be the main reason why conventional therapies fail. The second messenger cyclic AMP (cAMP) controls cell proliferation, differentiation, and apoptosis. Downregulation of cAMP sensitizes tumor cells for anti-cancer treatment. Opioid receptor agonists triggering opioid receptors can activate inhibitory Gi proteins, which, in turn, block adenylyl cyclase activity reducing cAMP. In this study, we show that downregulation of cAMP by opioid receptor activation improves the effectiveness of anti-cancer drugs in treatment of glioblastoma. The µ-opioid receptor agonist D,L-methadone sensitizes glioblastoma as well as the untreatable glioblastoma stem cells for doxorubicin-induced apoptosis and activation of apoptosis pathways by reversing deficient caspase activation and deficient downregulation of XIAP and Bcl-xL, playing critical roles in glioblastomas' resistance. Blocking opioid receptors using the opioid receptor antagonist naloxone or increasing intracellular cAMP by 3-isobutyl-1-methylxanthine (IBMX) strongly reduced opioid receptor agonist-induced sensitization for doxorubicin. In addition, the opioid receptor agonist D,L-methadone increased doxorubicin uptake and decreased doxorubicin efflux, whereas doxorubicin increased opioid receptor expression in glioblastomas. Furthermore, opioid receptor activation using D,L-methadone inhibited tumor growth significantly in vivo. Our findings suggest that opioid receptor activation triggering downregulation of cAMP is a promising strategy to inhibit tumor growth and to improve the effectiveness of anti-cancer drugs in treatment of glioblastoma and in killing glioblastoma stem cells

Active bacterial core surveillance of the emerging infections program network.

Active Bacterial Core surveillance (ABCs) is a collaboration between the Centers for Disease Control and Prevention and several state health departments and universities participating in the Emerging Infections Program Network. ABCs conducts population-based active surveillance, collects isolates, and performs studies of invasive disease caused by Streptococcus pneumoniae, group A and group B Streptococcus, Neisseria meningitidis, and Haemophilus influenzae for a population of 17 to 30 million. These pathogens caused an estimated 97,000 invasive cases, resulting in 10,000 deaths in the United States in 1998. Incidence rates of these pathogens are described. During 1998, 25% of invasive pneumococcal infections in ABCs areas were not susceptible to penicillin, and 13.3% were not susceptible to three classes of antibiotics. In 1998, early-onset group B streptococcal disease had declined by 65% over the previous 6 years. More information on ABCs is available at www.cdc.gov/ncidod/dbmd/abcs. ABCs specimens will soon be available to researchers through an archive