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

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

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

Cell death sensitization of leukemia cells by opioid receptor activation

Cyclic AMP (cAMP) regulates a number of cellular processes and modulates cell death induction. cAMP levels are altered upon stimulation of specific G-protein-coupled receptors inhibiting or activating adenylyl cyclases. Opioid receptor stimulation can activate inhibitory Gi-proteins which in turn block adenylyl cyclase activity reducing cAMP. Opioids such as D,L-methadone induce cell death in leukemia cells. However, the mechanism how opioids trigger apoptosis and activate caspases in leukemia cells is not understood. In this study, we demonstrate that downregulation of cAMP induced by opioid receptor activation using the opioid D,L-methadone kills and sensitizes leukemia cells for doxorubicin treatment. Enhancing cAMP levels by blocking opioid-receptor signaling strongly reduced D,L-methadone-induced apoptosis, caspase activation and doxorubicin-sensitivity. Induction of cell death in leukemia cells by activation of opioid receptors using the opioid D,L-methadone depends on critical levels of opioid receptor expression on the cell surface. Doxorubicin increased opioid receptor expression in leukemia cells. In addition, the opioid D,L-methadone increased doxorubicin uptake and decreased doxorubicin efflux in leukemia cells, suggesting that the opioid D,L-methadone as well as doxorubicin mutually increase their cytotoxic potential. Furthermore, we found that opioid receptor activation using D,L-methadone alone or in addition to doxorubicin inhibits tumor growth significantly in vivo. These results demonstrate that opioid receptor activation via triggering the downregulation of cAMP induces apoptosis, activates caspases and sensitizes leukemia cells for doxorubicin treatment. Hence, opioid receptor activation seems to be a promising strategy to improve anticancer therapies

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

An essential role of acetylcholine-glutamate synergy at habenular synapses in nicotine dependence

A great deal of interest has been focused recently on the habenula and its critical role in aversion, negative-reward and drug dependence. Using a conditional mouse model of the ACh-synthesizing enzyme choline acetyltransferase (Chat), we report that local elimination of acetylcholine (ACh) in medial habenula (MHb) neurons alters glutamate corelease and presynaptic facilitation. Electron microscopy and immuno-isolation analyses revealed colocalization of ACh and glutamate vesicular transporters in synaptic vesicles (SVs) in the central IPN. Glutamate reuptake in SVs prepared from the IPN was increased by ACh, indicating vesicular synergy. Mice lacking CHAT in habenular neurons were insensitive to nicotine-conditioned reward and withdrawal. These data demonstrate that ACh controls the quantal size and release frequency of glutamate at habenular synapses, and suggest that the synergistic functions of ACh and glutamate may be generally important for modulation of cholinergic circuit function and behavior

In situ radiographic investigation of de lithiation mechanisms in a tin electrode lithium ion battery.

The lithiation and delithiation mechanisms of multiple Sn particles in a customized flat radiography cell were investigated by in amp; 8197;situ synchrotron radiography. For the first time, four de lithiation phenomena in a Sn electrode battery system are highlighted 1 amp; 8197;the de lithiation behavior varies between different Sn particles, 2 amp; 8197;the time required to lithiate individual Sn particles is markedly different from the time needed to discharge the complete battery, 3 amp; 8197;electrochemical deactivation of originally electrochemically active particles is reported, and 4 amp; 8197;a change of electrochemical behavior of individual particles during cycling is found and explained by dynamic changes of de lithiation pathways amongst particles within the electrode. These unexpected findings fundamentaly expand the understanding of the underlying de lithiation mechanisms inside commercial lithium ion batteries LIBs and would open new design principles for high performance next generation LIB