Transformation of Rat-1 fibroblasts with the v-src oncogene increases the tyrosine phosphorylation state and activity of the alpha subunit of Gq/G11.

Two major intermediaries in signal transduction pathways are pp60v-sre family tyrosine kinases and heterotrimeric guanine nucleotide-binding proteins. In Rat-1 fibroblasts transformed by the v-src oncogene, endothelin-1 (ET-1)-induced inositol 1,4,5-trisphosphate accumulation is increased 6-fold, without any increases in the numbers of ET-1 receptors or in the response to another agonist, thrombin. This ET-1 hyperresponse can be inhibited by an antibody directed against the carboxyl terminus of the Gq/G11 alpha subunit, suggesting that the Gq/G11 protein couples ET-1 receptors to phospholipase C (PLC). While v-src transformation did not increase the expression of the Gq/G11 alpha subunit, immunoblotting with anti-phosphotyrosine antibodies and phosphoamino acid analysis demonstrated that the Gq/G11 alpha subunit becomes phosphorylated on tyrosine residues in v-src-transformed cells. Moreover, when the Gq/G11 protein was extracted from control and transformed cell lines and reconstituted with exogenous PLC, AIF*4-stimulated Gq/G11 activity was markedly increased in extracts from v-src-transformed cells. Our results demonstrate that the process of v-src transformation can increase the tyrosine phosphorylation state of the Gq/G11 alpha-subunit in intact cells and that the process causes an increase in the Gq/G11 alpha-subunit's ability to stimulate PLC following activation with AIF-4.

Regulation of Sticholysin II-Induced Pore Formation by Lipid Bilayer Composition, Phase State, and Interfacial Properties

Sticholysin II (StnII) is a pore-forming toxin that uses sphingomyelin (SM) as the recognition molecule in targeting membranes.After StnII monomers bind to SM, several toxin monomers act in concert to oligomerize into a functional pore. The regulation of StnII binding to SM, and the subsequent pore-formation process, is not fully understood. In this study, we examined how the biophysical properties of bilayers, originating from variations in the SM structure, from the presence of sterol species, or from the presence of increasingly polyunsaturated glycerophospholipids,affected StnII-induced pore formation. StnII-induced pore formation, as determined from calcein permeabilization, was fastest in the pure unsaturated SM bilayers. In 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/saturated SM bilayers (4:1 molar ratio), pore formation became slower as the chain length of the saturated SMs increased from 14 up to 24 carbons. In the POPC/palmitoyl-SM (16:0-SM) 4:1 bilayers, SM could not support pore formation by StnII if dimyristoyl-PC was included at 1:1 stoichiometry with 16:0-SM, suggesting that free clusters of SM were required for toxin binding and/or pore formation. Cholesterol and other sterols facilitated StnII-induced pore formation markedly, but the efficiency did not appear to correlate with the sterol structure. Benzyl alcohol was more efficient than sterols in enhancing the pore-formation process, suggesting that the effect on pore formation originated from alcohol-induced alteration of the hydrogen-bonding network in the SM-containing bilayers. Finally, we observed that pore formation by StnII was enhanced in the PC/16:0-SM 4:1 bilayers, in which the PC was increasingly unsaturated. We conclude that the physical state of bilayer lipids greatly affected pore formation by StnII. Phase boundaries were not required for pore formation, although SM in a gel state attenuated pore formation.

Representing Clients in the Marijuana Industry: Navigating State and Federal Rules

As the number of states legalizing medicinal and recreational marijuana increases and marijuana emerges as a growing lawful industry, lawyers find themselves in an awkward position. In most states, lawyers who represent clients in the marijuana industry risk discipline for assisting clients in the commission of a (federal) crime. Even in jurisdictions like Colorado, where the rules of professional conduct have been amended to permit lawyers to assist clients who comply with marijuana state laws, lawyers who are admitted to practice in federal courts risk being disciplined by these tribunals for assisting clients in the commission of a crime pursuant to the courts’ local rules of conduct. This short article explores the thorny issue of navigating state and federal rules of professional conduct while representing clients in the marijuana industry.

Quiescent state and outburst evolution of SGR 0501+4516

We report on the quiescent state of the soft gamma repeater SGR 0501+4516 observed by XMM–Newton on 2009 August 30. The source exhibits an absorbed flux ∼75 times lower than that measured at the peak of the 2008 outburst, and a rather soft spectrum, with the same value of the blackbody temperature observed with ROSAT back in 1992. This new observation is put into the context of all existing X-ray data since its discovery in 2008 August, allowing us to complete the study of the timing and spectral evolution of the source from outburst until its quiescent state. The set of deep XMM–Newton observations performed during the few years time-scale of its outburst allows us to monitor the spectral characteristics of this magnetar as a function of its rotational period, and their evolution along these years. After the first ∼10 d, the initially hot and bright surface spot progressively cooled down during the decay. We discuss the behaviour of this magnetar in the context of its simulated secular evolution, inferring a plausible dipolar field at birth of 3 × 1014 G, and a current (magnetothermal) age of ∼10 kyr.

Active edible films: Current state and future trends

Active edible films represent one of the current and future trends in the development of new polymers for selected applications, particularly food packaging. Some biopolymers show excellent performance as carriers for active compounds extracted from natural sources and are able to be released at a controlled rate to packaged food. In this review we aim to present, in a comprehensive way, the most recent advances and updates in this subject, where much research is currently ongoing and new studies are reported very often. This review focuses on innovative biopolymer matrices, their processing to obtain edible active films, and present and future applications.