H-ras, K-ras, and inner plasma membrane raft proteins operate in nanoclusters with differential dependence on the actin cytoskeleton

Plasma membrane compartmentalization imposes lateral segregation on membrane proteins that is important for regulating signal transduction. We use computational modeling of immunogold spatial point patterns on intact plasma membrane sheets to test different models of inner plasma membrane organization. We find compartmentalization at the nanoscale level but show that a classical raft model of preexisting stable domains into which lipid raft proteins partition is incompatible with the spatial point patterns generated by the immunogold labeling of a palmitoylated raft marker protein. Rather, approximate to 30% of the raft protein exists in cholesterol-dependent nanoclusters, with approximate to 70% distributed as monomers. The cluster/monomer ratio (number of proteins in clusters/number of proteins outside clusters) is independent of expression level. H-rasG12V and K-rasG12V proteins also operate in nanoclusters with fixed cluster/monomer ratios that are independent of expression level. Detailed calibration of the immunogold imaging protocol suggests that radii of raft and RasG12V protein nanoclusters may be as small as 11 and 6 nm, respectively, and shows that the nanoclusters contain small numbers (6.0-7.7) of proteins. Raft nanoclusters do not form if the actin cytoskeleton is disassembled. The formation of K-rasG12V but not H-rasG12V nanoclusters also is actin-dependent. K-rasG12V but not H-rasG12V signaling is abrogated by actin cytoskeleton disassembly, which shows that nanoclustering is critical for Ras function. These findings argue against stable preexisting domains on the inner plasma membrane in favor of dynamic actively regulated nanoclusters similar to those proposed for the outer plasma membrane. RasG12V nanoclusters may facilitate the assembly of essential signal transduction complexes.

A graphical specification of model transformations with triple graph grammars

Models and model transformations are the core concepts of OMG's MDA (TM) approach. Within this approach, most models are derived from the MOF and have a graph-based nature. In contrast, most of the current model transformations are specified textually. To enable a graphical specification of model transformation rules, this paper proposes to use triple graph grammars as declarative specification formalism. These triple graph grammars can be specified within the FUJABA tool and we argue that these rules can be more easily specified and they become more understandable and maintainable. To show the practicability of our approach, we present how to generate Tefkat rules from triple graph grammar rules, which helps to integrate triple graph grammars with a state of a art model transformation tool and shows the expressiveness of the concept.

Language, respect and the power of protest: Linguistic stereotyping and people with disabilities in Japan

Discriminatory language became an important social issue in the west in the late twentieth century, when debates on political correctness and minority rights focused largely on the issue of respect in language. Japan is often criticized for having made only token attempts to address this issue. This paper investigates how one marginalized group—people with disabilities—has dealt with discriminatory and disrespectful language. The debate has been played out in four public spaces: the media, the law, literature, and the Internet. The paper discusses the kind of language, which has generated protest, the empowering strategies of direct action employed to combat its use, and the response of the media, the bureaucracy, and the literati. Government policy has not kept pace with social change in this area; where it exists at all, it is often contradictory and far from clear. I argue that while the laws were rewritten primarily as a result of external international trends, disability support groups achieved domestic media compliance by exploiting the keen desire of media organizations to avoid public embarrassment. In the absence of language policy formulated at the government level, the media effectively instituted a policy of self-censorship through strict guidelines on language use, thereby becoming its own best watchdog. Disability support groups have recently enlisted the Internet as an agent of further empowerment in the ongoing discussion of the issue.