Superoxide perturbation of the organization of vascular endothelial cell membranes

Cultured porcine thoracic aorta endothelial cells were covalently labeled with 4-maleimido-2,2,6,6-tetramethylpiperidinooxyl. Electron paramagnetic resonance spectrometry revealed two major binding environments representing strongly and weakly immobilized species. The disorder parameter of weak/strong, determined from the respective peak amplitudes, was irreversibly elevated following incubation of endothelial cells with a superoxide-generating system, indicating increased membrane fluidity. The rate of increase in membrane disorder was dependent upon superoxide generation rates. Incorporation of the spin-label at concentrations less than 250 microM had no effect on cell viability. The cellular proteins reacting with the spin-label were predominantly membrane proteins, characterized by immunoblotting using a rabbit anti-4-maleimido-2,2,6,6-tetramethylpiperidinooxyl IgG, following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophorectic transfer to nitrocellulose.     

On the fisher and the cubic-polynomial equations for the propagation of species properties

For precise boundary conditions of biological relevance, it is proved that the steadily propagating plane-wave solution to the Fisher equation requires the unique (eigenvalue) velocity of advance 2(Df)^1/2, whereD is the diffusivity of the mutant species andf is the frequency of selection in favor of the mutant. This rigorous result shows that a so-called “wrong equation”, i.e. one which differs from Fisher's by a term that is seemingly inconsequential for certain initial conditions, cannot be employed readily to obtain approximate solutions to Fisher's, for the two equations will often have qualitatively different manifolds of exact solutions. It is noted that the Fisher equation itself may be inappropriate in certain biological contexts owing to the manifest instability of the lowerconcentration uniform equilibrium state (UES). Depicting the persistence of a mutantdeficient spatial pocket, an exact steady-state solution to the Fisher equation is presented. As an alternative and perhaps more faithful model equation for the propagation of certain species properties through a homogeneous population, we consider a reaction-diffusion equation that features a cubic-polynomial rate expression in the species concentration, with two stable UES and one intermediate unstable UES. This equation admits a remarkably simple exact analytical solution to the steadily propagating plane-wave eigenvalue problem. In the latter solution, the sign of the eigenvelocity is such that the wave propagates to yield the “preferred” stable UES (namely, the one further removed from the unstable intermediate UES) at all spatial points ast→∞. The cubic-polynomial equation also admits an exact steady-state solution for a mutant-deficient or mutant-isolated spatial pocket. Finally, the perpetuating growth of a mutant population from an arbitrary localized initial distribution, a mathematical problem analogous to that for ignition in laminar flame theory, is studied by applying differential inequality analysis, and rigorous sufficient conditions for extinction are derived here.     

Immunohistochemical evaluation of ras oncogene expression in pulmonary and pleural neoplasms

We undertook an immunohistochemical analysis of human bronchopulmonary epithelial neoplasms and pleural mesotheliomas using a monoclonal antibody which recognizes ras oncogene products (p21ras). The monoclonal antibody, RAP-5, recognizes both unaltered and certain mutated p21ras. Formalin fixed and paraffin embedded tissue samples of 187 lung epithelial tumors and 27 pleural mesotheliomas were investigated; normal and bronchiectatic lungs were similarly studied. Normal lung and pleural tissue did not immunostain except for occasional type II pneumocytes. Reactive type II pneumocytes adjacent to carcinomas and bronchiectasis immunostained consistently. Twenty four/34 (71%) squamous carcinomas immunostained. Only 8/50 (16%) adenocarcinomas immunostained focally and weakly whereas 19/24 (79%) bronchioloalveolar carcinomas immunostained. Eleven/18 (61%) large cell carcinomas immunostained with variable intensity. Eleven/13 (85%) carcinoids, 6/7 (85%) well differentiated neuroendocrine carcinomas, and 18/21 (86%) intermediate cell neuroendocrine carcinomas immunostained while none of 20 small cell neuroendocrine carcinomas immunostained. Only a few mesotheliomas were immunostained focally. Two/14 (14%) epithelial type and 1/9 (11%) biphasic type mesotheliomas immunostained weakly; none of 4 spindle cell mesotheliomas immunostained. We conclude that while at least occasional cases of most types of pulmonary epithelial neoplasms express p21ras, the frequency and intensity of the expression are distinctly greater in certain tumor types such as squamous, bronchioloalveolar, and neuroendocrine neoplasm except for the small cell type. Contrary to these lung epithelial neoplasms, most mesotheliomas did not immunostain for p21ras. Whether the enhanced p21ras expression may point to a different mechanism of transformation or may merely reflect differentiation features remains undetermined.     

A divergent lineage among Octopus minor (Sasaki, 1920) populations in the Northwest Pacific supported by DNA barcoding

Octopus minor (Sasaki, 1920) is an important economic fishery resource in China. In order to explore the stock information and the phylogeographic status of O. minor, mitochondrial DNA cytochrome c oxidase subunit I (COI, 565?bp) and 16S rRNA (493?bp) genes were amplified from 11 different sampling locations. Genetic diversity evaluated by haplotypic and nucleotidic diversity implied high diversity in Lianjiang, and relatively low diversity in Rongcheng, which suggests that effective measures to protect the O. minor resource in this area are urgently required. Private haplotypes and remarkable higher pairwise Φ_ST in Yilan are responsible for the deep genetic divergence between Yilan and the 10 other populations. Haplotypes networks and two clusters’ topological structure also support the distinct subgroups (lineages A and lineages B), which apparently possess smaller genetic variation than mean interspecies distance. Taiwan island and its strait may act as a natural barrier that restricts the gene flow from the mainland. Deep genetic divergence between mainland and Taiwanese east coasts suggests different genetic stock, indicating that different management strategies are required.     

Caspase-2 Short Isoform Interacts with Membrane-Associated Cytoskeleton Proteins to Inhibit Apoptosis

Caspase-2 (casp-2) is the most conserved caspase across species, and is one of the initiator caspases activated by various stimuli. The casp-2 gene produces several alternative splicing isoforms. It is believed that the long isoform, casp-2L, promotes apoptosis, whereas the short isoform, casp-2S, inhibits apoptosis. The actual effect of casp-2S on apoptosis is still controversial, however, and the underlying mechanism for casp-2S-mediated apoptosis inhibition is unclear. Here, we analyzed the effects of casp-2S on DNA damage induced apoptosis through “gain-of-function” and “loss-of-function” strategies in ovarian cancer cell lines. We clearly demonstrated that the over-expression of casp-2S inhibited, and the knockdown of casp-2S promoted, the cisplatin-induced apoptosis of ovarian cancer cells. To explore the mechanism by which casp-2S mediates apoptosis inhibition, we analyzed the proteins which interact with casp-2S in cells by using immunoprecipitation (IP) and mass spectrometry. We have identified two cytoskeleton proteins, Fodrin and α-Actinin 4, which interact with FLAG-tagged casp-2S in HeLa cells and confirmed this interaction through reciprocal IP. We further demonstrated that casp-2S (i) is responsible for inhibiting DNA damage-induced cytoplasmic Fodrin cleavage independent of cellular p53 status, and (ii) prevents cisplatin-induced membrane blebbing. Taken together, our data suggests that casp-2S affects cellular apoptosis through its interaction with membrane-associated cytoskeletal Fodrin protein.     

Topography of carpal bone - An experimental model for carpal tunnel syndrome

By preparing a new experimental model for the carpal tunnel syndrome, the authors evaluated the differences of the human and rabbit carpal tunnels using a comparative anatomical study. A nearly identical situation-regarding the osseous and connective tissue formations in the carpal channel--was found. Therefore, the carpal tunnel of the rabbit is recommended for a model of chronic nerve compression, which is now planned by the authors.