High-resolution neutron-diffraction measurements to 8 kbar

We describe the capability to measure high-resolution neutron powder diffraction data to a pressure of at least 8 kbar. We have used the HRPD instrument at the ISIS neutron source and a piston-cylinder design of pressure cell machined from a null-scattering titanium zirconium alloy. Data were collected under hydrostatic conditions from an elpasolite perovskite La(Formula presented.)NiMnO(Formula presented.); by virtue of a thinner cell wall on the incident-beam side of the cell, it was possible to obtain data in the instrument's highest resolution back-scattering detector banks up to a maximum pressure of 8.5 kbar

A comparison of the performance of the ice and ocean tracking modes of the ERS‐1 radar altimeter over non‐ocean surfaces

The European Space Agency's ERS-1 radar altimeter is the first to include separate operating modes to optimise performance over both ocean and non-ocean surfaces. As part of the ERS-1 commissioning activities, we have carried out a study of the tracking performance of this instrument over non-ocean surfaces. Statistics for land ice, sea ice, arid lands, and inland water are presented. Performance in both operating modes is shown to be better than that of previous missions

Phosphorylation of p130Cas initiates Rac activation and membrane ruffling

<p>Abstract</p> <p>Background</p> <p>Non-receptor tyrosine kinases (NTKs) regulate physiological processes such as cell migration, differentiation, proliferation, and survival by interacting with and phosphorylating a large number of substrates simultaneously. This makes it difficult to attribute a particular biological effect to the phosphorylation of a particular substrate. We developed the Functional Interaction Trap (FIT) method to phosphorylate specifically a single substrate of choice in living cells, thereby allowing the biological effect(s) of that phosphorylation to be assessed. In this study we have used FIT to investigate the effects of specific phosphorylation of p130Cas, a protein implicated in cell migration. We have also used this approach to address a controversy regarding whether it is Src family kinases or focal adhesion kinase (FAK) that phosphorylates p130Cas in the trimolecular Src-FAK-p130Cas complex.</p> <p>Results</p> <p>We show here that SYF cells (mouse fibroblasts lacking the NTKs Src, Yes and Fyn) exhibit a low level of basal tyrosine phosphorylation at focal adhesions. FIT-mediated tyrosine phosphorylation of NTK substrates p130Cas, paxillin and FAK and cortactin was observed at focal adhesions, while FIT-mediated phosphorylation of cortactin was also seen at the cell periphery. Phosphorylation of p130Cas in SYF cells led to activation of Rac1 and increased membrane ruffling and lamellipodium formation, events associated with cell migration. We also found that the kinase activity of Src and not FAK is essential for phosphorylation of p130Cas when the three proteins exist as a complex in focal adhesions.</p> <p>Conclusion</p> <p>These results demonstrate that tyrosine phosphorylation of p130Cas is sufficient for its localization to focal adhesions and for activation of downstream signaling events associated with cell migration. FIT provides a valuable tool to evaluate the contribution of individual components of the response to signals with multiple outputs, such as activation of NTKs.</p

Development and testing of the BONES physical activity survey for young children

<p>Abstract</p> <p>Background</p> <p>Weight-bearing and high intensity physical activities are particularly beneficial for stimulating bone growth in children given that bone responds favorably to mechanical load. While it is important to assess the contribution and impact of weight-bearing physical activity on health outcomes, measurement tools that quantify and provide information on these activities separately from overall physical activity are limited. This study describes the development and evaluation of a pictorial physical activity survey (PAS) that measures children's participation and knowledge of high-intensity, weight-bearing ("bone smart") physical activity.</p> <p>Methods</p> <p>To test reliability, two identical sets of the PAS were administered on the same day to 41 children (mean age 7.1 ± 0.8 years; 63% female) and compared. To test validity, accelerometry data from 40 children (mean age 7.7 ± 0.8 years; 50% female) were compared to data provided by the PAS. Agreements between categorical and ordinal items were assessed with Kappa statistics; agreements between continuous indices were assessed with Spearman's correlation tests.</p> <p>Results</p> <p>The subjects produced reliable results in all 10 physical activity participation items (κ range: 0.36-0.73, all p < 0.05), but less reliable in answering if the physical activities were "bone smart" (κ range: -0.04-0.66). Physical activity indices, including metabolic equivalent time and weight-bearing factors, were significant in test-retest analyses (Spearman's <it>r </it>range: 0.57-0.74, all p < 0.001). Minutes of very vigorous activity from the accelerometer were associated with the self-reported weight-bearing activity, moderate-high, and high activity scores from the PAS (Spearman's <it>r </it>range: 0.47-0.48, all p < 0.01). However, accelerometer counts, counts per minute, and minutes of moderate-vigorous and vigorous activity were not associated with the PAS scores.</p> <p>Conclusions</p> <p>Together, the results of these studies suggest that the PAS has acceptable test-retest reliability, but limited validity for early elementary school children. This survey demonstrates a first step towards developing a questionnaire that measures high intensity, weight-bearing activity in schoolchildren.</p

Deleted in Liver Cancer 1 (DLC1) Utilizes a Novel Binding Site for Tensin2 PTB Domain Interaction and Is Required for Tumor-Suppressive Function

Background: Deleted in liver cancer 1 (DLC1) is a Rho GTPase-activating protein (RhoGAP) frequently deleted and underexpressed in hepatocellular carcinoma (HCC) as well as in other cancers. Recent independent studies have shown interaction of DLC1 with members of the tensin focal adhesion protein family in a Src Homology 2 (SH2) domain-dependent mechanism. DLC1 and tensins interact and co-localize to punctate structures at focal adhesions. However, the mechanisms underlying the interaction between DLC1 and various tensins remain controversial. Methodology/Principal Findings: We used a co-immunoprecipitation assay to identify a previously undocumented binding site at 375-385 of DLC1 that predominantly interacted with the phosphotyrosine binding (PTB) domain of tensin2. DLC1-tensin2 interaction is completely abolished in a DLC1 mutant lacking this novel PTB binding site (DLC1ΔPTB). However, as demonstrated by immunofluorescence and co-immunoprecipitation, neither the focal adhesion localization nor the interaction with tensin1 and C-terminal tensin-like (cten) were affected. Interestingly, the functional significance of this novel site was exhibited by the partial reduction of the RhoGAP activity, which, in turn, attenuated the growth-suppressive activity of DLC1 upon its removal from DLC1. Conclusions/Significance: This study has provided new evidence that DLC1 also interacts with tensin2 in a PTB domain-dependent manner. In addition to properly localizing focal adhesions and preserving RhoGAP activity, DLC1 interaction with tensin2 through this novel focal adhesion binding site contributes to the growth-suppressive activity of DLC1. © 2009 Chan et al.published_or_final_versio

The distinct roles of the nucleus and nucleus-cytoskeleton connections in three-dimensional cell migration

Cells often migrate in vivo in an extracellular matrix that is intrinsically three-dimensional (3D) and the role of actin filament architecture in 3D cell migration is less well understood. Here we show that, while recently identified linkers of nucleoskeleton to cytoskeleton (LINC) complexes play a minimal role in conventional 2D migration, they play a critical role in regulating the organization of a subset of actin filament bundles – the perinuclear actin cap - connected to the nucleus through Nesprin2giant and Nesprin3 in cells in 3D collagen I matrix. Actin cap fibers prolong the nucleus and mediate the formation of pseudopodial protrusions, which drive matrix traction and 3D cell migration. Disruption of LINC complexes disorganizes the actin cap, which impairs 3D cell migration. A simple mechanical model explains why LINC complexes and the perinuclear actin cap are essential in 3D migration by providing mechanical support to the formation of pseudopodial protrusions

Epigenetic regulation of RhoB loss of expression in lung cancer

<p>Abstract</p> <p>Background</p> <p>RhoB is down-regulated in most lung cancer cell lines and tumor tissues when compared with their normal counterparts. The mechanism of this loss of expression is not yet deciphered.</p> <p>Methods</p> <p>Since no mutation has been reported in the RhoB sequence, we investigated the epigenetic regulation of RhoB expression by analyzing the effect of HDAC inhibitors and methyltransferase inhibitors, by direct sequencing after bisulfite treatment and by methylation specific PCR.</p> <p>Results</p> <p>We first showed that histone deacetylase (HDAC) inhibitors induce a significant RhoB re-expression in lung cancer cell lines whereas only a slight effect was observed with methyl transferase inhibitors. As promoter methylation is the most common epigenetic process in lung cancer, we performed methylation specific PCR and sequence analysis after bisulfite treatment and demonstrated that RhoB was methylated neither in lung cancer cell lines nor in tumor tissues. We also showed that a variable number of tandem repeats sequences in the 5' region of the RhoB gene was involved in HDAC response.</p> <p>Conclusion</p> <p>We thus propose that RhoB regulation of expression occurs mainly by histone deacetylation rather than by promoter hypermethylation and that this process can be modulated by specific 5' sequences within the promoter.</p

Pilot testing of a sampling methodology for assessing seed attachment propensity and transport rate in a soil matrix carried on boot soles and bike tires

Land managers of natural areas are under pressure to balance demands for increased recreation access with protection of the natural resource. Unintended dispersal of seeds by visitors to natural areas has high potential for weedy plant invasions, with initial seed attachment an important step in the dispersal process. Although walking and mountain biking are popular nature-based recreation activities there are few studies quantifying propensity for seed attachment and transport rate on boot soles and none for bike tires. Attachment and transport rate can potentially be affected by a wide range of factors for which field testing can be time-consuming and expensive. We pilot tested a sampling methodology for measuring seed attachment and transport rate in a soil matrix carried on boot soles and bike tires traversing a known quantity and density of a seed analog (beads) over different distances and soil conditions. We found % attachment rate on boot soles was much lower overall than previously reported but that boot soles had a higher propensity for seed attachment than bike tires in almost all conditions. We believe our methodology offers a cost-effective option for researchers seeking to manipulate and test effects of different influencing factors on these two dispersal vectors

A separated vortex ring underlies the flight of the dandelion

Wind-dispersed plants have evolved ingenious ways to lift their seeds1,2. The common dandelion uses a bundle of drag-enhancing bristles (the pappus) that helps to keep their seeds aloft. This passive flight mechanism is highly effective, enabling seed dispersal over formidable distances3,4; however, the physics underpinning pappus-mediated flight remains unresolved. Here we visualized the flow around dandelion seeds, uncovering an extraordinary type of vortex. This vortex is a ring of recirculating fluid, which is detached owing to the flow passing through the pappus. We hypothesized that the circular disk-like geometry and the porosity of the pappus are the key design features that enable the formation of the separated vortex ring. The porosity gradient was surveyed using microfabricated disks, and a disk with a similar porosity was found to be able to recapitulate the flow behaviour of the pappus. The porosity of the dandelion pappus appears to be tuned precisely to stabilize the vortex, while maximizing aerodynamic loading and minimizing material requirements. The discovery of the separated vortex ring provides evidence of the existence of a new class of fluid behaviour around fluid-immersed bodies that may underlie locomotion, weight reduction and particle retention in biological and manmade structures