New mutations in flagellar motors identified by whole genome sequencing in Chlamydomonas

BACKGROUND: The building of a cilium or flagellum requires molecular motors and associated proteins that allow the relocation of proteins from the cell body to the distal end and the return of proteins to the cell body in a process termed intraflagellar transport (IFT). IFT trains are carried out by kinesin and back to the cell body by dynein. METHODS: We used whole genome sequencing to identify the causative mutations for two temperature-sensitive flagellar assembly mutants in Chlamydomonas and validated the changes using reversion analysis. We examined the effect of these mutations on the localization of IFT81, an IFT complex B protein, the cytoplasmic dynein heavy chain (DHC1b), and the dynein light intermediate chain (D1bLIC). RESULTS: The strains, fla18 and fla24, have mutations in kinesin-2 and cytoplasmic dynein, respectively. The fla18 mutation alters the same glutamic acid (E(24)G) mutated in the fla10-14 allele (E(24)K). The fla18 strain loses flagella at 32?C more rapidly than the E(24)K allele but less rapidly than the fla10-1 allele. The fla18 mutant loses its flagella by detachment rather than by shortening. The fla24 mutation falls in cytoplasmic dynein and changes a completely conserved amino acid (L(3243)P) in an alpha helix in the AAA5 domain. The fla24 mutant loses its flagella by shortening within 6 hours at 32?C. DHC1b protein is reduced by 18-fold and D1bLIC is reduced by 16-fold at 21?C compared to wild-type cells. We identified two pseudorevertants (L(3243)S and L(3243)R), which remain flagellated at 32?C. Although fla24 cells assemble full-length flagella at 21?C, IFT81 protein localization is dramatically altered. Instead of localizing at the basal body and along the flagella, IFT81 is concentrated at the proximal end of the flagella. The pseudorevertants show wild-type IFT81 localization at 21?C, but proximal end localization of IFT81 at 32?C. CONCLUSIONS: The change in the AAA5 domain of the cytoplasmic dynein in fla24 may block the recycling of IFT trains after retrograde transport. It is clear that different alleles in the flagellar motors reveal different functions and roles. Multiple alleles will be important for understanding structure-function relationships

Turbulent transport of material particles: An experimental study of finite size effects

We use an acoustic Lagrangian tracking technique, particularly adapted to measurements in open flows, and a versatile material particles generator (in the form of soap bubbles with adjustable size and density) to characterize Lagrangian statistics of finite sized, neutrally bouyant, particles transported in an isotropic turbulent flow of air. We vary the size of the particles in a range corresponding to turbulent inertial scales and explore how the turbulent forcing experienced by the particles depends on their size. We show that, while the global shape of the intermittent acceleration probability density function does not depend significantly on particle size, the acceleration variance of the particles decreases as they become larger in agreement with the classical scaling for the spectrum of Eulerian pressure fluctuations in the carrier flow

Scintillator counters with WLS fiber/MPPC readout for the side muon range detector (SMRD)of the T2K experiment

The T2K neutrino experiment at J-PARC uses a set of near detectors to measure the properties of an unoscillated neutrino beam and neutrino interaction cross-sections. One of the sub-detectors of the near-detector complex, the side muon range detector (SMRD), is described in the paper. The detector is designed to help measure the neutrino energy spectrum, to identify background and to calibrate the other detectors. The active elements of the SMRD consist of 0.7 cm thick extruded scintillator slabs inserted into air gaps of the UA1 magnet yokes. The readout of each scintillator slab is provided through a single WLS fiber embedded into a serpentine shaped groove. Two Hamamatsu multi-pixel avalanche photodiodes (MPPC's) are coupled to both ends of the WLS fiber. This design allows us to achieve a high MIP detection efficiency of greater than 99%. A light yield of 25-50 p.e./MIP, a time resolution of about 1 ns and a spatial resolution along the slab better than 10 cm were obtained for the SMRD counters.Comment: 7 pages, 4 figures; talk at TIPP09, March 12-17, Tsukuba, Japan; to be published in the conference proceeding

Design of micromixers using CFD modelling

The effect of various geometrical parameters of a grooved staggered herringbone micromixer on the mixing performance has been investigated using Computational Fluid Dynamics. Mixing quality has been quantified with spatial data statistics, maximum striation thickness and residence time analyses. The results show that the number of grooves per mixing cycle does not affect the mixing quality in an important way. On the other hand, a larger groove depth and width allow the maximum striation thickness to be rapidly reduced, without increasing the pressure drop across the mixer. Wide grooves, however, create significant dead zones in the microchannel, whereas deep grooves improve the spatial mixing quality

The T2K Side Muon Range Detector

The T2K experiment is a long baseline neutrino oscillation experiment aiming to observe the appearance of {\nu} e in a {\nu}{\mu} beam. The {\nu}{\mu} beam is produced at the Japan Proton Accelerator Research Complex (J-PARC), observed with the 295 km distant Super- Kamiokande Detector and monitored by a suite of near detectors at 280m from the proton target. The near detectors include a magnetized off-axis detector (ND280) which measures the un-oscillated neutrino flux and neutrino cross sections. The present paper describes the outermost component of ND280 which is a side muon range detector (SMRD) composed of scintillation counters with embedded wavelength shifting fibers and Multi-Pixel Photon Counter read-out. The components, performance and response of the SMRD are presented.Comment: 13 pages, 19 figures v2: fixed several typos; fixed reference

Perceptions of Respite Care Benefits and the Measures of Leisure Satisfaction and Life Satisfaction among Informal Caregivers

The purpose of the proposed study was to determine the perceived benefits of respite care by informal caregivers who are actively pursuing respite services, and to evaluate their expectations after receiving respite services. In addition the study also was undertaken to better understand the roles of “leisure satisfaction” and “Perceived Stress Levels” in the lives of informal caregivers who are currently receiving respite services versus those who are not currently receiving services, in both a pre and post format for both groups of caregivers. The study group of caregivers was given a total of four surveys in the pre-respite package and three surveys in the post-respite package. The comparison group was also given four surveys in their initial package and two surveys in their second package. The findings indicate an increase in leisure satisfaction among the study group, with all caregivers exhibiting value for respite services. The implications from this study indicate that although there were no significant changes noted in state stress levels or overall leisure satisfaction levels, caregivers receiving respite confirmed the positive impact of respite services on their lives

Suppression of Phase Separation in LiFePO4 Nanoparticles During Battery Discharge

Using a novel electrochemical phase-field model, we question the common belief that LixFePO4 nanoparticles separate into Li-rich and Li-poor phases during battery discharge. For small currents, spinodal decomposition or nucleation leads to moving phase boundaries. Above a critical current density (in the Tafel regime), the spinodal disappears, and particles fill homogeneously, which may explain the superior rate capability and long cycle life of nano-LiFePO4 cathodes.Comment: 27 pages, 8 figure

Methods and Compositions Based on Culturing Microorganisms in Low Sedimental Fluid Shear Conditions

The benefits of applying a low sedimental fluid shear environment to manipulate microorganisms were examined. Microorganisms obtained from a low sedimental fluid shear culture, which exhibit modified phenotypic and molecular genetic characteristics, are useful for the development of novel and improved diagnostics, therapeutics, vaccines, and bio-industrial products. Furthermore, application of low sedimental fluid conditions to microorganisms permits identification of molecules uniquely expressed under these conditions, providing a basis for the design of new therapeutic targets

Comparison of X-ray and gamma-ray dose-response curves for pink somatic mutations in Tradescantia clone 02

Microdosimetric data indicate that the mean specific energy,zeta, produced by individual charged particles from X rays and gamma rays is different for the two radiation qualities by nearly a factor of two. In order to test whether this influences the initial, linear component in the dose-effect relations, a comparison was made between dose-response curves for pink somatic mutations inTradescantia clone 02 stamen hairs following X and gamma irradiations. Absorbed doses ranged from 2.66 to 300 rad. The results are in agreement with predictions made on the basis of microdosimetric data. At low doses gamma rays are substantially less effective than X rays. The RBE of gamma rays vs. X rays at low doses was approximately 0.6, a value lower than those usually reported in other experimental systems