Citations

A novel algorithm based on the variational formulation is presented for the determination of the material parameters (λ+2μ)/ρ and μ/ρ throughout inhomogeneous media given volumetric displacement information. The performance of the inversion algorithm is subsequently investigated when applied to measured displacement data throughout an inhomogeneous test phantom. The vector displacement components throughout the test phantom subject to monochromatic shear excitation measured in time using MRI (Magnetic Resonance Imaging) were temporally Fourier transformed to extract the components of monochromatic excitation, and the data were delivered to the inversion algorithm. A sequence of studies is presented on the basis of subsequent wavenumber filtering, polarization selection, and variation in the size of the volume elements of resolution. The resulting performance is assessed and recommendations for future efforts are discussed. [Work supported by ONR and NIH.]

A technique for measuring instantaneous three-dimensional pressure in a wideband ultrasound pulse is presented. A collimated laser pulse impinges on the ultrasound field, and the optical intensity in a diffraction plane subsequent to this scattering event is imaged onto a CCD camera and stored. Timing is controlled precisely, allowing the acousto-optic interaction to occur at the same point in the temporal evolution of the sound field each time the experiment is performed. After interaction with the ultrasound pulse, the optical phase is approximately proportional to the ray sum of instantaneous pressure in the pulse (taken in the direction of optical propagation). Optical phase immediately subsequent to passage through the ultrasound pulse is computed from recorded optical intensity via a phase retrieval algorithm. The experiment is performed at several angles as the ultrasound transducer is rotated about an axis normal to the optical axis of the imaging system. Tomographic reconstruction algorithms are applied to the retrieved optical phase data, computing pressure at all points in the ultrasound pulse. The resulting pulse may be propagated forward or backward in time under a source-free model. Reconstructions of the field from a 2.5-MHz array transducer compare well with hydrophone measurements.

This investigation evaluated a possible source of reduced intelligibility in hypokinetic dysarthric speech, namely the mismatch between listeners’ perceptual strategies and the acoustic information available in the dysarthric speech signal. A paradigm of error analysis was adopted in which listener transcriptions of phrases were coded for the presence and type of word boundary errors. Seventy listeners heard 60 phrases produced by speakers with hypokinetic dysarthria. The six-syllable phrases alternated strong and weak syllables and ranged in length from three to five words. Lexical boundary violations were defined as erroneous insertions or deletions of lexical boundaries that occurred either before strong or before weak syllables. A total of 1596 lexical boundary errors in the listeners’ transcriptions was identified unanimously by three independent judges. The pattern of errors generally conformed with the predictions of the Metrical Segmentation Strategy hypothesis [Cutler and Norris, J. Exp. Psychol. 14, 113–121 (1988)] which posits that listeners attend to strong syllables to identify word onsets. However, the strength of adherence to this pattern varied across speakers. Comparison of acoustic evidence of syllabic strength to lexical boundary error patterns revealed a source of intelligibility deficit associated with this particular type of dysarthric speech pattern.

Many methods are available for transferring foreign genes into mammalian cells (transfection). The advantages of ultrasound-mediated transfection are that it can be controlled in both space, by focusing, and time, by exposure. It was tested whether ultrasound-mediated gene transfection in cultured immortalized human chondrocytes would be enhanced using artificial cavitation nuclei in the form of Albunex®. Here 1.0 MHz ultrasound transmitted through the bottoms of six-well culture plates containing media with green fluorescent protein reporter gene plasmids at a concentration of 40 μg/ml and Albunex® at 50×106 bubbles/ml produced a peak transfection efficiency of about 50% of the living cells when exposure was 4×105 P spatial average peak pressure (SAPP) for 20 mins. Using these parameters, transfection efficiency increased linearly with ultrasound exposure pressure with a transfection threshold observed at a SAPP of 105 P. Adding fresh Albunex® at 50×106 bubbles/ml prior to sequential one second 3.2 or 4.0×105 P exposures increased transfection efficiency by 15% for each of three consecutive exposures. Efficient in vitro and perhaps in vivo transfection appears possible with ultrasound providing both temporal and spatial control over the transfection process.

An optical spin micromotor operated by optical tweezers, described in the present study, converts radiant energy directly to mechanical energy. This conversion is of great practical and fundamental interest. The optical spin micromotor consists of a two-bead linkage. The large bead (2.0 μm in diameter) is trapped by the optical tweezers and rotates around the laser beam axis. The small bead (0.94 μm in diameter) is partially coated with gold/palladium for generating the spin torque through the change of momentum from gradient radiation pressure of the same laser applied on the coated small bead. With a laser power of 29 mW at the sample plane, the spin speed averaged 158.8±155.5 rpm (mean±standard deviation) and the torque was estimated as 101.9±99.8 pN nm. This optical spin micromotor will be useful in (1) providing a fundamental step toward developing optical-operated mechanical devices, (2) twisting macromolecules, and (3) generating vortex and shear force in medium at nanoscale.

The assembly of inflammatory lesions in rheumatoid arthritis is highly regulated and typically leads to the formation of lymphoid follicles with germinal center (GC) reactions. We used microdissection of such extranodal follicles to analyze the colonizing T cells. Although the repertoire of follicular T cells was diverse, a subset of T cell receptor (TCR) sequences was detected in multiple independent follicles and not in interfollicular zones, suggesting recognition of a common antigen. Unexpectedly, the majority of shared TCR sequences were from CD8 T cells that were highly enriched in the synovium and present in low numbers in the periphery. To examine their role in extranodal GC reactions, CD8 T cells were depleted in human synovium-SCID mouse chimeras. Depletion of synovial CD8 T cells caused disintegration of the GC-containing follicles. In the absence of CD8 T cells, follicular dendritic cells disappeared, production of lymphotoxin-α1β2 markedly decreased, and immunoglobulin (Ig) secretion ceased. Immunohistochemical studies demonstrated that these CD8 T cells accumulated at the edge of the mantle zone. Besides their unique localization, they were characterized by the production of interferon (IFN)-γ, lack of the pore-forming enzyme perforin, and expression of CD40 ligand. Perifollicular IFN-γ+ CD8 T cells were rare in secondary lymphoid tissues but accounted for the majority of IFN-γ+ cells in synovial infiltrates. We propose that CD8+ T cells regulate the structural integrity and functional activity of GCs in ectopic lymphoid follicles.

The dynamins comprise an expanding family of ubiquitously expressed 100-kD GTPases that have been implicated in severing clathrin-coated pits during receptor-mediated endocytosis. Currently, it is unclear whether the different dynamin isoforms perform redundant functions or participate in distinct endocytic processes. To define the function of dynamin II in mammalian epithelial cells, we have generated and characterized peptide-specific antibodies to domains that either are unique to this isoform or conserved within the dynamin family. When microinjected into cultured hepatocytes these affinity-purified antibodies inhibited clathrin-mediated endocytosis and induced the formation of long plasmalemmal invaginations with attached clathrin-coated pits. In addition, clusters of distinct, nonclathrin-coated, flask-shaped invaginations resembling caveolae accumulated at the plasma membrane of antibody-injected cells. In support of this, caveola-mediated endocytosis of labeled cholera toxin B was inhibited in antibody-injected hepatocytes. Using immunoisolation techniques an anti-dynamin antibody isolated caveolar membranes directly from a hepatocyte postnuclear membrane fraction. Finally, double label immunofluorescence microscopy revealed a striking colocalization between dynamin and the caveolar coat protein caveolin. Thus, functional in vivo studies as well as ultrastructural and biochemical analyses indicate that dynamin mediates both clathrin-dependent endocytosis and the internalization of caveolae in mammalian cells.

The tumor suppressor p53 binding protein 1 (53BP1) binds to the DNA-binding domain of p53 and enhances p53-mediated transcriptional activation. 53BP1 contains two breast cancer susceptibility gene 1 COOH terminus (BRCT) motifs, which are present in several proteins involved in DNA repair and/or DNA damage–signaling pathways. Thus, we investigated the potential role of 53BP1 in DNA damage–signaling pathways. Here, we report that 53BP1 becomes hyperphosphorylated and forms discrete nuclear foci in response to DNA damage. These foci colocalize at all time points with phosphorylated H2AX (γ-H2AX), which has been previously demonstrated to localize at sites of DNA strand breaks. 53BP1 foci formation is not restricted to γ-radiation but is also detected in response to UV radiation as well as hydroxyurea, camptothecin, etoposide, and methylmethanesulfonate treatment. Several observations suggest that 53BP1 is regulated by ataxia telangiectasia mutated (ATM) after DNA damage. First, ATM-deficient cells show no 53BP1 hyperphosphorylation and reduced 53BP1 foci formation in response to γ-radiation compared with cells expressing wild-type ATM. Second, wortmannin treatment strongly inhibits γ-radiation–induced hyperphosphorylation and foci formation of 53BP1. Third, 53BP1 is readily phosphorylated by ATM in vitro. Taken together, these results suggest that 53BP1 is an ATM substrate that is involved early in the DNA damage–signaling pathways in mammalian cells.

We present results of a strong laser field interacting with a two-center system. The two centers consist of a donor and acceptor pair which are coupled by Fœrster–Dexter resonance energy transfer. The donor molecule is the indole chromophore in indolylethylenediaminetetraacetic acid (indolyl-EDTA) which chelates the lanthanide ion acceptor holmium (Ho3+). The changes in the donor lifetime consequent upon excitation of the acceptor were examined. Excitation of the lanthanide acceptor with a second laser field resulted in the appearance of a substantially increased lifetime component. This additional (new) lifetime component is on the order of a second, compared with the original nanosecond lifetime of the donor in its complex with the acceptor. This new component is not attributable to the spontaneous lifetime of the acceptor. Rather, we propose that the large lifetime is due to the interaction of the donor with the excited state acceptor.