Perfluorocarbons in Chemical Biology

Perfluorocarbons, saturated carbon chains in which all the hydrogen atoms are replaced with fluorine, form a separate phase from both organic and aqueous solutions. Though perfluorinated compounds are not found in living systems, they can be used to modify biomolecules to confer orthogonal behavior within natural systems, such as improved stability, engineered assembly, and cell-permeability. Perfluorinated groups also provide handles for purification, mass spectrometry, and ¹⁹F NMR studies in complex environments. Herein, we describe how the unique properties of perfluorocarbons have been employed to understand and manipulate biological systems.     

Ultrawideband radar images of the surface disturbance produced by a submerged, mine-like object

This paper describes an ultrawideband, imaging radar designed to image the fine structure of the surface disturbances caused by submerged objects and presents images of the surface wake generated by a sphere immersed in a current. The system was deployed on a research pier on the Outer Banks of North Carolina as part of the Mine Surface Effects (MISE) program, a program designed to investigate the feasibility of using microwave radar to detect submerged mines. The radar achieves extremely high range resolution, approximately 4 cm, through the use of a short pulse with a 3 dB bandwidth of 3 GHz and a center frequency of 9 GHz. Azimuthal imaging is achieved through the use of a linear array of receive antennas and a time-domain beam-forming technique. Radar images from the MISE program are presented which show the surface disturbance generated by a 91 cm-diameter sphere immersed in a 40-60 cm/s current. A surface wake is clearly visible in the imagery, both during the low phase of the tide, when the sphere broached the surface, and during high tide, when the sphere was completely submerged. Closer examination of the imagery reveals that when the target was fully submerged, the backscatter from the wake was primarily composed of long-lived (>1.5 s), isolated echoes that translated downstream at a velocity close to that of the measured surface current. Photographs of the wake suggest that these echoes are generated by sharp-crested surface features that form where the slow moving water of the turbulent, subsurface wake impinges the surface.     

Projection pursuit classification of multiband polarimetric SARland images

Results are presented for an experiment utilizing a pastoral land scene with a variety of eight classes, imaged by the NRL dual band (X and L) polarimetric synthetic aperture radar (NUWSAR) at a spatial resolution of 1.2 m. Projection pursuit (PP) statistical analysis tools were applied to a set of simultaneous L-band and X-band fully polarized images (six independent channels) to demonstrate the utility of land classification at high spatial resolution from a light aircraft using SAR. The statistical confusion matrix was used as a quantitative optimization measure of classification. Samples of eight classes from a portion of the scene were used to define a training set, then PP tools were used for classification. It is clear that L-band and X-band fully polarized data view the classes in a significantly different manner, and each brings independent information to the analysis. These results are not meant to be exhaustive at this time but to demonstrate the utility of applying PP tools to multiband and polarization SAR data and to give an indication of the quality of classification one can achieve with moderately high spatial resolution SAR data using a light plane platform     

Sea surface velocity vector retrieval using dual-beam interferometry: first demonstration

The dual-beam interferometer consists of two interferometric synthetic aperture radars (InSARs), one squinted at 20/spl deg/ forward of broadside, and the other 20/spl deg/ aft, to allow measurement of vector surface velocity with only a single aircraft pass. Estimates of surface velocity vectors in the coastal region during high tidal flow are presented. The data were gathered over the barrier islands west of Fort Myers, Florida, as part of a March 2004 deployment. Whereas no detailed bathymetry data were available, high-quality aerial photography appears to be a useful tool in inferring bottom topography and possible current obstructions. The retrieved velocity field clearly follows the expected outflow pattern. While comparisons with tidal current magnitudes predicted by the U.S. National Ocean Service do reveal discrepancies of up to 0.5 m/s, these differences are most likely due to the contribution of ocean surface waves to the overall InSAR velocity measurement. Velocity retrievals for the same area based on the data from different tracks show good consistency. The results constitute the first demonstration of vector retrieval of the surface velocity field with a single-pass InSAR system and confirm the robustness of the dual-beam interferometry principle.     

A glycosylated Bence Jones protein and its autologous amyloid light chain containing potentially amyloidogenic residues

Amyloidosis is characterized by deposition of protein fibrils in various tissues. The wide variety of sequences of both amyloidogenic and non-amyloidogenic immunoglobulin light chains makes them a unique tool for addressing the importance of primary structure in the formation of insoluble fibrils. In this study, we have determined the primary structure of the kappa I immunoglobulin light chain from both the urinary Bence Jones protein and the deposited amyloid fibrils of a patient (MH) with primary amyloidosis. The sequence identity of urinary-excreted and tissue-deposited light chains excluded biclonality and somatic mutations and confirmed that the light chain existed in both a soluble and an insoluble form. Several residues have been previously reported to be significantly associated with amyloidogenic kappa chains. Many of these were found in the MH sequence, including Asp31, Asn45, Phe49, Gln55, His70, Asn/Gly93 and ProN/Val96, thereby supporting their potential role in fibrillogenesis. In addition, Asn20 and Pro60 of protein MH replaced the normally conserved Thr20 and Ser60. Asn20 was glycosylated in both the Bence Jones and the amyloid fibril protein MH. Cumulative effects of amyloid-associated residues and glycosylation might have rendered the immunoglobulin light chain MH prone to fibril formation.     

Phase signature radars

The identification and classification of radar targets is facilitated by a newly developed technique based on measurements of the differential phase shift of the target scattering obtained at harmonic phase-locked frequencies. These phase differences are independent of target range and relative motion. It is mathematically proved that for a smooth conducting body in the Rayleigh region the copolarized terms of the scattering matrix have zero phase shift. Based on this, the absolute target phase can be measured. The differential phase shift between two harmonic frequencies has been measured for various types of targets. The experimental results indicate that the differential phase shift is a useful parameter for classifying or identifying targets, and one that can be readily measured in practical radars.     

Experimental Perspectives on Direct Visualization of Endosomal Rupture

Endosomal escape continues to be a limiting factor in the therapeutic use of nanomaterials. Assays to visualize endosomal escape often do not decouple the endosomal/lysosomal disruption from the release of payload into the cytosol. Here, we discuss three approaches to directly probe endosomal/lysosomal rupture: calcein dye dilution, lysosome size quantification and endosome/lysosome membrane integrity visualized with a genetically engineered cell line. We apply the three assays to endosomes/lysosomes ruptured via osmotic pressure and photochemical internalization.     

Extensive intimal apolipoprotein A1-derived amyloid deposits in a patient with an apolipoprotein A1 mutation

With the aim of identifying molecules that are expressed specifically in the brain during neurogenesis, we tried to generate monoclonal antibodies which recognize molecules showing unique temporal expression patterns and molecular characteristics. We used a homogenate of the rat fetal forebrain (day 12 of fetal life, E12) as an immunogen, and antibodies which reacted with this preparation were screened by immunoblotting. One of the antibodies, Mab3C8, recognized a 100-kDa antigen that is enriched in fetal brain. This 100-kDa antigen was constantly expressed during fetal life (from E12 to E20) and became scarcely detectable two days after birth. The antigen was detected in the insoluble fraction of fetal brain and its isoelectric point ranged from 6 to 7, suggesting that it was a membrane-coupled glycoprotein. Analysis by glycosidase treatment and lectin blotting suggested that it was an O-linked glycoprotein with an alpha2,6 sialyl linkage. Thus, a molecule unique to the fetal brain, an O-linked sialoglycoprotein with a molecular mass of 100 kDa (FOG100), was found by generating an antibody.     

Wastewater lagoon sludge characteristics

Biological and physical characteristics of four wastewater treatment lagoon sludges were investigated. The four lagoon sludges possessed total solids (TS), volatile solids (VS) and pH values within the range of values reported in the literature for primary sludges. The lagoon sludges VS concentrations were between 49 and 80%: relatively high values considering a possible sludge age of several months or years. In addition, heavy metal concentrations in the four lagoon sludges were reported.