Synthesis and biological testing of aminoxyls designed for long-term retention by living cells

Owing to recent advances in electron paramagnetic resonance (EPR) imaging methodologies, it is now potentially possible to track and image, in real time in vivo, cells that had been tagged with aminoxyl spin probes. We had previously reported that living cells can accumulate 3-carboxy-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl [1] to high (millimolar) intracellular concentrations through passive incubation with the corresponding acetoxymethyl (AM) ester [2]. In the present study, we show that under physiological conditions aminoxyl [1] is rapidly extruded by cells through an organic anion transport mechanism, resulting in an intracellular exponential lifetime (t(1/e) or tau) of just 9.84 min at 37 degree C. Through successive rational structural modifications, we arrived at (2,2,5,5-tetramethylpyrrolidin-1-oxyl-3-ylmethyl)amine-N,N-diacetic acid [10], which can still be accumulated by cells to high intracellular concentrations, but which, with an intracellular exponential lifetime of tau]= 114 min, is well retained by cells for long periods of time, where one expects 14% retention even after 5 h. These results suggest that it should be feasible to use EPR imaging to perform in vivo tracking of populations of cells that have accumulated high intracellular levels of aminoxyls.     

Critical behavior of light in mode-locked lasers

Light is shown to exhibit critical and tricritical behavior in passively mode-locked lasers with externally injected pulses. It is a first and unique example of critical phenomena in a one-dimensional many-body light-mode system. The phase diagrams consist of regimes with continuous wave, driven parapulses, spontaneous pulses via mode condensation, and heterogeneous pulses, separated by phase transition lines that terminate with critical or tricritical points. Enhanced non-Gaussian fluctuations and collective dynamics are present at the critical and tricritical points, showing a mode system analog of the critical opalescence phenomenon. The critical exponents are calculated and shown to comply with the mean field theory, which is rigorous in the light system.     

Simultaneous optical coherence tomography--Indocyanine Green dye fluorescence imaging system for investigations of the eye's fundus

We have developed a dual-channel optical coherence tomography-Indocyanine Green dye (OCT-ICG) fluorescence system based on a previously reported ophthalmic OCT confocal imaging system. The confocal channel is tuned to the fluorescence wavelength range of the ICG, and light from the same optical source is used to generate the OCT image and to excite the ICG fluorescence. The system enables the clinician to visualize simultaneously en face OCT slices and corresponding ICG angiograms of the ocular fundus, displayed side by side. C-scan (constant depth) and B-scan (cross section) images are collected by a fast en face scan (T scan). The pixel-to-pixel correspondence between the OCT and angiography images allows the user to capture OCT B scans precisely at selected points on the ICG confocal images.     

Method for the rapid synthesis of highly functionalized 2-hydroxy-1-naphthoates. Syntheses of the naphthoic acid components of neocarzinostatin chromophore and N1999A2

We describe a four-step sequence for the synthesis of complex 2-hydroxy-1-naphthoic acids involving Z-selective olefination of benzaldehyde derivatives with a novel dioxolenone-containing phenyl phosphonate reagent, followed by dioxolenone cleavage with alkaline trifluoroethanol and oxidative cyclization (Mn(OAc)(3)) of the resultant trifluoroethyl beta-keto esters. [reaction: see text]     

Experimental and theoretical characterization of arsenite in water: insights into the coordination environment of As-O

Long-term exposure to arsenic in drinking water has been linked to cancer of the bladder, lungs, skin, kidney, nasal passages, liver, and prostate in humans. It is therefore important to understand the structural aspects of arsenic in water, as hydrated arsenic is most likely the initial form of the metalloid absorbed by cells. We present a detailed experimental and theoretical characterization of the coordination environment of hydrated arsenite. XANES analysis confirms As(III) is a stable redox form of the metalloid in solution. EXAFS analysis indicate, at neutral pH, arsenite has a nearest-neighbor coordination geometry of approximately 3 As-O bonds at an average bond length of 1.77 A, while at basic pH the nearest-neighbor coordination geometry shifts to a single short As-O bond at 1.69 A and two longer As-O bonds at 1.82 A. Long-range ligand scattering is present in all EXAFS samples; however, these data could not be fit with any degree of certainty. There is no XAS detectable interaction between As and antimony, suggesting they are not imported into cells as a multinuclear complex. XAS results were compared to a structural database of arsenite compounds to confirm that a 3 coordinate As-O complex for hydrated arsenite is the predominate species in solution. Finally, quantum chemical studies indicate arsenite in solution is solvated by 3 water molecules. These results indicate As(OH)3 as the most stable structure existing in solution at neutral pH; thus, ionic As transport does not appear to be involved in the cellular uptake process.     

NOISE 2 imaging system: seeing through scattering tissue with a reference point

We propose a fly-eye-like imaging system for seeing objects embedded in scattering media. Objects are recovered from many speckled images observed by a digital camera through a microlens array. Each microlens in the array generates a speckle image of the object buried between two layers of chicken breast tissue. In the computer each image is Fourier transformed jointly with an image of the speckled pointlike source captured under the same conditions. A set of the squared magnitudes of the Fourier-transformed pictures is accumulated to form a single average picture. This final picture is again Fourier transformed, resulting in the reconstruction of the hidden object.     

Sequential optical coherence tomography and confocal imaging

We report a system capable of sequentially acquiring two en-face images of different depth resolutions. The two images are generated by use of different principles, optical coherence tomography (OCT) and confocal microscopy, and have depth resolutions, at present, of better than 20 microm and over 0.12 mm, respectively. The lower-depth-resolution image is ideal for target positioning before collection of stacks of en-face OCT images. Switching between the two types of image by flipping an opaque screen in the reference arm, coupled with self-adjusting gain operation of avalanche photodiodes in the receiver. We illustrate the usefulness of the system by imaging a leaf and an optic nerve in vivo.     

Whole-cell biodetection of halogenated organic acids

A whole-cell bacterial sensor system for short-chain halo-organic acids was constructed, using 2-chloropropionic acid (2-CPA) as a model pollutant. An Escherichia coli host was transformed with a moderate-copy plasmid containing a fusion of two foreign genetic elements: (a) a promoter-containing segment of the Pseudomonas DL-DEX (DL-2-haloacid dehalogenase) encoding gene and (b) bioluminescence (luxCDABE) genes of Photorhabdus luminescens. The resulting construct, named MT1, responded to the presence of 2-CPA by dose-dependent light emission, in a highly specific albeit a very insensitive manner. Thus, while the desired concept was successfully demonstrated, further genetic work is needed in order to make such a construct practical for environmental monitoring purposes.     

Thin points for Brownian motion

Let Θ(x,r) denote the occupation measure of the ball of radius r centered at x for Brownian motion {W_t}_0≤t≤1 in . We prove that for any analytic set E in [0,1], we have

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, where dim_P(E) is the packing dimension of E. We deduce that for any a≥1, the Hausdorff dimension of the set of “thin points” x for which

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, is almost surely 2−2/a; this is the correct scaling to obtain a nondegenerate “multifractal spectrum” for the “thin” part of Brownian occupation measure. The methods of this paper differ considerably from those of our work on Brownian thick points, due to the high degree of correlation in the present case. To prove our results, we establish general criteria for determining which deterministic sets are hit by random fractals of ‘limsup type' in the presence of long-range correlations. The hitting criteria then yield lower bounds on Hausdorff dimension. This refines previous work of Khoshnevisan, Xiao and the second author, that required decay of correlations.     

Accurate Solution to Overdetermined Linear Equations with Errors Using L1 Norm Minimization

It has been known for many years that a robust solution to an overdetermined system of linear equations Ax b is obtained by minimizing the L1 norm of the residual error. A correct solution x to the linear system can often be obtained in this way, in spite of large errors (outliers) in some elements of the (m × n) matrix A and the data vector b. This is in contrast to a least squares solution, where even one large error will typically cause a large error in x. In this paper we give necessary and sufficient conditions that the correct solution is obtained when there are some errors in A and b. Based on the sufficient condition, it is shown that if k rows of [A b] contain large errors, the correct solution is guaranteed if (m – n)/n 2k/, where > 0, is a lower bound of singular values related to A. Since m typically represents the number of measurements, this inequality shows how many data points are needed to guarantee a correct solution in the presence of large errors in some of the data. This inequality is, in fact, an upper bound, and computational results are presented, which show that the correct solution will be obtained, with high probability, for much smaller values of m – n.