Validation of 64Cu-ATSM damaging DNA via high-LET Auger electron emission.

Radioactive copper (II) (diacetyl-bis N4-methylthiosemicarbazone) (Cu-ATSM) isotopes were originally developed for the imaging of hypoxia in tumors. Because the decay of a (64)Cu atom is emitting not only positrons but also Auger electrons, this radionuclide has great potential as a theranostic agent. However, the success of (64)Cu-ATSM internal radiation therapy would depend on the contribution of Auger electrons to tumor cell killing. Therefore, we designed a cell culture system to define the contributions to cell death from Auger electrons to support or refute our hypothesis that the majority of cell death from (64)Cu-ATSM is a result of high-LET Auger electrons and not positrons or other low-LET radiation. Chinese hamster ovary (CHO) wild type and DNA repair-deficient xrs5 cells were exposed to (64)Cu-ATSM during hypoxic conditions. Surviving fractions were compared with those surviving gamma-radiation, low-LET hadron radiation, and high-LET heavy ion exposure. The ratio of the D(10) values (doses required to achieve 10% cell survival) between CHO wild type and xrs5 cells suggested that (64)Cu-ATSM toxicity is similar to that of high-LET Carbon ion radiation (70 keV/µm). γH2AX foci assays confirmed DNA double-strand breaks and cluster damage by high-LET Auger electrons from (64)Cu decay, and complex types of chromosomal aberrations typical of high-LET radiation were observed after (64)Cu-ATSM exposure. The majority of cell death was caused by high-LET radiation. This work provides strong evidence that (64)Cu-ATSM damages DNA via high-LET Auger electrons, supporting further study and consideration of (64)Cu-ATSM as a cancer treatment modality for hypoxic tumors.

Filamentous Escherichia coli cells swimming in tapered microcapillaries.

This study analyzed the swimming characteristics of filamentous Escherichia coli cells inside tapered capillaries with a diameter decreasing from 700 µm to 4 µm and a mean body length of 27.8 µm ± 11.9 µm. Cells that were pre-oriented towards the narrower diameter section of the tapered capillary swam with high directional persistence, following conical-helix trajectories along the capillary wall. The confinement of the tapered capillary significantly diminished the mean swimming speed of filamentous cells when compared to their unrestricted mean swimming speed. The cell body rotation of individual filamentous bacteria decreased along the tapered direction, likely due to increased steric interactions with the capillary wall. Filamentous cells that swam under imposed flow rates ranging from 0.2 µl/min to 0.8 µl/min showed positive rheotaxis inside the 150 µm-350 µm diameter region of the tapered capillary. Depending on the imposed flow rate, none of the bacteria could advance beyond a critical diameter in the tapered capillary. This critical diameter is likely to be the position of the maximum shear rate they can tolerate without being flushed away. This work showed experimental evidence of how a simple flow constriction such as a tapered tube forms a hydrodynamic barrier that can deter the advance of bacterial rheotaxis.

Parameter Determination for Singlet Oxygen Modeling of BPD-Mediated PDT.

Photodynamic therapy (PDT) offers a cancer treatment modality capable of providing minimally invasive localized tumor necrosis. To accurately predict PDT treatment outcome based on pre-treatment patient specific parameters, an explicit dosimetry model is used to calculate apparent reacted 1O2 concentration ([1O2]rx) at varied radial distances from the activating light source inserted into tumor tissue and apparent singlet oxygen threshold concentration for necrosis ([1O2]rx, sd) for type-II PDT photosensitizers. Inputs into the model include a number of photosensitizer independent parameters as well as photosensitizer specific photochemical parameters ξ, σ, and ß. To determine the specific photochemical parameters of benzoporphyrin derivative monoacid A (BPD), mice were treated with BPD-PDT with varied light source strengths and treatment times. All photosensitizer independent inputs were assessed pre-treatment and average necrotic radius in treated tissue was determined post-treatment. Using the explicit dosimetry model, BPD specific ξ, σ, and ß photochemical parameters were determined which estimated necrotic radii similar to those observed in initial BPD-PDT treated mice using an optimization algorithm that minimizes the difference between the model and that of the measurements. Photochemical parameters for BPD are compared with those of other known photosensitizers, such as Photofrin. The determination of these BPD specific photochemical parameters provides necessary data for predictive treatment outcome in clinical BPD-PDT using the explicit dosimetry model.

The infrared spectrum of the matrix-isolated phenyl radical.

We have measured the infrared absorption spectrum of C(6)H(5), /X (2)A(1), in an Ar matrix at 10 K. The experimental frequencies (cm(-)(1)) and polarizations follow. a(1) modes: 3086, 3072, 3037, 1581, 1441, 1154, 1027, 997, 976, 605; b(1) modes: 972, 874, 706, 657, 416; b(2) modes: 3071, 3060, 1624, 1432, 1321, 1283, 1159, 1063, and 587. Three different methods have been used for the production of the phenyl radicals. Infrared absorption spectra of five deuterated isotopomers, C(6)D(5), p-C(6)H(4)D, p-C(6)HD(4), o-C(6)H(4)D, and m-C(6)H(4)D, were recorded to compare experimental frequency shifts with calculated (UB3LYP/cc-pVDZ) harmonic frequency shifts. The use of CO(2) or NO as internal standards enabled the experimental determination of absolute infrared intensities. The linear dichroism was measured with photooriented samples to establish experimental polarizations of each vibrational band. True gas-phase vibrational frequencies were estimated by considering the gas-to-matrix shifts and matrix inhomogeneous line broadening. The phenyl radical matrix frequencies listed above are within +/-1% of the gas-phase vibrational frequencies. The C(6)H(5) frequencies from this paper supersede our earlier values reported in J. Am. Chem. Soc. 1996, 118, 7400-7401. See also: http://ellison.colorado.edu/phenyl.

Characterization of biomass pyrolysis vapors with molecular beam, single photon ionization time-of-flight mass spectrometry.

A single photon ionization, molecular beam sampling, reflectron time-of-flight mass spectrometer (SPI/MBTOFMS) has been developed and used to study pyrolysis products from a selection of biomass materials. Spectra are characterized by high resolution and decreased fragmentation compared to electron-impact ionization mass spectra from related molecular beam mass spectrometer systems equipped with quadrupole mass analyzers.

Characterization and control of a multielement dual-frequency liquid-crystal device for high-speed adaptive optical wave-front correction.

Multielement nematic liquid-crystal devices have been used by others and ourselves for closed-loop adaptive control of optical wave-front distortions. Until recently the phase retardance of available devices could be controlled rapidly in only one direction. The phase retardance of the dual-frequency device can be controlled rapidly in both directions. Understanding the dynamics of the phase retardance change is critical to the development of a high-speed control algorithm. We describe measurements and experiments leading to the closed-loop control of a multielement dual-frequency liquid-crystal adaptive optic.

On the use of dual frequency nematic material for adaptive optics systems: first results of a closed-loop experiment.

The use of liquid crystal devices for wavefront control has been suggested and implemented by several authors. In this paper we report some preliminary results on the use of Nematic based liquid crystal devices. Several experimental efforts have been carried out in the past few months. One of the main aims was to characterize a new device that uses dual frequency nematic material in a closed loop arrangement.

Theory and laboratory demonstrations on the use of a nematic liquid-crystal phase modulator for controlled turbulence generation and adaptive optics.

We discuss the use of liquid-crystal phase modulators (LCPM's) both as a repeatable disturbance test source and as an adaptive optics corrector. LCPM's have the potential to induce controlled, repeatable, dynamic aberrations into optical systems at low cost, low complexity, and high flexibility. Because they are programmable and can be operated as transmissive elements, they can easily be inserted into the optical path of an adaptive optics system and used to generate a disturbance test source. When used as wave-front correctors they act as a piston-only segmented mirror and have a number of advantages. These include low operating power requirements, relatively low cost, and compact size. Laboratory experiments with a Meadowlark LCPM are presented. We first describe use of the LCPM as a repeatable disturbance generator for testing adaptive optics systems. We then describe a closed-loop adaptive optics system using the LCPM as the wave-front corrector. The adaptive optics system includes a Shack-Hartmann wave-front sensor operated with a zonal control algorithm.

Experimental measurements of estimator bias and the signal-to-noise ratio for deconvolution from wave-front sensing.

Deconvolution from wave-front sensing (DWFS) has been proposed as a method for achieving high-resolution images of astronomical objects from ground-based telescopes. The technique consists of the simultaneous measurement of a short-exposure focal-plane speckled image, as well as the wave front, by use of a Shack-Hartmann sensor placed at the pupil plane. In early studies it was suspected that some problems would occur in poor seeing conditions; however, it was usually assumed that the technique would work well as long as the wave-front sensor subaperture spacing was less than r(0) (L/r(0) < 1). Atmosphere-induced phase errors in the pupil of a telescope imaging system produce both phase errors and magnitude errors in the effective short-exposure optical transfer function (OTF) of the system. Recently it has been shown that the commonly used estimator for this technique produces biased estimates of the magnitude errors. The significance of this bias problem is that one cannot properly estimate or correct for the frame-to-frame fluctuations in the magnitude of the OTF but can do so only for fluctuations in the phase. An auxiliary estimate must also be used to correct for the mean value of the magnitude error. The inability to compensate for the magnitude fluctuations results in a signal-to-noise ratio (SNR) that is less favorable for the technique than was previously thought. In some situations simpler techniques, such as the Knox-Thompson and bispectrum methods, which require only speckle gram data from the focal plane of the imaging system, can produce better results. We present experimental measurements based on observations of bright stars and the Jovian moon Ganymede that confirm previous theoretical predictions.

Adaptive optics using a liquid crystal phase modulator in conjunction with a Shack-Hartmann wave front sensor and zonal control algorithm.

Multi-segment liquid crystal spatial light modulators have received much attention recently for use as high-precision wavefront control devices for use in astronomical and non-astronomical applications. They act much like piston only segmented deformable mirrors. In this paper we investigate the use of these devices in conjunction with a Shack-Hartmann wave-front sensor. Previous investigators have considered Zernike modal control algorithms. In this paper we consider a zonal algorithm in order to take advantage of high speed matrix multiply hardware which we have in hand.

Suitability of fetal tissues from spontaneous abortions and from ectopic pregnancies for transplantation. Human Fetal Tissue Working Group.

OBJECTIVE: To assess the potential availability and utility of fetal tissues obtained from spontaneous abortions and from ectopic pregnancies for human transplantation therapy. DESIGN: Tissue collection and analysis by personnel skilled in tissue banking. SETTING: Procurement programs in five tissue banks located in diverse geographical areas that are funded by the National Institutes of Health. PATIENTS: All women entering obstetric clinics during 1993 who consented to participate in the study. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Evaluation of the products of conception by standard developmental, histological, microbiological, and cytogenetic criteria. RESULTS: From 22,235 obstetric admissions, 1250 spontaneously aborted embryos and 247 products of ectopic pregnancies were obtained. Of these, seven embryos (0.5%) were potentially useful for human transplantation therapy. CONCLUSION: Fetal tissues from spontaneous abortions and from ectopic pregnancies are quite limited as feasible sources for human transplantation therapy.

Atmospheric structure function measurements with a Shack-Hartmann wave-front sensor.

The phase structure function has been used as a convenient way to characterize aberrations introduced on optical propagation by the atmosphere. It forms the theoretical basis for the calculation of such things as the long- and short-exposure atmospheric transfer function. The structure function is difficult to measure directly and is usually assumed to follow Kolmogorov statistics. We present here a technique for direct measurement of the structure function through the use of a Shack-Hartmann wave-front sensor. Experiments confirm that the atmosphere behaves according to Kolmogorov theory most of the time. However, some instances of non-Kolmogorov behavior have been noted.

Experimental measurements of the statistical properties of a monochromatic speckle pattern.

An experimental approach to the measurement of phase of a coherent optical field with a Shack-Hartmann sensor is presented. We demonstrate the usefulness of this sensor by verifying a theoretical model for the distribution of ray directions in a monochromatic speckle pattern.

Coherent image synthesis from wave-front sensor measurements of a nonimaged laser speckle field: a laboratory demonstrations.

We report what are to our knowledge the first coherent images recovered in the laboratory from measurements made with a Shack-Hartmann wave-front sensor of the phase and amplitude of a laser speckle wave front. We discuss the design of our wave-front sensor, which can obtain the phase and amplitude of an optical field with a single intensity measurement, and we point out a particular type of phase jump that cannot be detected by the Shack-Hartmann sensor. We also discuss implementations of this technique that may permit near-diffraction-limited imaging through turbulent media.

First astronomical application of postdetection turbulence compensation: images of alpha Aurigae, nu Ursae Majoris, and alpha Geminorum using self-referenced speckle holography.

We describe a postdetection turbulence compensation technique for obtaining high resolution imagery through the atmosphere. We present preliminary results fromfield experiments.

Morphogenesis and malformations of the skin NICHD/NIADDK research workshop.

Developmentally caused skin malformations constitute a spectrum of birth defects, some of which can be recognized prenatally by morphologic or biochemical means. The number of prenatally diagnosable skin diseases could be greatly expanded with an increased understanding of the molecular and cellular bases of skin development and the mechanisms that result in the generation of skin defects. The National Institute of Child Health and Human Development and the National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases, therefore, sponsored a workshop that recommended basic biologic studies combined with clinical investigations of normal and abnormal cutaneous development set forth in this article. Investigations resulting from these research recommendations are intended to contribute to the knowledge that should aid in the prevention of developmentally caused skin deformities.

Neural tube defects: a review of human and animal studies on the etiology of neural tube defects.

Although neural tube defects are a common congenital anomaly, their etiology is not known. Human studies have emphasized the pathology and epidemiology of the defects and suggest that in the majority of cases the etiology is multifactorial. Factors which appear possibly to be important are genetic predisposition, maternal illness, and fetal drug exposure. Animal studies have utilized naturally occurring neural tube defects and teratologically induced lesions. No animal model has been convincingly established as the equivalent of human neural tube defects. However, animal models have allowed investigation of the mechanisms of suggested human teratogens and determination of the pathogenesis of naturally occurring animal defects. Their most important contribution has been in furthering the understanding of the normal mechanisms of neural tube closure. It may be through this understanding that the etiology of human neural tube defects will be determined.