Mutational spectrum induced by chromium(III) in shuttle vectors replicated in human cells: relationship to Cr(III)-DNA interactions

Trivalent chromium (Cr(III)), the ultimate species of chromium (VI) intracellular reduction, can associate with DNA forming Cr(III) monoadducts and DNA-DNA cross-links. However, the mutational specificity of Cr(III) has not been determined partly because Cr(III) has difficulty entering cells. In this study, we have characterized the types of Cr(III)-induced DNA lesions in two buffer systems and the mutational spectrum of Cr(III)-treated shuttle vectors replicated in human 293 cells. Plasmids were treated with Cr(III) in buffers consisting of either 10 mM potassium phosphate, pH 7.5 (designated as KP), or 0.2 mM Tris-HCl and 20 microM EDTA, pH 7.4 (designated as TE/50). The amounts of Cr(III) bound to DNA increased as Cr(III) concentration increased in both buffers; these Cr(III)-DNA associations were stable in both buffers during a 24-h dialysis. The electrophoretic mobility of supercoiled DNA was markedly retarded in samples treated with Cr(III) in TE/50 but not KP buffer, suggesting that Cr(III)-mediated DNA-DNA cross-links were generated in TE/50 but did not form in KP. Polymerase-stop assay showed that DNA polymerases were mostly blocked at the 3' adjacent bases of guanines on templates treated with Cr(III) in TE/50 but were not observed on those treated in KP. The signals of Cr(III)-mediated cross-links generated in TE/50 buffers were reduced when they were dialyzed against KP buffers. Similarly, Cr(III)-DNA monoadducts formed in KP were converted to primer-template cross-links by dialysis against TE/50. The mutation frequency of Cr(III) in the supF gene of pSP189 or pZ189 shuttle vectors replicated in human cells increased as Cr(III) concentration increased in both buffers. DNA sequencing analysis showed that single-base substitutions (61-68%), two-base substitutions (3-5%), and deletions (21-34%) were induced in similar frequencies in plasmids treated with Cr(III) in either TE/ 50 or KP. The Cr(III)-induced base-substitution hot spots are different from those occurring spontaneously. Cr(III) enhances G.C base substitutions, particularly G.C-->C.G transversions, at 5'GA, 5'CG, and 5'AG sites. Base-substitution hot spots did not correlate with strong polymerase-stop sites, suggesting that base substitutions are derived from Cr(III) monoadducts, not from DNA-DNA cross-links.     

A New Method for Microlens Fabrication by a Heating Encapsulated Air Process

This letter reports a new method to fabricate thin microlens array on a silicon substrate by a heating encapsulated air process. We use silicon bulk micromachining, wafer-to-wafer bonding, and photoresist (PR) spin coating to achieve the air sealing process. Under a heating process, the PR filled in the micro-through-hole of cap wafer is compressed by the thermal expansion of the sealed air to form a thin microlens with out-of-plane sphere shape. By adjusting the heating temperature and the sealed air volume, the curvature and size of the lens are controllable. A typical microlens with a diameter of 1475 mum and sag height of 486 mum was fabricated. The calculated radius of curvature and focal length are about 800 and 1200 mum, respectively. The fabrication provides an alternative way to manufacture thin microlens or microlens mold serve as master elements for replication     

Metastatic thyroid carcinoma arising from congenital goiter due to mutation in the thyroperoxidase gene

A very large cervical tumor that extended to the upper mediastinum was seen in a newborn after an uneventful pregnancy. The computed axial tomography scan confirmed the presence of a solid mass with precise limits and scattered foci of calcifications situated in the anterolateral region of the neck. The infant underwent thyroidectomy on the seventh day after birth. Pathological examination revealed a follicular carcinoma of the thyroid and probable dyshormonogenetic hyperplastic goiter. At 5 months of age, whole body scans indicated the presence of lung and bone metastases, which were treated with therapeutic doses of radioiodine. Genomic DNA was obtained from the newborn, her parents, her paternal aunt, and her paternal grandparents. Denaturing gradient gel electrophoresis analysis of PCR fragments corresponding to exon 14 of the thyroid peroxidase (TPO) gene indicated the presence of a mutant TPO allele present in the propositus, her father, and her paternal grandmother. Sequencing of the TPO gene demonstrated a mutation resulting from an insertion of a single extra cytosine in a stretch of seven cytosines at positions 2505-2511. The insertion caused a frame shift and a stop signal in exon 16. This sequence would translate into a structurally modified and probably inactive TPO protein. We conclude that the aggressive thyroid metastatic carcinoma arose from a dyshormonogenetic goiter caused by a defective TPO protein.     

Evaluation of Organometallic Compounds for Gamma Detection in Plastic Scintillators

A series of copolymer of vinyl toluene containing various organometallic compounds have recently been synthesized. By taking advantage of the weak fluorescent characteristics of the organometallic compounds and the better understanding of the energy transfer and quenching process (J. Phys. Chem. 68:300 (1964)), it is possible to prepare organometallic scintillation plastics that may be useful for gamma detection. A 20% triphenyltinstyrene and vinyl toluene copolymer containing 3% PBD and 0.05% POPOP have now been evaluated at the Health and Safety Laboratory, New York Operation Office. The sample size was only 1 ?" by 1" thick cylinder. In general, the pulse height is 45-50% of that observed from anthracene, and the cross section is still less than that observed for sodium iodide. However, the photopeaks are definitely resolvable when evaluated for Mercury-203, Cadmium-109, and Cesium-137. In the last case, the energy resolution for 662 MeV photons lies between 16-19%.     

Exploratory investigations of hypervelocity intact capture spectroscopy

The ability to capture hypervelocity projectiles intact opens a new technique available for hypervelocity research. A determination of the reactions taking place between the projectile and the capture medium during the process of intact capture is extremely important to an understanding of the intact capture phenomenon, to improving the capture technique, and to developing a theory describing the phenomenon. The intact capture of hypervelocity projectiles by underdense media generates spectra, characteristic of the material species of projectile and capture medium involved. Initial exploratory results into real-time characterization of hypervelocity intact capture techniques by spectroscopy include ultra-violet and visible spectra obtained by use of reflecting gratings, transmitting gratings, and prisms, and recorded by photographic and electronic means. Spectrometry proved to be a valuable real-time diagnostic tool for hypervelocity intact capture events, offering understanding of the interactions of the projectile and the capture medium during the initial period and providing information not obtainable by other characterizations. Preliminary results and analyses of spectra produced by the intact capture of hypervelocity aluminum spheres in polyethylene (PE), polystyrene (PS), and polyurethane (PU) foams are presented. Included are tentative emission species identifications, as well as gray body temperatures produced in the intact capture process.     

Sensory recoding via neural synchronization: integrating hue and luminance into chromatic brightness and saturation

If neural spike trains carry information in the frequency and timing of the spikes, then neural interactions--such as oscillatory synchronization--that alter spike frequency and timing can alter the encoded information. Using coupled oscillator theory, we show that synchronization-based processing can be used to integrate sensory information, resulting in new second-order sensory percepts signaled by the compromise frequency of the coupled system. If the signals to be coupled are nonlinearly compressed, the coupled system behaves as if it signals the product or ratio of the uncoupled signals, e.g., chromatic brightness can be signaled by the compromise frequency of coupled neurons responding to hue and luminance, and chromatic saturation can be signaled by the coupled frequency of neurons responding to hue and brightness, with a power- (Stevens's) law scaling like that observed psychophysically. These emergent properties of coupled sensory systems are intriguing because multiplicative processing and power-law scaling are fundamental aspects of sensory processing.