Library

Notes: Abstract: Mice carrying a mutation in the gene encoding the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) have recently been produced to provide an animal model for Lesch-Nyhan disease. The current-studies were conducted to characterize the consequences of the mutation on the expression of HPRT and to characterize potential changes in brain purine content in these mutants. Our results indicate that the mutant animals have no detectable HPRT-immunoreactive material on western blots and no detectable HPRT enzyme activity in brain tissue homogenates, confirming that they are completely HPRT deficient (HPRT-). Despite the absence of HPRT-mediated purine salvage, the animals have apparently normal brain purine content. However, de novo purine synthesis, as measured by [14C]formate incorporation into brain purines, is accelerated four- to fivefold in the mutant animals. This increase in the synthesis of purines may protect the HPRT- mice from potential depletion of brain purines despite complete impairment of HPRT-mediated purine salvage.

Notes: Abstract: Rat-1 fibroblasts were transduced to express Drosophila choline acetyltransferase. The presence of an active enzyme in these cells (Rat-1/dChAT) was confirmed using various methods. Rat-1/dChAT fibroblasts released acetylcholine (ACh) into the culture medium. Moreover, intra-and extracellular levels of ACh could be increased by adding exogenous choline chloride. In addition, serum starvation or confluence-induced quiescence caused an 80% decrease in recombinant choline acetyltransferase activity (compared with actively growing cells). ACh release was also repressed in quiescent fibroblast cultures. Exogenous choline could mitigate the decrease in ACh release. These results indicate that Rat-1 fibroblasts can be genetically modified to produce ACh and that ACh release can be controlled by introducing choline into the culture medium. Furthermore, these data demonstrate that the expression of the retroviral promoter used in this study decreases with the onset of quiescence; however, exogenous choline can increase the amount of ACh released by quiescent fibroblasts.

Notes: Abstract : Lesch-Nyhan disease is a neurogenetic disorder caused by deficiency of the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT). Affected individuals exhibit a characteristic pattern of neurological and behavioral features attributable in part to dysfunction of basal ganglia dopamine systems. In the current studies, striatal dopamine loss was investigated in five different HPRT-deficient strains of mice carrying one of two different HPRT gene mutations. Caudoputamen dopamine concentrations were significantly reduced in all five of the strains, with deficits ranging from 50.7 to 61.1%. Mesolimbic dopamine was significantly reduced in only three of the five strains, with a range of 31.6-38.6%. The reduction of caudoputamen dopamine was age dependent, emerging between 4 and 12 weeks of age. Tyrosine hydroxylase and aromatic amino acid decarboxylase, two enzymes responsible for the synthesis of dopamine, were reduced by 22.4-37.3 and 22.2-43.1%, respectively. These results demonstrate that HPRT deficiency is strongly associated with a loss of basal ganglia dopamine. The magnitude of dopamine loss measurable is dependent on the genetic background of the mouse strain used, the basal ganglia sub-region examined, and the age of the animals at assessment.

Notes: Ion-assisted pulsed laser deposition has been used to produce films containing (approximately-greater-than)85% sp3-bonded cubic boron nitride (c-BN). By ablating from a target of hexagonal boron nitride (h-BN), BN films have been deposited on heated (50–800 °C) Si(100) surfaces. The growing films are irradiated with ions from a broad beam ion source operated with Ar and N2 source gasses. Successful c-BN synthesis has been confirmed by Fourier transform infrared (FTIR) spectroscopy, high-resolution transmission electron microscopy (TEM), selected-area electron diffraction, electron energy-loss spectroscopy, and x-ray diffraction. The films are polycrystalline and show grain sizes up to 300 A(ring). In addition, Rutherford backscattering, elastic recoil detection, and Auger electron spectroscopies have been used to further characterize the samples. The effects of varying ion current density, substrate growth temperature, growth time, and ion energy have been investigated. It is found that stoichiometric films with a high c-BN percentage can be grown between 150 and 500 °C. Below ∼150 °C, the c-BN percentage drops dramatically, and the deposited film is completely resputtered at the current densities and ablation deposition rates used. As the deposition temperature rises above ∼500 °C the c-BN percentage also drops, but less dramatically than at low temperatures.In addition, the IR-active c-BN mode narrows considerably as the deposition temperature increases, suggesting that the c-BN material has fewer defects or larger grain size. It is found that films with a high c-BN percentage are deposited only in a narrow window of ion/atom arrival values that are near unity at beam energies between 800 and 1200 eV. Below this window the deposited films have a low c-BN percentage, and above this window the deposited film is completely resputtered. Using FTIR analysis, it is found that the c-BN percentage in these samples is dependent upon growth time. The initial deposit is essentially all sp2-bonded material and sp3-bonded material forms above this layer. Consistently, cross-section TEM samples reveal this layer to consist of an amorphous BN layer (∼30 A(ring) thick) directly on the Si substrate followed by highly oriented turbostratic BN (∼300 A(ring) thick) and finally the c-BN layer. The h-BN/t-BN interfacial layer is oriented with the 002 basal planes perpendicular to the plane of the substrate. Importantly, the position of the c-BN IR phonon changes with growth time. Initially this mode appears near 1130 cm−1 and decreases with growth time to a constant value of 1085 cm−1. Since in bulk c-BN the IR mode appears at 1065 cm−1, a large compressive stress induced by the ion bombardment is suggested. Possible mechanisms are commented on for the conversion process to c-BN based upon the results.

Notes: We examine the crystallographic texture exhibited by cubic boron nitride (cBN) in thin films grown by ion-assisted deposition. Our analysis indicates that the cBN is preferentially oriented such that individual crystallites have at least one [111] direction lying in the plane of the film but are otherwise randomly oriented about (1) the substrate normal and (2) the in-plane cBN [111] axis. This preferential orientation is consistent with an alignment between the cBN {111} planes and the basal planes of the layer of highly oriented graphitic boron nitride that forms in the initial stages of film growth. ©1996 American Institute of Physics.

Notes: A microstructural study of boron nitride films grown by ion-assisted pulsed laser deposition is presented. Fourier transform infrared spectroscopy, electron-energy-loss spectroscopy, and electron-diffraction measurements indicate that within the ion-irradiated region on the substrate, the film consists of a high fraction of the cubic phase (cBN) with a small amount of the turbostratic phase; outside the irradiated region, only the turbostratic phase is detected. Conventional and high-resolution electron microscopic observations show that the cBN is in the form of twinned crystallites, up to 40 nm in diameter. Particulates, formed by the laser ablation process, reduce the yield of cBN in the irradiated regions by shadowing local areas from the ion beam. The films exhibit a layered structure with an approximately 30-nm-thick layer of oriented turbostratic material forming initially at the silicon substrate followed by the cBN. The observations of oriented turbostratic material and twinned cBN crystallites are discussed in relation to a previously proposed compressive stress-induced mechanism for cBN synthesis by ion-assisted film deposition.

Notes: Electron emission characteristics combined with in situ scanning electron microscope images have been measured on a series of amorphous carbon films grown by pulsed laser deposition. Uniform, reproducible current–voltage characteristics without morphological damage are only observed with sequential voltage ramps ≤5 V/s for anode-cathode gaps of 10–200 μm. The field threshold and emission barrier increase with laser energy density used during film growth. This dependence of emission parameters on film growth conditions appears to be correlated with the presence of conducting filaments extending through the film thickness. © 1997 American Institute of Physics.

Notes: We have grown diamond films on films of cubic boron nitride (cBN). The cBN films were grown on Si(100) substrates using ion-assisted pulsed laser deposition. Fourier transform infrared (FTIR) spectroscopy indicated that the BN films contained ∼75% sp3-bonded cBN. The as-grown cBN films were inserted with no surface pretreatment (e.g., abrading or scratching) into a conventional hot filament diamond reactor. In situ Raman spectroscopy was used to confirm diamond synthesis during growth. The nucleation density of the diamond films was estimated at 1×109/cm2, equivalent to or higher than the best values for scratched silicon substrates. In addition, we found that the cBN films were etched in the diamond reactor; a film thickness (approximately-greater-than)1500 A(ring) was required to prevent total film loss before diamond nucleation occurred. The presence of cBN under the diamond was established using FTIR spectroscopy and Auger electron spectroscopy.

Notes: We are studying the boron nitride system by using a pulsed excimer laser to ablate from hexagonal BN(hBN) targets to form BN films. We have deposited BN films on heated (600 °C) and room-temperature silicon (100) surface in an ambient background gas of N2. Fourier transform infrared (FTIR) reflection spectroscopy indicates that the films grown at high temperature have short-range sp2 (hexagonal-like) order, whereas films grown at room temperature are a mixture of sp3-bonded BN and sp2-bonded BN. Electron diffraction confirms the presence of cubic BN (cBN) material in the films grown at low temperature and the corresponding TEM lattice images show a grain size of ∼200 A(ring). The presence of cBN in the films correlates with laser energy density, with cubic material appearing around 2.4 mJ/cm2. Auger electron spectroscopy (AES) indicates that the films are nitrogen deficient.

Notes: High-resolution transmission electron microscopy (HRTEM) shows that amorphous-tetrahedral diamondlike carbon (a-tC) films grown by pulsed-laser deposition on Si(100) consist of three-to-four layers, depending on the growth energetics. We estimate the density of each layer using both HRTEM image contrast and Rutherford backscattering spectrometry. The first carbon layer and final surface layer have relatively low density. The bulk of the film between these two layers has higher density. For films grown under the most energetic conditions, there exists a superdense a-tC layer between the interface and bulk layers. The density of all four layers, and the thickness of the surface and interfacial layers, correlate well with the energetics of the depositing carbon species. © 1998 American Institute of Physics.