Genetic variation in captive koalas (Phascolarctos cinereus): parentage determination and individual identification

Highly repeatable randomly amplified polymorphic DNA (RAPD) markers were developed for parentage studies in the koala (Phascolarctos cinereus). Of the 25 RAPD primers screened, 5 (20.0%) produced 32 repeatable polymorphic RAPD bands (average/primer = 6.4 +/- 4.2). A high level of polymorphism was observed for each group of koalas (Featherdale, 71.9%; Lone Pine, 84.4%). All 25 koalas could be uniquely identified using either RAPD or microsatellite markers. Of the 32 RAPD markers generated in koalas, 25 were informative for parentage analyses. These RAPD markers successfully determined both parents to three offspring and a male parent to a fourth offspring. Paternity analysis (where the female parent is known) succeeded in assigning the correct male parent to seven offspring. Our RAPD-PCR method generates informative genetic markers that are useful for parentage determination and individual identification of captive koalas. This would provide genetic analysis to zoos and wildlife parks as a low-cost alternative to the more expensive microsatellite markers.     

Use of phenylalanine-to-tyrosine ratio determined by tandem mass spectrometry to improve newborn screening for phenylketonuria of early discharge specimens collected in the first 24 hours

We compared the screening interpretation of fluorometric analytical results for phenylketonuria (PKU) with tandem mass spectrometry (MS/MS) in filter paper blood spots collected from newborns <24 h of age. In MS/MS, both Phe and Tyr are quantified. Two hundred and eight blood spots collected from infants <24 h of age were retrieved from storage from the California newborn screening program. These samples had been categorized on the basis of fluorometric analysis as initial negative, initial positive for hyperphenylalaninemia with negative determination on recall, or initial positive for hyperphenylalaninemia and confirmed on follow up as PKU or variant hyperphenylalaninemia. The retrieved samples were analyzed in a blinded fashion using MS/MS. Correlation analysis of fluorometry vs MS/MS for Phe concentration was high, with a Pearson correlation coefficient of 0.817. When 180 micromol/L was used as the cutoff Phe concentration for MS/MS and 258 micromol/L was used as the cutoff for fluorometry, all infants with confirmed classical PKU and variant hyperphenylalaninemia were detected. MS/MS analysis reduced the number of false-positive results from 91 to 3. Simultaneous quantification of Phe and Tyr by MS/MS with the use of a cutoff Phe/Tyr molar ratio of 2.5 further reduced the number of false positives to 1. Samples from affected infants showed a discernible trend of increasing Phe concentration and Phe/Tyr molar ratio with age of collection. These results demonstrate the utility of MS/MS in the routine PKU screening of early-discharge newborns. MS/MS reduces the false-positive rate of fluorometric screening almost 100-fold because of the improved accuracy and precision of Phe measurement and simultaneous confirmation with the Phe/Tyr molar ratio. In addition to the detection of PKU, MS/MS can also detect other aminoacidopathies and disorders of fatty acid and organic acid metabolism with lower false-positive rates than other methods currently used in newborn screening programs.     

Static and dynamic properties of the water/amorphous silica interface: a model for the undissociated surface

Amorphous silica–water interfaces are found ubiquitously in nanoscale devices, including devices fabricated from silica as well as from silicon that acquire a surface oxide layer. The surface silanol groups serve as hydrogen-bonding sites for a variety of chemical species, and their reactivity enables convenient chemical modification, making silica surfaces strategic in bio-sensing applications. We have extended the popular BKS and SPC/E models for bulk silica and water to describe the hydrated, hydroxylated amorphous silica surface. The parameters of our model were determined using ab initio quantum chemical studies on small fragments. Our model will be useful in empirical potential studies, and as a starting point for ab initio molecular dynamics calculations. At this stage, we present a model for the undissociated surface. Our calculated value for the heat of immersion, 0.6Jm⁻², falls within the range of reported experimental values of 0.2–0.8Jm⁻². We also study the perturbation of water properties near the silica–water interface. The disordered surface is characterized by regions that are hydrophilic and hydrophobic, depending on the statistical variations in silanol group density. We report non-equilibrium molecular dynamics simulations of Poiseuille flow of water near an amorphous silica surface.     

Radiation sterilization compatibility of medical packaging materials

We have examined gamma sterilization compatibility for many of medical packaging materials. Techniques used include thermal analysis, physical testing, optical property evaluations, and failure analysis. For a variety of PVC formulations, gamma radiation often leads to discoloration. This results from conjugated poly-ene sequences formed through dehydrohalogenation degradation during ionizing radiation. In addition to unacceptable color formation, excessive pH shifts and high extractables are often observed. For most polylefins, radiations was found to deplete the antioxidant package and lead to discoloration and/or mechanical failures. For polypropylenes, the degradation can lead to the well-publicized catastrophic failures during post radiation shelf life storage. These examples and methods to ensure radiation compatibility and shelf-life stability are presented.     

An all-implanted, self-aligned, GaAs JFET with a nonalloyedW/p+-GaAs ohmic gate contact

We describe a self-aligned, refractory metal gate contact, enhancement mode, GaAs junction field effect transistor (JFET) where all impurity doping was done by ion implantation. Processing conditions are presented for realizing a high gate turn-on voltage (~1.0 V at 1 mA/mm of gate current) relative to GaAs MESFET's. The high gate turn-on voltage is the result of optimizing the p⁺-gate implant and anneal to achieve a nonalloyed ohmic contact between the implanted p⁺-GaAs and the sputter deposited tungsten gate contact. Initial nominally 1.0 μm×50 μm n-JFET's have a transconductance of 85 mS/mm and f_t of 11.4 GHz     

Comparison of Mg and Zn gate implants for GaAs n-channel junction field effect transistors

Zinc and magnesium implants into GaAs were profiled with secondary ion mass spectroscopy and etching capacitance-voltage to measure the as-implanted and annealed profiles for the eventual formation of shallow p⁺/n junction gates for junction field effect transistors (JFETs). The larger mass of the zinc ions results in shorter projected range with significantly less tailing than magnesium implants. High dose, shallow zinc implants annealed under tungsten gate metal showed good activation with negligible diffusion. The improved profile of the zinc implant, as compared to a similar magnesium implant, allowed a tighter JFET design with increased performance. Zn gated n-channel enhancement mode GaAs JFETs with 0.9 μm gate lengths showed transconductances up to 200 mS/ mm with a f_t of 18 GHZ and a f_max of 37 GHz. The performance of these self-aligned fully implanted JFETs compare favorably with comparably sized implanted MESFETs.     

A motility function for the paraflagellar rod of Leishmania parasites revealed by PFR-2 gene knockouts

We demonstrate a functional role for the paraflagellar rod (PFR) in motility of Leishmania mexicana. The PFR is a complex cytoskeletal structure running parallel to the axoneme in the flagella of kinetoplastid protozoa. The PFR is composed of a latticework of protein filaments whose major constituents are two related proteins (PFR-1 and PFR-2 in Leishmania). The molecular details of their assembly into PFR filaments are unknown as is the biological function of the PFR. As an approach to understanding the structure and function of the PFR in Leishmania, we made L. mexicana null mutants of PFR-2. PFR-2 minus parasites grow and divide normally in culture and still express the PFR-1 protein. They lack most of the PFR structure demonstrating that the PFR-2 protein is an essential constituent of the PFR. Detailed ultrastructural analysis of the PFR-2 null mutant reveals the presence of a residual inner substructure of the PFR which contains PFR-1 protein, indicating that PFR-1 can polymerize in the absence of PFR-2. The PFR-2 null mutant displays pronounced changes in flagellar beat waveform and forward swimming velocity, compared to wild type parasites consistent with decreased internal elastic bending resistance in PFR-lacking flagella, and indicating a functional role for the PFR in the motility of Leishmania.     

On discontinuous Galerkin methods

Discontinuous Galerkin methods have received considerable attention in recent years for problems in which advection and diffusion terms are present. Several alternatives for treating the diffusion and advective fluxes have been introduced. This report summarizes some of the methods that have been proposed. Several numerical examples are included in the paper. These present discontinuous Galerkin solutions of one‐dimensional problems with a scalar variable. Results are presented for diffusion–reaction problems and advection–diffusion problems. We discuss the performance of various formulations with respect to accuracy as well as stability of the method. Copyright © 2003 John Wiley & Sons, Ltd.     

High performance GaAs JFET with shallow implanted Cd-gate

Shallow p⁺-regions in GaAs, formed by Cd ion implantation, have been used as the gate region for GaAs JFETs. 0.7 μm gate length JFETs demonstrated a transconductance of 165 mS/mm a saturation current of 130 mA/mm, an f_t of 26 GHz, and an f _max of 42 GHz. These frequency metrics are superior to previous Zn-gate JFETs of similar dimensions     

A simple general model for echelon overhaul and repair

In echelon overhaul and repair, failed or time-expired machines are removed and replaced by overhauled ones. The removed machines are sent to the rear echelon, typically the maker's works, to be overhauled to ‘good-as-new’. The paper presents a simple model for determining economic overhaul intervals in the presence of minor and major failures, PM and inspection of accessible and inaccessible components, and then goes on to consider how many machines should, for maximum long-term economy, be owned by the operator. The economic implications of an echelon policy for both the owners and the makers are briefly discussed. It is argued that simple models promply applied are often adequate for comparing competing systems and policies. An example involving gas-turbines in off-shore oil-rigs demonstrates the method generally, and in particular shows that the advantages may be dissipated if the overhaulers take too long to process overhauled machines.