Evaluation of sieving polymers for fast, reproducible electrophoretic analysis of short tandem repeats (STR) in capillaries

Efficient capillary electrophoretic STR analysis requires rapid, reproducible and robust separation of DNA fragments with reasonable capillary longevity--this is currently accomplished using proprietary commercial polymeric sieving matrices specifically developed for this separation. These matrices, while effective, are costly and do not provide adequate resolution of STR DNA fragments in capillaries with shorter effective separation lengths, increasing the time required to accomplish the separation and minimizing the potential extrapolation to other miniaturized platforms. As the forensic community looks toward next generation microchip technology as a means of processing casework more rapidly, new sieving polymers need to be evaluated for utilization in this platform. The research presented here describes the assessment of commercially-available polymeric sieving matrices for STR analysis, with consideration given to feasibility of incorporation into a microdevice. Polymer composition, molecular weight, and concentration were evaluated, along with an assessment of the effects of buffer composition, separation temperature, and capillary length. These variables were evaluated individually or collectively on the ability to resolve STR DNA fragments and the reproducibility of the separations and the results compared to a proprietary commercial product. A 600,000 Da MW poly(ethylene oxide) (PEO) solution at a 3% (w/v) concentration was determined to be the most suitable matrix for these separations. This polymer, in coated capillaries, provided highly robust and reproducible separations, with near baseline resolution of fragments having single base differences. Reductions in the temperature of the separation, from 60 degrees C to 40 degrees C, and the urea concentration of the buffer, from 7 M to 3.5 M, provided increased longevity of the PEO polymer for repeated separations. Comparison of this polymer with currently specified commercial products used for STR analysis showed that the optimized PEO matrix provided superior separations under all conditions tested. In addition, PEO could be utilized in shorter capillary systems, with a concurrent decrease in analysis time, highlighting its potential for use in shortened capillary or microdevice systems.     

Proficiency testing (external quality assessment) of drug detection in oral fluid

Eighteen external quality assessment (proficiency testing) samples were prepared from client specimens collected with the Intercept oral fluid collection device and by spiking drug-free oral fluid. Samples were circulated in pairs at quarterly intervals to 13 UK and USA based laboratories for analysis by a panel of OraSure micro-plate Intercept enzyme immunoassay kits and hyphenated mass spectrophotometric techniques. During the survey, there was a single case of non-specificity in a false report for methadone. The major errors were of lack of sensitivity relative to the concentration thresholds specified for the immunoassays. The sensitivity for overall "present"/"not found" reports calculated as true positives/(true positives+false negatives) were for the amphetamine specific assay 50%, methyl-amphetamines 93%, barbiturates 64%, cannabinoids 73%, cocaine and metabolites 100%, benzodiazepines 69%, methadone 95%, opiates 79% (opiates excluding oxycodone 93%), phencyclidine 93% and human gamma-globulin 97%. A small number of the sensitivity errors were attributable to errors in chromatographic confirmation techniques.     

Mandarins and kibitzers:men in and around the trenches of political power in Ottawa*

Book reviewed in this article: My Life and Work, A Happy Partnership: Memoirs of J. A. Corry. A Man of Influence: Norman A. Robertson and Canadian Statecraft, 1929–1968. The Ottawa Men: The Civil Service Mandarins 1935–1957.     

POLITICS, POLICY MAKING AND PRESSURE GROUPS: SOME SUGGESTIONS FOR REFORM OF THE AUSTRALIAN POLITICAL SYSTEM

Abstract: In Australia, current policy making is characterized by immobilism, drift and "pluralistic stagnation". The problem is not lack of technical expertise, but the mobilization of political consent in a polity in which pressure groups have proliferated. In the post-war period, the welfare state/managed economy has transformed the role of government and the potential power of pressure groups. New networks of interdependency have developed. Some of the principal groups are beneficiaries of redistributive programs, producer groups, state and local government agencies and professionals in government. Their potential power has been made actual by a new participative ethic. The post-materialist values of which this is part also result in the growth of groups focusing on quality of life and moral issues. In this context the political parties are losing their central place in the polity and the centralized organs of executive government appear over-secretive and aloof. Reforms are needed to realign institutions with the new cultural and structural realities. Broader interest group involvement is needed. Parliament, through a committee system, could play a major role and its historical function as custodian of the public interest should be revived. Departments must find new ways to involve more groups in policy formulation and implementation. Information must be shared more widely and policy analysts and advisors should work more closely with groups in defining issues, negotiating alternatives and encouraging the development of supportive coalitions. New "intelligence" and "outreach" functions within the public service are required, and the existing approaches of policy units and research bureaux should be remodelled.     

Differentiation of black gel inks using optical and chemical techniques

Gel ink pens have become a common writing instrument in the United States. Questioned document examiners often attempt to optically differentiate gel inks from each other and from other non-ballpoint ink writings (e.g., those from roller-ball pens). Since early formulations were primarily pigment-based, they do not elute when analyzed by thin-layer chromatography. However, recent gel ink formulations (i.e., within the past five years) include dye-based inks that can be easily separated. This study differentiates black gel inks using optical and chemical techniques. The techniques include: microscopy, visible and near infrared reflectance, near infrared luminescence, thin-layer chromatography (TLC), spot tests, and gas chromatography/mass spectrometry (GC/MS). As a result of this study a flow chart has been developed allowing for a systematic determination of a questioned ink. In addition, an analysis of volatile compounds found in gel inks revealed that there are some unique ingredients that may be found in gel inks that are not typically found in other non-ballpoint inks.