Microchip-based purification of DNA from biological samples

A microchip solid-phase extraction method for purification of DNA from biological samples, such as blood, is demonstrated. Silica beads were packed into glass microchips and the beads immobilized with sol-gel to provide a stable and reproducible solid phase onto which DNA could be adsorbed. Optimization of the DNA loading conditions established a higher DNA recovery at pH 6.1 than 7.6. This lower pH also allowed for the flow rate to be increased, resulting in a decrease in extraction time from 25 min to less than 15 min. Using this procedure, template genomic DNA from human whole blood was purified on the microchip platform with the only sample preparation being mixing of the blood with load buffer prior to loading on the microchip device. Comparison between the microchip SPE (microchipSPE) procedure and a commercial microcentrifuge method showed comparable amounts of PCR-amplifiable DNA could be isolated from cultures of Salmonella typhimurium. The greatest potential of the microchipSPE device was illustrated by purifying DNA from spores from the vaccine strain of Bacillus anthracis, where eventual integration of SPE, PCR, and separation on a single microdevice could potentially enable complete detection of the infectious agent in less than 30 min.     

Velocity fields of granular flows down a rough incline: a DEM investigation

This paper describes a statistical method to explore the velocity profiles of granular flows down rough inclines. Using 3D Discrete Element Method (DEM), granular material is released from a box onto a slope and allowed to flow indefinitely. Fluctuating velocity fields are observed, as particle motions become more dynamic and agitated. Linear regression is used to decompose the fluctuating velocity field into a best-fit velocity profile and a fluctuating component. Analysis shows that the slope inclination has a considerable influence on the rheology, in terms of both the fluctuating velocity and the shear rate of the flow.     

Controlled Nucleation of GaN Nanowires Grown with Molecular Beam Epitaxy

The location of GaN nanowires is controlled with essentially perfect selectivity using patterned SiN_x prior to molecular beam epitaxy growth. Nanowire growth is uniform within mask openings and absent on the mask surface for over 95% of the usable area of a 76 mm diameter substrate. The diameters of the resulting nanowires are controlled by the size of the mask openings. Openings of approximately 500 nm or less produce single nanowires with symmetrically faceted tips.     

A global collocation technique for the solution of opening mode crack problems

A review of various experimental and numerical techniques for determination of fracture mechanics calibration functions (i.e., the variation of K ₁ and CMOD with crack length) revealed that neither technique, employed independently, can determine K ₁, CMOD, and full field stresses in closed form over a wide range of crack lengths. To fill this void, a combined experimental/numerical collocation technique based on a series expansion of the modified Westergaard functions was developed. This technique uses both boundary conditions, known a priori, and interior stress field conditions, determined using a suitable experimental technique, for analysis of two dimensional, finite body, opening mode crack problems. This paper reports on an investigation of the accuracy of this technique and its sensitivity to errors in experimental data for a sample problem of practical interest.     

Private Justice: Losing Our Day in Court

In the early days of the dispute resolution movement—the ‘80s and ‘90s—the concern was often expressed that we were developing a system of “second‐class justice” for those who couldn't afford the courts. The wealthy, of course, would continue to have access to the “first‐class justice” that the public courts provided.     

Prevalence and numbers of Salmonella and Campylobacter spp. on raw,whole chickens in relation to sampling methods

Salmonella and Campylobacter continue to be major foodborne pathogens and raw poultry is considered to be an important source of these bacteria. In this study, the prevalence and numbers of Salmonella and Campylobacter spp. in relation to isolation/sampling methods were determined in 241 whole raw chickens purchased from retail outlets in England during the winters of 1998/1999 (101 chickens) and 1999/2000 (140 chickens). The packaging of the 140 chickens was also examined for the presence of the above pathogens. The prevalence and numbers of enterococci were examined in 21 of the 101 chickens. In total, Salmonella and Campylobacter spp. were present in 25% and 83% of the chickens, respectively. Salmonella were isolated from a sample representing both the inside and outside of the packaging in 19% of the chickens, while the corresponding figure for Campylobacter spp. was 56%. Both of these pathogens were isolated from the outside of the packaging in 6% of the chickens. Salmonella was more frequently isolated from samples containing chicken skin in comparison with those containing carcass-rinse fluid only. Two chickens (0.8%) were positive for Salmonella by direct enumeration methods with contamination levels of log10 3.8 and 4.5 colony forming units (cfu) per carcass, respectively. The most prevalent serotypes were S. Hadar, S. Enteritidis and S. Indiana and two different serotypes were identified in 5/20 salmonella-positive chickens. Resistance to at least one antibiotic was found in 70% of the strains, 46% were multiresistant (resistant to > or = four drugs) and 52% showed a lowered susceptibility to ciprofloxacin. The likelihood of isolating Campylobacter spp. from neck-skin, carcass-rinse or carcass-rinse plus whole skin samples was similar, Campylobacter spp. were found in higher levels in carcass-rinse or carcass-rinse plus whole skin samples than in neck-skin. The log10 cfu of Campylobacter spp. were 2.70-4.99 in 18% of the chickens and 5.00-6.99 in 20%. Campylobacter isolates (425) comprised Campylobacter jejuni (98%) and C. coli (2%) and 98 different sero/phagetypes of these two species were identified. Resistance to at least one antibiotic was found in 73% of the strains and 13% were multiresistant. Thirteen percent of the strains showed lowered susceptibility to ciprofloxacin, while 4.9% were resistant to erythromycin. Vancomycin-resistant enterococci (VRE), able to grow on agar containing 15 mg l(-1) vancomycin (VRE15), were present in 19 chickens. The log10 cfu of VRE15 was 2.90-3.99 in 10 chickens and between 4.00 and 4.99 in two chickens. The data presented here contribute to risk assessment and highlight the need to continue to emphasise the safe handling of raw retail poultry.     

Mycotoxin Detection in Human Samples from Patients Exposed to Environmental Molds

The goal of this study was to determine if selected mycotoxins (trichothecenes, aflatoxins, and ochratoxins) could be extracted and identified in human tissue and body fluids from patients exposed to toxin producing molds in their environment. Human urine and methanol extracted tissues and sputum were examined. Trichothecenes were tested using competitive ELISA techniques. Aflatoxins B1, B2, G1, and G2, and ochratoxin A were tested by using immunoaffinity columns and fluorometry. Test sensitivity and specificity were determined. Levels of detection for the various mycotoxins varied from 0.2 ppb for trichothecenes, 1.0 ppb for aflatoxins, and 2.0 ppb for ochratoxins. Trichothecene levels varied in urine, sputum, and tissue biopsies (lung, liver, brain) from undetectable (<0.2 ppb) to levels up to 18 ppb. Aflatoxin levels from the same types of tissues varied from 1.0 to 5.0 ppb. Ochratoxins isolated in the same type of tissues varied from 2.0 ppb to > 10.0 ppb. Negative control patients had no detectable mycotoxins in their tissues or fluids. These data show that mycotoxins can be detected in body fluids and human tissue from patients exposed to mycotoxin producing molds in the environment, and demonstrate which human tissues or fluids are the most likely to yield positive results.     

The consequences of recovery for the analysis of creep in steel structures

Creep strain and recovery data for ferritic steels are used to construct a simple model that separates permanent strain from visco-elastic, recoverable strain. The model is shown to be consistent with data from tests under gradually varying stress. The implications of this model are examined for the design analysis of representative structures. It is shown that the modelling of recovery is important in some circumstances and not in others.