Real-time PCR with molecular beacons provides a highly accurate assay for detection of Tay-Sachs alleles in single cells

The results presented here provide the first single-cell genetic assay for Tay-Sachs disease based on real-time PCR. Individual lymphoblasts were lysed with an optimized lysis buffer and assayed using one pair of primers that amplifies both the wild type and 1278 + TATC Tay-Sachs alleles. The resulting amplicons were detected in real time with two molecular beacons each with a different colored fluorochrome. The kinetics of amplicon accumulation generate objective criteria by which to evaluate the validity of each reaction. The assay had an overall utility of 95%, based on the detection of at least one signal in 235 of the 248 attempted tests and an efficiency of 97%, as 7 of the 235 samples were excluded from further analysis for objective quantitative reasons. The accuracy of the assay was 99.1%, because 228 of 230 samples gave signals consistent with the genotype of the cells. Only two of the 135 heterozygous samples were allele drop-outs, a rate far lower than previously reported for single-cell Tay-Sachs assays using conventional methods of PCR. Copyright © 2002 John Wiley & Sons, Ltd.     

Preparing quality audit reports: Ten steps (and some leaps) to improve auditing

Good writing is never easy, but it is critical to the effectiveness of environmental audit reports. This article discusses ten issues to consider while producing audit reports that will enhance a company's quality effort.     

Rapid Screening and Identification of Airborne Carcinogens of Greatest Concern in California

This paper describes a method for establishing a priority list of airborne carcinogens within a state Jurisdiction. In this case it was necessary to identify, from among hundreds of potential candidates, the five to ten materials of greatest potential concern in California as airborne carcinogens.

Because no previous inventory of carcinogens in California existed, published lists, rankings, and assessments of national scope were used to identify candidates. By systematic manipulation and comparison of these data sources, 47 materials of some notoriety were chosen for closer scrutiny. This selection was pared to 22 candidates largely by eliminating those which had very little production and use in California. (Substances primarily used as pesticides were excluded from the scope of this study.) The remaining candidates were then ranked by additive . and multiplicative algorithms and by a panel of experts. The results of these rankings were combined to produce a single selection of 11 priority candidates. In alphabetical order, they are arsenic, asbestos, benzene, cadmium, carbon tetrachloride, chloroform, ethylene dibromide, ethylene dichloride, N-nitrosoamines, perchloroethylene, and polycyclic aromatic hydrocarbons. In continuing studies, a baseline emissions inventory is being prepared, and a source testing program is being designed.     

A short scale for measuring safety climate

A 6-item measure that assesses global work safety climate was validated using multiple samples each from a hospital and a nuclear energy population. Across all 14 samples the 6-item measure had acceptable internal consistency. The measure was associated with better adherence to safe work practices, reduced exposure to environmental stressors, the presence of more safety policies and procedures, a positive general organizational climate, and decreased accidents. As evidence for discriminant validity, safety climate was unrelated to most demographic measures and had relatively small relationships with sleeping problems and negative mood. Evidence suggests that this measure is a reliable and valid way to assess global safety climate.     

Cranial biomechanics of Diplodocus (Dinosauria, Sauropoda): testing hypotheses of feeding behaviour in an extinct megaherbivore

Sauropod dinosaurs were the largest terrestrial herbivores and pushed at the limits of vertebrate biomechanics and physiology. Sauropods exhibit high craniodental diversity in ecosystems where numerous species co-existed, leading to the hypothesis that this biodiversity is linked to niche subdivision driven by ecological specialisation. Here, we quantitatively investigate feeding behaviour hypotheses for the iconic sauropod Diplodocus. Biomechanical modelling, using finite element analysis, was used to examine the performance of the Diplodocus skull. Three feeding behaviours were modelled: muscle-driven static biting, branch stripping and bark stripping. The skull was found to be ‘over engineered’ for static biting, overall experiencing low stress with only the dentition enduring high stress. When branch stripping, the skull, similarly, is under low stress, with little appreciable difference between those models. When simulated for bark stripping, the skull experiences far greater stresses, especially in the teeth and at the jaw joint. Therefore, we refute the bark-stripping hypothesis, while the hypotheses of branch stripping and/or precision biting are both consistent with our findings, showing that branch stripping is a biomechanically plausible feeding behaviour for diplodocids. Interestingly, in all simulations, peak stress is observed in the premaxillary–maxillary ‘lateral plates’, supporting the hypothesis that these structures evolved to dissipate stress induced while feeding. These results lead us to conclude that the aberrant craniodental form of Diplodocus was adapted for food procurement rather than resisting high bite forces.     

Combined effect of adsorption and biodegradation of biological activated carbon on H2S biotrickling filtration

In order to evaluate the combined effect of adsorption and biodegradation of H(2)S on activated carbon surface in biotrickling filtration, four laboratory-scale biofiltration columns were operated simultaneously for 120h to investigate the mechanisms involved in treating synthetic H(2)S streams using biological activated carbon (BAC). The first three columns (A, B, C) contained a mixture of activated carbon and glass beads, with the carbons (BAC or virgin activated carbon (VAC)) and conditions (with or without liquid medium recirculation) differentiated. The last column (D) used 100% glass beads with liquid medium recirculation. Air streams containing 45ppmv H(2)S were passed through the columns at 4s of gas retention time (GRT) and liquid flow rate was set at 0.71mlmin(-1). Column D got its breakthrough in 3min of operation, indicating a negligible contribution of glass beads to the adsorption of H(2)S. The removal efficiency (RE) of Columns B and C using VAC dropped quickly to 30% within the first 8h, and afterwards continued to drop further but slowly. Column A using BAC stayed at 25% of RE throughout the operation time. A thorough investigation of the H(2)S oxidation products, i.e., various S species in both aqueous (recirculation media) and solid phases (BAC and VAC), was conducted using ICP-OES, IC, XRF, and CHNS elemental analyzer. BAC demonstrated a better performance than columns with adsorption only. Water film was found to enhance H(2)S removal. The percentage of sulphate in the total sulphur of the BAC system improved to twice of that of VAC system, indicating sulphate is the main product of H(2)S biofiltration. The observed pH drop in BAC system double confirmed that the presence of biodegradation in the biofilm over carbon surface did profound effect on the oxidation of H(2)S, compare to the systems with adsorption only.     

Arm regeneration in the field in Ophiocoma echinata (Echinodermata: Ophiuroidea): effects on body composition and its potential role in a reef food web

Eighteen percent and 47% of two populations of Ophiocoma echinata in the Florida Keys were undergoing regeneration. An individual would take approximately 720 days to completely regenerate three arms. Regeneration of three arms had a greater effect on gonad production in females than in males, and reduced storage material in the stomach of both. Regeneration in O. echinata was estimated to be 0.07 kJ m^-2 day^-1, which could provide the equivalent of 0.07% of primary production on a reef to higher trophic levels per day.     

Measurement of particle concentrations in a dental office

Particles in a dental office can be generated by a number of instruments, such as air-turbine handpieces, low-speed handpieces, ultrasonic scalers, bicarbonate polishers, polishing cups, as well as drilling and air sprays inside the oral cavity. This study examined the generation of particles during dental drilling and measured particle size, mass, and trace elements. The air sampling techniques included both continuous and integrated methods. The following particle continuous measurements were taken every minute: (1) size-selective particle number concentration (Climet); (2) total particle number concentration (PTRAK), and; (3) particle mass concentration (DustTrak). Integrated particle samples were collected for about 5 h on each of five sampling days, using a PM_2.5 sampler (ChemComb) for elemental/organic carbon analysis, and a PM₁₀ sampler (Harvard Impactor) for mass and elemental analyses. There was strong evidence that these procedures result in particle concentrations above background. The dental procedures produced number concentrations of relatively small particles (<0.5 μm) that were much higher than concentrations produced for the relatively larger particles (>0.5 μm). Also, these dental procedures caused significant elevation above background of certain trace elements (measured by X-ray fluorescence) but did not cause any elevation of elemental carbon (measured by thermal optical reflectance). Dental drilling procedures aerosolize saliva and products of drilling, producing particles small enough to penetrate deep into the lungs. The potential health impacts of the exposure of dental personnel to such particles need to be evaluated. Increased ventilation and personal breathing protection could be used to minimize harmful effects.