A comparison of different lysis buffers to assess allele dropout from single cells for preimplantation genetic diagnosis

Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999 ). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (ΔF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. Copyright © 2001 John Wiley & Sons, Ltd.     

Greening the Fife Economy: Ecological Modernization as a Pathway for Local Economic Development

Progress in the greening of UK local government has been hampered by uncertainty about the relationship between resource use and environmental protection. Ecological modernizationmay offer the most appropriateparadigm to reconcile economic, social and environmentalinterpretationsof sustainability. The transition from corporate to strategic environmental management currently being attempted by some 'green' authorities can be construed as an attempt to promote ecological modernization in the form of institutional learning. Fife Council in Scotland provides an interesting case study. Progress with corporate environmental management has become bound up with a radical devolution of decision making. Efforts to link the authority's Sustainable Development Policy to a Local Agenda 21 programme have been initiated with the piloting of sustainability indicators.     

Volatiles of the honey bee larva initiate oogenesis in the parasitic mite Varroa destructor

Summary. Varroa reproduction is closely synchronized to the development of its host. In this study we present a new bioassay for field and laboratory tests to evaluate host factors triggering Varroa oogenesis. Female mites deprived of feeding activated oogenesis when perceiving larval volatiles. In laboratory assays the living L5-larva and pentane extracts of the larval cuticle had a clear activating effect. Wax and larval food did not elicit Varroa oogenesis. The activating components apparently are in the polar fraction of the cuticular volatiles. The consequences of this regulative mechanism for the host parasite relationship and prospects for further research are discussed.     

Upflow reactors for riparian zone denitrification

We used permeable reactive subsurface barriers consisting of a C source (wood particles), with very high hydraulic conductivities ( approximately 0.1-1 cm s(-1)), to provide high rates of riparian zone NO3-N removal at two field sites in an agricultural area of southwestern Ontario. At one site, a 0.73-m3 reactor containing fine wood particles was monitored for a 20-mo period and achieved a 33% reduction in mean influent NO3-N concentration of 11.5 mg L(-1) and a mean removal rate of 4.5 mg L(-1) d(-1) (0.7 g m(-2) d(-1)). At the second site, four smaller reactors (0.21 m3 each), two containing fine wood particles and two containing coarse wood particles, were monitored for a 4-mo period and were successful in attenuating mean influent NO3-N concentrations of 23.7 to 35.1 mg L(-1) by 41 to 63%. Mean reaction rates for the two coarse-particle reactors (3.2 and 7.8 mg L(-1) d(-1), or 1.5 and 3.4 g m(-2) d(-1)) were not significantly different (p > 0.2) than the rates observed in the two fine-particle reactors (5.0 and 9.9 mg L(-1) d(-1), or 1.8-3.5 g m(-2) d(-1)). A two-dimensional ground water flow model is used to illustrate how permeable reactive barriers such as these can be used to redirect ground water flow within riparian zones, potentially augmenting NO3- removal in this environment.     

Methods for treating soils contaminated with polychlorinated dibenzo-p-dioxins, dibenzofurans, and other polychlorinated aromatic compounds

This review provides a summary of methods for treating soils contaminated with polychlorinated aromatic compounds, especially polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). Unlike many other soil pollutants, PCDD/Fs bind tightly to the soil, which severely reduces the efficiency of most aqueous treatment procedures and leaves few realistic alternatives besides the traditional containment techniques (landfill, solidification/stabilization, and in situ vitrification). Incineration has long been, and still is, the most efficient destruction technique, with a removal efficiency of >99.9999%. However, supercritical water oxidation, base-catalyzed decomposition, steam distillation, and various extraction techniques, such as solvent and liquefied gas and subcritical water extraction, may provide removal efficiencies of >95%. Many of the alternative techniques are expected to be cheaper than incineration and may therefore be attractive for moderately polluted soils. However, some of them are at an early stage of development and need to be further tested before their true potential can be assessed.     

Spatial modelling of the probability of obtaining a detectable concentration of ammonia in Moreton Bay, Australia

Ammonia is an important water quality variable, which in excess, can be detrimental to waterways and their ecosystems. In the Ecosystem Health Monitoring Program in South-east Queensland ammonia is monitored monthly, however, often more than 50% of the ammonia observations in Moreton Bay are below detection limit, making it difficult to draw useful inferences. In this paper a clipped Gaussian random field is used to spatially model and map the probability of detectable concentration of ammonia. The methodology is applied to the Moreton Bay samples collected in February 2005. The results suggest that for this month the oceanic impacted areas have higher probability of detectable ammonia concentration than the areas closer to the main sources of anthropogenic inputs.     

Sorbents for the removal of airborne heavy metals from incinerator effluent

The effectiveness of inorganic alumino-silicate sorbents (alumina and kaolinite) to adsorb airborne lead and cadmium from the effluent stream of a simulated waste incinerator was studied. A 20 kW (68,000 BTU/h) flow reactor was used to achieve the temperature and residence times typical of a waste incinerator. Solutions containing lead or cadmium were introduced yielding airborne metals concentrations between 15 and 150 ppm. Gas samples were drawn into a particle impactor that collected the airborne particles and condensed phase metal aerosols, separating them into ten size ranges from 0.2 μm to greater than 10 μm. Metals to sorbent mass ratios between 0.03 and 1.56 were investigated. Scavenging efficiency increased as the ratio of sorbent to injected metal mass was increased. The scavenging efficiencies were as high as 76% for lead scavenged by kaolinite to as low as 14% for cadmium scavenged by alumina.