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.     

Exposure to radiation from global system for mobile communications at 1,800 MHz significantly changes gene expression in rat hippocampus and cortex

We have earlier shown that radio frequency electromagnetic fields can cause significant leakage of albumin through the blood–brain barrier of exposed rats as compared to non-exposed rats, and also significant neuronal damage in rat brains several weeks after a 2 h exposure to a mobile phone, at 915 MHz with a global system for mobile communications (GSM) frequency modulation, at whole-body specific absorption rate values (SAR) of 200, 20, 2, and 0.2 mW/kg. We have now studied whether 6 h of exposure to the radiation from a GSM mobile test phone at 1,800 MHz (at a whole-body SAR-value of 13 mW/kg, corresponding to a brain SAR-value of 30 mW/kg) has an effect upon the gene expression pattern in rat brain cortex and hippocampus—areas where we have observed albumin leakage from capillaries into neurons and neuronal damage. Microarray analysis of 31,099 rat genes, including splicing variants, was performed in cortex and hippocampus of 8 Fischer 344 rats, 4 animals exposed to global system for mobile communications electromagnetic fields for 6 h in an anechoic chamber, one rat at a time, and 4 controls kept as long in the same anechoic chamber without exposure, also in this case one rat at a time. Gene ontology analysis (using the gene ontology categories biological processes, molecular functions, and cell components) of the differentially expressed genes of the exposed animals versus the control group revealed the following highly significant altered gene categories in both cortex and hippocampus: extracellular region, signal transducer activity, intrinsic to membrane, and integral to membrane. The fact that most of these categories are connected with membrane functions may have a relation to our earlier observation of albumin transport through brain capillaries.     

Barriers Beyond the Partners: Bureaucratic and Political Constraints to Implementing Joint Forest Management in Tamil Nadu,India

Whereas past research has treated co-management of common pool resources as if villagers and project implementing authorities were the only relevant actors, numerous external factors beyond the control of these two partners create barriers to successful co-management. This paper draws on discussions with Forest Department officials to examine the influence of these forces on the outcomes of Joint Forest Management (JFM) in Tamil Nadu, India. An empirical inquiry into the operational aspects of JFM indicates the important roles of political parties, powerful people, and other state institutions and functionaries as well as the flow of foreign funding. Further, the strong demand by local people for socio-economic development interventions as opposed to improvement of degraded forests belittles the role of the Forest Department relative to other departments. Numerous other conditioning factors and relationships are explored. The authors call for reforms in public governance to allow better participation of all the actors involved for this participatory management approach to succeed and sustain.     

Perceptions toward stewardship among residents living near U.S. Department of Energy Nuclear Facilities

Although stewardship has been widely defined and used in environmental management and planning, there is a dearth of studies that describe how the lay public perceives this concept. A national sample of residents in 14 states who live near DOE nuclear facilities were interviewed to delineate public understanding and awareness of the stewardship program of the U.S. Department of Energy (DOE). This study discusses the findings of the survey and discusses how institutional trust influences public participation and resident’s choices of potential stewards. Almost 40% of the respondents could not define stewardship; those who did, believed that ‘responsibility,’ ‘management,’ and ‘accountability’ are key elements of stewardship. In addition, about a third of the respondents identified Federal groups and the DOE as potential stewards. Readers should send their comments on this paper to: BhaskarNath@aol.com within 3 months of publication of this issue