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.     

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     

Effects of ocean temperature on the southern range limits of two understory kelps, <Emphasis Type="Italic">Pterygophora californica</Emphasis> and <Emphasis Type="Italic">Eisenia arborea</Emphasis>, at multiple life-stages

Environmental factors have long been shown to influence species distributions, with range limits often resulting from environmental stressors exceeding organism tolerances. However, these abiotic factors may differentially affect species with multiple life-history stages. Between September 2004 and January 2006, the roles of temperature and nutrient availability in explaining the southern distributions of two understory kelps, Pterygophora californica and Eisenia arborea (Phaeophyceae, Laminariales), were investigated along the coast of California, USA and the Baja California Peninsula, Mexico, by limiting either: (a) tissue nitrogen uptake and storage by adult sporophytes during periods of elevated temperature, and/or (b) production of embryonic sporophytes by microscopic gametophytes. Results suggest that while adult sporophytes of both species are tolerant of high temperatures and low nutrients, reproduction by their microscopic stages is not. Specifically, while E. arborea produced embryonic sporophytes at both 12 and 18°C, temperatures commonly observed throughout the southern portion of its range, P. californica produced sporophytes at 12 but not at 18°C. As a result, it appears that the southern distribution of P. californica, which ends in northern Baja California, Mexico, may be limited by temperature acting on its microscopic stages. In contrast, the ability of E. arborea’s microscopic and adult stages to tolerate elevated temperatures allows it to persist in the warmer southern waters of Baja California, as well as to the north along the California coast where both species co-occur.     

GCM-based analysis of water availability along the Tone River and Tokyo Metropolitan Area under climatic changes

Tone River supplies most of the water requirements of the Tokyo Metropolitan Area (TMA). Lowering of Tone flow and yearly fluctuation, however, is causing water shortage along TMA nowadays. This study investigated the future water availability scenarios under climatic changes. A state-of-the-art approach to utilize the output of several GCM has been demonstrated to investigate the future water availability scenarios for TMA from the Tone River. An integrated modeling approach for water balance considering several hydrological risk indices was adopted to quantify the future changes in this case. It is observed that the future summer precipitation along the Tone basin is going to be increased considerably, while an almost constant or decreasing trend is observed for winter season. Natural flow availability for winter or spring seasons thus can be crucial under future scenarios. After reservoir routing, the hydrological risk indices estimated, however, were not found to be changed significantly due to the presence of a robust reservoir system at the upstream.     

SUSTAINABILITY AT THE LOCAL LEVEL: MANAGEMENT TOOLS AND MUNICIPAL TAX INCENTIVE MODEL

The implementation of sustainable development may seem a simple concept when written on paper. However to carry-out long term actions put forward by the Agenda 21 (AG21) at the local level represents one of the main challenges as municipal governments in general do not have the capacity to effectively implement the process. Regional environmental assessment (REA) has shown to be effective in supporting decision-making not only to correct environmental problems due to past unsustainable social-economic developments but also help local governments to implement sustainable actions. However this requires long-term investments of AG21 plans and projects. The allocation of regular and consistent financial resources is one of the main ingredients for the sustainable development process. But traditional plans and projects financed by national and/or international funds may not be sustainable in the long-term because they become dependent on external funding. Research demonstrate that innovative economic instruments such as ecotaxes represent a feasible alternative to sponsor local sustainability because taxes are collected permanently by the government and could be invested in continuous actions. Ecotaxes experiences have provided important reference to structure a municipal incentive model (MIM) to sponsor AG21’s environmental plans and projects on a long-term (permanent) basis. However sustainable development cannot be solely through economic investments. A comprehensive municipal environmental management scheme (MEMS) has been established to support the incentive model. The scheme seeks not only to improve local institutional framework but also incentive continuous participation of local stakeholders at all levels of society. Participatory events and the provision of incentives (educational and financial) are key to motivate society to protect the environment and support actively the sustainable development process as emphasised in the RIO-92 Conference.     

Optimal range of plug-in electric vehicles in Beijing and Shanghai

Both China’s national subsidy policies for plug-in electric vehicles (PEVs) purchasers and passenger cars corporate average fuel consumption and new vehicle credit regulation (dual-credit policy) favor long-range 300+ km battery electric vehicles (BEVs) and 80+ km plug-in hybrid electric vehicles (PHEVs). However, these electric vehicles tend to have lower energy efficiency and higher purchase and operation costs. Vehicle with larger batteries can also be less equitable because the subsidies are often provided to more expensive vehicles and wealthier owners. This study takes advantage of a novel dataset of daily driving data from 39,854 conventional gasoline vehicles in Beijing and 4999 PHEVs in Shanghai to determine the optimal range of BEVs and PHEVs within their respective cities. We simulate a model to explore ranges with which PEVs emit less GHGs than that of a baseline hybrid and conventional gasoline vehicle while ensuring that all daily travel demands are met. Our findings indicate that in both cities, the optimal ranges to balance cost and travel demand for BEVs are 350 km or less and for PHEVs are 60 km or less in Beijing and 80 km or less in Shanghai. We also find that to minimize carbon dioxide (CO₂) emissions, the ranges are even lower 10 km in Beijing and 30 km in Shanghai. Our study suggests that instead of encouraging long-range PEVs, governments should subsidize PEV models with shorter ranges. Parallel efforts should also be made to both increase renewable energy over fossil fuels and expand charging facilities. Although individual mobility demand varies, the government could reduce occasional long-distance driving by subsidizing alternative transportation choices. Providing week-long driving trials to consumers before their purchases may help decrease the demand of very long range PEVs by alleviating the range anxiety through a learning process.

     

Guppies and the TIT FOR TAT strategy: preference based on past interaction

Summary The evolution of cooperation requires either (a) nonrandom interactions, such that cooperators preferentially interact with other cooperators, or (b) conditional behaviors, such that individuals act cooperatively primarily towards other cooperators. Although these conditions can be met without assuming sophisticated animal cognition, they are more likely to be met if animals can remember individuals with whom they have interacted, associate past interactions with these individuals, and base future behavior on this information. Here we show that guppies (Poecilia reticulata), in the context of predator inspection behavior, can identify and remember (for at least 4 h) the more cooperative among two conspecifics and subsequently choose to be near these individuals in future encounters. Offprint requests to: L.A. Dugatkin     

Analysing the sensitivity behaviour of two hydrology models

Recently, the New Morris Method has been presented as an effective sensitivity analysis tool for mathematical models. The New Morris Method estimates the sensitivity of an output parameter to a given set of input parameters (first-order effects) and the extent these parameters interact with each other (second-order effects). This method requires the specification of two parameters (runs and resolution) that control the sampling of the output parameter to determine its sensitivity to various inputs. The criteria for these parameters have been set on the analysis of a well-behaved analytical function (see Cropp and Braddock, Reliab. Eng. Syst. Saf. 78:77–83, 2002), which may not be applicable to other physical models that describe complex processes. This paper will investigate the appropriateness of the criteria from (Cropp and Braddock, 2002) and hence the effectiveness of the New Morris Method to determine the sensitivity behaviour of two hydrologic models: the Soil Erosion and Deposition System and Griffith University Representation of Urban Hydrology. In the first case, this paper will separately analyse the sensitivity of an output parameter on a set of input parameters (first- and second-order effects) for each model and discuss the physical meaning of these sensitivities. This will be followed by an investigation into the sampling criteria by exploring the convergence of the sensitivity behaviour for each model as the sampling of the parameter space is increased. By comparing these trends to the convergence behaviour from Cropp and Braddock (2002), we will determine how well the New Morris Method estimates the sensitivity for each model and whether the sampling criteria are appropriate for these models. It will be shown that the New Morris Method can provide additional insight into the functioning of these models, and that, under a different metric, the sensitivity behaviour of these models does converge confirming the sampling criteria set by Cropp and Braddock.