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.     

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.     

Dissolved and Total Copper in a Coal Ash Effluent and Receiving Stream: Assessment of In Situ Biological Effects

An in situ chemical and biological study was conducted in the lower Muskingum River (southeast Ohio, U.S.A.) to evaluate potential effects of copper (Cu) discharged from a coal ash effluent. Effluent total Cu, dissolved Cu, TSS and pH measurements were performed monthly during January-December 1995. Benthic macroinvertebrates were sampled at five river locations using artificial substrate samplers, and in situ Cu analyses were conducted. Effluent Cu (total) ranged from 8 to 142 microg L(-1) (mean = 58 microg L(-1)), but dissolved Cu never exceeded 78 microg L(-1) (mean = 20 microg L(-1)). The mean ratio of dissolved Cu to total Cu in these samples was 32%. Total Cu concentrations at the biological sampling sites adjacent to the effluent discharge were higher than levels at ambient sites, but dissolved Cu levels were similar among all sites. The macroinvertebrate community proximal to the coal ash effluent had the highest number of taxa and total number of individuals; a high number of mayfly and caddis fly taxa; and the highest Invertebrate Community Index score. The high water velocity of the discharge (which likely contained particulate organic matter) apparently created a favorable microhabitat that, combined with Cu-complexing constituents in the discharge, superceded potential adverse effects of high Cu levels. This study emphasizes the importance of instream biological data when obtained in conjunction with chemical analyses.     

Environmental quality control and environmental games

Conclusion  In this paper we have considered a specific environmental game emphasizing both control-prevention efforts and the propensity to pollute by a firm which adopts a given pollution abatement technology. A random payoff game was constructed and solved under a risk neutral assumption and quadratic utilities for both the firm and the environmental controller. The game thus defined, provides a wide range of interpretations and potential approaches for selecting a control-inspection policies to prevent environmental risks. There are of course many facets to this problem, which could be considered and have not been considered in sufficient depth. For example, more complex control mechanisms and liabilities, the effects of insurance and risk sharing, the application of cooperative efforts and subvention of pollution abatement investments (through tax incentives and their like), etc. have not been considered [5,7]. These are topics for further research. The basic presumption of this paper is that it is very difficult to fully enforce pollution prevention by firms, as a result, some controls are needed to ensure that firms be controlled so that appropriate efforts are carried.     

Integrating disciplinary research into an interdisciplinary framework: A case study in sustainability research

The complexity of today’s research problems increasingly demands that scientists move beyond the confines of their own discipline. In this special issue, the basics of a transdisciplinary framework are established and problems analyzed in a specific discipline are successfully integrated in this transdisciplinary network. The intent has been to go beyond only statements about the importance of enabling early stage researchers to work across disciplinary boundaries, and to show that important discoveries are being made at the intersection of disciplines. Embedding various research projects in a more global framework can meet the demands of an interdisciplinary or transdisciplinary approach (especially for early stage researchers) and contribute significantly to sustainable research.