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.     

The effects of job demands and control on employee attendance and satisfaction

We examined the impact of stressful job demands on employee attitudes and attendance. Using Karasek's (1979) theory of job decision latitude as the conceptual foundation, we hypothesized that mental and physical work demands would interact with employee beliefs of personal control. Survey data from 90 male manufacturing employees regarding their control beliefs were combined with objective job analysis data concerning mental and physical demands and one year's worth of archival data regarding unexcused absences, sick days, and days tardy. There were significant interactions between control and objective psychological demands that indicated that these demands were associated with higher levels of tardiness and sick days only under conditions of low perceived control. In contrast, subjective workload ratings showed no relationship with tardiness and sick days, but, in interaction with control, predicted work satisfaction and voluntary absence. We discussed these results in terms of a stress process that affects health-related attendance independent of employee attitudes.     

Review of present European environmental policies and legislation involving cultural heritage

Cultural heritage is being addressed by a number of charters and conventions and it is clear that its consideration within decision-making processes is progressively becoming a real challenge, both for developers and public authorities. Against this background, this paper reviews the environmental assessment framework developed by the European Community, as this should increasingly influence decision making about cultural heritage in an urban setting. The legislative framework for access to environmental information is also reviewed because of its relevance to the decision-making process.The Directives on Environmental Impact Assessment and Strategic Environmental Assessment both require a consideration of cultural heritage in decision making. This requirement can go some way to addressing the paradox in the European Community's position whereby the community wishes to conserve and enhance its own cultural identity whilst, at the same time, cultural heritage is usually defined at a local level. The present paper suggests that, given their flexibility, the two European Community directives on environmental assessment constitute a promising opportunity to address this seeming paradox, but that there is a significant gap between legal obligations and the methodological tools to meet them.     

New birds on the block: new neighbors increase defensive costs for territorial male willow ptarmigan

Previous researchers have hypothesized that site-faithful animals may benefit from the presence of familiar neighbors. This study compares the relative costs of territorial defense against new and former neighbors by male willow ptarmigan (Lagopus lagopus). Territorial defense against new neighbors appeared to require a greater expenditure of both time and effort than did defense against former neighbors. Territorial males that had several new neighbors spent a higher proportion of time fighting than did males with fewer new neighbors, and males with both new and former neighbors spent a greater amount of time fighting with their new neighbors, on average, than with their former neighbors. In addition, fights with new neighbors occurred relatively more frequently and were longer than fights with former neighbors. Finally, fights involving new neighbors tended to escalate to higher levels than fights between former neighbors. Reduced defensive costs for site-faithful, territorial males may provide one explanation for the tendency of males to be more site-faithful than females in many species.     

Small queens in the ant Ectatomma tuberculatum: a new case of social parasitism

Social parasites exploit the worker force of colonies of other social insects to rear their own young. Social parasitism occurs in several Hymenoptera and is particularly common in several tribes of the ant subfamilies Myrmicinae and Formicinae. Here, we document the occurrence of miniaturized queens (microgynes) in colonies of Ectatomma tuberculatum, an ant belonging to the subfamily Ectatomminae. Behavioral observations and genetic analyses show that microgynes concentrate their reproductive efforts almost exclusively on the production of sexual offspring (microgynes and males), whereas the regular, large queens (macrogynes) produce workers in addition to sexuals. According to mitochondrial and nuclear markers, gene flow between microgynes and macrogynes is extremely limited. Whereas the co-occurrence of microgynes and macrogynes in the related species Ectatomma ruidum constitutes an intraspecific polymorphism associated with alternative dispersal tactics, microgynes found in colonies of E. tuberculatum appear to be a distinct species and to represent the first case of social parasitism in the poneromorph subfamilies of ants.