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.     

Visualizing mushroom body response to a conditioned odor in honeybees

Combining differential conditioning with optophysiological recordings of bee brain activity allows the investigation of learning-related changes in complex neural systems. In this study we focused on the mushroom bodies of the bee brain. Presenting different odors to the animal leads to significant activation of the mushroom body lips. After differential conditioning, the rewarded odor leads to stronger activation than it did before training. Activation by the unrewarded odor remains unchanged. These results resemble findings in the bee's antennal lobes, which are the first olfactory relay station in the insect brain. As an integrative neural network, enhanced activation of the mushroom body lip may carry additional information, i.e., for processing odor concentrations.     

Ammonia production and kinetic properties of glutamate dehydrogenase in the sipinculid Phascolosoma arcuatum exposed to anoxia

The amounts of total NH ₄ ⁺ detected in the external media in which Phascolosoma arcuatum had been exposed to various periods of anoxia were significantly greater than those in which the worms were exposed to normoxia for a similar period. The increased NH ₄ ⁺ production by P. arcuatum during anoxic exposure was unlikely to be due to an increased catabolism of adenine nucleotides or urea. In contrast, there were significant decreases in the concentrations of several free amino acids in the coelomic plasma and body tissues of individuals during the 48 h of anoxic exposure. The amount of NH ₄ ⁺ produced by the anoxic P. arcuatum could be accounted for by the decreases in the concentrations of aspartate or glycine. Increases in the catabolism of free amino acids (FAA), leading to the increased production of NH ₄ ⁺ , in P. arcuatum during anoxia were supported by the detection of significant changes in the kinetic properties of glutamate dehydrogenase (GDH), in the deaminating direction, from worms exposed to anoxia for 48 h. The apparent increase in the affinity of GDH from the anoxic worm to glutamate would bring about a greater deaminating activity at physiological concentrations of ths substrate. P. arcuatum used in these experiments were collected from the mangrove swamp at Mandai, Singapore between 1990 and 1993.     

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.     

THE INFLUENCE OF SETTINGS AND ENVIRONMENTAL EVENTS ON METAMOTIVATIONAL STATE AND AROUSAL: FURTHER TESTS OF REVERSAL THEORY ON ASIAN SAMPLES

The two studies reported further test reversal theory hypotheses concerning human psychological responses to different types of settings and environmental events on an Asian sample. In study 1, individuals in four settings representing different combinations of metamotivational state and arousal level (telic and paratelic/low arousal; telic and paratelic/high arousal) completed the telic state measure (TSM; Svebak & Murgatroyd, 1985). Significant differences between participants' mean scores across the four settings were obtained for TSM items serious–playful, planning–spontaneous, felt arousal, preferred arousal, effort and arousal discrepancy. In study 2 different environmental events in the same setting (changes in reading tasks), used in a crossover design, were used to induce reversals in individuals' operative metamotivational states. The results obtained from TSM scores pretask 1, between tasks and posttask 2. indicated significant time-of-testing interaction effects on the TSM serious–playful, planning–spontaneous and effort items. In addition, a main effect for TSM felt arousal was found. These results showed that reversals in metamotivational states were induced by the environmental event and that felt arousal scores increased across the three administrations of the TSM.     

The Potential of Public Participation Geographic Information Systems in UK Environmental Planning: Appraisals by Active Publics

The paper draws on an empirical study of two workshops in which the issues that arise from the use of geographic information systems (GIS) as a planning tool in public participation settings were explored by local residents who take an active interest in local planning matters in their London borough. The paper demonstrates how issues concerned with the democratization of GIS and public participation GIS (PPGIS) informed the structure and conduct of the workshops and the qualitative analysis of the workshop discussions. Key themes raised by participants included: the potential of PPGIS as a means of extending knowledge networks; issues of data ownership and the responsiveness of data providers to public concerns; and the role that institutional norms and practices play in democratizing information availability and the transparency of the decision-making process. The paper concludes that the potential of PPGIS as a planning tool cannot be separated from public concerns about the legitimacy of the planning process or local government.     

Nest-building by crested tit Parus cristatus males: an analysis of costs and benefits

In a population of crested tits in Belgium, one out of three breeding males participated in nest-building. Since variation in the amount of nest-building among 11 building males was low, analysis of male nest-building allowed a logistic regression design. The probability of building increased with male condition and for late nests. The effect of male condition decreased with date, and date was the only factor predicting nest-building for repeat broods. The effect of male condition on nest-building suggests that building entails important costs for males. Based on correlative data, we argue that building by males shortens the interval between the start of nest-building and the onset of egg-laying by about 5 days. Since clutch size, incubation and nestling time did not differ between early and late broods, male nest-building apparently affects fledging date. Since early fledgers (early dispersers) are more likely to settle in future winter flocks, building behaviour by crested tit males should be favoured by natural selection. So far, male nest-building in hole-breeding birds might have been overlooked due to the widespread use of artificial nest-boxes in behavioural studies.     

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.