Use of the chemical cleavage of mismatch method for prenatal deficiency diagnosis of alpha-1-antitrypsin

The most common mutation in alpha-1-antitrypsin deficiency, conversion of a G to an A at base 9989 (PI-Z), was detected with the chemical cleavage of mismatch method, demonstrating the power of the method for prenatal diagnosis. Exon V of the gene was amplified using the polymerase chain reaction and heteroduplexes were formed to test for the presence of the mutation. The predicted C mismatch was readily detectable with hydroxylamine, and by making the probe from the chorionic villus sample it was possible to determine that the fetus was heterozygous, not homozygous, for the mutation.     

The evolutionary position of turtles revised

Consensus on the evolutionary position of turtles within the amniote phylogeny has eluded evolutionary biologists for more than a century. This phylogenetic problem has remained unsolved partly because turtles have such a unique morphology that only few characters can be used to link them with any other group of amniotes. Among the many alternative hypotheses that have been postulated to explain the origin and phylogenetic relationships of turtles, a general agreement among paleontologists emerged in favoring the placement of turtles as the only living survivors of the anapsid reptiles (those that lack temporal fenestrae in the skull). However, recent morphological and molecular studies have radically changed our view of amniote phylogenetic relationships, and evidence is accumulating that supports the diapsid affinities of turtles. Molecular studies favor archosaurs (crocodiles and birds) as the living sister group of turtles, whereas morphological studies support lepidosaurs (tuatara, lizards, and snakes) as the closest living relatives of turtles. Accepting these hypotheses implies that turtles cannot be viewed any longer as primitive reptiles, and that they might have lost the temporal holes in the skull secondarily rather than never having had them.     

Forest ecosystem development in post-mining landscapes: a case study of the Lusatian lignite district

The restoration of surface mining landscapes requires the (re)creation of ecosystems. In Lusatia (eastern Germany), large-scale open-cast lignite mining operations generated spoil dumps widely consisting of acidified, phytotoxic substrates. Amelioration and rehabilitation measures have been developed and applied to these substrates since the 1950s. However, it is still not clear whether these approaches are sustainable. This paper reports on collaborative research work into the ecological potential of forest ecosystem development on typical minesites in the Lusatian lignite district. At first sight, pine stands on minesites along a chronosequence comprising about 35 years did not show differences when compared with stands on non-mined sites of the general region. Furthermore, with some modification, conceptual models for flora and fauna succession in forest stands on non-mined sites seem to be applicable, at least for the early stages of forest ecosystem development. For example, soil organism abundance and activity at minesites had already reached levels typical of non-mined sites after about 20-30 years. In contrast, mine soils are very different from non-mined soils of the test region. Chemically, mine soil development is dominated by processes originating from pyrite oxidation. Geogenic, i.e. lignitic, soil organic carbon was shown to substitute for some functions of pedogenic soil organic matter. Rooting was hampered but not completely impeded in strongly acidified soil compartments. Roots and mycorrhizae are apparently able to make use of the characteristic heterogeneity of young mine soils. Considering these recent results and the knowledge accumulated during more than 30 years of research on minesite rehabilitation internationally, it can be stated that minesite restoration might be used as an ideal case study for forest ecosystem development starting at "point zero" on "terra nova".     

Body condition and immune response in wild zebra finches: effects of capture, confinement and captive-rearing

Behavioural ecologists attempt to predict fitness in birds from estimates of body condition and immune capacity. We investigated how the stresses associated with capture, confinement and captive-rearing of wild zebra finches (Taeniopygia guttata) affected different elements of the immune system and body condition. Wild birds had higher heterophil:lymphocyte ratios and total leucocyte counts than aviary birds, presumably an outcome of mounting specific resistance to pathogens, but this response diminished significantly within 10 days of confinement. Wild birds had lower phytohaemagglutinin-A (PHA) responses than their aviary-bred counterparts possibly because energetic costs limited a general resistance response. Wild birds were heavier and had higher haematocrits than their aviary counterparts, but had less fat, although just 10 days of captivity significantly increased fat levels. Measures of body condition were of limited use for predicting immune responsiveness. We conclude that the different elements of the immune system and body condition respond independently, and often unpredictably, to many ecological and behavioural stressors.     

Forestry Mitigation Options for Mexico: Finding Synergies between National Sustainable Development Priorities and Global Concerns

We examine carbon (C) reference and mitigation scenarios for the Mexicanforest sector between the year 2000 and 2030. Estimates are presentedseparately for the period 2008–2012.Future C emissions and capture are estimated using a simulation modelthat: a) allocates the country land use/land cover classes among differentfuture uses and categories using demand-based scenarios for forestryproducts; b) estimates the total C densities associated to each land usecategory, and c) determines the net carbon implications of the process ofland use/cover change according to the different scenarios.The options analyzed include both afforestation/reforestation, such ascommercial, bionenergy and restoration plantations, and agroforestrysystems, and forest conservation, through the sustainable management ofnative forests and forest protection.The total mitigation potential, estimated as the difference between the totallong-term carbon stock in the reference and the mitigation scenario reaches300 × 10⁶ Mg C in the year 2012 and increases to 1,382 × 10⁶ Mg C in 2030. The average net sequestration in the 30 year period is 46 × 10⁶ Mg C yr^-1, or 12.5 × 10⁶ Mg C yr^-1 within the period 2008 to 2012. The costs of selected mitigation options range from 0.7–3.5 Mg C^-1 to 35 Mg C^-1. Some options are cost effective.     

Carbon mitigation potential and costs of forestry options in Brazil,China, India,Indonesia, Mexico,the Philippines and Tanzania

This paper summarizes studies of carbon (C) mitigation potential and costs of about 40 forestry options in seven developing countries. Each study uses the same methodological approach – Comprehensive Mitigation Assessment Process (COMAP) – to estimate the above parameters between 2000 and 2030. The approach requires the projection of baseline and mitigation land-use scenarios. Coupled with data on a per ha basis on C sequestration or avoidance, and costs and benefits, it allows the estimation of monetary benefit per Mg C, and the total costs and carbon potential. The results show that about half (3.0 Pg C) the cumulative mitigation potential of 6.2 Petagram (Pg) C between 2000 and 2030 in the seven countries (about 200× 10⁶ Mg C yr^-1) could be achieved at a negative cost and the remainder at costs ranging up to $100 Mg C^-1. About 5 Pg C could be achieved, at a cost less than $20 per Mg C. Negative cost potential indicates that non-carbon revenue is sufficient to offset direct costs of these options. The achievable potential is likely to be smaller, however, due to market, institutional, and sociocultural barriers that can delay or prevent the implementation of the analyzed options.     

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.