Denaturing high-performance liquid chromatography (DHPLC)-based prenatal diagnosis for tuberous sclerosis

Tuberous sclerosis (TSC) is a frequent autosomal-dominant condition (affecting 1 in 6000 individuals) caused by various mutations in either the hamartin (TSC1) or the tuberin gene (TSC2). This allelic and non-allelic heterogeneity makes genetic counseling and prenatal diagnosis difficult, especially as a significant proportion of TSC cases are due to de novo mutations. For this reason the identification of the disease causing mutation is mandatory for accurate counseling, yet current mutation detection methods such as single-strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) are labor intensive with limited detection efficiency. Denaturing high-performance liquid chromatography (DHPLC) is a high-throughput, semi-automated mutation detection system with a reported mutation detection rate close to 100% for PCR fragments of up to 800 bp. We used a recently described DHPLC assay allowing the efficient detection of mutations in TSC1 to analyze the DNA extracted from a chorion villus sample in order to perform a prenatal diagnosis for TSC. The fetus was found not to have inherited the deleterious mutation and the DHPLC diagnosis was confirmed by haplotype analysis. This represents the first DHPLC-based prenatal diagnosis of a genetic disease. Copyright © 2001 John Wiley & Sons, Ltd.     

Prenatal exclusion of X-linked hydrocephalus-stenosis of the aqueduct of sylvius sequence using closely linked DNA markers

X-linked hydrocephalus-stenosis of the aqueduct of Sylvius sequence (H-SAS, MIM number 307 000) is a rare genetic disorder characterized by hydrocephalus, macrocephaly, adducted thumbs, spasticity, mental retardation, and cerebral malformations. This regularly lethal condition is usually diagnosed at birth or prenatally by ultrasound, but hydrocephalus may be moderate or even undetectable on fetal ultrasound examination. Moreover, since heterozygous women are asymptomatic, carrier detection is at present impossible before the birth of an affected son. Therefore, mapping the H-SAS locus to distal Xq (Xq28) was of primary importance for genetic counselling and prenatal diagnosis. Here, we report prenatal exclusion of H-SAS with a probability of 97.6 per cent in two male fetuses with a 50 per cent a priori risk of being affected using closely linked Xq28 DNA markers.     

Genetic effects of air pollution on forest tree species of the Carpathian Mountains

The effects of air pollution on the genetic structure of Norway spruce, European silver fir and European beech were studied at four polluted sites in Slovakia, Romania and Czech Republic. In order to reduce potential effects of site heterogeneity on the health condition, pair-wise sampling of pollution-tolerant and sensitive trees was applied. Genotypes of sampled trees were determined at 21 isozyme gene loci of spruce, 18 loci of fir and 15 loci of beech. In comparison with Norway spruce, fewer genetic differences were revealed in beech and almost no differentiation between pollution-tolerant and sensitive trees was observed in fir. In adult stands of Norway spruce, sensitive trees exhibited higher genetic multiplicity and diversity. The decline of pollution-sensitive trees may result thus in a gradual genetic depletion of pollution-exposed populations of Norway spruce through the loss of less frequent alleles with potential adaptive significance to altered stressing regimes in the future. Comparison of the subsets of sensitive and tolerant Norway spruce individuals as determined by presence or absence of discolorations ("spruce yellowing") revealed different heterozygosity at 3 out of 11 polymorphic loci.     

Selection effects of air pollution on gene pools of Norway spruce, European silver fir and European beech.

The effects of industrial pollution on allelic and genotypic structures of Norway spruce. European silver fir and European beech were investigated by means of isozyme analysis. In a mixed Norway spruce-silver fir forest stand in an area heavily polluted by sulphur dioxide and heavy metals in the region of Spis (eastern Slovakia), pairs of neighbouring damaged and apparently healthy trees were selected in two replicates (44 and 69 pairs in a heavily and moderately damaged stand, respectively). Pairwise sampling of trees with contrasting vitality was applied to reduce potential effects of site heterogeneity on the vitality of sampled trees. No significant differences in allelic and genotypic frequencies were found between sets of healthy and declining trees. There were differences in the single-locus heterozygosities, but these were not consistent between the replicates. However, the set of damaged trees exhibited higher levels of genetic multiplicity and diversity, possibly due to the deleterious effect of rare alleles under the conditions of air pollution. Consequently. following the decline of pollutant-sensitive trees, the remaining stand will be depleted of a part of alleles with unknown adaptive value to future selection pressures.     

Effects of impervious cover on the surface water quality and aquatic ecosystem of the Kyeongan stream in South Korea

The extent of impervious cover in a watershed has been linked to the quality of an urban aquatic environment. The Kyeongan watershed in South Korea was investigated to evaluate the relationship between the total impervious area (TIA) and the aquatic ecosystem of the watershed, including water quality and aquatic life using a relatively high-resolution (0.4 m) image. The TIA was found to be approximately 12% of the watershed, which indicates that the quality of its environment was being adversely affected by it. For water quality, Pearson correlation analyses showed that all water quality parameters studied were found to be positively correlated with TIA at p < 0.01, except for nitrate (NO3-). In addition, the zone with a higher TIA was found to have worse water quality. Some water quality parameters, such as nitrite (NO2-), total phosphorus, and phosphate (PO4(3-)) were highly affected by discharges from wastewater treatment plants. Water quality data suggest that TIA could be used to predict the water quality of streams. For ecological parameters, the diatom index for organic pollution and trophic diatom index were found to be highly correlated with TIA, whereas physical habitat and benthic macroinvertebrates were poorly correlated with TIA. However, the results indicate that the extent of impervious cover can be a useful indicator for predicting the status of specific ecosystem of streams.     

Prenatal diagnosis of respiratory chain deficiency by direct mutation screening

Respiratory chain deficiency (RCD) is responsible for a clinically heterogeneous group of early-onset untreatable disorders. Enzymological prenatal diagnosis (PD) can only be offered to a fraction of families. Moreover, due to the two-fold genetic origin of the respiratory chain (nuclear and mitochondrial DNA) and owing to the large number of nuclear genes involved in the respiratory chain assembly, maintenance and functioning, the identification of the disease causing gene in a given family remains challenging. Here, we report on PD of RCD by direct screening of NDUFV1, SDH-Fp, SCO1 and SURF1 mutations in five unrelated families with complex I, II and IV deficiency, respectively. The identification of the disease-causing gene in a given family with RCD is a major issue to provide both adequate genetic counselling and early, reliable PD. Copyright © 2001 John Wiley & Sons, Ltd.     

Pleistocene Refugia for <Emphasis Type="Italic">Calluna vulgaris</Emphasis> (L.) Hull Populations in the European Atlantic Region

The distribution of nine chloroplast DNA haplotypes in four insular North-Atlantic and four European coastal Atlantic populations of Calluna vulgaris in the glacial zone of the range has been analyzed in comparison with that in six marginal southern populations in the nonglacial zone of the Atlantic and Mediterranean regions. As a result, two hypothetical Pleistocene refugia (HPRs) for this C. vulgaris population group have been revealed, one in the Cévennes mountain range and the other in the Southern Alps (Trento). Judging from the 1–F_ST value, it has been found that the group of populations in the glacial Atlantic zone and adjacent European coastal Atlantic region is genetically similar to the HPR in the Cévennes at a highly significant level (p ≥ 0.999) and less similar to the HPR in Trento; however, it differs significantly from other Mediterranean and Atlantic populations. It has been concluded that the most probable hypothetical Pleistocene refugium for the recent C. vulgaris populations of the northeastern Atlantic and European coastal Atlantic regions was in the west of the Mediterranean, in the Cévennes, while the additional refugium was in the Southern Alps. Possible directions of the postglacial dispersal and recolonization of habitats by C. vulgaris populations from the western Mediterranean to the northeast of the Atlantic and to Scandinavia have been revealed.     

Ecotoxicology,revisiting its pioneers

Environmental Science and Pollution Research - Ecotoxicology is a discipline resulting from pollution events that harmed human and environmental health by the mid-twentieth century. Environmental...