Rapid karyotyping in prenatal diagnosis: A comparative study of the ‘pipette method’ and the ‘in situ’ technique for chromosome harvesting

Rapid karyotyping in the second and third trimesters has important implications for the management of pregnancies at risk. From September 1985 to March 1992, 735 amniotic fluid samples sent to our laboratory for rapid karyotyping from 64 different diagnostic centres of the Federal Republic of Germany were included in a comparative study on harvesting for chromosome analysis using the ‘pipette method’ or the ‘in situ’ technique. The average time between preparation of the amniotic fluid and verbal notification of the analysed karyotype was 5·41 days. The ‘pipette method’ needed on average 4·65 days, and the ‘in situ’ technique 5·97 days. In comparison with other more invasive techniques available for rapid karyotyping such as cordocentesis and placental biopsy, amniocentesis and subsequent chromosome harvesting using the ‘pipette method’ and/or the ‘in situ’ technique proved very useful and efficient. The overall incidence of chromosome aberrations was 15·3 per cent. The high rate of structural chromosome aberrations and uncommon aneuploidies found in our investigation (12 per cent) indicates that for rapid karyotyping in the second and third trimesters, conventional cytogenetic techniques cannot be replaced by faster techniques based on fluorescent in situ hybridization on interphase cells in the near future.     

Evaluating and expanding the European Union's protected-area network toward potential post-2020 coverage targets

The Convention on Biological Diversity's (CBD) strategic plan will expire in 2020, but biodiversity loss is ongoing. Scientists call for more ambitious targets in the next agreement. The nature-needs-half movement, for example, has advocated conserving half of Earth to solve the biodiversity crisis, which has been translated to protecting 50% of each ecoregion. We evaluated current protection levels of ecoregions in the territory of one of the CBD's signatories, the European Union (EU). We also explored the possible enlargement of the Natura 2000 network to implement 30% or 50% ecoregion coverage in the EU member states’ protected area (PA) network. Based on the most recent land-use data, we examined whether ecoregions have enough natural area left to reach such high coverage targets. We used a spatially explicit mixed integer programing model to estimate the least-cost expansion of the PA network based on 3 scenarios that put different emphasis on total conservation cost, ecological representation of ecosystems, or emphasize an equal share of the burden among member states. To realize 30% and 50% ecoregion coverage, the EU would need to add 6.6% and 24.2%, respectively, of its terrestrial area to its PA network. For all 3 scenarios, the EU would need to designate most recommended new PAs in seminatural forests and other semi- or natural ecosystems. Because 15 ecoregions did not have enough natural area left to implement the ecoregion-coverage targets, some member states would also need to establish new PAs on productive land, allocating the largest share to arable land. Thirty percent ecoregion coverage was met by protecting remaining natural areas in all ecoregions except 3, where productive land would also need to be included. Our results support discussions of higher ecoregions protection targets for post-2020 biodiversity frameworks.     

Dental Fluorosis in Wild Deer: Its Use as a Biomarker of Increased Fluoride Exposure

The relationships between age (range: 3 to 14 yr), mandibular bone fluoride level (marker of accumulated internal dose, range: 597 to 4680 mg F^- kg^-1> dry wt) and intensity of dental fluorosis (reflecting fluoride exposure during enamel formation) were studied in a sample of 53 fluorosed red deer from a fluoride-polluted area in the Czech-German border region (Ore mountains and their southern foreland). Assessment of the severity of dental fluorosis was performed for the 3 permanent premolars and 3 molars of one hemimandible per animal by using an ordinal measurement scale. For statistical analysis, the maximum tooth score of fluorosis (MTS) and the dental lesion index of fluorosis (DLI, sum of the six tooth scores per individual) were used. In the sample, both MTS (r_s = 0.850) and DLI (r_s = 0.813) were highly significantly (p <0.00001) correlated with bone fluoride content. A weaker correlation existed between age and bone fluoride content (r_s = 0.322, p <0.05). The results demonstrate that in case of regional, long-term fluoride pollution, dental fluorosis (measured as MTS or DLI) can be used as a sensitive biomarker of fluoride exposure in deer and thus as an indicator of the level of environmental contamination by fluorides. In many countries, skulls and mandibles of wild deer are regularly and continuously collected by hunters. Assessment of the prevalence and severity of dental fluorosis in this material offers the opportunity for an efficient large-scale biomonitoring of environmental pollution by fluorides at very low cost.     

Mild hydrolysis of 2-trifluoromethylphenol: kinetics, mechanism and environmental relevance

2-Trifluoromethylphenol was hydrolysed in a phosphate buffer at neutral pH. At mild temperatures ranging from 34 degrees C to 69 degrees C this compound liberates consecutively fluorine anions to form salicylic acid. This process is energetically driven by the hydration of the fluorine anions. No intermediates have been detected by HPLC and (19)F-NMR and this was confirmed by computer calculations which favor the first step in the whole reaction sequence being rate-limiting. Accordingly, the reaction energy of the first dehalogenation of the trifluoromethyl anion is 28.4 kcal mol(-1) higher than for the second dehalogenation. The pseudo-first-order kinetic was determined and from an Arrhenius diagram an activation energy of E(a)=25.1 kcal mol(-1) has been estimated. At 37 degrees C and a pH of 7.4 the half-life was 6.9 h. The rate of hydrolysis was favored at higher pH and it was not influenced by oxygen, sunlight or trace elements found in natural water. The latter was shown by incubations with lake water instead of distilled water.     

Transport and anaerobic biodegradation of propylene glycol in gravel-rich soil materials

Continued input of airplane de-icing/anti-icing fluids (ADAF) to runway adjacent soils may result in the depletion of soil-borne terminal electron acceptors. We studied the transport and transformation of propylene glycol (PG), the major constituent of many ADAF, in topsoil and subsoil samples using saturated column experiments at 4 degrees C and 20 degrees C. The export of soil-borne DOC was generally high, non-exhaustive and rate limited. Retardation of added PG was negligible. Rapid PG degradation was observed only in topsoil materials high in organic matter at 20 degrees C. At 4 degrees C, no significant degradation was observed. Thus, under unfavorable, i.e., wet and cold conditions typical for winter de-icing operations, PG and its metabolites will be relocated to deeper soil horizons or even to the groundwater. In subsoil materials, PG degradation was very slow and incomplete. We found that subsoil degradation depended on the import of active microorganisms originating from the organic-rich topsoil material. The degradation efficiency is strongly influenced by the flow velocity, i.e., the residence time of PG in the soil column. Poorly crystalline iron(III) and manganese(IV) (hydr)oxides are used during microbial respiration acting as terminal electron acceptors. This results in the formation and effective relocation of reduced and mobile Fe and Mn species. Long-term application of ADAF to runway adjacent soil as well as the lasting consumption of Fe and Mn will tend to decrease the soil redox potential. Without proper counteractive measures, this will eventually favor the development of methanogenic conditions.     

Lac Dufault sediment core trace metal distribution, bioavailability and toxicity to Hyalella azteca

To determine changes in metal distribution, bioavailability and toxicity with sediment depth, two 20-cm-long replicate cores were collected from a lake historically subjected to the influence of metal mining and smelting activity. The vertical distribution of Pb, Cd and Cu in sediment was similar for all three metals, with the surface layers showing enrichment and the deeper (pre-industrial) layers showing lower concentrations. Toxicity of each sediment core section was determined in laboratory tests with the freshwater amphipod Hyalella azteca. Bioavailable metal in each sediment slice was estimated from metal concentrations in overlying water in these toxicity tests and, for Cd, also from metal bioaccumulation. The profile for Cd in tissue was comparable to Cd in sediment and overlying water, but relative Cd bioavailability from sediment increased with sediment depth. Survival increased with increasing sediment depth, suggesting that surface sediments were probably less or non-toxic before industrialization.     

Performance of an in situ passive sampling system for metals in stormwater

A passive sampler has been developed and is demonstrated in situ for urban runoff. The passive sampler is compared to conventional composite (time-dependent and flow-weighted) bottle sampling during and between storm events. The sampling was carried out at established stormwater stations; before and after a stormwater detention pond. In situ deployment of the passive sampler provides the metal concentrations, corresponding to the electrochemically available fraction of total metal, for time-dependent samples collected in parallel. The sampler provides improved accuracy compared to bottle sampling because contamination during sample transport and handling is minimised. Laboratory handling is reduced by direct analysis of the accumulated metals on the receiving membrane by laser ablation inductively coupled plasma mass spectrometry. Passive sampling also solves the problem of metal speciation change during transport to the laboratory, which is a potential problem for bottle samples. The low cost and convenience of the passive sampler and subsequent analysis should allow significantly more extensive spatial and temporal monitoring of metals in the aquatic environment than has previously been possible.     

The European Water Framework Directive: Water Quality Classification and Implications to Engineering Planning

The European Water framework directive (WFD) is probably the most important environmental management directive that has been enacted over the last decade in the European Union. The directive aims at achieving an overall good ecological status in all European water bodies. In this article, we discuss the implementation steps of the WFD and their implications for environmental engineering practice while focusing on rivers as the main receiving waters. Arising challenges for engineers and scientists are seen in the quantitative assessment of water quality, where standardized systems are needed to estimate the biological status. This is equally of concern in engineering planning, where the prediction of ecological impacts is required. Studies dealing with both classification and prediction of the ecological water quality are reviewed. Further, the combined emission–water quality approach is discussed. Common understanding of this combined approach is to apply the most stringent of either water quality or emission standard to a certain case. In contrast, for example, the Austrian water act enables the application of only the water quality based approach - at least on a temporary basis.