Rapid prenatal diagnosis of aneuploidy from uncultured amniotic fluid cells using five-colour fluorescence in situ hybridization

In this paper we describe the use of five-colour fluorescence in situ hybridization for prenatal diagnosis of aneuploidy using uncultured amniotic fluid cells. The analysis is based on ratio mixing of dual-labelled probes and digital imaging for the detection and visualization of five different probes specific for the five target chromosomes, 13, 18, 21, X, and Y. A retrospective blind analysis of 30 coded uncultured amniotic fluid samples correctly detected fetal sex and five trisomy 21 cases. Multicolour fluorescence in situ hybridization used in this way allows rapid and simultaneous detection of the most frequent aneuploidies.     

Continuous subcutaneous insulin infusion (CSII) in pregnant diabetic patients

The efficacy of the insulin infusion pump (CSII) in pregnancy was examined in 12 diabetic patients and compared with intermittent insulin therapy (IIT). In patients poorly controlled on IIT constant and rapid equilibrium was achieved with CSII (mean of glucose levels: CSII versus IIT=84 versus 137 mg/dl; S.D. = 36 versus 63 mg/dl; mean amplitude of glycemic excursion (MAGE)=65 versus 112 mg/dl. In patients well controlled on IIT, CSII led to a reduction in the variation of glucose excursions (S.D.= 29 versus 36 mg/dl; MAGE=48 versus 76 mg/dl). CSII generally produced a reduction of 20–37 per cent of daily insulin dose (in three cases there was an increase of dose with the achievement of glycemic control). Furthermore in CSII treated-patients amniotic glucose, insulin and C-peptide concentrations werefound tobeinthenormalrange(22.1±10.1 mg/dl; 5.2±2.7μ/ml; 1.25±0.71 ng/ml, respectively). All infants were horn at or near-term, had no macrosomia or neonatal problems. It is concluded that CSII is a highly efficient way to achieve normal glucose levels in pregnancy, not only in type I, hut also in type II or gestational diabetes.     

Prenatal diagnosis of β-thalassaemia in Mediterranean populations by dot blot analysis with DNA amplification and allele specific oligonucleotide probes

In this study, we describe a simple strategy to detect β-thalassaemia mutations in prospective parents and to make prenatal diagnosis in pregnancies at risk in the Mediterranean population. Screening of prospective parents is carried out by dot blot analysis on enzymatically amplified DNA with a set of oligonucleotide probes complementary to the most common mutations in this population. Prenatal diagnosis is accomplished by the same procedure on enzymatically amplified amniocyte or trophoblast DNA. The main advantages of this procedure are the simplicity, sensitivity (0.05 μg of DNA), and rapidity (12–24 h). Further simplification is obtained by amplification of the DNA from crude amniotic cell lysate. The very low amount of fetal material necessary for this analysis eliminates the need to culture amniotic fluid cells and may decrease the fetal loss rate associated with trophoblast sampling. The number of specific DNA sequences obtained by the amplification procedure allowed us to use non-radioactive labelled oligonucleotide probes, which have several advantages compared to radioactive probes.     

Transabdominal chorionic villus sampling: Analysis of 350 consecutive cases

We report a series of 350 patients submitted to transabdominal chorionic villus sampling (CVS). A technique using two ultrasound-guided needles and a suction pump was used. In most cases, the procedure was performed between 9 and 13 weeks. Twenty-one pregnancies were selectively terminated; nine spontaneous abortions followed the procedure and one fetal loss after 28 weeks was recorded; 153 pregnancies are in progress and 169 delivered fetuses are alive and well. Transabdominal biopsy is a feasible and effective technique for CVS.     

Landscape Structure and Historical Processes Along Diked European Valleys: A Case Study of the Arc/Isère Confluence (Savoie, France)

/ The valleys of European piedmonts constitute changing narrow corridors within which water, matter, nutrients, energy, and species flow. The dispersion patterns of these flows have been significantly disturbed since the end of the 18th century. Thus, western European valleys have been changed into complex mosaics by implementation of socioeconomical programs. In order to define future actions allowing the preservation of this "ecocomplex" (Blandin and Lamotte 1988), it is necessary to gather precise information of the landscape dynamics. Hence, the study of the European river valleys must be based upon two major steps: (1)the analysis of the present landscape with suitable remote sensing techniques, allowing us to map the complex mosaic of narrow corridors; and (2) the analysis of temporal landscape development patterns since the first engineering works that have transformed the braided channel system. In this paper, the efficiency of the addition of two techniques is highlighted: (1) the "wavelet merging method" from multispectral and panchromatic SPOT images for the floodplain land-cover mapping, and (2) the historical reconstruction techniques from old maps and archive documents in order to analyze the cumulative impacts of engineering works on landscape diversity. To illustrate the method, a particularly complex case study is chosen: the Arc/Isere confluence (downstream from Albertville, Savoie, France). Remote sensing, field survey, and historical reconstruction allowed us to distinguish two types of spatial units: (1) the "functional sets" characterized by independent state factors (edaphic, hydrological, and topographic) and supporting a limited number of vegetation types (spontaneous or cultivated), and (2) the functional unit (= ecotope), which corresponds to a unique combination of vegetation type/functional set.KEY WORDS: Landscape ecology; Floodplains; Vegetation; Land use; Remote sensing; Historical reconstruction; Mapping; Merging methods; Human impacts; Alps; Isere river; France     

Cholinesterase characterization and effects of the environmental contaminants chlorpyrifos and carbofuran on two species of marine crabs,Carcinus maenas and Pachygrapsus marmoratus

Among the most frequent targets for toxic effects of modern pesticides, namely organophosphates and carbamates, one may find cholinesterases (ChEs). ChEs exist in a wide variety of animals and have been used actively to discriminate among the environmental effects of different pollutant groups, including the aforementioned pesticides. This study had three purposes, namely (i) identifying the ChE forms present in tissues (eyes and walking legs muscle) of two crab species, Carcinus maenas and Pachygrapsus marmoratus; to (ii) determine the in vitro toxicological effects, and (iii) compare the sensitivity of such enzymatic forms towards commonly used anti-ChE pesticides, namely the organophosphate chlorpyrifos and the carbamate carbofuran. Our results showed that there was not a clear preference for any of the tested substrates in any of the tissues from both species. Furthermore, the ChE activity was almost completely suppressed following incubation with eserine and with the specific inhibitor BW284C51 in all tissues from both species. In vitro exposure to chlorpyrifos promoted a significant decrease in ChE activity in both species. Furthermore, the ChE activity was completely suppressed following incubation with carbofuran and chlorpyrifos. These results suggest that the major ChE forms present in tissues of both crab species show intermediate structural properties and activity patterns, halfway between classic acetylcholinesterase and pseudocholinesterases. However, the sensitivity of the found forms towards ChE inhibitors was established, and the responsiveness of such forms towards common anti-ChE chemicals was established. Both tested species seem to be promising test organisms to be used in marine and coastal scenarios of putative contaminations by anti-ChE chemicals, considering the here reported patterns of response.

     

Seasonality and microhabitat selection in a forest-dwelling salamander

Many small terrestrial vertebrates exhibit limited spatial movement and are considerably exposed to changes in local environmental variables. Among such vertebrates, amphibians at present experience a dramatic decline due to their limited resilience to environmental change. Since the local survival and abundance of amphibians is intrinsically related to the availability of shelters, conservation plans need to take microhabitat requirements into account. In order to gain insight into the terrestrial ecology of the spectacled salamander Salamandrina perspicillata and to identify appropriate forest management strategies, we investigated the salamander’s seasonal variability in habitat use of trees as shelters in relation to tree features (size, buttresses, basal holes) and environmental variables in a beech forest in Italy. We used the occupancy approach to assess tree suitability on a non-conventional spatial scale. Our approach provides fine-grained parameters of microhabitat suitability and elucidates many aspects of the salamander’s terrestrial ecology. Occupancy changed with the annual life cycle and was higher in autumn than in spring, when females were found closer to the stream in the study area. Salamanders showed a seasonal pattern regarding the trees they occupied and a clear preference for trees with a larger diameter and more burrows. With respect to forest management, we suggest maintaining a suitable number of trees with a trunk diameter exceeding 30 cm. A practice of selective logging along the banks of streams could help maintain an adequate quantity of the appropriate microhabitat. Furthermore, in areas with a presence of salamanders, a good forest management plan requires leaving an adequate buffer zone around streams, which should be wider in autumn than in spring.     

Runoff of pesticides from rice fields in the Ile de Camargue (Rhône river delta, France): field study and modeling

A field study on the runoff of pesticides was conducted during the cultivation period in 2004 on a hydraulically isolated rice farm of 120 ha surface with one central water outlet. Four pesticides were studied: Alphamethrin, MCPA, Oxadiazon, and Pretilachlor. Alphamethrin concentrations in runoff never exceeded 0.001 microg L(-1). The three other pesticides were found in concentrations between 5.2 and 28.2 microg L(-1) in the runoff water shortly after the application and decreased thereafter. The data for MCPA compared reasonably well with predictions by an analytical runoff model, accounting for volatilization, degradation, leaching to groundwater, and sorption to soil. The runoff model estimated that runoff accounted for as much as 18-42% of mass loss for MCPA. Less runoff is observed and predicted for Oxadiazon and Pretilachlor. It was concluded that runoff from rice paddies carries important loads of dissolved pesticides to the wetlands in the Ile de Camargue, and that the model can be used to predict this runoff.     

Improvements on Flood Alleviation in Germany: Lessons Learned from the Elbe Flood in August 2002

The increase in damage due to natural disasters is directly related to the number of people who live and work in hazardous areas and continuously accumulate assets. Therefore, land use planning authorities have to manage effectively the establishment and development of settlements in flood-prone areas in order to avoid the further increase of vulnerable assets. Germany faced major destruction during the flood in August 2002 in the Elbe and Danube catchments, and many changes have been suggested in the existing German water and planning regulations. This article presents some findings of a “Lessons Learned” study that was carried out in the aftermath of the flood and discusses the following topics: 1) the establishment of comprehensive hazard maps and flood protection concepts, 2) the harmonization of regulations of flood protection at the federal level, 3) the communication of the flood hazard and awareness strategies, and 4) how damage potential can be minimized through measures of area precaution such as resettlement and risk-adapted land use. Although attempts towards a coordinated and harmonized creation of flood hazard maps and concepts have been made, there is still no uniform strategy at all planning levels and for all states (Lae nder) of the Federal Republic of Germany. The development and communication of possible mitigation strategies for “unthinkable extreme events” beyond the common safety level of a 100-year flood are needed. In order to establish a sustainable and integrated flood risk management, interdisciplinary and catchment-based approaches are needed.