First-trimester diagnosis of menkes disease: Intermediate copper values in chorionic villi from three affected male fetuses

Chorionic villus samples with copper contents of 1·91, 4·2, 5·6, and 6·3 ng/mg were observed in four cases with male karyotypes. These values were outside the range for unaffected males (0·30–0·85 ng/mg), and three of them were outside the control range (0·20–2·39 ng/mg). But these three values were below the values previously observed for affected Menkes fetuses (12·0–24·8 ng/mg). Follow-up by ⁶⁴Cu uptake studies on the amniotic fluid cells was performed in three of these cases. A combination of ⁶⁴Cu uptake and chase experiments on the amniotic fluid cells showed more convincingly than ⁶⁴Cu uptake per se the direct copper values of 4·2 and 5·6 ng/mg to correspond to affected fetuses. Amniotic fluid cells from the male fetus with the CV copper value of 1·9 ng/mg showed normal results. The CV copper value of 6·3 ng/mg was considered pathognomonic for Menkes disease. The pregnancy was terminated, and the diagnosis was confirmed on fetal fibroblasts. Maternal deciduum prepared from the placentae showed in one of the cases with an affected fetus copper values ranging from 1·5 to 5·7 ng/mg. In six additional diagnostic cases, the copper content was determined in both CV samples and maternal deciduum. In three of these cases with normal CV sample copper, maternal decidua values of 4·85–7·8 ng/mg copper were observed. These results show that maternal deciduum contamination of a CV sample could cause a false-positive diagnosis.     

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".     

Identifying water resources management strategies in adaptation to climate change under uncertainty

In this study, an integrated simulation-based allocation modeling system (ISAMS) is developed for identifying water resources management strategies in response to climate change. The ISAMS incorporates global climate models (GCMs), a semi-distributed land use-based runoff process (SLURP) model, and a multistage interval-stochastic programming (MISP) approach within a general framework. The ISAMS can not only handle uncertainties expressed as probability distributions and interval values but also reveal climate change impacts on water resources allocation under different projections of GCMs. The ISAMS is then applied to the Kaidu-kongque watershed with cold arid characteristics in the Tarim River Basin (the largest inland watershed basin in China) for demonstrating its efficiency. Results reveal that different climate change models corresponding to various projections (e.g., precipitation and temperature) would lead to changed water resources allocation patterns. Variations in water availability and demand due to uncertainties could result in different water allocation targets and shortages. A variety of decision alternatives about water allocations adaptive to climate change are generated under combinations of different global climate models and ecological water release plans. These findings indicate that understanding the uncertainties in water resources system, building adaptive methods for generating sustainable water allocation patterns, and taking actions for mitigating water shortage problems are key adaptation strategies responding to climate change.     

First Detection of Hepatitis E Virus in Shellfish and in Seawater from Production Areas in Southern Italy

Shellfish samples (n = 384) from production areas, water samples from the same areas (n = 39) and from nearby sewage discharge points (n = 29) were analyzed for hepatitis E virus (HEV) by real-time and nested RT-PCR. Ten shellfish samples (2.6%) and five seawater samples (12.8%) tested positive for HEV; all characterized strains were G3 and showed high degree of sequence identity. An integrated surveillance in seafood and waters is relevant to reduce the risk of shellfish-associated illnesses.     

Utilisation of bubble resonance phenomena to improve gas–liquid contact

 In several branches of science and technology a gaseous phase is dispersed into a liquid in the form of bubbles, a gaseous component then dissolves into the liquid and subsequently undergoes chemical reaction. The overall process performance can be improved substantially when the area of gas–liquid contact is increased. By subjecting the liquid phase to low frequency vibrations, the bubbles are shown to suffer significant breakage, induced by resonance. When the vibration is properly tuned, the interfacial area is found to increase by a factor of 1.8–2.4, depending on the properties of the liquid. Resonance-induced bubble breakage phenomena have a great potential for improving the rates of chemical processes involving fast reactions, with minimal energy input. Received: 7 July 2000 / Accepted in revised form: 28 August 2000     

Optimisation of a PMAxx™-RT-qPCR Assay and the Preceding Extraction Method to Selectively Detect Infectious Murine Norovirus Particles in Mussels

Human noroviruses are a major cause for gastroenteritis outbreaks. Filter-feeding bivalve molluscs, which accumulate noroviruses in their digestive tissues, are a typical vector for human infection. RT-qPCR, the established method for human norovirus detection in food, does not allow discrimination between infectious and non-infectious viruses and can overestimate potentially infectious viral loads. To develop a more accurate method of infectious norovirus load estimation, we combined intercalating agent propidium monoazide (PMAxx™)-pre-treatment with RT-qPCR assay using in vitro-cultivable murine norovirus. Three primer sets targeting different genome regions and diverse amplicon sizes were used to compare one-step amplification of a short genome fragment to three two-step long-range RT-qPCRs (7 kbp, 3.6 kbp and 2.3 kbp amplicons). Following initial assays performed on untreated infectious, heat-, or ultraviolet-inactivated murine noroviruses in PBS suspension, PMAxx™ RT-qPCRs were implemented to detect murine noroviruses subsequent to their extraction from mussel digestive tissues; virus extraction via anionic polymer-coated magnetic beads was compared with the proteinase K-dependent ISO norm. The long-range RT-qPCR process detecting fragments of more than 2.3 kbp allowed accurate estimation of the infectivity of UV-damaged murine noroviruses. While proteinase K extraction limited later estimation of PMAxx™ pre-treatment effects and was found to be unsuited to the assay, magnetic bead-captured murine noroviruses retained their infectivity. Genome copies of heat-inactivated murine noroviruses differed by 2.3 log₁₀ between RT-qPCR and PMAxx™-RT-qPCR analysis in bivalve molluscs, the PMAxx™ pre-treatment allowing a closer approximation of infectious titres. The combination of bead-based virus extraction and PMAxx™ RT-qPCR thus provides a more accurate model for the estimation of noroviral bivalve mollusc contamination than the conjunction of proteinase K extraction and RT-qPCR and has the potential (once validated utilising infectious human norovirus) to provide an added measure of security to food safety authorities in the hazard assessment of potential bivalve mollusc contamination.