Trophoblast-like cells sorted from peripheral maternal blood using flow cytometry: A multiparametric study involving transmission electron microscopy and fetal dna amplification

Three monoclonal antibodies (MAbs) against trophoblast (GB17, GB21, and GB25) and flow cytometry were used to sort trophoblast-like cells (TLCs) from peripheral blood of pregnant women. Sorted TLCs were processed for electron microscopy and fetal DNA amplification of the Y-specific sequences from mothers carrying male fetuses. At the ultra-structural level, most of the nucleated cells had the morphology of leucocytes, suggesting maternal contaminants, and we did not find the characteristic features of the free inter-villous trophoblast cells. Nevertheless, polymerase chain reaction (PCR) analysis showed an amplification of Y-specific sequences in two out of three samples of sorted TLCs. These results suggest that besides the maternal leucocytes, sufficient trophoblast nucleated fetal cells can be obtained using cell enrichment by sorting. This sensitive method holds promise for non-invasive prenatal diagnosis of fetal sex and if sufficient Y(positive) nuclei are found, for the diagnosis of selected numerical chromosome abnormalities.     

Human induced salinisation and the use of quantitative methods

Sustainable development of the earth's limited water and land resources is of paramount importance because of rising world population and existing conflicting demands for these resources. Enormous capital investment has been made in developing these resources, but now there is irrefutable evidence that such developments have led to major resource degradation. This includes problems of salinisation and damages to ecosystems. The countries predominantly affected by human induced salinisation are located in arid and semi-arid regions of the world and include Australia, China, Egypt, India, Pakistan, USSR, and USA. This paper describes the processes of salinisation, its impacts and the use of quantitative methods in salinity investigation and management. Australia is used as a case study of typical salinity problems and as a demonstration of the fruitful application of quantitative methods. The paper concludes that quantitative methods such as surface water and groundwater models are powerful design, management and predictive tools in salinity investigation. However, application of some models, such as those for unsaturated flow and transport and groundwater solute transport, are not widespread due to uncertainties in describing the complexity of the processes and the lack of hydrodispersive data.     

Prenatal diagnosis of femur-fibula-ulna complex by ultrasonography in a male fetus at 24 weeks of gestation

We present a case of ultrasonographic prenatal diagnosis at 24 weeks of femur—fibula—ulna (FFU) complex. To our knowledge, this is the first report of an early prenatal diagnosis of FFU.     

In situ assessment of modification of sediment properties by burrowing invertebrates

Benthic organisms can significantly alter the physical properties of marine sediments, but it has hitherto been difficult to assess and quantify the effects of bioturbation. In situ geophysical techniques offer new methods for measuring these effects: measurement of acoustic shear-wave velocity and electrical resistivity allows nondestructive assessment of the properties of the grain framework and pore-fluid matrix, respectively, of the seabed sediment. The influence of burrowing invertebrates on the structural properties of sandy sediments at intertidal locations on the coast of Wales (UK) was investigated during the periol 1986–1987 using these techniques. Three species (Arenicola marina, Corophium arenarium and Lanice conchilega) were selected on the basis of their contrasting styles of burrow construction. All three species produced measurable and significant, although different, changes in bed properties. They modified shear-wave propagation through the bed by changing bed rigidity: while A. marina and C. arenarium decreased rigidity by creating open burrows, L. conchilega increased rigidity by building shell-lined tubes. All produced a decrease in electrical resistivity by altering porosity and/or tortuosity, which implies an increase in permeability; these changes were attributable not only to the presence of the burrows but also to modification of the between-burrow sediment texture and bed properties.     

Toxic equivalent concentrations (TEQs) for baseline toxicity and specific modes of action as a tool to improve interpretation of ecotoxicity testing of environmental samples

The toxic equivalency concept is a widely applied method to express the toxicity of complex mixtures of compounds that act via receptor-mediated mechanisms such as induction of the arylhydrocarbon or estrogen receptors. Here we propose to extend this concept to baseline toxicity, using the bioluminescence inhibition test with Vibrio fischeri, and an integrative ecotoxicity endpoint, algal growth rate inhibition. Both bioassays were validated by comparison with literature data and quantitative structure-activity relationships (QSARs) for baseline toxicity were developed for all endpoints. The novel combined algae test, with Pseudokirchneriella subcapitata, allows for the simultaneous evaluation of specific inhibition of photosynthesis and growth rate. The contributions of specific inhibition of photosynthesis and non-specific toxicity could be differentiated by comparing the time and endpoint pattern. Photosynthesis efficiency, measured with the saturation pulse method after 2 h of incubation, served as indicator of specific inhibition of photosynthesis by photosystem II inhibitors. Diuron equivalents were defined as toxicity equivalents for this effect. The endpoint of growth rate over 24 h served to derive baseline toxicity equivalent concentrations (baseline-TEQ). By performing binary mixture experiments with reference compounds and complex environmental samples from a sewage treatment plant and a river, the TEQ concept was validated. The proposed method allows for easier interpretation and communication of effect-based water quality monitoring data and provides a basis for comparative analysis with chemical analytical monitoring.     

Gas chromatographic methods for monitoring of wastewater chlorophenol degradation in anaerobic reactors

Wastewater samples from an anaerobic reactor were extracted with hexane and derivatized with diazomethane (method 1) and with acetic anidride (method 2). Gas chromatography with electron-capture detection (ECD) was employed for separating the parent compound and intermediates trichlorophenols (TCP) and dichlorophenols (DCP) which originated from the penta chlorophenol (PCP) degradation process. The relations between concentrations of PCP, TCP and DCP areas were linear in the range of concentrations of 0.2 to 8 mg/L and 0.025 mg/L to 5 mg/L for methods 1 and 2, respectively. The repeatability of the extraction methods was satisfactory, with variation coefficients lower than 11%. For method 1, at the fortification level of 0.2 mg/L, recovery of PCP, TCP, and DCP was 112%, 74% and 45%, respectively. For method 2, the corresponding recovery values at the fortification level of 0.1 mg/L were 91%, 93% and 103%, respectively. Storage of the frozen samples did not alter their PCP determination properties. The chromatographic methods adapted for chlorophenol determination in wastewater were suitable with relatively simple manipulation techniques. The obtained results were reproducible and allowed identification of intermediates formed during the PCP degradation process.     

Water quality effects of clearcut harvesting and forest fertilization with best management practices

Nine small (2.5 ha) and four large (70-135 ha) watersheds were instrumented in 1999 to evaluate the effects of silvicultural practices with application of best management practices (BMPs) on stream water quality in East Texas, USA. Two management regimes were implemented in 2002: (i) conventional, with clearcutting, herbicide site preparation, and BMPs and (ii) intensive, which added subsoiling, aerial broadcast fertilization, and an additional herbicide application. Watershed effects were compared with results from a study on the same small watersheds in 1981, in which two combinations of harvesting and mechanical site preparation without BMPs or fertilization were evaluated. Clearcutting with conventional site preparation resulted in increased nitrogen losses on the small watersheds by about 1 additional kg ha(-1) each of total Kjeldahl nitrogen (TKN) and nitrate-nitrogen (NO(3)-N) in 2003. First-year losses were not significantly increased on the large watershed with a conventional site preparation with BMPs. Fertilization resulted in increased runoff losses in 2003 on the intensive small watersheds by an additional 0.77, 2.33, and 0.36 kg ha(-1) for NO(3)-N, TKN, and total phosphorus, respectively. Total loss rates of ammonia nitrogen (NH(4)-N) and NO(3)-N were low overall and accounted for only approximately 7% of the applied N. Mean loss rates from treated watersheds were much lower than rainfall inputs of about 5 kg ha(-1) TKN and NO(3)-N in 2003. Aerial fertilization of the 5-yr-old stand on another large watershed did not increase nutrient losses. Intensive silvicultural practices with BMPs did not significantly impair surface water quality with N and P.