Enrichment of fetal cells from maternal blood by high gradient magnetic cell sorting (double MACS) for PCR-based genetic analysis

For simple and effective isolation of fetal cells from peripheral maternal blood, we combined depletion of maternal cells and enrichment of fetal cells by high-gradient magnetic cell separation (MACS). First CD45⁺ and CD14⁺ cells were depleted from maternal peripheral blood mononuclear cells by MACS. From the depleted fraction, CD71⁺ erythroid cells were enriched up to 80 per cent by MACS. This ‘double-MACS’ procedure yielded an average depletion rate of 780-fold and an average enrichment rate of 500-fold, with approximate recovery rates of 40–55 per cent. For paternity testing, cells from unseparated blood and the various fractions were analysed for polymorphism of the HLA-DQ-A1 locus and D1S80 locus by the polymerase chain reaction (PCR). In CD45⁻/CD71⁺ sorted cells from maternal blood, but not in unfractionated cells from maternal blood or CD45⁻/CD14⁻ cells, paternal alleles could be detected. In the CD45⁻/CD71⁺ fraction, the relative frequency of paternal alleles compared with maternal alleles ranged from 1 in 20 to 1 in 200 (determined by titration and depending on the quality of separation and biological variation). In 7 out of 11 cases, between weeks 12 and 25 of gestation, we could identify paternal alleles by PCR, either HLA-DQ-A1 or D1S80. This double-MACS procedure is simple, fast, efficient, and reliable for non-invasive prenatal diagnosis.     

Ökotoxikologie am Beispiel der Rheinverschmutzung durch den Chemie-Unf all bei Sandoz in Basel

A fire at a chemical manufacturing plant at Schweizerhalle (near Basel) in November 1986 and the subsequent release of toxic agrochemicals into the Rhine river is taken as a basis to discuss some problems and needs in ecotoxicological research. Especially serious is the lack of knowledge about chronic effects of mixtures of chemicals to individuals and whole ecosystems. There is an urgent need for generally applicable principles and concepts. A discussion of the relationship between toxic effects to fish, exposure time, and concentration provides some hints about the direction towards which research should proceed. Finally, eight theses pertaining to ecotoxicology summarize what can be learned from the analysis of a chemical spill.     

Elektronisches Publizieren: Möglichkeiten zu wenig genutzt

Naturwissenschaften Aktuell

Innovative chemische Grundlagenforschung     

Die Regulation der Blutgerinnung

The patency of the vascular tree as well as thrombus formation are achieved by the interactions of several components of the circulating blood and the vessel wall. Circulating glycoproteins (coagulation factors), cellular elements of the blood (platelets), the fibrinolytic system, the endothelium of the vasculature and inhibitors that are present in the blood stream are involved in the maintainance of the subtle balance between anti-coagulant and pro-coagulant forces. Regulation of the activity of thrombin represents a key event in this process.