Detection of fetal cells in transcervical samples and prenatal diagnosis of chromosomal abnormalities

Transcervical samples collected by lavage, aspiration, and cytobrush from women between 6 and 13 weeks of gestation were tested for the presence of fetal cells using fluorescence in situ hybridization (FISH) with probes for chromosomes X, Y, 1, and 21, and by polymerase chain reaction (PCR) amplification of DNA sequences derived from chromosomes X, Y, and 21. With a few exceptions, a good correlation was observed between the results of sexing the fetuses using FISH or PCR on transcervical cell (TCC) samples retrieved by lavage and those obtained by testing fetal (placental) tissue. In a comparative study between TCC samples collected by lavage or cytobrush, the sex of the fetus was correctly diagnosed by PCR amplification of a Y-derived DNA sequence. Variable results were observed with samples obtained by aspiration, mainly because this procedure was found to be more prone to failure to remove thick mucus without previous injection of physiological saline. Chromosome 21-derived small tandem repeats (STRs) of fetal origin were successfully detected in about 40 per cent of TCC samples recovered by lavage. Two cases of chromosomal abnormalities, one of trisomy 21 and one of triploidy, were detected in TCC samples in the course of our investigations.     

Dystrophin gene analysis and prenatal diagnosis of Duchenne muscular dystrophy in Russia

Of 126 families referred for counselling of Duchenne muscular dystrophy (DMD), DNA analysis has been suggested to 119 families with at least one affected child or with an affected close male relative of the woman at risk of being a DMD carrier. A large proportion (about 80 per cent) of the families were represented by sporadic cases (only one affected individual). By means of multiplex polymerase chain reactions with different sets of oligoprimers providing amplification of 10–11 different exons, altogether 49 dystrophin gene deletions were identified (41 per cent). Eighteen deletions clustered in the 5′ ‘hot spot’ region of DMD cDNA and 36 in the distal half of the central rod domain around exons 43–53. An unusually high frequency (18 per cent) of deletions involving exons 17–19 was discovered. Large deletions extending through both ‘hot spot’ regions and thus occupying over 30–40 exons were recorded in five cases (10 per cent). Seventy-six of 94 families were found to be informative by RFLP analysis for intragenic or extragenetic DNA probes. Carrier status was ascertained in 20 and rejected in 32 female relatives in 40 DMD families. Eight DMD-affected fetuses were diagnosed prenatally by direct deletion testing or by RFLP analysis. Feasible interpopulation variations in the dystrophin gene deletion pattern are discussed. The prospects for more effective prenatal diagnosis and carrier detection in high-risk DMD families in Russia are briefly outlined.     

Assessment of compost quality for its environmentally safe use by means of an ecotoxicological test on a soil organism

A research project was carried out to evaluate toxicological effects of compost addition to agricultural soil using the earthworm Eisenia foetida (Annellida) as a representative organism of the soil fauna. Moreover, the correlation between compost biochemical stabilization and toxicity at different phases of the composting process was assessed. Samples were collected from three composting plants at three different maturation levels (beginning of the composting process, intermediate compost after bio-oxidation, and mature refined compost). Two tests were performed: a standard chronic solid-phase test and an acute solid-phase test (developed originally by the authors). In the first test, the measured end-points were mortality, growth and reproduction; while in the second test earthworms’ behavior was evaluated. The chosen compost concentrations in soil ranged from 2.5 to 100 %, with the aim of obtaining the toxicological parameters (LC50) and to mimic real agricultural dosages for the lower concentrations. Results indicated an increase in compost toxicity with greater compost concentrations; in particular, agricultural compost dosage below 10 % showed no toxicity. Moreover, toxicity did not decrease during composting; intermediate compost showed the highest LC50 values. As a consequence, no correlation was ascertained between the results of ecotoxicological analysis and waste biochemical stability parameters during the composting process.     

Ecology of the metahaline pool of Di Zahav,Gulf of Elat,with notes on the siphonocladacea and the typology of near-shore marine pools

The hydrobiology of a metahaline near-shore pool, at Di Zahav on the Sinai coast of the Red Sea, has been studied. Salinity of the pool fluctuates between 45 and 60, and winter temperatures are as low as 10°C. The shore consists partly of beach rock, some of it still in statu nascendi. Serpulid reef-boulders also occur, covered with typical marine species. Three siphonocladacean green algae (Siphonocladus rigidus, Dictyosphaeria cavernosa, and Valonia utricularis), form abundant crusts on the boulders. Several peculiar faunal elements are discussed. Traits in common with the Suez Canal biota are discerned. A typology for the nearshore pools of Sinai is proposed.     

The re-use of derelict land

Conclusions Despite these problems there are a number of positive cases, where exemplary solutions have been found after long processes of negotiation and compensation and which can stand as examples of what can be achieved in a re-organisation of the environment and housing structure between the Ruhr and the Lippe.The transformation of the old Rosenblumendelle pit in Mülheim into a shopping centre, the create of a modern industrial park on the land of the old Friedrich Ernestine pit in Essen (from the pit closure to the establishment of the park the whole process only took four and a half years!), and the transformation of the old Dahlhauser Tiefbau pit in Bochum into a leisure and recreation area, which will soon be completed. These and other examples show that reclamation, given favourable circumstances, can be very successful indeed.     

First report of prenatal diagnosis of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency in a pregnancy at risk

Prenatal diagnosis of long-chain 3-hydroxyacyl-CoA dehydrogenase (3-HAD) deficiency was performed in a family at risk. The diagnosis of an affected fetus was carried out by enzyme assay in cultured chorionic villus cells.     

First trimester studies of A fetus at risk for triose phosphate isomerase deficiency

A first trimester prenatal diagnosis was offered to a mother whose child had died of haemolytic anaemia and multisystem disease caused by TPI deficiency. The deficiency state was characterized by greatly reduced TPI activity in both erythrocytes and peripheral lymphocytes. Specific activity of TPI in trophoblast homogenates from the index fetus was about 30 per cent less than in the controls, but the heat stability test showed overlap. These data were confirmed in uncultured and cultured amniotic cells, where glycolytic intermediate concentrations DHAP, GAP and FDP fell in the range of controls. These results suggested that the fetus was a TPI heterozygote. This prenatal prediction was confirmed by RBC and haematological studies at birth.     

Methods for the transcervical collection of fetal cells during the first trimester of pregnancy

Before termination of pregnancy, four techniques for retrieving fetal cells transcervically were investigated: uterine lavage, endocervical lavage, cytobrush and mucus aspiration. The yield of fetal cells in these samples was studied and found to be somewhat better after uterine lavage. A preliminary assessment of the safety of mucus aspiration was carried out before transcervical chorionic villus sampling (CVS) in continuing pregnancies. No difference in outcome to a control group having only CVS was found.