Detection of fetal HLA-DQα sequences in maternal blood: A gender-independent technique of fetal cell identification

The objective of this study was to detect fetal HLA-DQα gene sequences in maternal blood. HLA-DQα genotypes of 70 pregnant women and their partners were determined for type A1. We specifically sought couples where the father, but not the mother, had genotype A1. In 12 women, maternal blood samples were flow-sorted. Candidate fetal cells were isolated and amplified by using PCR primers specific for a paternal HLA-DQα A1 allele. Fetal HLA-DQα A1 genotype was predicted from sorted cells; amniocytes or cheek swabs were used for confirmation. Six of twelve sorted samples had amplification products indicating the presence of the HLA-DQα A1 allele; 6/12 did not. Prediction of the fetal genotype was 100 per cent correct, as determined by subsequent amplification of amniocytes or cheek swabs. We conclude that paternally inherited uniquely fetal HLA-DQα gene sequences can be identified in maternal blood. This system permits the identification of fetal cells independent of fetal gender, and has the potential for non-invasive prenatal diagnosis of paternally inherited conditions.     

Expanding multiple marker screening for Down's syndrome to include Edward'S syndrome

Information on maternal age and maternal serum alpha-fetoprotein, unconjugated oestriol (uE₃), and human chorionic gonadotrophin (hCG) levels was used to investigate retrospectively the effect of estimating Edward's syndrome risk in women having multi-marker screening for Down's syndrome. The screened population comprised 15 pregnancies affected by Edward's syndrome, 15 with Down's syndrome and 5472 unaffected pregnancies. The use of all three markers to estimate Edward's syndrome risk would have led to the detection of 10–12 (67–80 per cent) cases with a false-positive rate of 0.3–0.6 per cent depending on the risk cut-off. A further case would have been detected as a result of screening for Down's syndrome alone. Similar results were obtained when the Edward's syndrome risk was based on uE₃ and hCG only. These data suggest that extending Down's syndrome screening to include Edward's syndrome risk will yield a high detection rate with only a small increase in the false-positive rate.     

Carbon isotope ratios in logged and unlogged boreal forests: Examination of the potential for determining wildlife habitat use

Due to assimilation of recycled CO₂ from litter decomposition and photosynthetic changes in carbon fractionation at low light levels, the foliage at the base of a forest is often more depleted in¹³C compared to that exposed to the atmosphere in either the canopy or in open clearings. This is referred to as the canopy effect. African research has indicated that these habitat differences in foliar ¹³C can be substantial enough to affect the carbon isotope ratios of resident fauna. Previous work documenting a 30-year chronology on moose teeth from Isle Royale National Park indicated a progressive depletion in¹³C and suggested that this could be due to forest regrowth following extensive burning. The present study examined the assumption implicit in this hypothesis that foliar ¹³C varies between open and closed boreal forest sites. I found a marginal canopy effect of 2 ¹³C difference between upper canopy and ground flora for a forest in northwestern Ontario and an average difference of 1.2 in under- and mid-story vegetation between closed forests and open clear-cuts. Because of these small differences, the utility of carbon isotope analysis in quantifying temporally integrated exploitation of deforested habitats will be low for northern boreal locations. In denser forests, such as those in the tropics or western North American where the canopy effect can be expected to be much greater, ¹³C analysis may still offer some promise for determining selection by wildlife of disturbed habitats.     

Larval development in the Antarctic nemertean Parborlasia corrugatus (Heteronemertea: Lineidae)

Embryonic and larval development were followed from fertilisation to settlement in the Antarctic heteronemertean Parborlasia corrugatus (McIntosh, 1876). The first cleavage occurred 10 to 15 h after fertilisation, and the second at 17 h. Larvae hatched at the gastrula stage, between 170 and 200 h post-fertilisation, and were 150 m in diameter. Early larval stages aggregated in dense groups near the surface of incubation vessels and were positively phototactic. Early pilidium larvae were recognisable 435 h post-fertilisation. They were 155×152 m in size, and possessed a complete apical tuft of cilia and a full marginal band of locomotory cilia. At this stage, the gust was visible through the body wall, and the mouth was open and was 40 m in diameter. Late pilidia, 222×193 m in size, were helmet-shaped. They had an apical tuft over 100 m long, and possessed a lobed marginal band of locomotory cilia. Pilidia were observed aggregating close to the bottom of incubation vessels 1200 to 1350 h (50 to 56 d) after fertilisation, and this was interpreted as settlement behaviour. At this stage, the apical tuft had been lost and they were highly contractile, being capable of compressing their bodies. However, neither developing juveniles within the larval envelope nor hatched juveniles were observed. Pilidia consumed the microalgae Tetraselmis suecica, Thalassiosira pseudonana and Isochrysis galbana. They also fed on particulate organic material < 1 m in size, as shown by the presence of material in the guts of larvae offered filtered extracts of algal cultures. There was some indication that larvae could use dissolved organic material, since pilidia held in seawater with organic material removed did not survive as long as those in filtered seawater or in filtered water with added amino acids. However, the only larvae to exhibit settlement behaviour in the feeding experiments were those offered Tetraselmis succica and Thalassiosira pseudonana, and these required a longer development time to reach this stage than pilidia in the standard cultures, where a mixed algal diet was offered.     

Relation between heavy metal concentrations in salt marsh plants and soil

The aim of the research reported here was to investigate the relation between heavy metal concentrations in salt marsh plants, extractability of the metals from soil and some soil characteristics. In April 1987, Spartina anglica and Aster tripolium plants and soil were collected from four salt marshes along the Dutch coast. The redox potential of the soil between the roots of the plants and at bare sites was measured. Soil samples were oven-dried and analyzed for chloride concentration, pH, fraction of soil particles smaller than 63 microm (f < 63 microm), loss on ignition (LOI) and ammonium acetate and hydrochloric acid extractable Cd, Cu and Zn concentrations. The roots and shoots of the plants were analyzed for Cd, Cu and Zn. Because drying of the soil prior to chemical analysis might have changed the chemical speciation of the metals, and therefore the outcome of the ammonium acetate extraction, a second survey was performed in October 1990. In this survey A. tripolium plants and soil were collected from two salt marshes. Fresh and matched oven-dried soil samples were analyzed for water, ammonium acetate and diethylene triaminepentaacetic acid (DTPA) extractable Cd, Cu and Zn concentrations. The soil samples were also analyzed for f < 63 microm, LOI and total (HNO(3)/HCl digestion) metal concentrations. Soil metal concentrations were correlated with LOI. Drying prior to analysis of the soil had a significant effect on the extractability of the metals with water, ammonium acetate or DTPA. Plant metal concentrations significantly correlated only with some extractable metal concentrations determined in dried soil samples. However, these correlations were not consistently better than with total metal concentrations in the soil. It was concluded that extractions of metals from soil with water, ammonium acetate or DTPA are not better predictors for metal concentrations in salt marsh plants than total metal concentrations, and that a major part of the variation in metal concentrations in the plants cannot be explained by variation in soil composition.     

Population dynamics of Mytilicola intestinalis in Mytilus edulis in south west England

The population dynamics of Mytilicola intestinalis Steuer in mussels (Mytilus edulis L.) from the River Lynher, Cornwall, England, have been studied over 3 years. By transplanting uninfested mussels from the River Erme, South Devon, into the Lynher mussel bed, the study was extended to the growth and development of new infestations under natural conditions. Female Mytilicola intestinalis are shown to breed twice, and two generations of parasites coexist for most of the year, with recruitment taking place in summer and autumn. One generation contributes its first brood to the autumn recruits before overwintering and contributing its second brood to the following summer's recruits. The other generation overwinters as juvenile and immature stages to contribute its two broods successively to the summer and autumn recruits. Environmental temperatures are believed to control the rates of development at all stages rather than acting as triggers in the onset or cessation of breeding at specific times. There is no evidence to support the contention that heavily infested mussels are killed, and parasite mortality is shown to be density-independent.     

Cost-effective selection of sampling frequencies for regulatory water quality monitoring

A dynamic programming code was formulated for the purpose of assigning sampling frequencies throughout a regulatory water quality monitoring network in order to optimize the statistical performance of the network while operating within a fixed budgetary constraint. The statistical objective is to achieve the greatest possible station to station uniformity in confidence interval widths about annual geometric means of the measured water quality variables and to keep the average confidence width reasonably small. The objective function is the sum (over several selected variables and all stations) of the normalized positive deviations of the predicted confidence interval widths from preselected design confidence interval widths. The code was designed to account for the effects of deterministic seasonal variation and serial correlation of the water quality observations by incorporating the results of the time series analysis of historical quality data. The economic constraint ensures that the annual operating cost of the system, including direct costs of travel and laboratory analysis, will not exceed the allowable budget. As an example situation, the dynamic programming code was used to assign sampling frequencies to the nine stations in Illinois from which historical quality data had been obtained and analyzed. Using five design quality constituents and representative travel and laboratory costs, an “optimal” design was produced. The optimal design achieved a 10% improvement in uniformity (standard deviation) of confidence interval widths when compared to a more traditional design based on the same budget and using identical sampling frequencies at every station.