A simple and effective approach for detecting maternal cell contamination in molecular prenatal diagnosis

The presence of maternal cells in fetal samples constitutes a serious potential source for prenatal misdiagnosis. Here we present our approach for detecting maternal cell contamination (MCC) at prenatal diagnosis for eight monogenic disorders (autosomal recessive: β-thalassaemia, sickle-cell anaemia, cystic fibrosis, prelingual deafness; autosomal dominant: achondroplasia, Huntington disease, myotonic dystrophy, neurofibromatosis type I; X-linked: spinobulbar muscular atrophy). Our aim was to apply a simple and low-cost approach, which would easily and accurately provide information on the fetal tissue MCC status. MCC testing was applied to cases of recessive inheritance where the primary mutation screening of the fetus revealed the presence of the maternal mutation, to cases concerning dominant inheritance and to cases of multiple gestation. The potential presence of maternal cells was determined by the amplification of the 3′-HVR/APO B, D1S80, THO1 and VNTRI of vWf polymorphic loci, which have previously demonstrated high heterozygosity in Caucasians. Among 135 prenatal diagnoses, 44 finally needed to be tested for MCC (32.6%). MCC was detected in four cases, where DNA was isolated directly from chorionic villi samples (CVS), and in one case with DNA isolated directly from amniotic fluid (AF). In almost 90% of cases a simple test of one polymorphic locus provided sufficient information about MCC. The choice of the appropriate locus is therefore essential, while the simultaneous screening of both parents provides the means for distinguishing non-informative sites about MCC. Copyright © 2002 John Wiley & Sons, Ltd.     

Variability in larval development of the tropical reef fish Pomacentrus amboinensis (Pomacentridae): the parental legacy

The effects of food availability, female size, and social interactions on the quality of Pomacentrus amboinensis larvae at hatching were examined using two field-based experiments. In Experiment 1, food availability and female size significantly influenced size, eye diameter and levels of yolk reserves of larvae at hatching. Small females (47 to 52 mm standard length, SL) whose diets were not supplemented, produced the longest larvae (3.0 ± 0.01 mm total length, TL) with the least yolk reserves (50.1 ± 1.04 μm²). Irrespective of female size, those that received additional food produced larvae with the largest yolk-sacs (large females: 87.60 ± 1.53 μm²; small females: 80.14 ± 1.24 μm²). In Experiment 2, interactions with conspecifics had a greater affect on the somatic development of larvae at hatching than food availability. Increased social interactions resulted in larvae that were ⋍3% longer, with 2% greater head depth, than larvae from females that spawned in isolation on the experimental reefs. Fed females produced larvae with ⋍20% more yolk than larvae from females whose diets were not supplemented. All three factors (food availability, female size, and intensity of social interactions) tested within these experiments vary spatially and temporally among reefs. There is the potential, therefore, for larvae at the onset of the planktonic stage to vary in quality, level of development, and probability of survival. Received: 12 August 1996 / Accepted: 26 August 1996     

Site characteristics,growth and nutrition of natural red pine stands in newfoundland

Red pine (Pinus resinosa Ait.) is rare (< 15 000 mature trees) in Newfoundland and is known from only 22 locations in the central region. Red pine occupies 3 major site types in Newfoundland: 1) red pine on medium-textured sands (RP1), 2) red pine on coarse-textured glaciofluvial deposits (RP2), and 3) red pine on Folisols over bedrock (RP3). The succession of red pine site types after cutting is from red pine to Kalmia-black spruce (Picea mariana (Mill.) B.S.P.) for RP1, and to Cladonia-Kalmia-black spruce for types RP2 and RP3. Succession after fire is usually to the pre-fire type, but this depends on the severity of the fire.Although occupying a relatively poor site, red pine at 60–70 years reaches heights in excess of 18 m, dbh in excess of 40 cm, and individual tree volumes greater than 1 m³ were recorded in 75 stemanalyzed fire-killed trees. Black spruce on that same site produces less than one-third that volume in 60 years. Merchantable volume of 140–280 m³ ha^-1 were recorded i.e., Canada Land Inventory (CLI) forest capability class 5 and class 4 ratings. This raises the CLI rating two capability classes if red pine were occupying these poor quality sites over black spruce. In terms of nutrition, even the best growing red pine are nitrogen (N) deficient as shown by foliar analysis. All natural stands have foliar N concentrations below 1.3% which is the critically low level shown in the literature. Immediately after fire, foliar concentrations reach this level but are usually about 1% or less. Most other nutrients are low but are within the generally reported adequate levels in testing for P, K, Ca and Mg.Fire influences soil nutrient availability as pH increases in the RP1 type. Burning temperature also affects soil pH and the understory vegetation. The RP2 type loses more N in hotter burns on this site type and more N is tied up in these ortstein hardpan soils. The pattern of regeneration following wildfire is related to slope, density, age and species mixture of the stand as well as the thickness and composition of the duff layer.     

Effects of high temperature on larval development and metamorphosis of Arachnoides placenta (Echinodermata: Echinoidea)

Adults of the sea urchin Arachnoides placenta (L.) were induced to spawn, and eggs were fertilized at 28°C in September 1989. After 5 min, eggs were transferred to 28, 31, 34, or 37°C and reared to metamorphosis. Embryos were observed at 20-min intervals during the first 2 h; larvae were observed daily. The cleavage was higher at higher temperatures. Embryos reared at 28°C were still at the 4th cleavage (16-cell stage) after 100 min, while those at 34°C had reached the 5th cleavage (32-cell stage). All embryos reared at 37°C died on the second day. Incidence of abnormality was 20 to 30% at 28 and 31°C, 48% at 34°C, and 77% at 37°C. The 8-arm stage was reached after 4 d at 28°C, 3 d at 31°C and 2 d at 34°C. Larvae displayed decreasing body length and arm length with increasing temperature. Larvae at 31°C have relatively long arms, as a result of a decrease in body length, not because of increased arm length. Incidence of metamorphosis was 43.9±1.7% (mean/plusmn;SD) at 28°C, 24.5±1.9% at 31°C, and 5.3% at 34°C. The size of metamorphosed juveniles was significantly larger at 28°C than at 31 and 34°C. Temperatures of 31°C negatively affect larvae and juveniles of the sand dollar.     

The effect of selenium on fluoride — clay interactions: Possible environmental health implications

Previous epidemiological studies have shown that dental fluorosis is endemic in the lowland, dry zone of Sri Lanka, which is considered to be an area in which excessive quantities of fluorides are present in the drinking water supplies. It has been found that kaolinitic clay forms a suitable raw material in the defluoridation of water.It is shown that there is a noticeable effect of selenium and media pH on the reactions involved in the interaction of fluoride with clay. In this study, 1 mM fluoride solutions containing SeO ₃ ^2– (selenite) concentrations of 0 mM, 0.1 mM, 0.5 mM and 1 mM were used in the reactions with kaolinitic clay. The effect of pH was monitored in the range 4 to 8. It was observed that fluoride adsorption was maximum at a pH of 5.6 without either SeO ₃ ^2– or SeO ₄ ^2– , the adsorption capacity being 15.2 mol F^– g^–1 clay. However, when the SeO ₃ ^2– concentration was increased up to 0.5 mM at this optimum pH, the adsorption capacity reduced to 12.8 mol F^– g^–1 clay. Monitoring of the effect of SeO ₄ ^2– and media pH on fluoride adsorption showed that when the SeO ₄ ^2– concentration increases from zero to 0.1 mM, there is a reduction of fluoride adsorption capacity. However, when the SeO ₄ ^2– concentration is further increased from 0.1 mM to 1.0 mM, there was an increase in the fluoride adsorption capacity, indicating a more consistent effect of SeO ₃ ^2– on fluoride-kaolinitic clay interaction than SeO ₄ ^2– .Fluoride concentrations in drinking water supplies have a marked effect on dental health and the geochemistry of selenium appears to play an important role in the geochemical mobility of fluoride ions.     

Orientation ofRhizophagus grandis (Coleoptera: Rhizophagidae) to oxygenated monoterpenes in a species-specific predator-prey relationship

Summary The predator,Rhizophagus grandis, is linked to its specific prey,Dendroctonus micans, by semiochemical signals regulating oviposition and long range orientation. A mixture of simple oxygenated monoterpenes [(–)—fenchone, (–)—pinocamphone, rac. camphor, terpinene-4-ol, borneol, fenchol and verbenone], identified from the frass ofD. micans has been found to be extremely active in a flight windtunnel as a long—range attractant for the predator. The mixture elicited 84 % of the response to larval frass of the prey. Excluding pinocamphone from the mixture did not influence its activity. Also, changing the absolute configurations of some of the components (fenchol, terpinene-4-ol and borneol) did not influence the predators' response. However, the addition of (–)—-terpineol increased the attractivity of the synthetic blend to almost that of larval frass of the prey. Ecological implications of the identified semiochemicals and their use in pest management are discussed.     

Effect of nickel on some aspects of protein metabolism in the gill and kidney of the freshwater fish, Cyprinus carpio L

Nickel has an adverse effect on some aspects of protein metabolism of the freshwater fish, Cyprinus carpio. The main changes observed were: (a) Decrease in soluble, structural and total proteins, AlAT and AAT activities with an increase in the levels of free amino acids, protease and GDH activities and ammonia in the gill and kidney at 1, 2, 3, and 4 days of exposure to a lethal concentration, 40 mg litre(-1) of nickel; (b) Increase in soluble, structural and total proteins, free amino acids and the activities of protease, AlAT, AAT and GDH with a decrease in ammonia and urea in these organs at 1, 5, 10 and 15 days of exposure to sublethal concentration, 8 mg litre(-1); (c) The magnitude in these changes increased over time with both concentrations of the metal, and was more marked in gill than in kidney.     

A test on social behavior as a cause of dispersal of spruce grouse

Summary Social behavior is one hypothesized cause for dispersal. We tested this hypothesis on spruce grouse (Dendragapus canadensis) in two years by removing adults in summer and monitoring emigration of juveniles in autumn. Despite the absence of adults, juveniles emigrated from the natal range at rates comparable to juveniles on control areas. Data on density and dispersal from earlier spruce grouse studies are reviewed, which show that emigration is constant despite substantial differences in density, and that a clear female sex bias occurs across a wide range of densities. Results do not support the hypothesis that social behavior influences dispersal nor do they support a prediction of the Oedipus hypothesis that no sex bias occurs in polygynous birds. Offprint requests to: D.M. Keppie     

Deposition of atmospheric ammonia to moorlands

Micrometeorological methods were applied to measure fluxes of atmospheric ammonia (NH3) to moorlands. Measurements were made in a wide variety of surface conditions and included both Calluna vulgaris (L.) Hull and Eriophorum vaginatum L. dominated sites. NH3 was found to deposit rapidly to all the sites investigated, providing large deposition velocities (Vd, typically 10-40 mm s(-1)) and usually minimal surface resistances (rc). A small number of measurements were made in frozen conditions and suggest a possible exception to this pattern with mean rc of 50-200 s m(-1). The effect of vegetation drying was also investigated and a possible increase in rc observed, though this was small (< 10 s m(-1)). The results are interpreted in terms of the processes controlling exchange; it is shown that NH3 deposition is predominantly to the leaf surfaces and that the net NH3 compensation point approaches zero. Annual estimates show that dry deposition of NH3 is a major source of atmospheric nitrogen to moorland ecosystems. For two typical UK sites subject to background air concentrations, NH3 dry deposition is of similar magnitude to equivalent NH4+ inputs in wet deposition. In the vicinity of emission sources, NH3 dry deposition is expected to dominate inputs of atmospheric nitrogen.