An investigation of methods for enriching trophoblast from maternal blood

Trophoblast deportation is known to occur in normal human pregnancy, but it is not yet clear whether these cells routinely enter the maternal peripheral circulation and are available as a source of fetal DNA for non-invasive prenatal diagnosis of genetic disorders. To resolve this issue requires an efficient method of enriching trophoblast from maternal blood combined with a means to confirm its identity. Five different techniques were tested on ten retroplacental blood samples to determine the most sensitive and operator-efficient method. Lysis of red cells alone gave the best recovery of trophoblast but had to be discounted, together with Ficoll density gradient centrifugation, due to the very low purity and the excessive time required. Fluorescence-activated cell sorting (FACS) of pre-enriched trophoblast resulted in the lowest recovery rate (8 per cent) despite a 3250-fold enrichment and a very high purity. Immunomagnetic beads (Dynabeads) coated with anti-CD 16 antibody proved to be the best method for the subsequent immunocytochemical characterization of deported trophoblast. However, IO beads coated with anti-CD45 antibody may be more useful for isolating trophoblast for prenatal diagnosis due to the high purity, enrichment (32-fold), and recovery rate (78 per cent) obtained with this method.     

COST EFFECTIVE ARSENIC REDUCTIONS IN PRIVATE WELL WATER IN MAINE1

ABSTRACT: This study was undertaken to investigate the cost effectiveness of selected arsenic avoidance methods. Annual costs of reverse osmosis (RO), activated alumina (AA), bottled water, and rented and purchased water coolers for various household sizes in Maine were compared. Relative ranking of systems shows that RO ($411 annually) is the most cost effective, followed by AA ($518) and one‐gallon jugs of water ($321 to $1,285), respectively, for households larger than one person. One‐gallon jugs ($321) followed by 2.5‐gallon jugs ($358) of water were found to be the most cost effective for households of one person or for households with arsenic III concentrations of 0.02 to 0.06 mg/L and arsenic V concentrations of 0.08 to 1.0 mg/L. Point‐of‐entry systems and water coolers were not found to be cost effective under any of the study's conditions. The research reported here will help states make more definitive treatment recommendations to households regarding the cost effectiveness of alternative treatment systems to reduce arsenic concentrations below 0.01 mg/L. While arsenic removal technologies are improving, which enhances removal rates and reduces costs, the major insights from this analysis appear to be reinforced by technological improvements.     

An approach to preimplantation diagnosis of β-thalassaemia

Using the polymerase chain reaction (PCR), it was possible to amplify a single copy fragment of the β-globin gene from 2–32 human embryonic cells obtained from arrested preimplantation embryos. For the detection of β-thalassaemia mutations, allele specific priming of the PCR using nested primers was employed using approximately 10 pg of DN A from individuals known to carry these mutations. This approach was successful in detecting the presence or absence of five Asian Indian β-thalassaemia mutations that were selected for this study. In spite of meticulous precautions against contamination, false-positive amplification was observed, a problem that will have to be overcome before this approach can be used in clinical practice.     

A study of the variations in lipid levels,lipid class composition and fatty acid composition in the first stages ofArtemia sp.

The lipid class composition and the fatty acid composition of total lipids of the cysts, newly hatched nauplii and 24-h-old metanauplii of a Spanish parthenogenetic diploid strain ofArtemia sp. were studied. Substantial differences in the total lipid level occurred among these stages, with a marked increase from the cyst to the nauplii being followed by a decrease in the metanaupliar stages. This variation affected the absolute levels (mg/g dry wt) of the total lipid classes and individual fatty acids, although the percent composition of the fatty acids in total lipid was essentially unchanged. An exception occurred during hatching in that the percentages of 16:0 and 16:1n-7 in total lipid decreased whereas that of 20: 5n-3 increased. The lipid classes showed higher variation than the fatty acids both in absolute and in relative terms, and in particular, the ratio of phosphatidylcholine:phosphatidylethanolamine decreased progressively from cysts to nauplii and metanauplii. The implications of these findings for the use ofArtemia sp. as a larval feed in aquaculture are considered.     

Parental investment decision rules and the Concorde fallacy

Summary Investment theory states that animals should base their parental investment decisions on expected benefits, and not on whether or not past investment will be wasted. Otherwise, they would comnit the Concorde fallacy. If reproduction has a cost, however, then past investment and expected benefits are necessarily confounded. Assuming a cost of reproduction, animals will be selected to maximize their remaining lifetime reproductive success, subject to a tradeoff between present and future reproduction (Williams' principle). We extend Williams' principle and develop an experimental design that would allow past investment and expected benefits to be varied independently. This design illustrates the importance of the value of the brood relative to the value of future reproduction.     

Allopaternal care in the fathead minnow,Pimephales promelas: females prefer males with eggs

Summary Some species of fishes with exclusive male parental care exhibit the phenomenon of allopaternal care; that is, some males acquire and care for other males' eggs. We conducted a series of experiments to investigate the dynamics and evolution of allopaternal care in one such species, the fathead minnow (Pimephales promelas). In choosing a nest site, a newly reproductive male tended to take over the nest site of a parental male by evicting the resident male, rather than occupy a physically identical empty nest site. The new male generally cared for the old male's eggs, and in most cases, daily egg survival improved under the new male's care. When males were given a choice among unguarded nest sites, they preferred to occupy nest sites already containing eggs. When eggs were randomly assigned to nesting males, females preferred to spawn with males who had eggs in their nest sites. Thus, it appears that female preference for males with eggs led to the evolution of allopaternal care in the fathead minnow.     

Effects of diet on fatty acid composition of body zones in larvae of the sea bass Dicentrarchus labrax: a chemometric study

Larvae of the sea bass Dicentrachus labrax were fed four Artemia sp. diets for 28 d. Three were nauplii enriched with emulsions of polyunsaturated fatty acids, and the fourth nauplii enriched with baker's yeast. At the end of the experimental period, the fatty acids of the bodies, heads and eyes of the larvae were analysed. A multivariate statistical method (discriminant analysis, DA) applied to the data revealed anatomical as well as dietary fatty acid pattern-discrimination. We propose here the use of discriminant analysis as a pattern-recognition method that will help to integrate the fatty acid information obtained in nutritional studies.