Rapid sexing of human embryos by non-radioactive in situ hybridization: Potential for preimplantation diagnosis of X-linked disorders

Sixty spare human embryos at various stages of preimplantation development were prepared for cytogenetic analysis. Fluorescent staining of those with metaphases allowed scoring for the presence of a Y chromosome. In situ hybridization was then performed using a biotinylated Y-specific sequence, and the probe was detected by a standard streptavidinlinked alkaline phosphatase system. This enabled comparison of the chromosomal sex with that obtained after in situ hybridization in 28 embryos, and the sexing result obtained by the two methods was concordant in all cases. A further 21 embryos in which no metaphase chromosomes were obtained were sexed by biotinylated in situ hybridization only. Overall, 66 per cent of male interphase nuclei demonstrated a Y-specific hybridization signal. Results were obtained in under 24 h, which may permit the sexing of an embryo biopsied during cleavage and the transfer of sexed embryos at the blastocyst stage to the mother's uterus in the same cycle as oocytes are collected for in vitro fertilization.     

Offspring sex ratio in relation to parental structural size and body condition in the long-lived wandering albatross (<Emphasis Type="Italic">Diomedea exulans</Emphasis>)

Sex ratio theory is one of the most controversial topics in evolutionary ecology. Many deviations from an equal production of males and females are reported in the literature, but few patterns appear to hold across species or populations. There is clearly a need to identify fitness effects of sex ratio variation. We studied this aspect in a population of a long-lived seabird, the wandering albatross (Diomedea exulans), using molecular sex-identification techniques. We report that parental traits affect both (1) fledgling traits in a sex-dependent way and (2) chick sex: Sons are overproduced when likely to be large at fledging and, to a lesser extent, daughters are overproduced when likely to be in good body condition at fledging. Because for the same population, a previous study reported that post-fledging survival was positively affected by size in males and by body condition in females, our results suggest that wandering albatrosses manipulate offspring sex to increase post-fledging survival.     

Sabaki River sediment load and coral stress: correlation between sediments and condition of the Malindi-Watamu reefs in Kenya (Indian Ocean)

Sediment discharges from rivers have a negative impact on coral reef ecosystems. Indicators of coral decline measured in the present study were: (1) injury to living stony corals; (2) soft coral cover; and (3) bare rocky substrate suitable for colonization by corals. The relationship between these indicators and the distribution of terrigenous sediment was studied for the Malindi-Watamu fringing reef complex along the Kenyan coast off East Africa during 1982 and 1983. Decline of this reef had been repeatedly noted during the preceding decade. The influence of terrigenous sediment from the Sabaki River appears to be strongest in the Watamu area in the south and in the northern-most part of the Malindi reef area. Correlations, between each of the above three coral stress response indicators, on the one hand, and quantitative indicators of sediment loading, on the other hand, were not clear. However, a combined coral stress indicator involving all three factors was shown to have a clear relationship with terrigenous sediment loading and provided a rapid means of field evaluation of the effects of sediment stress on stony corals. Values for the combined coral stress indicator were found to increase in proportion to increasing values of terrigenous sediment loads in both study areas. A higher coral stress indicator value means a high proportion of injured or algae infested corals, and/or a high soft coral cover, and/or a high proportion of rocky substrate suitable for, but unoccupied by, living corals.     

The role of microbial mats during primary succession in calcareous dune slacks: an experimental approach

Laminated microbial mats from a sandy beach plain were grown in water-saturated pots in a glass house for six months and then used to assess their effect on the establishment of juveniles of three plant species representing different successional stages in dune slack development. The selected species wereSamolus valerandi, characteristic of pioneer stages,Calamagrostis epigejos, characteristic of more productive, late successional stages, andJuncus alpinoarticulatus, which occurs in a wide range of successional stages. Juveniles of all three species that were placed on top of intact living microbial mats established themselves in the mat.C. epigejos andJ. alpinoarticulatus survived for several weeks but later on their numbers decreased and the total biomass production of the species after six months was poor.S. valerandi, in contrast, grew profusely in intact microbial and algal mats. Heating of the microbial mat by heat sterilization, prior to the experiment, did not improve the performance of the species. When the juveniles were planted in the microbial mats after breaking the surface of the mat, the survival of juveniles ofC. epigejos andJ. alpinoarticulatus was much higher and so was the biomass of surviving plants after six months. Planting ofSamolus in the mats had some positive effect on the survival percentage of the juveniles, but not on the total biomass at the end of the experiment. Slightly lower water tables had a negative effect on the performance of all species. Measurements of the pH in the pots revealed that there were no significant differences in the top layer. Sulphide concentrations were very low in all the pots where juveniles had been planted and also in the pots withS. valerandi. Relatively high concentrations (30–50 μmol/l) were found in pots with poor growth ofJuncus andCalamagrostis plants. These values may exceed toxic levels for these species. Although oxygen concentrations in the pots were generally low, no relation existed between plant biomass and oxygen content, indicating that plant growth was not primarily limited by oxygen stress. These experiments support the idea that microbial mats may assist in extending the life span of early pioneer stages during dune slack succession by inhibiting the growth of species of later successional stages.     

Biotransformation of phenylurea herbicides by a soil bacterial strain, Arthrobacter sp. N2: structure, ecotoxicity and fate of diuron metabolite with soil fungi

In order to assess the influence of the aromatic substitution on the ability of a soil bacterial strain, Arthrobacter sp. N2, to degrade phenylurea herbicides, biotransformation assays were performed in mineral medium with resting cells of this soil bacterial strain on three phenylurea herbicides (diuron, chlorotoluron and isoproturon). Each herbicide considered, led to the formation of only one metabolite detected by HPLC analysis. After isolation, the metabolites were identified by NMR and MS, as the corresponding substituted anilines. According to the Microtox test (realized on the bacterium Vibrio fischeri), these metabolites presented non-target toxicity far more important (up to 600 times higher for 4-isopropylaniline) than the parent molecule. For isoproturon and chlorotoluron, the amount of substituted anilines obtained at the end of the biotransformation was very low, whereas the biotransformation of diuron into 3,4-dichloroaniline was almost quantitative. In this last case, the degradation product accumulated in the medium. In soil, other microorganisms are present that might degrade it. So the biotransformation of 3,4-dichloroaniline was then tested with four fungal strains: Aspergillus niger, Beauveria bassiana, Cunninghamella echinulata var. elegans and Mortierella isabellina. The aniline was further transformed with all the microorganisms tested. Only one metabolite was detected by HPLC analysis and after isolation, it was identified to be 3,4-dichloroacetanilide. This acetylated compound led to biological effects less important on V. fischeri than 3,4-dichloroaniline. These results stress the importance of identifying the degradation products to assess the impact of a polluting agent. Indeed, the pollutant may undergo transformation yielding compounds more toxic than the parent molecule.     

Transthyretin (prealbumin) binding of PCBS, a model for the mechanism of interference with vitamin A and thyroid hormone metabolism

In this study a direct interaction of a metabolite of 3,4,3′,4′-tetrachlorobiphenyl with transthyretin (prealbumin) was observed, leading to reduced plasma levels of retinol, RBP and thyroxin in rats and mice.     

In vitro EROD induction equivalency factors for the 10 PAHs generally monitored in risk assessment studies in The Netherlands

The ethoxy resorufin dealkylase (EROD) inducing potency of 10 polycyclic aromatic hydrocarbons (PAHs) is measured in the H4IIE in vitro bioassay and the results are compared to those reported in literature. The selected PAHs varied considerably in their potency to induce EROD activity. Anthracene (Ant) and phenanthrene (Phe) showed consistently no response. Naphthalene (Nap) showed no or a very weak response on EROD activity. Fluoranthene (Fla) and benzo[g,h,i]perylene (BghiP) showed weak responses at the highest doses. The other PAHs, including indeno[1,2,3-cd]pyrene (IP), benz[a]anthracene (BaA), benzo[a]pyrene (BaP), chrysene (Chr) and benzo[k]fluoranthene (BkF), showed full bell shaped dose-response curves. BaP EROD induction equivalency factors (BaP-1EF) were calculated and increased in the order Ant approximately Phe < Fla < Nap < BghiP < IP < BaA < BaP < Chr < BkF. Comparison of BaP-IEFs based on 50% effect concentration (EC50) or lowest effect concentration (LEC), yielded a significant relationship between both methods described by the equation log(BaPIEF(EC50) = 0.55 x log(BaPIEF(LEC)) + 0.07 (r2 = 0.913). BaP-IEFs as derived from our measurements and as reported in literature and measured in other in vitro assays deviated up to a factor of 17 among the different studies, but the potency rankings were comparable. For the PAH mixture as on average present in the human diet an overall tetrachlorodibenzo-p-dioxin (TCDD)-IEF of 1 x 10(-4) was estimated. The total PAH based TCDD induction equivalents (IEQ) intake then was calculated 300 pg/day, which is approximately 2 times higher then the PHAH based TCDD-EQ intake reported for humans.     

Occurrence of organotin compounds in house dust in Berlin (Germany)

In a study in the year 2000 on the occurrence of hazardous environmental contaminants house dust samples from 28 Berlin apartments were measured for the presence and concentrations of six organotin compounds, monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), monooctyltin (MOT), dioctyltin (DOT) and triphenyltin (TPT). The concentrations of MBT and DBT determined ranged considerably from 0.01 mg kg-1 to 1.5 mg kg-1 (median: 0.05 mg kg-1) and 0.01 to 5.6 mg kg-1 (median: 0.03 mg kg-1), respectively. Maximum levels of TBT and MOT were only 0.08 mg kg-1 and 0.04 mg kg-1. The maximum total value of the organotins was 7.2 mg kg-1 (median: 0.11 mg kg-1). MBT was found in 86% and DBT in 82% of the samples above the limit of quantification, TBT and MOT only in 50% and DOT in 43%. The focus of ecotoxicology is on the risks arising from organotin compounds (especially butyltins) when used as biocides in antifouling paints. TBT acts as an endocrine disrupter in animals, inducing masculinization (imposex) in female gastropods of different species by increasing testosterone levels. The most critical organ site in experimental animals is the cellular immune system, where lymphocyte depletion in the thymus and peripheral lymphoid tissues takes place. Our study does not provide data on the basis of which population exposure could be estimated; house dust containing harmful organotins could, however, under some conditions, become a relevant intake possibility for young children.