Carrier detection and microsatellite analysis of duchenne and becker muscular dystrophy in spanish families

Duchenne and Becker muscular dystrophy (D/BMD) are usually problematical when trying to determine the carrier status of at-risk women, which usually has to be based on haplotype or dosage analysis on Southern blots. Using multiplex polymerase chain reaction (PCR) analysis, we have detected deletions in 20 out of 44 D/BMD families with living affected members (45·5 per cent), more often in sporadic cases of DMD (14/22 with detectable deletion) than in familial ones (4/15), the majority (15/20) occurring in the distal region of the D/BMD gene. Four highly informative short tandem repeat polymorphisms (STRPs), which lie within the distal deletion hot spot of the D/BMD gene, can show loss of heterozygosity in carrier females, providing direct evidence of their carrier status. These STRPs greatly improve informativity, with a combined heterozygosity of 100 per cent and with the majority of families informative for three of the four STRPs. In 14/15 (93 per cent) of the families with distal deletions, the STRPs provided direct information on carrier status, and in some cases, they provide valuable information on recombination breakpoints and non-paternity.     

Study of fractionation and potential mobility of metal in sludge from pyrite mining and affected river sediments: changes in mobility over time and use of artificial ageing as a tool in environmental impact assessment

Sludge from mining exploitation can be a source of land and water contamination in the adjacent zone. Accidents such as the break of waste mining pools in the Aznalcollar Mine (Seville, Spain) in 1998 produce important ecological disasters. In this work is presented a study of the evolution of aqua regia leachable concentration and mobility of metals in sediment samples of Guadiamar River basin from the accident date up to 2001. The application of BCR standard extraction procedures provides valuable information about the mobility and toxicity of the spill and the metal mobility in Guadiamar River polluted sediments. As a rule, themobility of several metals in the initial sludge (sulphides) is low, except for copper. Otherwise, the results of aqua regia leachable concentration and mobility of metal obtained for sediments samples indicate that the cleaning and inertisation works carried out in the zone have been adequate but insufficient, being the metal levels observed in the zone are higher than natural levels. To establish the initial impact of the spill and the mobility changes with time, mineralogical composition of the pyritic sludge and its evolution after the natural weathering and after the induction, of accelerated ageing processes by light and temperature was studied in the laboratory. Oxidation of initial sulphides to sulphates was observed. Both environmental and laboratory oxidation of the metallic sulphides increase the mobility of all metals, especially of copper, zinc and lead. The proposed laboratory procedure allows to predict the changes in mobility and therefore in toxicity that can occur at short or long term after exposure of sludge at environ-mental conditions. The mobility results in sludge and sediment samples are interpreted in terms of the mineral composition of the samples.     

Euglena gracilis as a model for the study of Cu2+ and Zn2+ toxicity and accumulation in eukaryotic cells

We have observed the effect of copper and zinc on the biology of Euglena gracilis. The cells displayed different sensitivities to these metals, as the apparent LC50 for Cu2+ was 0.22 mM, and for Zn2+ it was 0.88 mM. While Zn2+ was able to increase cell proliferation even at 0.1 mM, the minimal CuCl2 concentration tested (0.02 mM) was sufficient to impair cell division. Higher concentrations of these metals not only inhibited cell division in a concentration-dependent manner, but also interfered with the metabolism of E. gracilis. A higher accumulation of proteins and lipids per cell was observed at the DI50 concentration for metal-treated cells. These results suggest that the test concentration of both metals leads to a failure in completing cell division. Ultrastructural analysis indicated a chloroplast disorganization in copper-treated cells, as well as the presence of electron dense granules with different shapes and sizes inside vacuoles. Microanalysis of these granules indicated an accumulation of copper, thus suggesting a detoxification role played by the vacuoles. These results indicate that E. gracilis is an efficient biological model for the study of metal poisoning in eukaryotic cells. They also indicate that copper and zinc (copper being more poisonous) had an overall toxic effect on E. gracilis and that part of the effect can be ascribed to defects in the structure of chloroplast membranes.     

Heterospecific call recognition and phonotaxis in the orientation behavior of the marbled newt,Triturus marmoratus

The role of acoustic cues as reference cues for orientation by amphibians has been demonstrated in anurans, the only amphibian group that engages in acoustic communication, but not in urodeles. Orientation responses of marbled newts, Triturus marmoratus, were studied to determine whether heterospecific calls elicited positive phonotaxis. The orientation tests consisted in presenting either a familiar acoustic stimulus, the advertisement calls of natterjack toads (Bufo calamita), or a control stimulus, the advertisement calls of European green toads (Bufo viridis) that the newts would not be expected to recognize. Marbled newts and natterjack toads occur in simpatry, but T. marmoratus and B. viridis are allopatric species. Thus, T. marmoratus is distributed over the northern half of the Iberian Peninsula, whereas B. viridis occurs in the Balearic Islands, but not over the Iberian Peninsula. Newts were released in a circular arena while a recorded chorus of natterjack toads or European green toads played outside the arena to determine whether they displayed positive phonotactic orientation. Our results show that marbled newts performed positive phonotaxis when exposed to the breeding calls of natterjack toads, but not to those of European green toads. Newts chose a compass course in the direction of the advertisement calls of B. calamita. Acoustic information might improve orientation accuracy. This study is the first to provide evidence of heterospecific call recognition and positive phonotactic response in urodeles.Communicated by W. Wiltschko     

Monitoring concentrations of persistent organic pollutants in the general population: the international experience

Assessing the adverse effects on human health of persistent organic pollutants (POPs) and the impact of policies aiming to reduce human exposure to POPs warrants monitoring body concentrations of POPs in representative samples of subjects. While numerous ad hoc studies are being conducted to understand POPs effects, only a few countries are conducting nationwide surveillance programs of human concentrations of POPs, and even less countries do so in representative samples of the general population. We tried to identify all studies worldwide that analyzed the distribution of concentrations of POPs in a representative sample of the general population, and we synthesized the studies' main characteristics, as design, population, and chemicals analyzed. The most comprehensive studies are the National Reports on Human Exposure to Environmental Chemicals (USA), the German Environmental Survey, and the Arctic Monitoring and Assessment Programme. Population-wide studies exist as well in New Zealand, Australia, Japan, Flanders (Belgium) and the Canary Islands (Spain). Most such studies are linked with health surveys, which is a highly-relevant additional strength. Only the German and Flemish studies analyzed POPs by educational level, while studies in the USA offer results by ethnic group. The full distribution of POPs concentrations is unknown in many countries. Knowledge gaps include also the interplay of age, gender, period and cohort effects on the prevalence of exposures observed by cross-sectional surveys. Local and global efforts to minimize POPs contamination, like the Stockholm convention, warrant nationwide monitoring of concentrations of POPs in representative samples of the general population. Results of this review show how such studies may be developed and used.     

Effects of moderate pollution on toxic and trace metal levels in calves from a polluted area of northern Spain

This study evaluated the contribution of anthropogenic pollution to toxic metal residues (Cd, Pb and As) in cattle in an industrialized area of Asturias (northern Spain), and investigated possible implications of toxic metal exposure for metabolism of essential metal elements (Cu, Zn, Fe, Mn). Samples of liver, kidney, muscle and blood from animals aged 9-12 months were obtained from an industrialized area (78 calves) and from a rural area (92 calves). Samples were acid-digested, and levels of metals determined by atomic absorption spectrophotometry. Cadmium and lead contents in the liver and kidney were moderately and significantly higher in calves from the industrialized area (Cd: liver 29.6, kidney 161; Pb: liver 38.1, kidney 38.3 microg/kg wet weight) than in calves from the rural area (Cd: liver 22.9, kidney 96.4; Pb: liver 20.7, kidney 15.9 microg/kg kg wet weight). Although these toxic metals were only moderately raised, our results suggest that they may nevertheless have interfered with trace element metabolism, as reported previously for severely polluted regions. Notably, copper levels in calves from the industrialized area were generally low, and nearly half of these animals showed tissue copper levels indicating risk of copper deficiency.     

Superciliums in white-eared hummingbirds as badges of status signaling dominance

The role of badges as indicators of contest ability has been previously described. In hummingbirds, the exhibition of a badge is expected to save energy expenditure in agonistic interactions and to favor energy intake. Here, we investigate whether variable supercilium size in the white-eared hummingbird has a role in dominance status signaling. Firstly, 45 hummingbird males were captured and their superciliums were photographed to investigate variation in size and any possible allometric relationships. Secondly, 42 male birds were used to analyze whether the supercilium has a role in dominance status signaling in a dyadic contest. We found that supercilium size varied continuously but that despite variability between individuals, there was no relationship between supercilium size and body size. However, our dyad experiment indicated that birds with larger badges were able to make more visits to the feeders than individuals with smaller badges. We suggest a status signaling function of the supercilium.