Rapid molecular prenatal diagnosis of ataxia-telangiectasia by direct mutational analysis

Mutations of the ataxia-telangiectasia-mutated (ATM) gene are responsible for the autosomal recessive disorder ataxia-telangiectasia (A-T). This study reports the first A-T prenatal diagnosis performed in Spain by direct molecular analysis. The pregnant woman had a previous child suffering from A-T due to a deletion in the ATM gene. The ATM coding region was sequenced in the A-T patient and her parents. Then, a specific polymerase chain reaction (PCR) to detect the deletion was performed for prenatal diagnosis. Additionally, polymorphic HLA loci were examined in order to exclude the possible contamination by maternal DNA. In this family of Gypsy origin, we carried out a rapid molecular diagnosis of A-T. Then, a prenatal diagnosis was carried out, identifying the deletion in the fetal DNA. Additionally, we performed a population study in unrelated Spanish Gypsies and in unrelated controls, showing that the deletion described could be a hotspot in the Spanish Gypsy population. The size of the coding region and the genomic structure, together with the absence of hotspots, make the mutation screening of the ATM gene difficult. The ability to identify ATM mutations provides a tool that can be applied in confirmatory diagnosis, genetic counselling, carrier prediction and prenatal diagnosis. Copyright © 2007 John Wiley & Sons, Ltd.     

Investigation into antimony mobility in sewage sludge fermentation

Antimony is distributed in the environment in inorganic and organic species with different solubility and mobility characters. Here we investigate the transformation of antimony in view of biomethylation during sewage sludge fermentation as a case study for an anaerobic environment. Our approach was to identify if antimony methylation follows the Challenger pathway by using isotopically enriched antimonite (123Sb(v)). The antimony source was subjected to methylation in sewage sludge, an anaerobic dominant methanogenic Archaea community. The antimony species were determined in the gas phase using cryotrapping (CT)-GC-ICP-MS, and in the medium (sewage slude) by hydride generation (HG) prior CT-GC-ICP-MS. The determined 123/121Sb isotope ratios in the volatile trimethylstibine and non-volatile methylantimony species indicated that the methylation follows the proposed methylation pathway. With this approach we were able to quantify 123Sb incorporation into monomethyl-, dimethyl- and trimethylantimony, respectively. The incorporation decreased with further methylation from 91% to 82% and 73%. Volatilisation as trimethystibine was generally lower than 0.1%, however, up to 0.8% of added antimony was found methylated to methylantimony species and mainly accumulated in the cell. Moreover, antimony biomethylation was enhanced by stimulation of the anaerobic communities of methanogenic Archaea and sulfate reducing bacteria (SRB), with the methanogens showing a higher activity.     

Assessment of application-rate dependent effects of a long-term fire retardant chemical (Fire Trol 934) on Typha domingensis germination

Although long-term fire retardants (LTR) gain increasingly acceptance as effective tools for wildfire management, recent studies indicate their potential harmfulness in the environment, especially in aquatic ecosystems. This study comprises a first laboratory impact assessment of LTR action in temporal Mediterranean wetlands, using seed germination of Typha domingensis as indicator of impact. Our aim was to identify application rates upon which seed germination could be significantly affected. We tested for low (1 l m(-2)) and high (3 l m(-2)) application rates of Fire Trol 934 which are recommended by the manufacturers as a function of fuel characteristics. In addition, we simulated the impact of a higher application rate of 5 l m(-2) because inhomogeneous dispersal of the LTR during fire control and prevention operations can result in locally elevated applications. Results of a microcosm experiment indicate that application rates of 1 or 3 l m(-2) can impact Typha germination rates in the short-term via indirect LTR-mediated effects on water quality, which suppressed necessary cues for germination. However, a subsequent experiment with Petri dishes, using seeds isolated from the LTR treated sediments did not show significantly different germination rates between the control and the treatments with application rates of 1 or 3 l m(-2). This suggests that retardant pre-application germination success could be recovered in nature once the retardant is eliminated. By contrast, seeds almost completely failed to germinate in the microcosm experiment and the subsequent Petri dish essay when an application rate of 5 l m(-2) was used. This suggests a critical level upon which Typha seed germination may be perpetually limited. Research should be extended to other plant species to provide fire managers with guidelines for environmentally safe use of LTR in the Mediterranean region.     

Concentrations of trace elements in tissues of loggerhead turtles (Caretta caretta) from the Tyrrhenian and the Ionian coastlines (Calabria,Italy)

Environmental Science and Pollution Research - Toxic trace elements from both, natural and anthropogenic origin, pose a threat to aquatic environments and marine wildlife due to their long-range...     

The clinical impact of the first-trimester nuchal translucency between the 95th–99th percentiles

Objectives

To evaluate the clinical significance of nuchal translucency (NT) between the 95th–99th percentile in terms of typical and atypical chromosomal abnormalities (ACAs), associated fetal congenital defects and postnatal outcome.

Methods

A retrospective cohort study of fetuses with NT between the 95th–99th percentile. Data regarding the rate of associated fetal defects, genetic abnormalities and postnatal outcome were collected.

Results

A total of 306 cases of fetuses with an NT between the 95th–99th percentiles were included. The overall rate of genetic abnormalities was 12.1% (37/306). Chromosomal abnormalities were found in 10.1% (31/306) of cases and 2% were ACA (6/306). Within this group, two were pathogenic Copy Number Variants (CNVs) and four were single gene disorders. The overall rate of fetal congenital defects was 13.7% (42/306). All ACAs were found in fetuses with congenital defects. Postnatally, a new diagnosis of a single gene disorder was made in 0.85% of cases (2/236).

Conclusions

The presence of an NT between the 95th–99th percentiles carries a 10-fold increased risk of fetal defects, representing an indication for referral for a detailed fetal anatomy evaluation. The risk of ACA is mainly related to the presence of fetal defects, irrespective of the combined test risk.     

Biomarkers of internal dose for the assessment of environmental exposure to benzene

The urinary excretion of t,t-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA) and urinary benzene and the influence of a smoking habit and of exposure to urban traffic on the urinary excretion of these biomarkers were investigated in 137 male adults from the general population. All subjects were not occupationally exposed to benzene and resident in two cities in Puglia (Southern-Italy). Environmental exposure to benzene was measured using passive personal samplers. The biomarkers t,t-MA, SPMA and urinary benzene were determined in urine samples collected from each subject at the end of the environmental sampling. The percentage of cases above the limit of detection was higher for SPMA and urinary benzene in smokers than in non-smokers, and for airborne benzene and urinary benzene in subjects exposed to urban traffic. Airborne benzene was correlated with the time spent in urban traffic during the environmental sampling. Among the biomarkers, urinary benzene was found to be correlated with airborne benzene only in non-smokers, and with the time spent in urban traffic, both in smokers and non-smokers considered together, and in non-smokers only. Finally, multiple regression analysis showed that the urinary excretion of all the biomarkers was dependent on the number of cigarettes smoked per day and, for urinary benzene, also on the time spent in urban traffic. In conclusion, urinary benzene seems to be a more valid biomarker than t,t-MA and SPMA to assess environmental exposure to extremely low concentrations of benzene. Cigarette smoking prevailed over traffic exhaust fumes in determining the internal dose of benzene.     

Polychlorinated biphenyls in two salt marsh sediments of the Venice Lagoon

Polychlorinated biphenyls (PCBs) were measured in two dated salt marsh cores of the Venice Lagoon to assess their input chronology and to evaluate the importance of atmospheric deposition as a source. Sampling sites were chosen in order to evidence the differences between areas located leeward and windward with respect to inputs originating in both the city of Venice and the industrial area. Concentrations of PCB indicators (0.13-15.6 ng g?1) increased gradually from the 1930s, reached maxima from the 1950s to the late 1970s, and then decreased. PCB loadings to marshes are driven by both the atmospheric deposition and the resuspension of subtidal sediments, this latter being more important for heavier congeners. The downwind marsh recorded higher fluxes (0.06-9.72 ng cm?2 year?1) than the upwind one (0.01-0.53 ng cm?2 year?1). Recent fluxes are rather consistent with bulk deposition measurements. A higher contribution of CB-101 and CB-118 was detected in the intermediate layers of the downwind site, suggesting a different PCB source for the corresponding time interval. In the other marsh, PCBs showed a rather constant composition at all levels (mostly CB-153, CB-138 and CB-180), accounting for a regional influence. Deep layers showed an enrichment of higher chlorinated congeners at both sites, whereas recent samples conserve the patterns typical of surficial and subsurficial subtidal sediments. The scientific approach adopted in this research can be considered as a sort of methodological procedure for the determination of fluxes and pathways of PCBs through the study of marsh cores.     

Performance and water-use efficiency (single-leaf vs. whole-canopy) of well-watered and half-stressed split-root Lambrusco grapevines grown in Po Valley (Italy)

‘Lambrusco a foglia frastagliata’ grapevines (Vitis vinifera L.) were grown outdoors at Piacenza (44°55′N, 9°44′E, Po Valley, Italy) with the root system split between two 30-L pots and subjected from pre-veraison (17 July) to harvest (5 September) to soil drying of half of the root system (HS) induced by withholding water from one of the two pots as compared to well-watered (WW) vines (both pots daily recharged at field capacity). Volumetric soil-water content, pre-dawn and mid-morning leaf water potential, single-leaf gas-exchange as assimilation rate, stomatal conductance and transpiration were monitored throughout the trial. Whole-canopy gas-exchange as net CO₂ exchange rate (NCER) and transpiration were tracked from 31 August to 7 September on three vines per treatment on a 24-h basis using an enclosure method. Primary leaf carbon isotope (δ¹³C) composition, yield components and must composition were determined at harvest.Withdrawing water from one pot triggered a water stress response showing higher stomatal sensitivity to changes in air vapour pressure deficit, relatively low assimilation rates, high intrinsic and extrinsic water-use efficiency (WUE) and earlier cessation of shoot growth. Yet, mid-morning leaf water potential was consistently lower in HS treatment over stress as compared to WW, indicating an anisohydric adjustment. Canopy NCER given on a leaf-area basis showed mean daily rates ranging from 3.9 to 4.9 μmol m² s^?1 in WW canopies against 2.6–3.0 μmol m^?2 s^?1 in HS. Conversely, canopy transpiration rates varied from 0.915 to 1.157 mmol m^?2 s^?1 for WW to 0.630–0.714 mmol m² s^?1 in HS. Increased leaf-based intrinsic and extrinsic WUE in HS did not match the canopy response, which to some extent resulted in an opposite outcome, i.e. higher canopy WUE in well-watered vines especially in the morning hours. Likewise, δ¹³C did not differ between treatments. This suggests caution when point-time determinations of single-leaf-based WUE are extrapolated to the whole-canopy behaviour when assessing the water saving strategies of a given genotype. The stressed vines achieved no variation in yield level and components and had improved grape composition as to soluble solids and total anthocyanins. This optimal behaviour is likely due to earlier shoot growth cessation, enhanced maturity and a buffering leaf-to-fruit ratio (3.61 m² kg^?1) that mitigated the effects of post-veraison stress.