Denaturing high-performance liquid chromatography (DHPLC)-based prenatal diagnosis for tuberous sclerosis

Tuberous sclerosis (TSC) is a frequent autosomal-dominant condition (affecting 1 in 6000 individuals) caused by various mutations in either the hamartin (TSC1) or the tuberin gene (TSC2). This allelic and non-allelic heterogeneity makes genetic counseling and prenatal diagnosis difficult, especially as a significant proportion of TSC cases are due to de novo mutations. For this reason the identification of the disease causing mutation is mandatory for accurate counseling, yet current mutation detection methods such as single-strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) are labor intensive with limited detection efficiency. Denaturing high-performance liquid chromatography (DHPLC) is a high-throughput, semi-automated mutation detection system with a reported mutation detection rate close to 100% for PCR fragments of up to 800 bp. We used a recently described DHPLC assay allowing the efficient detection of mutations in TSC1 to analyze the DNA extracted from a chorion villus sample in order to perform a prenatal diagnosis for TSC. The fetus was found not to have inherited the deleterious mutation and the DHPLC diagnosis was confirmed by haplotype analysis. This represents the first DHPLC-based prenatal diagnosis of a genetic disease. Copyright © 2001 John Wiley & Sons, Ltd.     

Augmenting Watershed Model Calibration with Incorporation of Ancillary Data Sources and Qualitative Soft Data Sources

Watershed simulation models such as the Soil & Water Assessment Tool (SWAT) can be calibrated using “hard data” such as temporal streamflow observations; however, users may find upon examination of model outputs, that the calibrated models may not reflect actual watershed behavior. Thus, it is often advantageous to use “soft data” (i.e., qualitative knowledge such as expected denitrification rates that observed time series do not typically exist) to ensure that the calibrated model is representative of the real world. The primary objective of this study is to evaluate the efficacy of coupling SWAT‐Check (a post‐evaluation framework for SWAT outputs) and IPEAT‐SD (Integrated Parameter Estimation and Uncertainty Analysis Tool‐Soft & hard Data evaluation) to constrain the bounds of soft data during SWAT auto‐calibration. IPEAT‐SD integrates 59 soft data variables to ensure SWAT does not violate physical processes known to occur in watersheds. IPEAT‐SD was evaluated for two case studies where soft data such as denitrification rate, nitrate attributed from subsurface flow to total discharge ratio, and total sediment loading were used to conduct model calibration. Results indicated that SWAT model outputs may not satisfy reasonable soft data responses without providing pre‐defined bounds. IPEAT‐SD provides an efficient and rigorous framework for users to conduct future studies while considering both soft data and traditional hard information measures in watershed modeling.     

Urban tracer dispersion experiments during the second DAPPLE field campaign in London 2004

As part of the DAPPLE programme two large scale urban tracer experiments using multiple simultaneous releases of cyclic perfluoroalkanes from fixed location point sources was performed. The receptor concentrations along with relevant meteorological parameters measured are compared with a three screening dispersion models in order to best predict the decay of pollution sources with respect to distance. It is shown here that the simple dispersion models tested here can provide a reasonable upper bound estimate of the maximum concentrations measured with an empirical model derived from field observations and wind tunnel studies providing the best estimate. An indoor receptor was also used to assess indoor concentrations and their pertinence to commonly used evacuation procedures.     

Probability distributions and statistical inference for axial data

Observations on axes which lack information on the direction of propagation are referred to as axial data. Such data are often encountered in enviromental sciences, e.g. observations on propagations of cracks or on faults in mining walls. Even though such observations are recorded as angles, circular probability models are inappropriate for such data since the constraint that observations lie only in [0, π) needs to be enforced. Probability models for such axial data are argued here to have a general structure stemming from that of wrapping a circular distribution on a semi-circle. In particular, we consider the most popular circular model, the von Mises or circular normal distribution, and derive the corresponding axial normal distribution. Certain properties of this distribution are established. Maximum likelihood estimation of its parameters are shown to be surprisingly, in contrast to trigonometric moment estimation, numerically quite appealing. Finally we illustrate our results by several real life axial data sets. Received: September 2004/ Revised: December 2004     

“Protecting coastal zones in the Mediterranean: an economic and regulatory analysis”

This paper examines existing measures taken to protect the coastal zones of the Mediterranean Sea and assesses their success. A summary of the main pressures facing this region is given, followed by an analysis of the legislation covering coastal zone development in ten countries: Algeria, Croatia, Egypt, France, Israel, Italy, Malta, Spain, Tunisia and Turkey. The paper finds that not all of these States have legislation specifically covering coastal zones, but there is concern in all areas that existing legislation is not working. The costs and benefits of controlling coastal development are examined. Firstly, a literature review of valuation studies identifies a range of values placed on the developed and undeveloped coastline for both users and local property owners. These values were then used in a model to evaluate policy options to control development of a stretch of coastline. The model indicates that a stricter control regime of coastal development may provide significant benefits.

Antioxidant status, flight performance and sexual signalling in wild-type parrots

During mate choice, individuals are predicted to assess traits that honestly signal the quality of potential partners. Locomotor capacity may be such a trait, potentially signalling condition and ability to resist oxidative damage. In this study, we experimentally manipulated nutritional status: Male wild-type budgerigars, imported from Australia, were provided with either an enhanced (EQ) or reduced quality (RQ) diet varying in vitamin (particularly retinol and α-tocopherol) and mineral levels. Then, we assessed whether this influenced locomotor capacity, i.e. escape flight performance, and sexual attractiveness in male budgerigars Melopsittacus undulatus. Males in the EQ group showed significantly greater total antioxidant capacity and higher blood plasma concentrations of the dietary antioxidants retinol and α-tocopherol, but not carotenoids, than the RQ group. Over 8 weeks of flight training, males on the EQ diet showed significantly greater improvement on the most strenuous flight test than RQ males. In mate choice trials, females preferred EQ over RQ males. EQ males that were relatively fast in escape flight trials were more strongly preferred in the mate preference arena than their RQ competitors. Interestingly, males with high plasma carotenoid levels flew slower and were less attractive than males with low carotenoid levels. This might indicate that carotenoids are not effective antioxidants in birds. Overall, our results show that dietary-derived antioxidants can influence sexual attractiveness and other fitness-related traits through multiple pathways. Locomotor capacity appears to be an honest signal of male condition in birds.     

CO2 enrichment inhibits shoot nitrate assimilation in C3 but not C4 plants and slows growth under nitrate in C3 plants

The CO2 concentration in Earth's atmosphere may double during this century. Plant responses to such an increase depend strongly on their nitrogen status, but the reasons have been uncertain. Here, we assessed shoot nitrate assimilation into amino acids via the shift in shoot CO2 and O2 fluxes when plants received nitrate instead of ammonium as a nitrogen source (deltaAQ). Shoot nitrate assimilation became negligible with increasing CO2 in a taxonomically diverse group of eight C3 plant species, was relatively insensitive to CO2 in three C4 species, and showed an intermediate sensitivity in two C3-C4 intermediate species. We then examined the influence of CO2 level and ammonium vs. nitrate nutrition on growth, assessed in terms of changes in fresh mass, of several C3 species and a Crassulacean acid metabolism (CAM) species. Elevated CO2 (720 micromol CO2/mol of all gases present) stimulated growth or had no effect in the five C3 species tested when they received ammonium as a nitrogen source but inhibited growth or had no effect if they received nitrate. Under nitrate, two C3 species grew faster at sub-ambient (approximately 310 micromol/mol) than elevated CO2. A CAM species grew faster at ambient than elevated or sub-ambient CO2 under either ammonium or nitrate nutrition. This study establishes that CO2 enrichment inhibits shoot nitrate assimilation in a wide variety of C3 plants and that this phenomenon can have a profound effect on their growth. This indicates that shoot nitrate assimilation provides an important contribution to the nitrate assimilation of an entire C3 plant. Thus, rising CO2 and its effects on shoot nitrate assimilation may influence the distribution of C3 plant species.     

The Influence of Varying Algal Biomass On Contaminant Exposure in Benthic-Planktonic Mesocosms: Copper (Ii)

A series of mesocosms was exposed to a suite of light treatments and nutrient enrichment in order to generate algal communities of varying biomass. the influence of this biomass on the speciation of copper (II) was studied. Distribution coefficients (_Kd,Lkg-1) were relatively high (log_Kd = 5 to 7), indicative of robust trace metal sequestration, and were likely controlled by the particulate organic carbon content (foc). Differences in _Kd over time and among treatments were significant, as was the relationship between _Kd and foc. Fluorescence quenching was used to determine binding capacities (_Lt, M) and their associated binding constants (_Kcond,M^-1) in order to model the solid phase copper speciation. the _Kcond ranged between 2.1 and 5.2 × 10¹²M^-1, indicating a very strong copper-ligand complex, and was higher in mesocosms that received more light. the light _Lt increased over time, dramatically after the nutrient enrichment, but did not vary systematically among light treatments. _Lt ranged from 7.2 × 10- 7 to 4.9 × 10- 5 M. the large magnitudes of _Kd, _Kcond and _Lt ensured that greater than 97% of total copper in the mesocosms was complexed by organic matter. the total copper concentration ([Cu]_T, M) needed to reach a target dissolved copper concentration of 10^-12.5 M (pCu = 12.5) was determined for each mesocosm over time. [Cu]_T was between 8.02 × 10^-5 and 3.41 × 10^-2 M, and increased over time. the [Cu]_T normalized to the target pCu (Effective Dose Ratio, EDR) increased directly with increases in algal biomass, indicating a direct link between system productivity and copper exposure. Approximately 45% of the variance in EDR was explained by variance in total biomass, while the residual variance in EDR was due likely to differences in the strengths of particle associations and magnitude of binding capacities.     

Environmental photodegradation of mefenamic acid

Pharmaceuticals and personal care products are an emerging class of environmental pollutants. Photolysis is expected to be a major loss process for many of these compounds in surface waters, including the common non-steroidal anti-inflammatory drug mefenamic acid. The direct photolysis solar quantum yield of mefenamic acid was observed to be 1.5+/-0.3x10(-4). Significant photosensitization was observed in solutions of Suwanee River fulvic acid and Mississippi River water, as well as for the model photosensitization compounds 3'-methoxyacetophenone, 2-acetonaphthone and perinaphthenone. Quenching, sparging and light-filtering experiments suggested a direct reaction of mefenamic acid with excited triplet-state dissolved organic matter as the major photosensitization process. The persistence of the model photosensitizer suggests that the photosensitization by perinaphthenone occurs either by triplet-energy transfer or an electron transfer followed by rapid regeneration of the sensitizer. Due to its low quantum yield, the loss of mefenamic acid in sunlit natural waters is expected to depend on both direct and indirect photodegradation processes.