Miscanthus for Renewable Energy Generation: European Union Experience and Projections for Illinois

Mitigation and Adaptation Strategies for Global Change -     

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.     

Aseptically collected calf serum as an effective alternative to fetal calf serum in the culture of amniotic fluid cells

The growth-promoting activities of three different bovine sera have been compared in primary and secondary amniotic fluid cell cultures. In secondary amniotic fluid cell micro- cultures, aseptically collected calf serum (CS) was slightly, though not significantly more effective than fetal calf serum (FCS) in promoting DNA synthesis, while newborn calf serum (NCS) was significantly less effective than either CS or FCS. All three sera were optimally effective at concentrations of 10 per cent (v/v). Significant variation in quality occurred within four batches of each of CS and FCS, but not within four batches of NCS. A selected batch of CS was significantly more effective in promoting the growth of primary amniotic fluid cell cultures than were a number of batches of FCS then in routine laboratory use. It is suggested that CS may serve as an effective and economical alternative to FCS in the culture of amniotic fluid cells, thereby expanding the scope of serum batch testing. A possible explanation for the varying growth-promoting activities of different sera is discussed.     

Cost of parental care in the woodroach Cryptocercus punctulatus Scudder (Dictyoptera: Cryptocercidae)

Summary The objective of this study was to measure the effect of parental care on the reproductive capabilities and survival of adult Cryptocercus punctulatus Scudder. Brood removal studies show that the long term parental care exhibited by this woodroach can exact a cost in terms of future reproduction by delaying a second reproductive episode. If C. punctulatus is used as a model of the ancestral state of the termites, changes in responsibility for care giving behavior can account for all the higher components of eusociality (overlap of worker generations, brood care by workers, non-reproductive castes) by shifting the cost of parental care from the adults to the oldest offspring.     

Accuracy and reliability of dogs in surveying for desert tortoise (Gopherus agassizii).

The desert tortoise (Gopherus agassizii) is federally listed as "threatened" and is afforded protection in several U.S. states including California, Nevada, Utah, and Arizona. Numerous factors ranging from habitat destruction to disease are thought to contribute to the species' decline throughout its range. Data collection on desert tortoises in the wild is challenging because tortoises are secretive, and many age and size classes are virtually undetectable in the wild. Detection dogs have been used for decades to assist humans, and the use of dogs for wildlife surveys is of increasing interest to scientists and wildlife managers. To address the basic question of whether dogs could be used to survey for the desert tortoise, we quantified the reliability and efficacy of dogs trained for this purpose. Efficacy is the number of tortoises that dogs find out of a known population. Reliability is a measure of how many times a dog performs its trained alert when it has found a tortoise. A series of experimental trials were designed to statistically quantify these metrics in the field setting where dogs trained to locate live desert tortoises were tested on their ability to find them on the surface, in burrows, and in mark-recapture surveys. Results indicated that dogs are effective at and can safely locate desert tortoises with reliability on the surface and are capable of detecting tortoises in burrows under a range of environmental conditions. Dogs found tortoises at the same statistical rate at temperatures between 12 degrees and 27 degrees C, relative humidity from 16% to 87%, and wind speeds up to 8 m/s. In both surface and burrow trials, dogs found >90% of the experimental animals. In comparative studies with humans, dogs found tortoises as small as 30 mm, whereas the smallest tortoise located by human survey teams was 110 mm. Although not all dogs or dog teams meet the requirements to conduct wildlife surveys, results from this study show the promise in using dogs to increase our knowledge of rare, threatened, and endangered species through improved data collection methods.     

Enantiomeric separation of metolachlor and its metabolites using LC-MS and CZE

The stereoisomers of metolachlor and its two polar metabolites [ethane sulfonic acid (ESA) and oxanilic acid (OXA)] were separated using liquid chromatography-mass spectrometry (LC-MS) and capillary zone electrophoresis (CZE), respectively. The separation of metolachlor enantiomers was achieved using a LC-MS equipped with a chiral stationary phase based on cellulose tris(3,5-dimethylphenyl carbamate) and an atmospheric pressure chemical ionization source operated under positive ion mode. The enantiomers of ESA and OXA were separated using CZE with gamma-cyclodextrin (gamma-CD) as chiral selector. Various CZE conditions were investigated to achieve the best resolution of the ESA and OXA enantiomers. The optimum background CZE electrolyte was found to consist of borate buffer (pH=9) containing 20% methanol (v/v) and 2.5% gamma-CD (w/v). Maximum resolution of ESA and OXA enantiomers was achieved using a capillary temperature of 15 degrees C and applied voltage of 30 kV. The applicability of the LC-MS and CZE methods was demonstrated successfully on the enantiomeric analysis of metolachlor and its metabolites in samples from a soil and water degradation study that was set up to probe the stereoselectivity of metolachlor biodegradation. These techniques allow the enantiomeric ratios of the target analytes to be followed over time during the degradation process and thus will prove useful in determining the role of chirality in pesticide degradation and metabolite formation.