We have sequenced 14 introns from the ciliate Tetrahymena thermophila and include these in an analysis of the 27 intron sequences available from seven T. thermophila protein-encoding genes. Consensus 5' and 3' splice junctions were determined and found to resemble the junctions of other nuclear pre-mRNA introns. Unique features are noted and discussed. Overall the introns have a mean A + T content of 85% (21% higher than neighbouring exons) with smaller introns tending towards a higher A + T content. Approximately half of the introns are less than 100 bp. Introns from other organisms (approximately 30 of each) were also examined. The introns of Dictyostelium discoideum, Caenorhabditis elegans and Drosophila melanogaster, like those of T. thermophila, have a much higher mean A + T content than their neighbouring exons (greater than 20%). Introns from plants, Neurospora crassa and Schizosaccharomyces pombe also have a significantly higher A + T content (10%-20%). Since a high A + T content is required for intron splicing in plants (58), the elevated A + T content in the introns of these other organisms may also be functionally significant. The introns of yeast (Saccharomyces cerevisiae) and mammals (humans) appear to lack this trait and thus in some aspects may be atypical. The polypyrimidine tract, so distinctive of vertebrate introns, is not a trait of the introns in the non-vertebrate organisms examined in this study.

We have determined the nucleotide sequence of the cnjB gene from the ciliate Tetrahymena thermophila. This gene is transcriptionally active only during early conjugation, peaking in meiotic prophase. It contains 13 introns, four transcription start points and codes for a putative polypeptide (CnjB) of 1748 amino acids with a calculated molecular weight of 200 kilodaltons and a pl of 7.9. The coding region of cnjB has a low GC content (32% GC) and unusual codon usage. The C-terminal one-third of CnjB consists of three repetitive domains. Introns were absent in this region of cnjB. One of the repetitive domains consists of seven CCHC or retroviral-type zinc fingers, a motif found in one or two copies in retroviral nucleocapsid proteins. This motif has also been found recently in seven copies in the human nucleic-acid binding protein CNBP, in an apparent CNBP homologue in Schizosaccharomyces pombe and in one copy in a Xenopus gene active in early embryos. The other two domains are on either side of the zinc finger domain and contain a repeated glycine-rich motif seen in the heterogeneous nuclear ribonuclear proteins A1 and A2/B1 as well as other proteins. Both CCHC zinc fingers and glycine-rich repeats have been found in proteins with single-stranded nucleic acid-binding activity as well as strand-annealing activity. CnjB is, to our knowledge, the first protein found to contain both types of motifs.

Multiple introns have been found in a gene from a ciliated protozoan. This Tetrahymena thermophila gene (cnjB) is large (7.5 kb mRNA) and active only during conjugation, the organism's sexual cycle. Six introns ranging in size from 62 bp to 676 bp were found when we sequenced a 3.1 kb segment of the cnjB gene together with its corresponding cDNA. We estimate, by extrapolation of our current data, a total of approximately 30 introns in this gene with a total gene size (introns plus exons) of 15 kb or more. The number of introns is surprising given the scarcity of introns in ciliate genes examined to date. Our findings constitute the first example of multiple introns in a ciliate gene. Having the sequence of several introns has allowed us to construct consensus sequences for T. thermophila mRNA introns. The 5' and 3' intron junctions resemble those of general nuclear mRNA (GT/AG rule is followed) but differences are seen. In particular, stretches of 10 or more adenines and thymines are found adjacent to the conserved GT and AGs at the junctions. Unusual aspects of the coding region of this gene are discussed.

The development of techniques for the detection of water leaks from underground pipelines is seen as a high profile activity by water companies and regulators. This is due to increasing water demands and problems with current leak detection methods. In this paper optical reflectance and microwave backscatter models (SAIL + PROSPECT and RT2) were used to try and identify optimal indices for detecting water leaks amongst a variety of different land cover types at different growth stages. Results suggest that red/near infrared and red/middle infrared ratios show potential for leak detection. Given the sensitivity of L-band radar to moisture, and the ability to separate contributions from canopy and ground surface, it is possible to detect saturated soils through vegetation canopies. The results of both approaches are used to infer limits of detection in terms of season and meteorological conditions for a range of land covers. Preliminary findings suggest that leaks may be optimally detected when canopy height is low, surrounding soil is dry, and the leak has been present for more than 14 days. The modelled data is compared with L -band fully polarimetric E-SAR data, and 200 channel HYMAP hyperspectral airborne data which were acquired over an 8km section of the Vrynwy aqueduct (UK), which included a high concentration of leaks. Data was acquired as part of the British National Space Centre (BNSC) and Natural Environmental Research Council (NERC), SAR and Hyperspectral Airborne Campaign (SHAC) in June 2000. The results from this work suggest that remote sensing is both an effective and feasible tool for leak identification.

The traditional approach to delineating and extracting features from remotely sensed images relies predominantly on manual interpretation, a procedure that is often time consuming and expensive. Automation offers the potential for reduced costs and wider utilization of remote sensing within the business community, but involves difficulty in representing the expertise of remote sensing scientists within a series of decision rules. The objectives of this paper are two-fold: firstly, to produce a system for automated feature discrimination in remotely sensed images, using leaks from water supply networks as a case study; and secondly, to test whether the system is suitable for use with the next generation of satellite sensors. The automated system was calibrated by integrating HyMap and Airborne Thematic Mapper (ATM) images with context data from a variety of sources (such as ambient irradiance environment; topography; land use, and field boundaries). The automated system was assessed for its applicability to satellite remote sensing by testing the system on airborne data that were degraded to the resolutions of satellite images. It is proposed in this paper that automation, particularly with respect to satellite remote sensing, makes leak detection from water supply networks commercially viable.