Cell surface analysis of the basidiomycete yeast cryptococcus neoformans

Cell surface properties of the basidiomycete yeast Cryptococcus neoformans were investigated with a combination of novel and well proven approaches. Non-specific cell adhesion forces, as well as exposed carbohydrate and protein moieties potentially associated with specific cellular interaction, were analysed. Experimentation and analysis employed cryptococcal cells of different strains, capsular status and culture age. Investigation of cellular charge by particulate microelectrophoresis revealed encapsulated yeast forms of C. neoformans manifest a distinctive negative charge regardless of the age of cells involved; in turn, the neutral charge of acapsulate yeasts confirmed that the polysaccharide capsule, and not the cell wall, was responsible for this occurrence. Hydrophobicity was measured by MATH and HICH techniques, as well as by the attachment of polystyrene microspheres. All three techniques, where applicable, found C. neoformans yeast to be consistently hydrophilic; this state varied little regardless of strain and culture age. Cell surface carbohydrates and protein were investigated with novel fluorescent tagging protocols, flow cytometry and confocal microscopy. Cell surface carbohydrate was identified by controlled oxidation in association with biotin hydrazide and fluorescein-streptavidin tagging. Marked amounts of carbohydrate were measured and observed on the cell wall surface of cryptococcal yeasts. Furthermore, tagging of carbohydrates with selective fluorescent lectins supported the identification, measurement and observation of substantial amounts of mannose, glucose and N-acetyl-glucosamine. Cryptococcal cell surface protein was identified using sulfo-NHS-biotin with fluorescein-streptavidin, and then readily quantified by flow cytometry. Confocal imaging of surface exposed carbohydrate and protein revealed common localised areas of vivid fluorescence associated with buds, bud scars and nascent daughter cells. Carbohydrate and protein fluorescence often varied between strains, culture age and capsule status of cells examined. Finally, extension of protein tagging techniques resulted in the isolation and extraction of two biotinylated proteins from the yeast cell wall surface of an acapsulate strain of C.neoformans.

Establishment of a reedbed within a created surface water fed wetland nature reserve

In the early 1990's, outline designs for two wetland nature reserves were being prepared: the Teeside International Nature Reserve (TINR) and the Cardiff Bay Barrage Environmental Compensation Measures at Redwick, Gwent. The initial design for both proposals identified reedbed as a desirable habitat for establishment. The initial design works identified the importance of reedbed evapotranspiration [ET(Reed)] within the water budget, however, literature searches identified a paucity of information on this parameter. Field experiments for the measurement of ET(Reed) from Phragmites australis are described for three sites distributed across England and Wales. Reference Crop Evapotranspiration (ETo) was calculated using techniques recommended by the Food and Agriculture Organisation. A technique for the calculation of a reedbed crop coefficient [Kc(Reed)[, from ET(Reed) and ETo data is discussed. Kc(Reed) values produced in the project were found to be similar to those developed previously in continental Europe. Mean monthly and crop development stage Kc(Reed) values are presented which are applicable in the UK and possibly worldwide. A conceptual hydrological model of surface water fed reedbed systems is developed, and used to calculate the hydrological sustainability of reedbed creation areas in the UK. Finally, the water budget model is verified using data from a small clay catchment located on the TINR. In addition, a methodology is developed for the hydrological design of surface water fed reedbed systems, and recommendations required for the feasibility, design and establishment stage of reedbed creation sites. Further research needs are also identified.

Identification of industrial sources of airborne heavy metal pollution in urban areas

The research examines the deposition of airborne particles which contain heavy metals and investigates the methods that can be used to identify their sources. The research focuses on lead and cadmium because these two metals are of growing public and scientific concern on environmental health grounds. The research consists of three distinct parts. The first is the development and evaluation of a new deposition measurement instrument - the deposit cannister - designed specifically for large-scale surveys in urban areas. The deposit cannister is specifically designed to be cheap, robust, and versatile and therefore to permit comprehensive high-density urban surveys. The siting policy reduces contamination from locally resuspended surface-dust. The second part of the research has involved detailed surveys of heavy metal deposition in Walsall, West Midlands, using the new high-density measurement method. The main survey, conducted over a six-week period in November - December 1982, provided 30-day samples of deposition at 250 different sites. The results have been used to examine the magnitude and spatial variability of deposition rates in the case-study area, and to evaluate the performance of the measurement method. The third part of the research has been to conduct a 'source-identification' exercise. The methods used have been Receptor Models - Factor Analysis and Cluster Analysis - and a predictive source-based deposition model. The results indicate that there are six main source processes contributing to deposition of metals in the Walsall area: coal combustion, vehicle emissions, ironfounding, copper refining and two general industrial/urban processes. |A source-based deposition model has been calibrated using facctorscores for one source factor as the dependent variable, rather than metal deposition rates, thus avoiding problems traditionally encountered in calibrating models in complex multi-source areas. Empirical evidence supports the hypothesised associatlon of this factor with emissions of metals from the ironfoundry industry.

Assessment of the impact of discharges from surface water sewers on receiving water quality

A broad based approach has been used to assess the impact of discharges to rivers from surface water sewers, with the primary objective of determining whether such discharges have a measurable impact on water quality. Three parameters, each reflecting the effects of intermittent pollution, were included in a field work programme of biological and chemical sampling and analysis which covered 47 sewer outfall sites. These parameters were the numbers and types of benthic macroinvertebrates upstream and downstream of the outfalls, the concentrations of metals in sediments, and the concentrations of metals in algae upstream and downstream of the outfalls. Information on the sewered catchments was collected from Local Authorities and by observation of the time of sampling, and includes catchment areas, land uses, evidence of connection to the foul system, and receiving water quality classification. The methods used for site selection, sampling, laboratory analysis and data analysis are fully described, and the survey results presented. Statistical and graphical analysis of the biological data, with the aid of BMWP scores, showed that there was a small but persistent fall in water quality downstream of the studied outfalls. Further analysis including the catchment information indicated that initial water quality, sewered catchment size, receiving stream size, and catchment land use were important factors in determining the impact. Finally, the survey results were used to produce guidelines for the estimation of surface water sewer discharge impacts from knowledge of the catchment characteristics, so that planning authorities can consider water quality when new drainage systems are designed.

Surface waves in mesh synthetic photonic lattices

Eigenmodes and dispersion curves in different configurations of synthetic photonic lattices are studied numerically. Eigenmodes localized on borders between areas with different optical potential are found. Stability of these eigenmodes against potential disturbances of different type is studied.