Determination of Coniothyrium minitans conidial and germling lectin avidity by flow cytometry and digital microscopy

The avidity of conidia and 48-h-old germlings of Coniothyrium minitans for FITC-conjugated lectins was characterised by flow cytometry and digital microscopy. Six isolates of C. minitans representing three morphological types were compared. Binding of Con A, SBA and WGA by conidial populations varied markedly in extent and pattern between isolates, however, with increasing culture age, conidia from all isolates demonstrated a significant reduction in lectin avidity. Germling isolates bound significantly different amounts of lectins and lectin binding differed significantly with locality. Spore walls of all germlings from all isolates bound more ConA compared with hyphal apices and mature hyphal walls. In contrast, hyphal apices of the majority of germling isolates, readily bound SBA and mature hyphal walls of germling isolates bound more WGA than other regions of the germlings. Such differential lectin binding by conidia and germlings may influence their specific surface interactions and adherence characteristics.

Analysis of Acanthamoeba polyphaga surface carbohydrate exposure by FITC-lectin binding and flourescence evaluation

Aims: Characterization of the representative protozoan Acanthamoeba polyphaga surface carbohydrate exposure by a novel combination of flow cytometry and ligand-receptor analysis. Methods and Results: Trophozoite and cyst morphological forms were exposed to a panel of FITC-lectins. Population fluorescence associated with FITC-lectin binding to acanthamoebal surface moieties was ascertained by flow cytometry. Increasing concentrations of representative FITC-lectins, saturation binding and determination of K d and relative Bmax values were employed to characterize carbohydrate residue exposure. FITC-lectins specific for N-acetylglucosamine, N-acetylgalactosamine and mannose/glucose were readily bound by trophozoite and cyst surfaces. Minor incremental increases in FITC-lectin concentration resulted in significant differences in surface fluorescence intensity and supported the calculation of ligand-binding determinants, Kd and relative B max, which gave a trophozoite and cyst rank order of lectin affinity and surface receptor presence. Conclusions: Trophozoites and cysts expose similar surface carbohydrate residues, foremost amongst which is N-acetylglucosamine, in varying orientation and availability. Significance and Impact of the Study: The outlined versatile combination of flow cytometry and ligand-receptor analysis allowed the characterization of surface carbohydrate exposure by protozoan morphological forms and in turn will support a valid comparison of carbohydrate exposure by other single-cell protozoa and eucaryotic microbes analysed in the same manner.

Characterization of FITC-conjugated lectin binding to Candida albicans

Avidity of yeast and hyphal forms of Candida albicans for FITC-conjugated lectins was determined by flow cytometry and digital microscopy. Yeast phase cells bound Con A, a lectin with marked affinity for mannose, irrespective of growth phase, yet demonstrated little avidity for WGA and SBA. Yeast phase cell avidity for mannose-specific lectins was characterized through determination of FITC-conjugated Con A, LcH, PSA and GNA binding and subsequent calculation of Bmax, EC50 and Hn values. Such an approach, through comparison among FITC-conjugated lectins of differing specific activities, furnishes further insight into exposed outer cell wall mannose moieties. The rank order of lectin affinity as defined by EC50 values was GNA > Con A > LcH > PSA. Values for Hn suggest that lectins predominantly bind to a single receptor class, the relative abundance of which as defined by Bmax values was PSA > GNA > Con A > LcH. Hyphal surfaces in common with yeast phase cells demonstrated marked avidity for FITC-Con A, however, fluorescence of Candida morphological forms differed significantly, indicative of varying outer cell wall mannose exposure.

Measurements of polarimetric sensitivity to hydrostatic pressure, strain and temperature in birefringent dual-core microstructured polymer fiber

We experimentally characterized a birefringent microstructured polymer fiber of specific construction, which allows for single mode propagation in two cores separated by a pair of large holes. The fiber exhibits high birefringence in each of the cores as well as relatively weak coupling between the cores. Spectral dependence of the group and the phase modal birefringence was measured using an interferometric method. We have also measured the sensing characteristics of the fiber such as polarimetric sensitivity to hydrostatic pressure, strain and temperature. Moreover, we have studied the effect of hydrostatic pressure and strain on coupling between the cores.

Identifying the ICT challenges of the agri-food sector to define the architectural requirements for a future internet core platform

This paper discussed the specific challenges of the agri-food sector in the light of research carried out in the Smart-AgriFod project. Using questionnaires and focus groups, our research identified as number of business needs and drivers which enable the identification of suitable Future Internet technologies across the three sub-domains of Smart Farming, Smart Agri-logistics, and Smart Food Awareness. The universal need for information access and the importance of standards to enable this lead us to propose an integrated scenario for end to end information access from farm to fork. We conclude by discussing wider implications of such developments especially for climate change and urbanisation.

FITC-lectin avidity of Cryptococcus neoformans cell wall and capsular components

Flow cytometry and confocal microscopy were used to quantify and visualize FITC-lectin binding to cell-surface carbohydrate ligands of log and stationary phase acapsular and capsular Cryptococcus neoformans strains. Cell populations demonstrated marked avidity for terminal a-linked mannose and glucose specific FITC-Con A, mannose specific FITC-GNL, as well as N-acetylglucosamine specific FITC-WGA. Exposure to other FITC-lectins specific for mannose, fucose and N-acetylgalactosamine resulted in little cell-surface fluorescence. The nature of cell-surface carbohydrates was investigated further by measurement of the fluorescence from surfaces of log and stationary phase cell populations after exposing them to increasing concentrations of FITC-Con A and FITC-WGA. Cell fluorescence increased significantly with small increases in FITC-Con A and FITC-WGA concentrations attaining reproducible maxima. Measurements of this nature supported calculation of the lectin binding determinants EC 50, Hn, Fmax and relative Bmax values. EC50 values indicated that the yeast-cell surfaces had greatest affinity for FITC-WGA, however, relative Bmax values indicated that greater numbers of Con A binding sites were present on these same cell surfaces. Hn values suggested a co-operative lectin-carbohydrate ligand interaction. Imaging of FITC-Con A and FITC-WGA cell-surface fluorescence by confocal microscopy demonstrated marked localization of both lectins to cell surfaces associated with cell division and maturation, indicative of dynamic carbohydrate ligand exposure and masking. Some fluorescence was associated with entrapment of FITC-Con A by capsular components, but FITC-Con A and FITC-WGA readily penetrated the capsule matrix to bind to the same cell surfaces labelled in acapsular cells.

Polarimetric sensitivity to hydrostatic pressure and temperature in birefringent dual-core microstructured polymer fiber

We experimentally characterized a birefringent microstructured polymer fiber of specific construction, which allows for single mode propagation in two cores separated by a pair of large holes. The fiber exhibits high birefringence in each of the cores as well as relatively weak coupling between the cores. Spectral dependence of the group and the phase modal birefringence was measured using an interferometric method. We have also measured the sensing characteristics of the fiber such as the polarimetric sensitivity to hydrostatic pressure and temperature. © 2010 SPIE.

Laboratory focussed learning of core electronic engineering concepts in the first year of an honours degree programme

In common with most universities teaching electronic engineering in the UK, Aston University has seen a shift in the profile of its incoming students in recent years. The educational background of students has moved away from traditional Alevel maths and science and if anything this variation is set to increase with the introduction of engineering diplomas. Another major change to the circumstances of undergraduate students relates to the introduction of tuition fees in 1998 which has resulted in an increased likelihood of them working during term time. This may have resulted in students tending to concentrate on elements of the course that directly provide marks contributing to the degree classification. In the light of these factors a root and branch rethink of the electronic engineering degree programme structures at Aston was required. The factors taken into account during the course revision were:. Changes to the qualifications of incoming students. Changes to the background and experience of incoming students. Increase in overseas students, some with very limited practical experience. Student focus on work directly leading to marks. Modular compartmentalisation of knowledge. The need for provision of continuous feedback on performance We discuss these issues with specific reference to a 40 credit first year electronic engineering course and detail the new course structure and evaluate the effectiveness of the changes. The new approach appears to have been successful both educationally and with regards to student satisfaction. The first cohort of students from the new course will graduate in 2010 and results from student surveys relating particularly to project and design work will be presented at the conference. © 2009 K Sugden, D J Webb and R P Reeves.