Study of the active site residues of a glycoside hydrolase family 8 xylanase

in Journal of Molecular Biology (2005), 354(2), 425-435

Site-directed mutagenesis and a comparative characterisation of the kinetic parameters, pH dependency of activity and thermal stability of mutant and wild-type enzymes have been used in association with ... [more ▼]

Site-directed mutagenesis and a comparative characterisation of the kinetic parameters, pH dependency of activity and thermal stability of mutant and wild-type enzymes have been used in association with crystallographic analysis to delineate the functions of several active site residues in a novel glycoside hydrolase family 8 xylanase. Each of the residues investigated plays an essential role in this enzyme: E78 as the general acid, D281 as the general base and in orientating the nucleophilic water molecule, Y203 in maintaining the position of the nucleophilic water molecule and in structural integrity and D144 in sugar ring distortion and transition state stabilization. Interestingly, although crystal structure analyses and the pH-activity profiles clearly identify the functions of E78 and D281, substitution of these residues with their amide derivatives results in only a 250-fold and 700-fold reduction in their apparent k(cat) values, respectively. This, in addition to the observation that the proposed general base is not conserved in all glycoside hydrolase family 8 enzymes, indicates that the mechanistic architecture in this family of inverting enzymes is more complex than is conventionally believed and points to a diversity in the identity of the mechanistically important residues as well as in the arrangement of the intricate microenvironment of the active site among members of this family. (c) 2005 Elsevier Ltd. All rights reserved. [less ▲]

Study of the airflow patterns in healthy and diseased cattle

in Proceedings of the Sixth Veterinary Respiratory Symposium (1987, November)

A study of the anatomy and injection techniques of the ovine stifle by positive contrast arthrography, computed tomography arthrography and gross anatomical dissection

in Veterinary Journal (in press)

Study of the antifungal activity of lactic acid bacteria isolated from fermented cassava and mill flour

Poster (2009, April)

Study of the behaviour of welded joints between elliptical hollow sections

in Proceedings of the 10th International Symposium on Tubular Structures (2003)

Study of the Boettcher cells along their development: Junctions and expression of the urea-transporter B (UT-B)

Poster (2012, September 30)

The Boettcher cells (BC) lie on the sensory epithelium of the cochlea. Their function has never been clearly defined. However it has been suggested that they may influence the ionic composition of the ... [more ▼]

The Boettcher cells (BC) lie on the sensory epithelium of the cochlea. Their function has never been clearly defined. However it has been suggested that they may influence the ionic composition of the fluids of the inner ear, which play a central role in the conduction of the sensory information. In this context the compartimentating function of the BC around and after the onset of hearing may influence the subsequent refining of hearing. We collected ultrastructural and immunohistological data during the final maturation stage of the sensory epithelium. In particular the cell junctions were investigated to clarify the compartimentating function of the BC at early stages. As a potential actor in the ion flow in the sensory epithelium, the urea transporter-B (UT-B) was also immunolocalised during the development of the BC. At the mature stage (P25) the BC are linked to the adjacent cells by numerous adherens and non-adherens junctions. They rest on a basilar membrane to which they are attached by hemidesmosomes. They typically exhibit large basolateral interdigitations. We found that, at the 8th postnatal day, the BC are separated from the neighbouring cells by wide spaces entered by scarce cytoplasmic extensions. These spaces are interrupted by areas of close contact, where adherens and non-adherens junctions may be found. Thus, although there seems to be fewer interdigitations at P8, gap junctions probably still allow easy cell-to-cell exchanges. Moreover non-adherens junctions can systematically be identified apically. Although it was impossible to differenciate tight and gap junctions without specific labeling, we postulate that these non-adherens junctions correspond to tight junctions and seal the apex of the BC. This feature is necessary to enable the control of the ion concentrations surrounding the sensory epithelium. We also found that UT-B, known for water and urea transport in red blood cells, is present in the membranes of the BC from P12 (the earliest stage tested) to P25. Thus UT-B may play a role in the regulation of the ionic concentrations of the inner ear fluids. [less ▲]

Study of the Cape Ghir upwelling filament using variational data analysis and regional numerical model

Doctoral thesis (2011)

Aim: Upwelling filaments are shallow, elongated structures of cool waters often visible in the regions of coastal upwelling. The present thesis aims to improve our knowledge on the mechanisms responsible ... [more ▼]

Aim: Upwelling filaments are shallow, elongated structures of cool waters often visible in the regions of coastal upwelling. The present thesis aims to improve our knowledge on the mechanisms responsible for the generation of filaments. Location: the study is centered on the Canary Current Upwelling system, located off northwest Africa, in particular around Cape Ghir (30°38'N, 9°53'W), where these filaments are frequently observed. Methods: - We compiled historical in situ data to build a high-resolution climatology of temperature and salinity in the region of interest using the Data Interpolating Variational Analysis (\diva) method. - We ran the experiments with the numerical model ROMS to investigate the processes responsible for the filament generation. - We collected in situ measurements during a cruise in August-September 2009, in the frame of the CAIBEX project, in the filament area and analyzed them to get a detailed description of the Cape Ghir filament, its variability and its biogeochemical characteristics. - We computed anomalies of several variables with respect to a long-term mean to examine the time-evolution of the Canary Current Upwelling system. Results: The climatology showed improvements with respect to existing products, especially in the coastal area, and was a valuable asset to provide initial and boundary conditions for the numerical model. The results of the simulations underlined the role of the wind in the filament generation and confirmed our assumption of a mechanism based on the potential vorticity balance. Preliminary analysis of the cruise data allowed us to characterize the filament physical and biological properties, in particular we observed the signal of a subsurface (at a depth of 300~m) anticyclonic eddy on the northern flank of the filament, also present in the numerical model outputs. Anomalies for the year 2010 revealed a widespread warming over the tropical and subtropical Atlantic Ocean and a diminution of the upwelling activity. We attributed these anomalies to a weakening of the winds, itself related to a strongly negative North Atlantic Oscillation (NAO) index. [less ▲]

Study of the combined affect of solutes and water activities on growth rate of Botrytis cinerea, agent of grey mould on apple fruits

Poster (2004)

Study of the combined effect of 5-ALA-based photodynamic therapy and NF-kappaB inhibitors on human glioblastoma cell survival

Poster (2008, October)

Study of the combined effects of data assimilation and grid nesting in ocean models – application to the Gulf of Lions

in Ocean Science (2006), 2

Modern operational ocean forecasting systems routinely use data assimilation techniques in order to take observations into account in the hydrodynamic model. Moreover, as end users require higher and ... [more ▼]

Modern operational ocean forecasting systems routinely use data assimilation techniques in order to take observations into account in the hydrodynamic model. Moreover, as end users require higher and higher resolution predictions, especially in coastal zones, it is now common to run nested models, where the coastal model gets its open-sea boundary conditions from a low-resolution global model. This configuration is used in the "Mediterranean Forecasting System: Towards environmental predictions" (MFSTEP) project. A global model covering the whole Mediterranean Sea is run weekly, performing 1 week of hindcast and a 10-day forecast. Regional models, using different codes and covering different areas, then use this forecast to implement boundary conditions. Local models in turn use the regional model forecasts for their own boundary conditions. This nested system has proven to be a viable and efficient system to achieve high-resolution weekly forecasts. However, when observations are available in some coastal zone, it remains unclear whether it is better to assimilate them in the global or local model. We perform twin experiments and assimilate observations in the global or in the local model, or in both of them together. We show that, when interested in the local models forecast and provided the global model fields are approximately correct, the best results are obtained when assimilating observations in the local model. [less ▲]