Implementation of a sustainable project in the Tanbi Wetland Complex, Banjul (The Gambia)

The aim of this on-going study is creating a design for a boardwalk that is part of an ecotouristic project in the Tanbi Wetland National Park (TWNP). This mangrove area is located adjacent to the capital of The Gambia, Banjul. The nature reserve was listed as a Ramsar site in 2007 (an "Area of International Importance") but it is under great human pressure, such as through the dumping of municipal waste in the mangrove swamps, the erosion of banks by speedboats from tourist water sports, etc. In this way, the mangrove habitats are degraded and can even be destroyed.The entire project is supported by the City of Ostend, which has a city link with Banjul since 2003. As the City of Ostend co-finances the entire project, a cost calculation will be carried out.The boardwalk project aims at creating awareness, both for the people of Banjul and for tourists visiting Gambia. Eco-education also plays an important role in this process; local students can visit the TWNP and learn about the mangroves and their importance. The TWNP is located close to hotels and tourist areas in Banjul. The local boats could be used for excursions in the small creeks, combined with the boardwalk-excursion. Guided tours by local inhabitants make visitors aware of the problems affecting the nature reserve.The project includes a pedestrian walk-on bridge with a length of approximately one kilometer, two observation towers that can be used by birdwatchers, a platform where oyster women can prepare local products for visitors, a pontoon for moor canoes, etc. Another proposition is to build an openair museum, the “Sea Life Center”, located in seven different huts across the entire boardwalk. It portrays the different habitats and their ecological and socio-ecological components present in and around the mangrove.During the calculation of the wood structure the Belgian standards (NBN) and Eurocode (EC) were followed where possible. The Australian standards (AUS) were also consulted, in contrast to the Belgian standards, they have standards specifically for boardwalks. These standards are used when designing constructions. Design values were obtained from tests carried out according to the standard. We drew every boardwalk model using Google Sketchup Pro 7.1, a 3D software tool.During a 5-week stay in Banjul in January-February 2011, the mangrove area will be mapped using a surveying device. The Trimble Pathfinder Pro XRT equipped with an OmniSTAR license allows measurements with an accuracy up to 20 centimeters. Based on the maps that were made, implementation plans are composed, making construction possible by the inhabitants of Banjul

Recherche des déterminants de la stimulation de l'expression parvovirale précoce, induite par la transformation

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Lack of a detectable effect of capsid proteins on the cell-dependent activity of parvovirus MVMp promoters

Deletion of upstream elements from the early P4 promoter of parvovirus MVM (minute virus of mice, prototype strain MVMp) has a differential effect on its activity, depending on the host cell considered. This indicates that upstream motifs participate in the control of promoter P4 functioning and are responsive to factors which are, at least in part, cell-type specific. In contrast with other viral systems, the capsid proteins of MVMp had no detectable effect on gene expression driven by either the early P4 or late P38 promoter of MVMp in permissive and non-permissive cells. © 1994 Institut Pasteur/Elsevier.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Method for concentrating and purifying recombinant autonomous parvovirus vectors designed for tumour-cell-targeted gene therapy

Recent work has highlighted the use of parvoviruses as potential vectors for tumour-cell-targeted gene therapy. The oncotropic properties of the prototype strain of minute virus of mice (MVMp) suggest that this virus might be a useful vehicle for introducing selectively therapeutic genes, e.g. lymphokine or suicide genes, into tumour cells and preferentially expressing them. But the low titre of recombinant virus stocks (105-106 infectious units per ml) and their high level of contamination by cell proteins make it practically impossible to evaluate their efficacy in in vivo systems. A technique is described for producing cellular contaminant-free stocks of recombinant virus particles, with titres up to 5 x 108 IU/ml.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Programmed killing of human cells by means of an inducible clone of parvoviral genes encoding non-structural proteins.

Although its dependence on the target cell type is well established, the cytopathogenicity of parvoviruses has remained elusive to date as far as its mechanism is concerned. However, indirect evidence suggested that parvoviral non-structural (NS) proteins may be the cytotoxic effectors. In order to test this hypothesis, a molecular clone of parvovirus MVMp was modified, by replacing the P4 promoter of the NS transcription unit by the glucocorticoid-inducible promoter of the mouse mammary tumour virus. Clones of neoplastic human cells that had incorporated this construct and that were induced to produce NS proteins by dexamethasone, showed a cytopathic effect and eventually died. Our data strongly suggest that the intracellular accumulation of parvoviral NS products jeopardizes the survival of the cells, which cannot be detected unless a threshold protein concentration is reached. Interestingly, a cell variant could be isolated which resisted dexamethasone-induced killing, although it was fully inducible for the production of NS proteins. This variant was also unusually resistant to infection with MVMp virions, thus confirming the essential role played by the NS proteins in the parvoviral cytotoxicity and indicating that the cytocidal activity of the parvoviral NS products is modulated by cellular factors that may vary from one cell to another

Oncoselective Parvoviral Vector-Mediated Gene Therapy of Cancer

We replaced capsid genes by reporter genes and assessed expression in different types of human cancer cells and their normal counterparts, either at the level of whole cell population (CAT ELISA) or at the single cell level [FACS analysis of green fluorescent protein (GFP)]. CAT expression was substantial (up to 10,000 times background) in all infected tumor cells, despite variations according to the cell types. In contrast, no gene expression was detected in similarly infected normal cells (with the exception of an expression slightly above background in fibroblasts). FACS analysis of GFP expression revealed that most tumor cells expressed high level of GFP while no GFP-positive normal cells could be detected with the exception of very few (less than 0.1%) human fibroblast cells expressing high level of GFP. We also replace capsid genes by genes coding for the costimulatory molecules B7-1 and B7-2 and show that, upon infection with B7 recombinant virions, only tumor cells display the costimulatory molecules and their immunogenicity was increased without any effect on normal cells. Using a recombinant minute virus of mice (MVM) containing the herpes simplex thymidine kinase gene, we could get efficient killing of most tumor cell types in the presence of ganciclovir, without affecting normal proliferating cells. The prospects and limitations of these different strategies are discussed.SCOPUS: cp.jinfo:eu-repo/semantics/publishe

Initiation of transcription from the minute virus of mice P4 promoter is stimulated in rat cells expressing a c-Ha-ras oncogene.

Transformation of FR3T3 rat fibroblasts by a c-Ha-ras oncogene but not by bovine papillomavirus type 1 is associated with an increase in the abundance of mRNAs from prototype strain MVMp of infecting minute virus of mice, an oncosuppressive parvovirus. This differential parvovirus gene expression correlates with the reported sensitization of ras- but not bovine papillomavirus type 1-transformed cells to the killing effect of MVMp (N. Salomé, B. van Hille, N. Duponchel, G. Meneguzzi, F. Cuzin, J. Rommelaere, and J. Cornelis, Oncogene 5:123-130, 1990). Experiments were performed to determine at which level parvovirus expression is up-regulated in ras transformants. An MVMp "attenuation" sequence responsible for the premature arrest of RNA elongation was either placed or not placed in front of the chloramphenicol acetyltransferase gene and brought under the control of MVMp early promoter P4. Although the MVMp attenuator reduced P4-driven chloramphenicol acetyltransferase expression, the extent of attenuation was similar in normal and ras-transformed cells. Moreover, the analysis of P4-directed viral RNAs in MVMp-infected cultures by RNase protection and nuclear run-on assays also revealed a transcription elongation block of a similar amplitude in both types of cells. In addition, the stabilities of the three major parvoviral mRNAs did not vary significantly between normal and ras-transformed cells. Hence, it is concluded that the ras-induced increase in the accumulation of parvoviral mRNAs is mainly controlled at the level of transcription. Consistently, the TATA motif of the P4 promoter proved to have a differential photoreactivity when tested by in vivo UV footprinting assays in ras-transformed versus normal cells

Use of an autonomous parvovirus vector for selective transfer of a foreign gene into transformed human cells of different tissue origins and its expression therein

In this work, we report the transduction of a chloramphenicol acetyltransferase (CAT) reporter gene into a variety of normal and transformed human cells of various tissue origins. The vector used was MVM/P38cat, a recombinant of the prototype strain of the autonomous parvovirus minute virus of mice (MVMp). The CAT gene was inserted into the capsid-encoding region of the infectious molecular clone of MVMp genome, under the control of the MVM P38 promoter. When used to transfect permissive cells, the MVM/P38cat DNA was efficiently replicated and expressed the foreign CAT gene at high levels. By cotransfecting with a helper plasmid expressing the capsid proteins, it was possible to produce mixed virus stocks containing MVM/P38cat infectious particles and variable amounts of recombinant MVM. MVM/P38cat viral particles were successfully used to transfer the CAT gene and to express it in a variety of human cells. Both viral DNA replication and P38-driven CAT expression were achieved in fibroblasts, epithelial cells, T lymphocytes, and macrophages in a transformation-dependent way, but with an efficiency depending on the cell type. In transformed B lymphocytes, however, the vector was not replicated, nor did it express the CAT gene.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Sensitiztion of transformed rat fibroblasts to killing by parvovirus minute virus of mice correlates with an increase in viral gene expression

Cultures of established rat fibroblasts transformed by the avian erythroblastosis virus were more susceptible to the cytopathic effect of the autonomous parvovirus minute virus of mice, prototype strain (MVMp), than were their untransformed homologs. This effect could be ascribed to the presence of a greater fraction of cells that were sensitive to the killing action of MVMp in transformed cultures than in their normal parents. Yet, transformed and normal lines were similarly efficient in virus uptake, DNA amplification, and capsid protein synthesis. In contrast, transformants accumulated 2.5- to 3-fold greater amounts of all three major MVM mRNA species and nonstructural protein than did their normal progenitors. Thus, in this system transformation-associated sensitization of cells to MVMp appears to correlate primarily with an increase in their capacity for the expression of the viral transcription unit which encodes nonstructural proteins and is controlled by the P4 promoter. Consistently, a report gene was expressed at a higher level by transformed versus normal cultures, when placed under the control of the MVM P4 promoter. As infectious MVMp was produced in larger amounts by transformed cultures, a late step of the parvoviral cycle, such as synthesis, encapsidation of progeny DNA, or both, was also stimulated in the transformed cells.SCOPUS: NotDefined.jinfo:eu-repo/semantics/publishe