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Biomedical subjects

B Dupuy

Publications and source records attributed to B Dupuy.

At least 37 records · Page 2Linked to original sources
PubMed1996

Influence of the structure of three corals on their resorption kinetics.

The aim of this study was to investigate the influence of the structure of corals on their resorption kinetics after implantation in subcutaneous areas. Three types of coral (Porites astreoides, Montastrea annularis and Dichocoenia stokesi) identical in composition but different in structure were implanted for periods of 1 and 2 months in subcutaneous sites in OF1 mice. The resorption of the implants was studied by means of qualitative (histology, scanning electron microscopy, fluorochrome labelling method) and quantitative approaches (gravimetric method). The results of the qualitative study revealed a process of irregular deterioration of the coral, linked to the detachment of crystals at the surface of the implant. The results of the quantitative study showed that the speed of resorption increases with the implantation time and the open porosity of the coral. These reactions are explained by the increase of the surface exchange area in contact with factors responsible for resorption: biological medium and cells. When considering the choice of coral as a bone substitute, these factors must be taken into account to allow the in situ maintenance of the implant over a sufficiently long period of time according to the clinical situation.

Animals↗
PubMed1995

The resorption of bone-implanted corals varies with porosity but also with the host reaction.

Three different exoskeletons of coral species Porites asteroides (P), Montastrea annularis (M), and Dichocoenia stokesi (D) were implanted for 2-20 weeks in rabbits. At 2, 4, 8, or 20 weeks, the exoskeletons presented variations in their resorptions depending on the species. To understand the variations in the decreasing speed of the implants despite their similar chemical composition, a study of the surface and architecture of the coral was carried out using scanning electronic microscopy, porosity was evaluated, and growth and differentiation of osteogenic cells cultured in vitro were observed for more than 1 month. At the cellular level, the surface of the implants was identical. Three-dimensional structures of the implants were variable, but the porosity values [P = 42.7%, M = 40.7%, and D = 17.4%] could not completely account for the differences in the resorbing process of the species. Standard histologic studies performed at 2, 4, 8, and 20 weeks after implantation produced the same pattern with P or M, showing aspects of rapid resorption; however, with D there were images resembling those of a foreign-body reaction. It seems that when resorption is not quick enough, a foreign body reaction develops which further slows down the process. This work focuses on the importance of porosity when using coral as bone substitute.

Animals↗
PubMed1995

Cholate-induced disruption of calcitonin-loaded liposomes: formation of trypsin-resistant lipid-calcitonin-cholate complexes.

PURPOSE: The work was performed to obtain a better understanding why the oral administration of calcitonin (CT)-loaded liposomes to rats results in a hypocalcemia, while liposomes are normally disrupted in the gastro-intestinal tract and cannot protect the hormone from enzymatic digestion. METHODS: In vitro comparisons between the stability of calcein and CT-loaded liposomes in the presence of cholate solutions led to an interpretation of the results observed. By means of gel filtration, turbidimetry, and fluorescence measurements, the interactions between CT and lipids were studied after sonicated liposomes had been broken down by cholate. RESULTS: Experiments showed that CT in the external medium of a liposome suspension had no effect on the vesicles. Gel filtration of cholate-treated liposomes loaded with calcein and CT resulted in a total separation of calcein from the lipid fraction for detergent concentrations higher than 4 mM. However, 50% of the CT was reencapsulated even when the cholate-to-phospholipid molar ratio was increased up to 100. Incubation of cholate-solubilized liposomes with 1% trypsin resulted in a partial CT-breakdown. CONCLUSIONS: These results strongly suggest that during membrane solubilization by cholate, lipid-CT complexes are formed which retain most of the CT initially embedded in the liposomal membrane, and which offer some protection to CT under the action of trypsin. The existence of these complexes could be one of the reasons for the reported hypocalcemia in rats after oral administration of CT-loaded liposomes.

Animals↗
PubMed1995

Rapid expansion of the physical and genetic map of the chromosome of Clostridium perfringens CPN50.

The physical map of the 3.6-megabase chromosome of Clostridium perfringens CPN50 was extended by positioning sites for the endonucleases SfiI and I-CeuI, and in parallel, the gene map was expanded by using a genome scanning strategy. This involved the cloning and sequencing of random chromosomal fragments, identification of the functions of the putative genes by database searches, and then hybridization analysis. The current gene map comprises almost 100 markers, many of which encode housekeeping functions while others are involved in sporulation or pathogenesis. Strikingly, most of the virulence genes were found to be confined to a 1,200-kb segment of the chromosome near oriC, while the pleiotropic regulatory locus, virRS, was situated toward the putative replication terminus. A comparison of the gene maps of three endospore-forming bacilli, C. perfringens, Clostridium beijerinckii, and Bacillus subtilis, revealed a similar order and distribution of key sporulation and heat shock genes which might reflect an ancient evolutionary relationship.

Chromosome Mapping↗
PubMed1994

Calcitonin-loaded liposomes: stability under acidic conditions and bile salts-induced disruption resulting in calcitonin-phospholipid complex formation.

Calcitonin-loading in liposomes composed of phosphatidylcholine, cholesterol and stearylamine or dipalmitoyl phosphatidylglycerol was studied at low pH values and in the presence of bile salts to check whether liposomal entrapment could be a possible means of protecting the peptide against the aggressive conditions present in the gastrointestinal tract. The association of calcitonin with the lipidic vesicles was monitored using radioactive labelling of the peptide and gel-filtration separation of the free and liposome-associated fractions. The results show that for all phospholipid compositions tested, loading was preserved in light acidic or basic buffers, and that only a slight disruption was observed at pH 2.5. Cholate caused a significant but only partial release of calcitonin even when the cholate-to-phospholipid ratio was increased. To understand the mode of calcitonin entrapment in the vesicles, the release of liposome-entrapped calcein was monitored concomitantly and taken as a stability criterion. Liposome integrity appears to be resistant at low pHs but to be totally destroyed by 4 mM cholate in a manner quasi-independent of the phospholipid concentration. These results strongly suggest that bile salts induce a disruption of the liposomes which results in the formation of new lipidic structures involving calcitonin and probably cholate.

Bile Acids and Salts↗
PubMed1994

Type IV prepilin peptidase gene of Neisseria gonorrhoeae MS11: presence of a related gene in other piliated and nonpiliated Neisseria strains.

The assembly of type IV pili in Neisseria gonorrhoeae is a complex process likely to require the products of many genes. One of these is the enzyme prepilin peptidase, which cleaves and then N methylates the precursor pilin subunits prior to their assembly into pili. We have used a PCR amplification strategy to clone the N. gonorrhoeae prepilin peptidase gene, pilDNg. A single copy of the gene is shown to be present in the chromosome. Its product promotes correct cleavage of the gonococcal prepillin in Escherichia coli cells carrying both the prepilin peptidase gene and the pilin structural gene. PilDNg also cleaves prePulG, a type IV pilin-like protein of Klebsiella oxytoca. Moreover, PilDNg complements a mutation in the gene coding for the prepilin peptidase-like protein of K. oxytoca, pulO, partially restoring PulG-PulO-dependent extracellular secretion of the enzyme pullulanase. Finally, we show that genes homologous to pilDNg are present and expressed in a variety of species in the genus Neisseria, including some commensal strains.

Amino Acid Sequence↗
PubMed1994

FT-IR of membranes made with alginate/polylysine complexes. Variations with the mannuronic or guluronic content of the polysaccharides.

FT-IR spectra of polylysine/alginate membranes made with alginate containing various contents of mannuronic or guluronic acid residues have been recorded. The interpretation of the more important absorptions related to functional groups engaged in the complexes have been proposed and discussed using comparisons with spectra of cellulosic films and other published results. Mannuronnic rich alginate seemed to link stronger than guluronic rich alginate to the polylysine molecules which is illustrated by the continuum in absorption between 3000 cm-1 and 2000 cm-1. However, the analysis of the 2000-1000 cm-1 region prompted us to believe that the polymers were engaged in the same basic sort of molecular complexes. Therefore it is necessary that other parameters (either physical, as toughness, porosity, ...) other than variations in molecular structures are studied in order that the biological differences of the membranes may be explained.

Alginates↗
PubMed1994

[Multimodal or multisensorial agnosia?].

A 75 year-old right handed woman had persistent right homonymous hemianopia and alexia without agraphia caused by a haemorrhagic stroke of the left occipito-temporal region. Six months later she suffered sudden onset visual and auditory agnosia, following a second haematoma, contralateral to the first one, in the right occipito-temporal region including the lingual and fusiform gyri. None of the disorders concerned semantic representation, so that an asemantic agnosia was excluded. Her performance in naming and recognition tests, in both visual and auditory modalities, demonstrated a wide range of responses and errors. The pattern of visual symptoms suggested "associative visual agnosia narrow sense" (Farah, 1990); auditory agnosia concerned only the non verbal stimuli. These findings were discussed in terms of anatomical mechanisms subserving perceptual, semantical, visuo and auditory-verbal representation. In this case, visual and auditory, agnosia appears to be independent.

Aged↗
PubMed1994

[Accuracy and reproducibility of the Eyesys corneal topographic analysis system].

Corneal topographic analysis has become quite useful for corneal surgery. Quantitative data on the quality and accuracy of the available instruments are not well known. We studied the accuracy and the reproducibility of the Eyesys machine using 4 calibrated balls: 8.99 mm, 7.93 mm, 7.10 mm, 6.13 mm. The machine is accurate (precision less than 0.1D for all the balls) and reproducible (4 measurements) with a SD less than 0.05 diopter for all the balls except for the 55.04's ball (0.06 to 0.1). We also studied reproducibility with six normal corneas using the same method. SD was 0.26 diopter for keratometry and 0.50 diopter for the astigmatism value.

Astigmatism↗
PubMed1993

Study of in vitro and in vivo stability of liposomes loaded with calcitonin or indium in the gastrointestinal tract.

Factors affecting liposome transport to the blood compartment after oral administration to rats were evaluated. A high entrapment of calcitonin (CT) was obtained when the vesicles were prepared by sonication and were composed of egg phosphatidylcholine, cholesterol and stearylamine. In vitro tests showed that the liposomes were stable in light acidic or basic buffers, but that they were partly lysed in pH 2.5, 10 mM bile salts and pancreatin. Oral administration of liposomes entrapping calcitonin in fasting rats showed that the vesicles facilitate transport of the hormone to the general circulation and that they increase the lifetime of 125I-CT in blood. Oral administration of liposomes entrapping radioactive indium in fasting rats did not induce radioactivity in blood. This could be explained by disruption of most of the vesicles in the enterocytes.

Administration, Oral↗
PubMed1993

Association of polyacrylamide beads to polyethylene terephthalate prostheses.

A method for the coupling of polyacrylamide beads to polyethylene terephthalate (PET) vascular prostheses is described. The reactional procedure used is performed along several steps; acrylic acid grafting on PET textile fibres, in order to introduce reactive carboxylic groups, introduction of terminal primary amine groups onto the beads, and then, attachment method which consists in coupling carboxylic groups of prostheses with amine groups of modified beads. The relative weight increase of the samples before and after the coupling reaction and, microscopic observations of beads distribution onto the prostheses surface demonstrate the binding feasibility of polyacrylamide matrices to PET prostheses. In the near future, authors expect to replace these beads by microcapsules with polyacrylamide wall and containing active compounds to improve the vascular prostheses biocompatibility.

Acrylic Resins↗
PubMed1992

PulO, a component of the pullulanase secretion pathway of Klebsiella oxytoca, correctly and efficiently processes gonococcal type IV prepilin in Escherichia coli.

The PulO protein required for extracellular secretion of pullulanase by Klebsiella oxytoca is known to be highly homologous to two type IV prepilin peptidases, namely XcpA(PilD) (Pseudomonas aeruginosa) and TcpJ (Vibrio cholerae). The predicted prepilin peptidase activity of PulO was confirmed by showing that it could correctly process the product of the cloned pilE.1 type IV pilin structural gene from Neisseria gonorrhoeae in Escherichia coli. The P. aeruginosa prepilin peptidase and another putative prepilin peptidase, ComC from Bacillus subtilis, also processed prePilE. Subcellular fractionation showed that the pilE gene product that had been processed by PulO remained associated with the cytoplasmic membrane, as did the unprocessed precursor. PulO was also shown to process three of the four prePilE-PhoA hybrids tested. Southern hybridization experiments suggest that a pulO homologue is present in the N. gonorrhoeae chromosome.

Bacterial Outer Membrane Proteins↗
PubMed1992

An enzyme with type IV prepilin peptidase activity is required to process components of the general extracellular protein secretion pathway of Klebsiella oxytoca.

The last gene (pulO) of the pulC-O pullulanase secretion gene operon of Klebsiella oxytoca codes for a protein that is 52% identical to the product of the pilD/xcpA gene required for extracellular protein secretion and type IV pilus biogenesis in Pseudomonas aeruginosa. The PilD/XcpA protein is known to remove the first six amino acids of the signal sequence of the type IV pilin precursor by cleaving after the glycine residue in the conserved sequence GF(M)XXXE (where X represents hydrophobic amino acids). This prepilin peptidase cleavage site is present in the products of four genes in the pulC-O operon (PulG, PulH, Pull and PulJ proteins). It is shown here that PulO processes the pulG gene product in vivo. Processing was maximal within 15 seconds, but experiments in which the expression of pulO was uncoupled from that of the other genes in the secretion operon suggest that processing can also occur post-translationally. The products of two pulG derivatives with internal inframe deletions were also processed by PulO, but the three PulG-PhoA hybrids, two PulJ-PhoA hybrids and the single PulH-PhoA hybrid tested did not appear to be processed. Sucrose gradient fraction experiments showed that both precursor and mature forms of PulG appear to be associated with low-density, outer membrane vesicles prepared by osmotic lysis of sphaeroplasts. Neither the xcpA gene nor the Bacillus subtilis gene comC, which is also homologous to pulO and codes for a protein with type IV prepilin peptidase activity, can correct the pullulanase secretion defect in an Escherichia coli strain carrying all of the genes required for secretion except pulO. Furthermore, neither XcpA nor ComC is able to process prePulG protein in vivo.

Amino Acid Sequence↗
PubMed1992

Role of pilA, an essential regulatory gene of Neisseria gonorrhoeae, in the stress response.

Sequence analysis has shown that PilA, a transcriptional regulator of pilin gene expression in Neisseria gonorrhoeae, has extensive homology with the 54-kDa protein of the signal recognition particle of eukaryotes and its receptor, as well as with two proteins of Escherichia coli, FtsY and Ffh, which have been proposed to be a part of a signal recognition particle-like apparatus. We tested the putative role of PilA in protein export in N. gonorrhoeae and did not find any effect. However, we did observe induction of a heat shock response and a previously described slow-growth phenotype when PilA function was impaired. We also examined the interference of pilA expression in E. coli with the function of the products of ftsY and ffh and observed an accumulation of pre-beta-lactamase. We argue against a direct role for PilA in protein export in gonococci and propose instead that PilA is involved in the modulation of cell growth rate in response to different environmental conditions.

Bacterial Outer Membrane Proteins↗
PubMed1992

Embedded adrenal cells graft reduced local and early nonspecific inflammatory phenomena which follow agarose beads implantation.

Microencapsulation of adrenal cells is proposed for reducing the nonspecific inflammatory reaction observed around polymer implants. This hypothesis was tested by comparing both host cellular reaction and the surrounding graft cell populations which appeared either when agarose embedded cells or when empty agarose beads were implanted. Our results showed that the fibrotic material that surrounded the implanted empty agarose microbeads was not as severe and important when adrenal cells were present. Similarly, T lymphocyte population surrounding the graft was considerably reduced together with the percentage of CD4 and CD8 positive cell subpopulations. The activation macrophage marker IaD disappeared. Our results support the hypothesis that embedded adrenal cells may be a suitable solution for reducing early inflammatory events due to microcapsules implantation.

Adrenal Cortex↗
PubMed1991

Microencapsulation of isolated pituitary cells by polyacrylamide microlatex coagulation on agarose beads.

Microlatex beads of homogenous size were made by polymerization of a mixture of acrylamide/bisacrylamide dispersed in a microemulsion. The microlatex was aggregated by dilution of the microemulsion in acrylamide solutions. The aggregates were then coagulated by polymerization at the interfaces of agarose beads circulating in a capillary tube containing paraffin oil. Biocompatibility was tested on isolated pituitary cells microencapsulated by this procedure.

Acrylamide↗