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Biomedical subjects

C M Lapière

Publications and source records attributed to C M Lapière.

At least 19 recordsLinked to original sources
PubMed1998

Gelatinases in drug-induced toxic epidermal necrolysis.

BACKGROUND: The matrix metalloproteinases (MMPs) MMP2 and MMP9 play a significant role in epidermal detachment, inflammation and re-epithelialization. We have evaluated their activity in toxic epidermal necrolysis (TEN). DESIGN: The level and pattern of activity of MMP2 and MMP9 were investigated by measuring the degradation of 3H-labelled gelatin and by zymography in blister fluid from six TEN patients and compared the results with three other blistering conditions: bullous pemphigoid (n = 6), second-degree burn (n = 13) or suction blister (n = 3). RESULTS: A higher amount of MMP2 was found in TEN blister fluid with the constant presence of a significantly larger proportion of the activated forms of MMP2, a particular feature of TEN, than the other blistering diseases studied. CONCLUSION: This study emphasizes the potential role of MMP2 in the specific inflammatory reaction and reparation process in TEN skin.

Adult↗
PubMed1998

In vitro modulation of human gingival epithelial cell attachment and migration by minocycline-HCL.

Although the influence of tetracyclines on periodontal connective tissue cells has been the topic of many in vitro and in vivo studies, data regarding their effects on gingival epithelial cells are scarce. The present in vitro study was designed to examine the influence of minocycline, a semi-synthetic analog of tetracycline, on human gingival keratinocyte (HGK) attachment and migration. Attachment tests were performed with HGK prelabeled by tritiated amino-acids. Increasing concentrations of minocycline (10, 50, 100 micrograms/ml) in the medium produced no significant modification of cell adhesion kinetics compared to control conditions, except for 100 micrograms/ml which statistically significantly (p < 0.05) reduced the number of attached cells beyond 6 h. A 24-h cell preincubation in 10 micrograms/ml of minocycline did not alter the kinetics of HGK attachment. Scanning electron microscopic observations of attached HGK showed that the presence of 10 micrograms/ml of minocycline in the "attachment medium" induced the production of multiple filopodial extensions. Migration tests in Boyden chambers for 40 h demonstrated that HGK preincubation for 24 h in a 10 micrograms/ml minocycline-HCl solution increased significantly (p < 0.005) cell migration towards a gradient of fetal calf serum. The presence of 10 micrograms/ml of minocycline in contact with the keratinocytes in the upper compartment of the migration chambers also produced a significant (p < 0.005) result. In contrast, the presence of minocycline in the lower compartments did not produce any chemoattractive effect. Within the limits of their significance, these results suggest that, at concentrations not beyond 50 micrograms/ml, minocycline could fasten the periodontal wound coverage by epithelial cells and allow the normal reformation of a junctional epithelium.

Adolescent↗
PubMed1997

Angiogenesis by fibroblast growth factor 4 is mediated through an autocrine up-regulation of vascular endothelial growth factor expression.

The infection of normal mouse mammary EF43 cells by a retroviral vector carrying either Fgf-3 (EF43.Fgf-3) or Fgf-4 (EF43.Fgf-4) cDNA resulted in the transformation of cells displaying different tumorigenic potentials in nude mice (A. Hajitou and C-M. Calberg-Bacq, Int. J. Cancer, 63: 702-709, 1995). EF43.Fgf-4 produced rapidly developing tumors at all sites of inoculation, whereas EF43.Fgf-3 produced slowly growing tumors only in the mammary fat pad. Cells infected with the vector carrying the selection gene alone (EF43.C) were not tumorigenic. The angiogenic properties of these cells were tested in an in vitro angiogenesis model using human umbilical vein endothelial cells (HUVECs) cultured at the surface of a type I collagen gel and their capacity to form tube-like structures on invasion of the gel. Only the conditioned medium (CM) of EF43.Fgf-4 induced an angiogenic morphotype in HUVECs. In parallel, the mRNA expression of matrix metalloproteinase 1 and c-ETS-1 was increased in the HUVECs displaying a differentiated phenotype, whereas the tissue inhibitor of matrix metalloproteinase 1 mRNA level was decreased. Recombinant human fibroblast growth factor 4 (FGF-4) did not induce an angiogenic phenotype in HUVECs by itself. By Western blot analysis, a high expression of vascular endothelial growth factor (VEGF) was detected in the EF43.Fgf-4 CM. This result was confirmed by Northern blot analysis of total RNA extracted from the three cell types; the steady-state level of VEGF mRNA was low and equivalent in EF43.C and EF43.Fgf-3, whereas it was strongly increased in EF43.Fgf-4. Culturing EF43 cells carrying only the selection gene with increasing concentrations of recombinant human FGF-4 resulted in a dose-dependent stimulation of VEGF. The induction of the angiogenic morphotype and the parallel modulations of the biosynthetic phenotype in HUVECs were completely suppressed by adding a neutralizing antibody directed against VEGF to EF43.Fgf-4 CM. Furthermore, inhibition of protein kinase C by bisindoylmaleimide suppressed the angiogenic phenotype induced by the CM of EF43.Fgf-4. Our results point to an indirect angiogenic activity of FGF-4 through the autocrine induction of VEGF secretion by EF43.Fgf-4 cells, an original signaling pathway that might be significant in tumor progression and metastasis.

Animals↗
PubMed1997

cDNA cloning and expression of bovine procollagen I N-proteinase: a new member of the superfamily of zinc-metalloproteinases with binding sites for cells and other matrix components.

Procollagen N-proteinase (EC 3.4.24.14) cleaves the amino-propeptides in the processing of type I and type II procollagens to collagens. Deficiencies of the enzyme cause dermatosparaxis in cattle and sheep, and they cause type VIIC Ehlers-Danlos syndrome in humans, heritable disorders characterized by accumulation of pNcollagen and severe skin fragility. Amino acid sequences for the N-proteinase were used to obtain cDNAs from bovine skin. Three overlapping cDNAs had an ORF coding for a protein of 1205 residues. Mammalian cells stably transfected with a complete cDNA secreted an active recombinant enzyme that specifically cleaved type I procollagen. The protein contained zinc-binding sequences of the clan MB of metallopeptidases that includes procollagen C-proteinase/BMP-1. The protein also contained four repeats that are homologous to domains found in thrombospondins and in properdin and that can participate in complex intermolecular interactions such as activation of latent forms of transforming growth factor beta or the binding to sulfatides. Therefore, the enzyme may play a role in development that is independent of its role in collagen biosynthesis. This hypothesis was supported by the observation that in some tissues the levels of mRNA for the enzyme are disproportionately high relative to the apparent rate of collagen biosynthesis.

Amino Acid Sequence↗
PubMed1997

[Mechanism of the growth and rupture of abdominal aortic aneurysm].

The relationship between atherosclerosis and abdominal aortic aneurysm development is well known. Atherosclerosis cannot explain the whole mechanism. Genetic characters of mechanisms leading to abdominal aortic development is obvious from this study and others. Our study evidences an increased metalloproteases activity in aortic wall proportionally to the size of the abdominal aortic aneurysm. A decrease of aortic wall elastin is evidenced proportionally to the AAA size. Extractable collagen is significantly increased in the aortic wall of patients operated on for aortic rupture.

Age Factors↗
PubMed1996

Activated forms of MMP2 and MMP9 in abdominal aortic aneurysms.

PURPOSE: This consistent observation of a reduction of the elastin concentration in abdominal aortic aneurysms (AAAs) has led us to investigate in AAA specimens two metalloproteinases that display elastase activity, MMP2 (gelatinase A/72kDa) and MMP9 (gelatinase B/92 kDa). METHODS: Samples of full-thickness aortic wall, adherent thrombus, and serum were collected in 10 patients with AAAs. Samples of normal aortic wall and serum were taken from 6 age-matched control patients. Quantitative gelatin-zymography and gelatinolytic soluble assays after acetyl-phenyl mercuric acid activation were performed on serum and tissue extracts, and the results were expressed in units on a comparative wet-weight basis. Histologic analysis was performed in parallel to score the inflammatory infiltrate. RESULTS: The luminal and parietal parts of the thrombus contained, respectively, 20- and 10-fold more gelantinolytic activity than the serum. The predominate form was MMP9. Although the total gelatinolytic activity was in the same range both in AAAs and in normal walls, a significantly higher proportion of MMP9 was found in the aneurysmal aortic walls. Furthermore, a significant proportion of MMP9 was under its processed active form, which was never observed in normal samples. A significantly higher proportion of MMP2 was also present as processed active form in AAA wall. This latter parameter positively correlated with the inflammatory score. CONCLUSIONS: The presence of activated MMP9 and MMP2 might contribute to the degradation of the extracellular matrix proteins that occurs during the development of aneurysms.

Aged↗
PubMed1996

Primary systemic amyloidosis. A report of 2 cases.

We report 2 cases of primary systemic amyloidosis. A monoclonal gammopathy was confirmed at the postmortem examination of the first patient. An extensive search for evidence of chronic infection, inflammation, neoplasms and paraproteinemia was conclusively negative in the other patient. The recognition of cutaneous signs of primary systemic amyloidosis is crucial to insure a rapid management aimed at postponing the fatal issue of the disease.

Aged↗
PubMed1996

[Low intensity electromagnetic fields produce a wave of calcium in the fibroblasts].

Human fibroblasts display a Ca2+ wave after irradiation with an electromagnetic field (EMF) of low intensity (100 to 900 microT) as seen by LASER confocal microscopy and excitation of Fluo 3. The number of excited cells is proportional to the intensity of EMF between 100 and 900 microT. Cellular activation by a dialysable serum factor is required to induce the Ca2+ wave. It also depends on extracellular Ca2+ and active tyrosine kinases and phospholipase C gamma.

Calcium↗
PubMed1995

Characterization and partial amino acid sequencing of a 107-kDa procollagen I N-proteinase purified by affinity chromatography on immobilized type XIV collagen.

Procollagen I N-proteinase (EC 3.4.24.14), the enzyme that specifically processes type I and type II procollagens to collagen, was isolated from extracts of fetal calf skin. After two chromatographic steps on concanavalin A-Sepharose and heparin-Sepharose, the semi-purified preparation was used to produce monoclonal antibodies. One reacting antibody was found to recognize not the enzyme itself but type XIV collagen on which the enzyme was bound. This binding, highly sensitive to ionic conditions (plH, salt concentrations) but not affected by non-ionic detergents, was used for affinity chromatography that strongly improved the purification procedure. The enzyme is extensively characterized: 1) it has a molecular mass of 107 kDa as determined by polyacrylamide gel electrophoresis in presence of SDS and of about 130 kDa when estimated by gel filtration on a Sephacryl-S300; 2) in standard assay (pH 7.5, 0.2 M NaCl, 35 degrees C), the activation energy for reaction with amino procollagen type I was 17,000 calories per mole. In the same conditions, Km and Vmax values were, respectively, 435 and 39 nM per hour but varied strongly with pH and salt concentration; 3) the enzyme cleaved the NH2-terminal propeptide of type I procollagen at the specific site, the Pro-Gln bond in the alpha 1 type I procollagen chain; 4) the enzyme contained a high proportion of Gly, Asx, and Glx residues but no Hyp or Hyl; 5) partial amino acid sequences obtained from internal peptides of the enzyme displayed no significant homology with known sequences. The association of procollagen I N-proteinase with a FACIT (fibril-associated collagens with interrupted triple helices) collagen as found here might be of physiological significance.

Amino Acid Sequence↗
PubMed1995

Investigation of the relationship between osteoporosis and the collagenase gene by means of polymorphism of the 5'upstream region of this gene.

Osteoporosis is a slowly progressing disease resulting from an imbalance between bone accretion and degradation. As interstitial collagenase is a key enzyme in the degradation of bone matrix, we investigated a possible relationship between the collagenase gene and osteoporosis. Analysis of an amplified genomic DNA fragment from -524 to +52 by denaturing gradient gel electrophoresis and sequencing allowed us to detect three dimorphic sites upstream of base -300, one of them leading to a BanI restriction site. None of the sites could be directly associated with osteoporosis. The allele frequencies of the three dimorphic sites were estimated. The interallelic ratios were high, thus providing new useful genetic markers for linkage analysis. When comparing these ratios in osteoporotic and nonosteoporotic subjects, no significant differences could be observed.

Adult↗
PubMed1995

Keratinocytes modulate the biosynthetic phenotype of dermal fibroblasts at a pretranslational level in a human skin equivalent.

In this study we investigated the influence of keratinocytes on the phenotype of fibroblasts in an in vitro human skin equivalent. Keratinocytes were seeded at the surface of fibroblast-populated mechanically restrained type I collagen gels (lattices). Lattices without keratinocytes were handled in parallel as controls. After 2 and 4 days in culture, the keratinocyte layer was removed and the steady-state level of the mRNA for the main extracellular matrix macromolecules and interstitial collagenase produced by the fibroblasts was measured by Northern and dot blot analysis. A 50% decrease in the amount of procollagen type I and type III mRNAs was observed after 2 and 4 days of coculture while collagenase gene expression was upregulated by 300% when compared with control lattices. No significant modulation of type IV and type VI collagen, elastin or laminin B1 mRNA levels was found. Fibronectin mRNA levels in fibroblasts were significantly increased only on day 4. All the observed changes could be reproduced using a conditioned medium collected from a lattice covered with keratinocytes added to a lattice containing fibroblasts alone. These results indicate that in an in vitro reconstituted skin, keratinocytes are able to modulate the biosynthetic phenotype of fibroblasts at a pretranslational level through a paracrine signalling pathway.

Adult↗
PubMed1994

Cultured myofibroblasts display a specific phenotype that differentiates them from fibroblasts and smooth muscle cells.

BACKGROUND: Myofibroblasts (MF) from human granulation tissue derive from perilesional cells and display similarities with fibroblasts (DF) and smooth muscle cells (SMC). OBJECTIVE: Determine additional markers to assess the phenotype of MF. METHODS: MF, DF and SMC derived from human tissues were amplified in culture. Their phenotype and response to various regulatory processes were compared in vitro. RESULTS: MF resemble SMC by their mechanical properties and cytoskeletal proteins. They resemble DF by their biosynthetic phenotype and their response to the regulation operated by the extracellular matrix and mechanical forces. CONCLUSION: In vitro MF display a phenotype that differentiates them from DF and SMC.

Cells, Cultured↗
PubMed1994

[Type VII C Ehlers-Danlos disease or human dermatosparaxia: the product of a fruitful union of so-called fundamental sciences with human and veterinary clinical medicine].

Ehlers-Danlos type VII C disease is a heritable disorder of the connective tissues and a disease similar to dermatosparaxia in bovine and other animal species. The determination of the pathomechanism of these diseases, the inactivity of the specific protease excising the aminoterminal extension of the type I collagen precursor, resulted of interactive basic and clinical research.

Ehlers-Danlos Syndrome↗
PubMed1994

[Anodontia and hidrotic ectodermal dysplasia, hyporeactive to heat. Effect of acitretin].

We observed an adult female patient presenting an autosomal recessive ectodermal dysplasia difficult to classify among the various reported forms of this disease. She had the major signs of the hidrotic type of the disease (palmo-plantar hyperkeratosis, nail and hair dystrophies, and typical sweat glands) and anodontia as reported in the hypohidrotic form of the syndrome. We evaluated the function of the sweat glands and found them hyporeactive to a heat stimulus. Most of the epithelial alterations have improved under therapy by acitretin.

Acitretin↗