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Biomedical subjects

F Bois

Publications and source records attributed to F Bois.

26 records · Page 2Linked to original sources

Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines.

In order to study the roles of the different alternatively spliced variants of human fibronectin (FN) as well as of its binding sites and structural domains in the processes of extracellular matrix assembly, cell adhesion, and cell migration, we constructed expression vectors coding for different human full-length FN polypeptides and deleted versions of these constructs. We expressed them transiently in mammalian cells by calcium phosphate transfection and microinjection techniques. While the deleted recombinants were expressed by both transfection and microinjection, the full-length recombinants could be only expressed by microinjection. Calcium phosphate transfection leads to the accumulation of recombinant FN in cytoplasmic vesicles of both matrix-forming and nonforming cells. On the contrary, microinjection leads to the accumulation of recombinant FN in cytoplasmic vesicles in cells that do not form a matrix, but to the rapid incorporation into the extracellular matrix of matrix-forming cells in addition to a cytoplasmic localization. Identical results were obtained when the FN signal and propeptides were replaced by those of E-cadherin.

Animals↗

Effect of glycosidases and proteinases on cobalamin binding and physicochemical properties of purified saturated haptocorrin and intrinsic factor.

The effect of exoglycosidase, N-glycanase, trypsin and chymotrypsin was studied on the binding capacity and physicochemical properties of intrinsic factor and of haptocorrin using Superose 6 gel filtration. Intrinsic factor was purified as recently described by us. Haptocorrin was purified 6000-fold from human saliva using thermolabile affinity chromatography and high-performance cationic exchange chromatography with a specific activity of 20.6 nmol of cobalamin (Cbl) per mg protein and a yield of 44.7%. Exoglycosidases provoked a decrease of 54.3 and 78.2% of the Cbl binding capacity of haptocorrin and intrinsic factor, respectively. The sequential incubation of haptocorrin and intrinsic factor wit exoglycosidases and proteinases provoked a decrease of, respectively, 100 and 92.7% of their Cbl binding capacity, whereas the incubation with proteinase decreased the Cbl binding capacity of, respectively, 67.9 and 7.9%. The result of the incubation of [3H]intrinsic factor or [3H]haptocorrin with chymotrypsin and trypsin gave, respectively, no change in the elution position and a shift corresponding to a decrease of 50% of the estimated molecular mass. The estimated molecular mass of Cbl-intrinsic factor and of Cbl-haptocorrin decreased, respectively, to 57.1 kDa and to 88.1 kDa after incubation with exoglycosidases. It was concluded that (1) the carbohydrate core of intrinsic factor protects the whole protein whereas the carbohydrate core of haptocorrin protects only half part of the protein and (2) the carbohydrates are implicated in the formation of the cobalamin binding site of haptocorrin and intrinsic factor.

Chemical Phenomena↗

Identification of an hyperglycemic factor from heated serum.

A fraction of in vitro heated serum (44 degrees C, 15 minutes) obtained by gel filtration chromatography and high performance size exclusion chromatography was found to induce an hyperglycemia when injected to Wistar rats. This fraction corresponded to a molecular weight range of 2,000-11,000. We speculate that this fraction contained a polypeptide responsible of hyperglycemia and induced by hyperthermia.

Animals↗

Sequestration of crystalline and endogenous cobalamin by R binders down to the distal ileum in exocrine pancreatic dysfunction.

It has been recently shown that crystalline cyanocobalamin in exocrine pancreatic insufficiency is sequestered by R binders down to the proximal jejunum, and that bile inhibits the binding of cobalamin to intrinsic factor. In freshly collected human bile, we have found a single type of apo R binder, with a relative molecular mass of 128 100, a molecular radius of 4.65 nm, and a mean isoelectric point of 3.72. Salivary and biliary holo R binders were incubated with normal human gastric juice and intestinal juice from healthy subjects and patients having exocrine pancreatic insufficiency. No degradation of these two holo R binders occurs with normal gastric juice and intestinal juice from patients after four hours incubation time at 37 degrees C, but a partial degradation of salivary holo R binders and a complete loss of biliary Cbl binding capacity were observed with normal intestinal juice in the same in vitro conditions. We have confirmed in vivo, using a triple-lumened tube, that a part of the salivary and biliary holo R binders remains undegraded down to the distal ileum in two patients with exocrine pancreatic insufficiency. These findings strongly suggest that the enterohepatic circulation of cobalamin is effective in healthy subjects, whereas it is partially interrupted in the patients. They provide a proof that a part of endogenous and crystalline exogenous cobalamin is sequestered to R binders down to the distal ileum, and confirm that the failure to degrade the digestive R binders is responsible for the malabsorption of crystalline cobalamin in exocrine pancreatic dysfunction.

Bile↗

Screening blood donations for hepatitis C in Central Africa: analysis of a risk- and cost-based decision tree.

Four screening strategies (no testing, HC Abbott, HC Pasteur, and a combined test) for the detection of hepatitis C virus (HCV) antibody in donated blood were considered in a formal decision tree. Decision criteria included residual risk of infection and overall monetary cost. Tree parameters were determined using data from the Central African Republic. The prevalences observed among blood donors for HIV infection, hepatitis B, syphilis, and hepatitis C varied between 6% and 15%. The current residual risk of transfusion-transmitted infections is very high (8.4%). Screening for HCV would bring that risk down to about 3% with either the HC Pasteur, the HC Abbott, or the combined test. Even though baseline analysis gives preference to the HC Abbott test (the combined test coming out last), Monte Carlo sensitivity and uncertainty analyses showed that Abbott's and Pasteur's tests are interchangeable, on the basis or either risk or cost considerations.

Blood Donors↗