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C Figarella

Publications and source records attributed to C Figarella.

155 records · Page 9Linked to original sources
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Histidylated polylysine as a synthetic vector for gene transfer into immortalized cystic fibrosis airway surface and airway gland serous cells.

BACKGROUND: We recently designed a cationic polymer called histidylated polylysine made of polylysine partially substituted with histidyl residues which become protonated at slightly acidic pH. This polymer is thought to induce the leakage of acidic vesicles containing plasmid/histidylated polylysine complexes. METHODS: and results Here, we have analyzed the ability of histidylated polylysine to transfer reporter or CFTR genes into immortalized cystic fibrosis airway surface epithelial cells (sigmaCFTE29o- cells) and airway gland serous cells (CF-KM4 cells) which are both important targets for cystic fibrosis gene therapy. The luciferase reporter gene expression measured after gene transfer with histidylated polylysine into both cell lines was quite high and similar to that obtained with commercially available vectors. In addition, the level of expression was not dependent on the presence of a membrane disrupting agent such as chloroquine. Histidylated complexes were present in slightly acidic non-lysosomal cellular compartments as shown by a cytological approach using biotinylated plasmid, lysosome-specific antibodies and confocal microscopy. Histidylated complexes appeared to be of small size when prepared at low ionic strength and formed aggregates upon increasing the ionic strength. However, aggregate formation was prevented by the addition of 10% fetal bovine serum. Gene transfer efficiency varied with the size of the complexes and decreased when small particles were used. CONCLUSIONS: These results suggest that histidylated polylysine may be an efficient non-viral vector for gene transfer into cystic fibrosis airway surface epithelial cells and airway gland serous cells.

Animals↗
PubMedDate not supplied

Cystic fibrosis proteins detected by electrophoretic techniques.

For the detection of the cystic fibrosis protein (CFP) in serum of cystic fibrosis (CF) carriers, thin-layer polyacrylamide gel isoelectric focusing proved inappropriate as a diagnostic test, but was useful for screening fractions on purification of CFP by chromatofocusing on a Mono P column. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis an Mr 12,000 protein (P12) was found in most CFP-positive sera, indicating good correlation between these two CF-associated proteins. Detection of the P12 protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was well reproducible and less delicate than IEF. The technique was also used to purify P12 from serum by two successive preparative electrophoresis steps in a 7.5-15% gradient and 15% homogeneous gel. The use of silver staining revealed that P12, which was present in all sera of CF patients and carriers with variable intensities, was also present in trace amounts in normal sera.

Blood Proteins↗
PubMedDate not supplied

Isolation and properties of prophospholipase A2 from human pancreatic juice.

Human prophospholipase A2 was purified from pancreatic juice. The protein has a molecular weight of 14500 and a free N-terminal residue identified as aspartic acid (or asparagine). The amino acid composition was determined. Partial immunological identity has been obtained between human and porcine prophospholipase A2. As other phospholipases, the human enzyme requires the presence of calcium for its activity. However, the activity of human phospholipase A2 on egg yolk emulsion is partially inhibited at 0.4 mM calcium concentration, which differs from the porcine homologous enzyme. Kinetics of activation of the two zymogens (human and porcine) by 4 different trypsins (bovine, porcine and human) indicate a difference between the two zymogens only when activated by human trypsins, which suggests a marked specificity of both human trypsins for human prophospholipase.

Animals↗