Search

[EN] This work explore the use of supercritical CO2 drying as alternative technique for the obtainment of pasteurized and high quality dried product. Several tests were conducted on animal, vegetable and fruit matrixes in order to investigate the effectiveness of SC-CO2 drying process at different process conditions. Design of experiment was performed to find the optimal process conditions for vegetable and fruit matrices, using the final water activity of the products as key indicator for the drying efficiency. The inactivation of naturally present microorganisms and inoculated pathogens demonstrated the capability of SC-CO2 drying process to assure a safe product. Moreover, retention of nutrients was compared with conventional drying methods. Results suggest that supercritical drying is a promising alternative technology for food drying.

[EN] This work explore the use of supercritical CO2 drying as alternative technique for the obtainment of pasteurized and high quality dried product. Several tests were conducted on animal, vegetable and fruit matrixes in order to investigate the effectiveness of SC-CO2 drying process at different process conditions. Design of experiment was performed to find the optimal process conditions for vegetable and fruit matrices, using the final water activity of the products as key indicator for the drying efficiency. The inactivation of naturally present microorganisms and inoculated pathogens demonstrated the capability of SC-CO2 drying process to assure a safe product. Moreover, retention of nutrients was compared with conventional drying methods. Results suggest that supercritical drying is a promising alternative technology for food drying.

[EN] This work explore the use of supercritical CO2 drying as alternative technique for the obtainment of pasteurized and high quality dried product. Several tests were conducted on animal, vegetable and fruit matrixes in order to investigate the effectiveness of SC-CO2 drying process at different process conditions. Design of experiment was performed to find the optimal process conditions for vegetable and fruit matrices, using the final water activity of the products as key indicator for the drying efficiency. The inactivation of naturally present microorganisms and inoculated pathogens demonstrated the capability of SC-CO2 drying process to assure a safe product. Moreover, retention of nutrients was compared with conventional drying methods. Results suggest that supercritical drying is a promising alternative technology for food drying.

[EN] This work explore the use of supercritical CO2 drying as alternative technique for the obtainment of pasteurized and high quality dried product. Several tests were conducted on animal, vegetable and fruit matrixes in order to investigate the effectiveness of SC-CO2 drying process at different process conditions. Design of experiment was performed to find the optimal process conditions for vegetable and fruit matrices, using the final water activity of the products as key indicator for the drying efficiency. The inactivation of naturally present microorganisms and inoculated pathogens demonstrated the capability of SC-CO2 drying process to assure a safe product. Moreover, retention of nutrients was compared with conventional drying methods. Results suggest that supercritical drying is a promising alternative technology for food drying.

[EN] This work explore the use of supercritical CO2 drying as alternative technique for the obtainment of pasteurized and high quality dried product. Several tests were conducted on animal, vegetable and fruit matrixes in order to investigate the effectiveness of SC-CO2 drying process at different process conditions. Design of experiment was performed to find the optimal process conditions for vegetable and fruit matrices, using the final water activity of the products as key indicator for the drying efficiency. The inactivation of naturally present microorganisms and inoculated pathogens demonstrated the capability of SC-CO2 drying process to assure a safe product. Moreover, retention of nutrients was compared with conventional drying methods. Results suggest that supercritical drying is a promising alternative technology for food drying.

BACKGROUND & AIMS: Progressive liver disease is a severe complication of cystic fibrosis, a genetic disease characterized by impaired epithelial adenosine 3',5'-cyclic monophosphate-dependent secretion caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). In the liver, CFTR is expressed in cholangiocytes and regulates the fluid and electrolyte content of the bile. Glibenclamide, a sulfonylurea and a known CFTR inhibitor, paradoxically stimulates cholangiocyte secretion. We studied the molecular mechanisms underlying this effect and whether glibenclamide could restore cholangiocyte secretion in cystic fibrosis. METHODS: NRC-1 cells, freshly isolated rat cholangiocytes, isolated rat biliary ducts, and isolated biliary ducts from CFTR-defective mice (Cftr tm1Unc ) were used to study fluid secretion (by video-optical planimetry), glibenclamide-induced secretion (by high-performance liquid chromatography in cell culture medium), intracellular pH and intracellular Ca 2+ concentration transients [2'7'-bis(2-carboxyethyl)-5,6,carboxyfluorescein-acetoxymethylester and Fura-2 f-AM (5-Oxazolecarboxylic acid, 2-(6-(bis(2-((acetyloxy)methoxy)-2-oxoethyl)amino)-5-(2-(2-(bis(2-((acetyloxy)methoxy)-2-oxoethyl)amino)-5-methylphenoxy)ethoxy)-2-benzofuranyl)-, (acetyloxy)methyl ester) microfluorometry], gene expression (by reverse-transcription polymerase chain reaction), and changes in membrane capacitance (by patch-clamp experiments). RESULTS: Stimulation of cholangiocyte secretion by glibenclamide and tolbutamide required Cl - and was mediated by the sulfonylurea receptor 2B. Glibenclamide-induced secretion was blocked by inhibitors of exocytosis (colchicine, wortmannin, LY294002, and N -ethylmaleimide) and by inhibitors of secretory granule acidification (vanadate, bafilomycin A1, and niflumic acid) but was Ca 2+ and depolarization independent; membrane capacitance measurements were consistent with stimulation of vesicular transport and fusion. Gli

Pathogenic strains of Helicobacter pylori produce a potent exotoxin, VacA, which intoxicates gastric epithelial cells and leads to peptic ulcer. The toxin is released from the bacteria as a high molecular mass homo-oligomer of a 95 kDa polypeptide which undergoes specific proteolytic cleavage to 37 kDa and 58 kDa subunits. We have engineered a strain of H. pylori to delete the gene sequence coding for the 37 kDa subunit. The remaining 58 kDa subunit is expressed efficiently and exported as a soluble dimer that is non-toxic but binds target cells in a manner similar to the holotoxin. A 3D reconstruction of the molecule from electron micrographs of quick-freeze, deep-etched preparations reveals the contribution of each building block to the structure and permits the reconstruction of the oligomeric holotoxin starting from individual subunits. In this model P58 subunits are assembled in a ring structure with P37 subunits laying on the top. The data indicate that the 58 kDa subunit is capable of folding autonomously into a discrete structure recognizable within the holotoxin and containing the cell binding domain., Journal Article, Research Support, Non-U.S. Gov't, SCOPUS: ar.j, info:eu-repo/semantics/published

Pathogenic strains of Helicobacter pylori produce a potent exotoxin, VacA, which intoxicates gastric epithelial cells and leads to peptic ulcer. The toxin is released from the bacteria as a high molecular mass homo-oligomer of a 95 kDa polypeptide which undergoes specific proteolytic cleavage to 37 kDa and 58 kDa subunits. We have engineered a strain of H. pylori to delete the gene sequence coding for the 37 kDa subunit. The remaining 58 kDa subunit is expressed efficiently and exported as a soluble dimer that is non-toxic but binds target cells in a manner similar to the holotoxin. A 3D reconstruction of the molecule from electron micrographs of quick-freeze, deep-etched preparations reveals the contribution of each building block to the structure and permits the reconstruction of the oligomeric holotoxin starting from individual subunits. In this model P58 subunits are assembled in a ring structure with P37 subunits laying on the top. The data indicate that the 58 kDa subunit is capable of folding autonomously into a discrete structure recognizable within the holotoxin and containing the cell binding domain., Journal Article, Research Support, Non-U.S. Gov't, SCOPUS: ar.j, info:eu-repo/semantics/published

The cell vacuolating activity of the protein toxin VacA, released by Helicobacter pylori, is strongly increased in vitro by exposure to acidic pH followed by neutralization. This short acid exposure does not increase significantly the binding of VacA to cell or to lipid membranes. However, membrane photolabeling with photoactivatable radioactive phospholipids and ANS binding studies show that VacA transiently exposed to pH equal or lower than 5 changes conformation and exposes on its surface hydrophobic segments. Both the 32 and the 58 kDa subunits of the toxin insert in the lipid bilayer and interact with the fatty acid chains of phospholipids. Membrane binding and penetration are enhanced by incubating target cells or liposomes with the toxin at mild acidic pH values, similar to those present around H. pylori on the stomach mucosa. These findings are discussed with respect to the critical step in cell intoxication consisting in the translocation of the active toxin domain into the cell cytosol. We suggest that membrane translocation takes place at the plasma membrane level., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published

The cell vacuolating activity of the protein toxin VacA, released by Helicobacter pylori, is strongly increased in vitro by exposure to acidic pH followed by neutralization. This short acid exposure does not increase significantly the binding of VacA to cell or to lipid membranes. However, membrane photolabeling with photoactivatable radioactive phospholipids and ANS binding studies show that VacA transiently exposed to pH equal or lower than 5 changes conformation and exposes on its surface hydrophobic segments. Both the 32 and the 58 kDa subunits of the toxin insert in the lipid bilayer and interact with the fatty acid chains of phospholipids. Membrane binding and penetration are enhanced by incubating target cells or liposomes with the toxin at mild acidic pH values, similar to those present around H. pylori on the stomach mucosa. These findings are discussed with respect to the critical step in cell intoxication consisting in the translocation of the active toxin domain into the cell cytosol. We suggest that membrane translocation takes place at the plasma membrane level., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published