1,721,159 research outputs found
Novel particulate systems for the delivery of an oligonucleotide decoy to Nuclear Factor-kappaB: a potential strategy for treating cystic fibrosis
No full textObjectives To develop safe and effective particulate systems for cystic fibrosis gene therapy by an oligonucleotide decoy to Nuclear Factor-kB. Background/Rationale Decoy oligonucleotide (decoy ODN) able to interfere with NF-kB pathway may be of great help in reducing NF-kB mediated pulmonary chronic inflammation, which is the primary cause of bronchioectasis, respiratory failure and death of CF patients. To overcome issues related to the use of oligonucleotide (ODN) in therapy, delivery through poly(lactide-co-glycolide) (PLGA) microparticles, able to protect ODN toward the biological environment and/or release the drug in a controlled way, has been suggested. Recent studies indicate that PLGA microparticles allow an increase of ODN biological stability, are able to maintain its hybridization capability and provide a sustained ODN pharmacological activity. Conceiving PLGA microspheres for CF treatment, another important challenge to be taken into account is their potential in ODN pulmonary delivery. On this matter, recent data indicate that adequately formulated PLGA particles may effectively transport high-molecular weight drugs to the lungs. Preliminary results In our previous studies, PLGA microspheres for ODN delivery have been developed. We investigated the potential of PLGA microspheres as delivery systems for a decoy phosphorotioate ODN against NF-kB in RAW 264.7 macrophages stimulated with lipopolysaccharide. In particular, in vitro incubation of macrophages with decoy ODN-releasing microspheres resulted in an inhibition of NF-kB activation eighty-times higher than that achieved with naked decoy ODN. We have also demonstrated that the use of polyethylenimine, a cationic polymer, can be of help in improving cell internalization and nuclear characterization of ODN. These findings suggest that due to their sustained release properties PLGA microspheres allow the full optimization of ODN pharmacokinetics. More recently, we have designed and developed PLGA-based dry powders intended for drug pulmonary delivery. In vitro/in vivo experimental results suggest that large porous particles (LPP) with flow properties and size suitable for aerosolization and deposition in deep regions of the lung following inhalation can be achieved. Project description (experimental plan, methods, timetable) The project will be carried out in two years. PLGA microspheres loaded with a decoy ODN against NFkB, eventually containing PEI, will be developed. Particular attention will be paid to particle for pulmonary delivery by evaluating in depth their aereodynamic properties which will affect their potential as pulmonary carriers. All the particles will be fully characterized for morphology, size, loading efficiency and in vitro release. The optimized systems will be tested on specific CF cell lines and animal models of CF. Anticipated output Research results such as patents, scientific publications and presentations at international symposia can be expected. We believe that this will occur in a reasonable short period (by the end of the 1st year) due to the advanced knowledge and expertise of the research teams involved in the project as well as the fact that the groups already collaborate on similar topics. Relevance to Italian CF Foundation The project could be of great interest for pharmaceutical and biotechnological industries involved in the development of novel therapeutic approaches for rare diseases. In fact, thanks to a specific EU legislation, industries which desire to sponsor an orphan drug can be the recipients of several economic incentives. Due to the high industrial applicability of the research, CF foundation could attract industrial attention and benefit of new opportunity of financial support
LIPOCORTIN AND VASOCORTIN: TWO SPECIES OF GLUCOCORTICOID INDUCED PROTEINS AFFECTING VASCULAR PERMEABILITY.
No full textCloricromene inhibits the induction of nitric oxide synthase.
No full textThe effect of cloricromene, a coumarin derivative, was investigated on the lipopolysaccharide-stimulated nitric oxide (NO) synthase induction in intact aortas from endotoxin shocked rats and in the murine macrophage cell line J774. Rings of thoracic aortas from lipopolysaccharide (4 mg/kg, i.v.)-shocked rats, contracted with phenylephrine, showed a progressive decrease in tone, that was of a greater magnitude than that of aortas from naive rats. Moreover, a decreased response to the constrictor effect of phenylephrine was observed in aortas from shocked rats. In vivo treatment with cloricromene (2 mg/kg, i.v.) 30 min before lipopolysaccharide administration partially prevented the loss in tone of aortic rings and improved their reactivity to phenylephrine. Murine J774 macrophages activated with lipopolysaccharide (100 ng/ml) produced significant amounts of nitrites (NO2-; 28.2 +/- 3.5 nmol/10(6) cells per 24 h). Cloricromene (2, 20 or 200 microM) added to the cells concomitantly with lipopolysaccharide inhibited NO2- production in a concentration-dependent manner. Maximum inhibition (84.0 +/- 8.0\%) was observed when cloricromene (200 microM) was added to the cells 6 h before lipopolysaccharide, whereas it was ineffective when given 6 h after endotoxin. These results demonstrate that cloricromene inhibits the expression but not the activity of the inducible NO synthase
Modulation of granuloma formation by endogenous nitric oxide.
No full textWe have studied the possible involvement of nitric oxide (NO) in granuloma formation induced by subcutaneous implantation in rats of carrageenin-soaked polyether sponges. Modulation of the L-arginine: NO pathway in rats was achieved by treating rats with the NO synthase inhibitor, NG-nitro-L-arginine methyl ester, as well as with L- or D-arginine. Granulomatous tissue formation, cell infiltration and NO2- production were reduced, in a dose-dependent manner, by NG-nitro-L-arginine methyl ester and increased by L-arginine but not by D-arginine. These results suggest that endogenous NO plays a modulating role in granuloma formation
Hydrocortisone-induced inhibitor of prostaglandin biosynthesis in rat leucocytes.
No full textRat peritoneal luecocytes incubated with hydrocortisone (10 micrograms/ml) release a factor which inhibits prostaglandin generation. The steroid-induced inhibitor, which mediates the anti-phospholipase effect of antiinflammatory steroids, may be a protein or a polypeptide since its formation is blocked by cycloheximide, a known inhibitor of protein synthesis
Characterization of vasocortin-like proteins induced by dexamethasone in endothelial cells.
No full textIn this study we have shown that dexamethasone induces vasocortin-like proteins in bovine endothelial cells as well as in the bovine aortic endothelial cell line, GM 7373. Vasocortin-like proteins have been characterized by their ability to mimic the glucocorticoid inhibition of rat dextran oedema and histamine release induced by concanavalin-A in rat mast cells. Following partial purification of these proteins by gel filtration, vasocortin activity was found to be associated to proteins with molecular weight between 20-35 kD. This study showed that dexamethasone induces vasocortin-like proteins in endothelial cells and suggests that endothelial cells are target cells of glucocorticoid activity in vascular tissue
LIPOCORTIN AND VASOCORTIN: TWO SPECIES OF GLUCOCORTICOID INDUCED PROTEINS AFFECTING VASCULAR PERMEABILITY.
No full textCyclolinteinone, a sesterterpene from sponge Cacospongia linteiformis, prevents inducible nitric oxide synthase and inducible cyclo-oxygenase protein expression by blocking nuclear factor-kappaB activation in J774 macrophages.
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