We have evaluated the ability of two carbohydrate biopolymers, chitosan and gellan, to enhance antibody responses to subunit influenza virus vaccines delivered to the respiratory tracts of mice. Groups of mice were vaccinated three times intranasally (i.n.) with 10 microg of purified influenza B/Panama virus surface antigens (PSAs), which consist of hemagglutinin (HA) and neuraminidase (NA), either alone or admixed with chitosan or gellan solutions. Separate groups were vaccinated subcutaneously (s.c.) with PSAs adsorbed to Alhydrogel or chitosan or gellan alone i.n. Serum antibody responses were determined by enzyme-linked immunosorbent assay (ELISA) for influenza virus-specific immunoglobulin G (IgG) and by HA inhibition (HAI) and NA inhibition (NAI) assays. The local respiratory immune response was measured by assaying for influenza virus-specific IgA antibody in nasal secretions and by enumerating nasal and pulmonary lymphocytes secreting IgA, IgG, and IgM anti-influenza virus-specific antibodies by enzyme-linked immunospotting (ELISPOT). When administered alone i.n., B/Panama PSA was poorly immunogenic. Parenteral immunization with B/Panama PSA with Alhydrogel elicited high titers of anti-B/Panama antibodies in serum but a very poor respiratory anti-B/Panama IgA response. In contrast, i.n. immunization with PSA plus chitosan stimulated very strong local and systemic anti-B/Panama responses. Gellan also enhanced the local and serum antibody responses to i.n. PSA but not to the same extent as chitosan. The ability of chitosan to augment the immunogenicity of influenza vaccines given i.n. was confirmed using PSA prepared from an influenza A virus (A/Texas H1N1). (+info)
Cholesterol reduction by glucomannan and chitosan is mediated by changes in cholesterol absorption and bile acid and fat excretion in rats.
(18/1030)
Glucomannan, a viscous polysaccharide, and chitosan, a derivative of chitin, have both been demonstrated to lower cholesterol in animals. However, the mechanism of cholesterol lowering has not been established for either material. This study was conducted to determine the effect of glucomannan (G), chitosan (CH), or an equal mixture of the two (G + CH) on cholesterol absorption and fat and bile acid excretion. Rats were fed a modified AIN-93G diet for 18 d containing 0.125 g/100 g cholesterol and initially 10 g/100 g of the test materials or cellulose (C) as the control. However, the concentration of test materials and cellulose was reduced to 7.5 g/100 g after 1 wk due to lower weight gain compared with controls. Total liver cholesterol was significantly reduced in G, CH and G + CH groups compared with the C group. The intestinal contents supernatant viscosity of the C and the CH groups was negligible, whereas both G and G + CH produced high viscosities. Cholesterol absorption, measured by the fecal isotope ratio method, was significantly reduced from 37.5% in the C group to 20.2% in G, 18.2% in G + CH and 9.4% in CH. Daily fecal fat excretion did not differ between the C and G groups, but was significantly greater in G + CH and CH compared with the C and G groups. Daily fecal bile acid excretion was significantly greater in the CH and G + CH groups compared with the C and G groups. These results suggest that G lowered liver cholesterol by a viscosity-mediated interference of cholesterol absorption. In contrast, CH appears to lower cholesterol through a different mechanism. (+info)
A comparison of the effects of DNA-damaging agents and biotic elicitors on the induction of plant defense genes, nuclear distortion, and cell death.
(19/1030)
Pea (Pisum sativum L. cv Alcan) endocarp tissue challenged with an incompatible fungal pathogen, Fusarium solani f. sp. phaseoli or fungal elicitors results in the induction of pathogenesis-related (PR) genes and the accumulation of pisatin, a phytoalexin. Essentially the same response occurs in pea tissue exposed to DNA-specific agents that crosslink or intercalate DNA. In this study, the effects of DNA-damaging agents were assessed relative to the inducible expression of several pea PR genes: phenylalanine ammonia lyase, chalcone synthase, and DRR206. Mitomycin C and actinomycin D mimicked the biotic elicitors in enhancing the expression of all three PR genes. The activities of these PR gene promoters, isolated from different plants, were evaluated heterologously in transgenic tobacco. It is remarkable that beta-glucuronidase expression was induced when plants containing the heterologous phenylalanine ammonia lyase, chalcone synthase, and DRR206 promoter-beta-glucuronidase chimeric reporter genes were treated by DNA-damaging agents. Finally, cytological analyses indicated that many of these agents caused nuclear distortion and collapse of the treated pea cells. Yet we observed that cell death is not necessary for the induction of the PR gene promoters assessed in this study. Based on these observations and previously published results, we propose that DNA damage or the associated alteration of chromatin can signal the transcriptional activation of plant defense genes. (+info)
Mechanical, bioadhesive strength and biological evaluations of chitosan films for wound dressing.
(20/1030)
PURPOSE: To investigate the suitability of chitosan films prepared using two different solvents, acetic acid (Chitosan-AA) and lactic acid (Chitosan-LA), for wound dressing, in comparison with a commercial preparation, Omiderm. METHODS: The mechanical and in-vitro bioadhesive strength properties of Chitosan-AA, Chitosan-LA, and Omiderm were investigated using texture analyzer equipment. The vapour permeability of chitosan films was determined using a method for evaluation of moisture permeability of containers and packaging material described in USP XXII. In addition, the biological evaluations were performed via primary skin irritation, intracutaneous, and systemic injection tests. RESULTS: The three preparations differed significantly in terms of the mechanical and bioadhesive strength properties. Chitosan-LA exhibited a lower tensile strength, but more flexible and bioadhesive than Chitosan-AA. Chitosan film was found to be permeable to water vapour. Chitosan-LA and Omiderm were non-irritant and did not cause any skin allergic reaction. In contrast, Chitosan-AA films inflicted adverse skin reactions. Nevertheless, no gross sign of toxicity was encountered from the systemic injection of the extracts of the three preparations. CONCLUSION: Chitosan films demonstrated significantly different mechanical and bioadhesive strength properties from Omiderm. Chitosan-LA was more soft, flexible, pliable and bioadhesive when compared to Chitosan-AA films. Furthermore, Chitosan-LA did not cause erythema, edema and systemic toxicity. Hence, Chitosan-LA film is suitable to be used in the management of wound healing and skin burn. (+info)
Biodegradation and drug release of chitosan gel beads in subcutaneous air pouches of mice.
(21/1030)
Chitosan (CS) gel beads were prepared in 10% amino acid solution (pH 9) and implanted into air pouches (AP) prepared subcutaneously on the dorsal surface of mice. No inflammatory response was observed, and degradation of the beads in the AP increased as their degree of deacetylation decreased. Degradation could be altered by changing the nature of the CS or by increasing the CS concentration. The release of prednisolone (PS) in vivo from CS gel beads was similar to the release in vitro. When a suspension of PS was injected into the AP, the PS had almost completely disappeared 24 h after injection. Retention of PS in the AP was not increased by using a viscous CS solution. Alginate (Alg) gel beads, which were not degraded, released PS slowly into the AP over 3d. The in vitro release profile of PS using 1% CS (deacetylation: 70% (7B) and 80% (8B)) and 1.5% CS (deacetylation: 90% (9B)) gel beads was similar to that with Alg gel beads. However, the in vivo release of PS was affected by the degradability of the gel beads. CS7B and 8B (1%) gel beads had released PS into the AP earlier than 3 d according to their rate of degradation. CS9B (1.5%) gel beads were not degraded after 3 d and went on to release PS into the AP for 3 d similar to the release profile of Alg gel beads. CS9B (2%) gel beads were also not degraded after 3 d and the release of PS from these beads into the AP was sustained; 76% and 27% of administered PS remained in the gel beads after 1 and 3 d, respectively. Therefore, degradation and drug release of CS gel beads can be controlled by changing the structure of the gel matrix, which appears to make these beads a promising biodegradable vehicle for sustained drug delivery. (+info)
Characteristics of a Streptomyces coelicolor A3(2) extracellular protein targeting chitin and chitosan.
(22/1030)
Upstream of the Streptomyces coelicolor A3(2) chitinase G gene, a small gene (named chb3) is located whose deduced product shares 37% identical amino acids with the previously described CHB1 protein from Streptomyces olivaceoviridis. The chb3 gene and its upstream region were cloned in a multicopy vector and transformed into the plasmid-free Streptomyces lividans TK21 strain. The CHB3 protein (14.9 kDa) was secreted by the S. lividans TK21 transformant during growth in the presence of glucose, N-acetylglucosamine, yeast extract, and chitin. The protein was purified to homogeneity using anionic exchange, hydrophobic interaction chromatographies, and gel filtration. In contrast to CHB1, CHB3 targets alpha-chitin, beta-chitin, and chitosan at pH 6.0 but does so relatively loosely. The ecological implications of the divergence of substrate specificity of various types of chitin-binding proteins are described. (+info)
Evaluation of properties microcrystalline chitosan as a drug carrier. Part 1. In vitro release of diclofenac from mictocrystalline chitosan hydrogel.
(23/1030)
The influence of microcrystalline chitosan hydrogel, alone (MCCh) as well as in combination with methylcellulose (MC) or Carbopol (CP), on the release of diclofenac free acid (DA) and its salt (DS) was studied in vitro. Commercial Olfen gel (Mepha Ltd., Switzerland) was applied as a reference preparation. The influence of hydrophilizing agents (1,2-propylene glycol and glycerol) and methycellulose hydrogel on the rheological properties of the vehicle and on the release of drug from modified MCCh hydrogel was studied. The quantity of the released substance was determined by UV-spectroscopy. The results confirmed that release was dependent on the chemical character of the drug and on the type of vehicle. The process of diclofenac release from MCCh hydrogels as well as from Carbopol hydrogels runs in two phases. The first phase is characterised by rapid release whereas in the second phase the release is much slower. The most suitable basis for diclofenac is microcrystalline chitosan hydrogel with addition glycerol, 1,2-propylene glycol, and methylcellulose hydrogel. (+info)
In vitro and in vivo evaluation of sustained release chitosan-coated ketoprofen microparticles.
(24/1030)
Simple ketoprofen microspheres (MS) were prepared by the dry-in-oil method using ethylcellulose (EC) as a matrix polymer. Further, the microspheres modified by addition of polyethylene glycol (PEG) and hydroxypropyl cellulose (HPC), called MS-P and MS-H, respectively, were prepared. The in vitro release from MS, MS-P and MS-H was examined in the JP XIII second fluid, pH 6.8, at 37 degrees C and 60 rpm. Chitosan-coated ketoprofen microparticles (Chi-MP) were prepared by the precipitation of droplets of chitosan solution containing MS, and their adhesion to the rat small intestinal mucosa was tested. The plasma concentrations after duodenal administration were investigated for ketoprofen powder suspension, MS and Chi-MP. The particle size was raised with the increase in amount of ketoprofen added. The drug content and addition of PEG or HPC affected the drug release rate. The microspheres with moderate drug content, prepared by addition of modest amount of PEG, exhibited better gradual drug release. Chi-MP showed a good mucoadhesion. The maximum plasma concentration of ketoprofen for Chi-MP was less than one-third of that for ketoprofen powder suspension. Chi-MP tended to show the higher and steadier plasma level than MS. (+info)