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The effects of lysozyme dimer (2 and 20 μg/kg) administered i.p. once and four times to mice on the phagocytic and killing ability of peritoneal macrophages, interleukin-1 (IL-1) production by murine macrophages stimulated in vitro with lipopolisaccharide of E. coli and expression of thymocyte, splenocyte and mesenteric lymphonode cell CD3+, CD4+ and CD8+ markers were studied. It was found that lysozyme dimer administered once or four times at doses of 2 (μg/kg and 20 μg/kg augments the phagocytic and killing activity of peritoneal macrophages. The strongest stimulating effect was noted after four injections of lysozyme dimer at a dose of 20 μg/kg. Moreover, lysozyme dimer is able to modulate the production of IL-1 by murine macrophages stimulated in vitro with LPS. Exposure to four doses of lysozyme dimer (20 μg/kg) enhances the synthesis and release of IL-1, but this drug administered once (2 μg/kg and 20 μg/kg) or four times (2 μg/kg) decreases IL-1 production by peritoneal macrophages. It was also found that administration of lysozyme dimer at a dose of 20 μg/kg, irrespective of the number of doses applied, increases the percentage of CD4+ thymocytes and splenocytes. Moreover, exposure to four doses of lysozyme dimer (2 and 20 μg/kg) increases the percentage of CD4+ and CD8+ mesenteric lymphonode cells.
The effect of lysozyme dimer (20 μg/kg) administered i.p. once or four times to mice immunized with sheep red blood cells (SRBC) on the secondary humoral response was studied with respect to the time of exposure to the drug in relation to priming and challenge. It has been found that lysozyme dimer potentiates secondary humoral response to SRBC resulting in an increased number of splenocytes producing hemolytic anti-SRBC antibodies (PFC) and anti-SRBC hemagglutinin titers (total and 2-mercaptoethanol resistant) when the drug is administered after the challenge. Both a single dose and four doses of lysozyme dimer administered after priming and challenge prolong its potentiating effect on anti-SRBC hemagglutinin titers as compared to the mice treated with lysozyme dimer only after challenge. A single dose of lysozyme dimer (20 μg/kg) administered only after priming does not affect secondary humoral response of SRBC-immunized mice. However, four doses of lysozyme dimer administered only after priming enhance the secondary humoral response to this antigen after challenge.
The effectiveness of antibacterial action of lysozyme modified by the membrane technique (ultrafiltration and reverse osmosis) against selected strains of bacteria was determined. Its bacteriostatic activity was dependent on modification conditions. Among lysozyme preparations modified by ultrafiltration the highest bacteriostatic activity against selected strains of Proteus mirabilis, Pseudomonas fluorescens and Staphylococcus epidermidis bacteria was noted in the preparation containing 53.3% polymeric forms. The modification procedure facilitates the extension of antibacterial spectrum of lysozyme, particularly against Pseudomonas fluorescens and Proteus mirabilis Gram (-) bacteria.
Lysozyme (EC 3.2.1.17, mucopeptyde N-acetylmuramic-hydrolase) exhibits the ability to destroy cell walls of Gram-positive bacteria. The range of activity for this enzyme may be extended through modifications, as a result of which polymerized forms of lysozyme are obtained. It was found that lysozyme dimer exhibits bacteriostatic properties towards both Gram-positive and Gram-negative bacteria. The aim of the study was to determine the conditions of thermal modification of lysozyme and to assess the antibacterial action of this enzyme after modification. The dimer content in the modified lysozyme preparations depended on the pH and concentration of a given solution. It was found that modified lysozyme was characterized by higher antibacterial activity towards Micrococcus luteus and Escherichia coli in comparison to the unmodified form of the enzyme.
The aim of the study was an attempt to apply selected analytical methods for the evaluation of preparations of modified lysozyme. Lysozyme isolated from hen egg white was modified using thermal, thermal-chemical, chemical and membrane methods and subsequently the obtained preparations were evaluated with the use of the spectrophotometric method, as well as electrophoresis and differential scanning calorimetry (DSC).
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