Membrane-bound DNA from Escherichia coli: extraction by freeze-thaw-lysozyme.
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Seminal plasmin (SPLN) a 47-residue peptide, isolated from bovine seminal plasma, exhibits antibacterial activity against Gram-positive and Gram-negative bacteria. Although SPLN strongly inhibits the transcription of various natural and synthetic templates by E. coli RNA polymerase in vitro, it also associates with model membranes of phosphatidylcholine and phosphatidic acid. We have undertaken experiments to ascertain whether SPLN permeabilizes the bacterial inner membrane and thereby exerts its antibacterial activity, as in the case of recently isolated antibacterial peptides from mammalian sources. Our results show that SPLN affects the permeability properties of the bacterial inner membrane which is reflected by increased uptake of ortho-nitrophenylgalactoside (ONPG), which can normally be translocated only by protein transporters. SPLN has also been shown to act on the outer membrane, since divalent cations inhibit antibacterial activity.
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Storage of intact macrophages would be a useful procedure for laboratories studying the biological functions of these cells. Oil-induced guinea pig peritoneal cells were harvested and frozen at an ultra-low temperature using a cryoprotective media. Studies of the biological properties of the frozen cells upon thawing indicated that the cells retained viability for a fairly long period of time, were metabolically active and comparable to normal cells in morphology. Indications of activation were obtained in enhanced phagocytic activity and significant decrease in migration of the stored cells.
Monocyte-macrophages are required for the development of cell mediated immunity to a variety of microorganisms and tumors. Quantitative assays of human monocyte-macrophage function would be most useful in the evaluation of cell mediated immune function in man. Five quantitative assays are described that provide a human monocyte-macrophage function profile. These assays parallel the physiologic steps necessary for monocyte-macrophages to function as phagocytes: 1) chemotaxis, 2) opsonization, 3) phagocytosis, 4) phagocytosis-induced metabolic stimulation and 5) destruction of foreign material. Application of these quantitative assays will allow detection and dissection of disorders of monocyte-macrophage function in man.
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