muramidase and sodium-carbonate

Washing soda, chemically identified as anhydrous sodium carbonate, is a popular cleaning agent among the rural and urban populations of India which often contaminates the freshwater ponds and lakes, the natural habitat of sponge Eunapius carteri. Present investigation deals with estimation of cellular aggregation, generation of ROS and activities of antioxidant enzymes, lysozyme and acetylcholinesterase in the cells of E. carteri under the environmentally realistic concentrations of washing soda. Prolonged treatment of washing soda inhibited the degree of cellular aggregation. Experimental exposure of 8 and 16mg/l of sodium carbonate for 48h elevated the physiological level of reactive oxygen species (ROS) generation in the agranulocytes, semigranulocytes and granulocytes of E. carteri, whereas, treatment of 192h inhibited the ROS generation in three cellular morphotypes. Activities of superoxide dismutase, catalase and glutathione-S-transferase were recorded to be inhibited under prolonged exposure of washing soda. Washing soda mediated inhibition of ROS generation and depletion in the activities of antioxidant enzymes were indicative to an undesirable shift in cytotoxic status and antioxidative defense in E. carteri. Inhibition in the activity of lysozyme under the treatment of sodium carbonate was suggestive to a severe impairment of the innate immunological efficiency of E. carteri distributed in the washing soda contaminated habitat. Washing soda mediated inhibition in the activity of acetylcholinesterase indicated its neurotoxicity in E. carteri. Washing soda, a reported environmental contaminant, affected adversely the immunophysiological status of E. carteri with reference to cellular aggregation, oxidative stress, antioxidative defense, lysozyme and acetylcholinesterase activity.

Topics: Acetylcholinesterase; Animals; Antioxidants; Biomarkers; Carbonates; Catalase; Cell Aggregation; Dose-Response Relationship, Drug; Environmental Monitoring; Fresh Water; Glutathione Transferase; India; Muramidase; Oxidative Stress; Porifera; Reactive Oxygen Species; Risk Assessment; Superoxide Dismutase; Time Factors; Water Pollutants, Chemical

beta 2-Adrenoceptors (beta 2-AR) have been purified from many mammalian tissues. Unfortunately, other beta-AR subtypes expressed in the same cells are usually copurified, contaminating the preparation and interfering with subsequent investigations such as receptor characterization, ligand binding studies, immunoprecipitation, or development of anti-receptor antibodies. The advent of molecular biology techniques has facilitated the expression of beta 2-AR in cells in which no other similar molecules are present; thus, receptor purification has been simplified. beta 2-AR expressed in Escherichia coli provides a convenient source of receptor without the need for specialized culture facilities required for eukaryotic cells. The greater complexity of the gram-negative cell wall structure, however, complicates the purification of membrane-bound receptor from this source. In this report, we describe a reliable method for the partial purification of membrane-bound beta 2-AR from transgenic E. coli. Spheroplast formation followed by cell disruption and a carbonate wash procedure provided beta 2-AR bound to bacterial inner membrane in high yield.

Topics: Carbonates; Cell Fractionation; Cell Membrane; Cell Wall; Centrifugation, Density Gradient; Escherichia coli; Muramidase; Peptidoglycan; Pressure; Receptors, Adrenergic, beta-2; Recombinant Fusion Proteins; Sonication; Species Specificity; Spheroplasts