CATALASE ACTIVITY OF A CRUDE ENZYME PREPARATION FROM IRON-CHLOROTIC BARLEY (HORDEUM VULGARIS) SEEDLINGS.
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Biomedical subjects
Publications and source records attributed to A P KRUEGER.
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Although gaseous ions possess no obvious physical properties which would predict a capability to influence living cells or tissues they can be shown to produce functional changes. The physiological effects so far demonstrated are limited in extent. In the mammalian trachea air ion action appears to depend upon the release of bound 5-hydroxytryptamine by CO(2) (+) and upon O(-)-induced acceleration of its metabolic destruction.
Exposure of Avena sativa seedlings to unipolar ionized atmospheres of either charge produced statistically significant stimulation of growth as measured by mean stem length, integral elongation, and dry weight. The extent of growth increase was related to the atmospheric ion density and this in turn determined the magnitude of current flow to ground. The minimal current measured in the ground circuit and capable of producing a measurable difference in growth was 4.3 to 4.6 x 10(-13) amp/plant.
Air pollutants seriously interfere with the maintenance of unipolar ionized atmospheres required in experimenting with the biological effects of gaseous ions. The construction and operation of an air purification unit designed to reduce air pollution to tolerable levels are described; it has functioned satisfactorily in conducting experiments with plants and animals.
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Intravenous administration of 5-hydroxytryptamine to rabbits and guinea pigs is shown to bring about changes very similar to those produced by (+) air ions, including (1) decreased ciliary rate, (2) contraction of the posterior tracheal wall, (3) exaggerated response of the tracheal mucosa to trauma, (4) marked vasoconstriction in the tracheal wall, and (5) increased respiratory rate. These effects are reversed by (-) air ions. Iproniazid, which raises 5-hydroxytryptamine levels in the animal by blocking monamine oxidase, produces similar but non-reversible effects. Reserpine, which depletes 5-hydroxytryptamine in the animal, causes changes that resemble those produced by (-) air ions, including (1) increased ciliary rate, (2) relaxed posterior sulcus, (3) hyperemia of the tracheal mucosa, (4) lowered respiratory rate, and (5) increased volume and rate of mucus flow. On the basis of these facts, the hypothesis is advanced that (+) air ion effects are mediated by the release of free 5-hydroxytryptamine, while (-) air ion effects depend on the ability of (-) ions to accelerate the enzymatic oxidation of 5-hydroxytryptamine.
Negative air ions are shown to decrease 5-hydroxytryptamine concentrations in extirpated strips of rabbit trachea and in the respiratory tracts of living mice. An initial exposure of guinea pigs to (-) air ions causes a transient rise in urinary 5-hydroxyindoleacetic acid excretion which is not observed upon subsequent exposures. These findings are compatible with the hypothesis advanced earlier that (-) air ion effects depend on the ability of (-) ions to accelerate enzymatic oxidation of 5-hydroxytryptamine.
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A. Duration of Effects Groups of mice exposed to high densities of unipolar light air ions for 72 hours exhibited persistent alterations in the functional efficiency of their tracheas. These effects lasted at least 4 weeks, and in the case of animals treated with (+) ions included diminished ciliary activity, pale and contracted tracheal mucosa, and enhanced vulnerability to trauma. Following treatment with (-) ions, animals displayed increased ciliary activity with no other detectable changes. It required at least 60 minutes of exposure to ions to induce such "permanent" functional changes. B. Minimal Effective Ion Densities The minimal ion densities producing changes in ciliary activity within an arbitrary period of 30 minutes were determined with extirpated tracheal strips from rabbits and guinea pigs. The threshold value for (-) ions was approximately 2.5 x 10(3) ions/cm.(2)/sec. and that for (+) ions was in the range between 1 x 10(4) and 2.5 x 10(5) ions/cm.(2)/sec.The minimal ion densities producing changes in ciliary activity within an arbitrary period of 30 minutes were determined with extirpated tracheal strips from rabbits and guinea pigs. The threshold value for (-) ions was approximately 2.5 x 10(3) ions/cm.(2)/sec. and that for (+) ions was in the range between 1 x 10(4) and 2.5 x 10(5) ions/cm.(2)/sec. The evidence indicates that ion-induced functional changes in the ciliated epithelium of the pulmonary tree are the results of direct contact of ions with surface cells and do not involve participation of the central nervous system or circulation. So far as ciliary activity is concerned, the number of ions required to produce a change in rate is very small.
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Studies on the effects of air ions on the functional efficiency of the extirpated tracheal strip have been extended to the trachea of the living rabbit, rat, and mouse. Animals exposed to high mobility (+) air ions administered via a tracheotomy aperture displayed: (a) Decreased ciliary activity. (b) Decline in mucus flow rate, sometimes reversed by prolonged exposure to (+) ions; a frequent drop in the volume of mucous secretion. (c) Contraction of the membranous posterior tracheal wall. (d) Increased vulnerability to trauma of cilia and mucosal blood vessels. Similar treatment with (-) air ions reversed (+) ion effects on ciliary activity, mucus flow, contraction of the tracheal smooth muscle. Continued (-) ion treatment raised the ciliary rate (invariably) and the mucus flow rate (often) above their initial levels. (+) Air ions administered to unoperated resting mice and rats increased the respiratory rate; (-) ions reversed this effect. Long exposure of unoperated ambulatory mice to (+) air ions produced: (a) Decreased ciliary activity. (b) No clear cut effect on mucus flow. (c) Contraction of the posterior tracheal wall. (d) Increased vulnerability of the mucosa to trauma. (-) Air ions increased ciliary activity but had no clear-cut effect on the mucus flow rate.
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LYSIS FROM WITHOUT (LFW) OCCURS IN TWO STEPS: (1) sensitization of cells by phage, which renders the cells susceptible to (2) destruction of an essential cell structure by an extracellular lytic enzyme. Virolysin, from phage-infected cells, was used in these studies. Normal cell autolysin is also effective. Evidence is presented that: 1. Neither phage nor lysin alone causes LFW. 2. Sensitization requires phage adsorption. 3. It can be caused by non-infectious particles. This establishes a new biological activity of the particle. 4. Heat, U.V., detergents, penicillin, and other damaging agents also sensitize cells. 5. Sensitization involves a non-lethal, reversible reaction. 6. Sensitization by phage prevents virus synthesis. Following adsorption, a cell can undergo sensitization or infection but not simultaneously. When only a few particles are adsorbed, infection can occur; when sufficient particles are adsorbed, sensitization takes place. 7. Quantitative aspects of LFW are described. Lysis proceeds logarithmically. The lysis end-point depends upon the phage concentration but is independent of the enzyme concentration.
Techniques have been devised for studying quantitatively the effects of air ions on microorganisms suspended in small drops. In smog-contaminated atmospheres moderate concentrations of positive and negative air ions exerted a protective effect on staphylococci by delaying the drop in pH customarily observed and by diminishing the rate of evaporation. In clean air higher concentrations of positive and negative air ions accelerated the rate of death of staphylococci apparently by direct action on the cells and by increasing the rate of evaporation. Air ion action in these experiments did not involve cell agglutination or direct radiation from the radioactive isotopes employed.
Virolysin is a lysin which appears in Staphylococcus aureus K(1) cells infected with phage P(14); together with phage, virolysin is released from phage-infected cells at the time of lysis. Autolysin is a lysin formed by uninfected cells of the K(1) strain; autolysin is released from uninfected cells by autolysis. They show the following similarities: Both agents act within the genus Micrococcus. They lyse cells only after the cell has been subjected to a damaging or "sensitizing" treatment, such as heat, bacteriophage, acetone, or ultraviolet irradiation. The course of lysis of heated cells by both lysins has been found to proceed in a similar manner. A constant percentage of cells is lysed, independent of the concentration of lysin; the residual cells remain resistant to either lysin. Lysis proceeds logarithmically with time, and the velocity constants K are proportional to the lysin concentration. K increases with increasing temperature. Both lysins are unaffected by antiserum to the phage. They are inhibited alike by a number of chemicals, including known enzyme inhibitors. Both agents are destroyed by proteolytic enzymes and are precipitated by 40 per cent saturation with (NH(4))(2)SO(4). Both lysins are very thermolabile. The two lysins differ with respect to their pH optimum, antigenic relationship and specificity for Micrococcus lysodeikticus. These results suggest that (1) both lysins have many properties associated with enzymes, (2) the lysis of heated cells, which they produce, has some of the characteristics of a chemical reaction, (3) the lysin from the phage-infected cell is clearly different from the lysin of the uninfected cell.