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Biomedical subjects

E M Fielden

Publications and source records attributed to E M Fielden.

At least 55 records · Page 3Linked to original sources
PubMed1981

Radiosensitization of hypoxic mammalian cells in vitro by some 5-substituted-4-nitroimidazoles.

The efficiencies of various 5-substituted-4-nitroimidazoles as radiation sensitizers have been determined in hypoxic Chinese hamster cells irradiated in vitro. Compared with published data on the sensitizing properties of substituted 2-nitro- and 5-nitroimidazoles, some of the 4-nitro derivatives show unusually high sensitizing efficiencies defined as the concentrations required to give an enhancement ratio of 1.6. The equilibrium one-electron reduction potentials of the compounds (E17) were measured by a pulse radiolysis technique and the results show that although sensitizing efficiencies are unexpectedly high, based on considerations of electron affinity, they still increase with increasing values of E17. Enhancement ratios were determined in two V79 cell lines for combinations of one of these compounds (a 4-nitroimidazole containing the group SO2.O.phenyl in the 5-position, NSC 38087) with various concentrations of misonidazole. The sensitization observed suggests that the two compounds may be operating by different mechanisms.

Animals↗
PubMed1980

The development of some nitroimidazoles as hypoxic cell sensitizers.

The most promising hypoxic cell sensitizer for use in radiotherapy is, at present, the 2-nitroimidazole, misonidazole, although neurotoxicity prevents its use at dosages necessary for optimum results. In the search for agents with better therapeutic ratios, we have investigated several classes of substituted 2-, 4-, and 5-nitroimidazoles. In general, 2-nitro derivatives are the most efficient in line with electron affinity considerations. However, notable exceptions include some 4-nitroimidazoles substituted in the 5-position by various sulphonamide or sulphonate groups. Some of these compounds are as efficient as oxygen on a concentration basis and display activity greater than predicted from their electron affinities. Cellular studies with these compounds will be described. Further, in the 2-nitroimidazole series the effect of the chain length of the N1 side chain terminated with a morpholine, piperidine, and pyrrolidine group has been studied. Notwithstanding the fairly constant electron affinities in these compounds, results show that systematic changes in sensitizing efficiency and cytotoxicity can occur with varying chain length, which indicates possible potential for increasing therapeutic ratio. These and other studies will be used as examples of the methods used in the search for better hypoxic cell sensitizers.

Cells, Cultured↗
PubMed1980

Respiratory effects and ascorbate reactions with misonidazole and other recently developed drugs.

A number of nitro compounds behave as electron acceptors with ascorbate in a non-enzyme-catalyzed reaction and with NADPH as electron donor in an enzyme-catalyzed reaction with liver homogenate. In both systems, the oxygen consumption is conveniently monitored by a Clark-type oxygen electrode. The oxygen consumption by S9 homogenate was found to be influenced by the electron affinity of the nitro compound. Detailed studies with misonidazole as the electron acceptor demonstrated that the oxygen consumption was influenced by protein and drug concentrations. Various inhibitors of electron transfer reactions, such as rotenone, antimycin A, and sodium azide, had no effect on misonidazole-stimulated oxygen utilization. However, cyanide was found to be stimulating in this system and this may be due to the inhibition of enzymes such as catalase and superoxide dismutase known to be present in S9. The oxygen consumption by S9 did not appear to be due to a hydroxylating reaction because metyrapone, a known inhibitor of this reactions, had no effect. Methional, a hydroxyl radical scavenger, was inhibitory as was glutathione. Clelands reagent had no effect. Additional studies with thiol-reactive agents suggest that a protein thiol may be important for the enzyme-catalyzed reaction between NADPH, misonidazole, and oxygen.

Animals↗
PubMed1978

The effect of the electron affinic sensitizers Ro 07-0582 and Ro 03-6156 on the survival of several E. coli mutants.

Concentration versus radiosensitizing effect curves have been determined for four E. coli strains with two nitroaromatic sensitizers. The data are consistent with a simple competition kinetic model of sensitization, and K values for Ro 07-0582 are reported. An empirical relationship, DMF=alpha o.e.r., relating the dose-modifying factor produced by a given concentration of sensitizer to the oxygen-enhancement ratio of a bacterium was found to hold for all the mutants tested for values of alpha greater than 1/o.e.r. The limiting value of alpha at high concentrations of sensitizer was 0.75. Some implications of this relationship and the limiting value of alpha are discussed.

Dose-Response Relationship, Drug↗
PubMed1978

Polyfunctional radiosensitizers IV. The effect of contact time and temperature on sensitization of hypoxic Chinese hamster cells in vitro by bifunctional nitroxyl compounds.

Three biradical nitroxyl compounds have been shown to modify the slope and shoulder region of the hypoxic cell survival curve of V79 cells to different degrees. The amount of sensitization produced by these compounds is dependent on the cell-drug contact time at both 20 degree and 37 degree C whereas sensitization by the monoradical nitroxyl, NPPN, is independent of these factors. The results suggest that biradicals may modify cellular biochemistry in such a way as to change the repair capacity of the cells. This could be responsible for changes in the shape of survival curves when cell-drug contact times are increased.

Alkaloids↗
PubMed1977

A pulse-radiolysis study of the manganese-containing superoxide dismutase from Bacillus stearothermophilus. A kinetic model for the enzyme action.

The enzymic reaction mechanism of a manganese-containing superoxide dismutase from Bacillus stearothermophilus was studied by using pulse radiolysis. During catalysis (pH 8.9; 25 degrees C), changes occurring in the kinetics of substrate disappearance and in the visible absorption of the enzyme at 480 nm established that the simple two-step mechanism found for copper- and iron-containing superoxide dismutases is not involved. At a low ratio (less than 15) of substrate concentration to enzyme concentration the decay of O2--is close to exponetial, whereas at much higher ratios (greater than 100) the observed decay is predominantly zero-order. The simplest interpretation of the results invokes a rapid one-electron oxidation-reduction cycle ('the fast cycle') and, concurrently, a slower reaction giving a form of the enzyme that is essentially unreactive towards O2-- but which undergoes a first-order decay to yield fully active native enzyme ('the slow cycle'). The fast cycle involves the native enzyme EA and a form of the enzyme EB which can be obtained also by treating the form EA with H2O2. Computer calculations made with such a simple model predict behaviour in excellent agreement with the observed results.

Catalysis↗
PubMed1977

A pulse-radiolysis study of the manganese-containing superoxide dismutase from Bacillus stearothermophilus.

In the preceding paper the mechanism of catalysis of the manganese-containing superoxide dismutase from Bacillus stearothermophilus was shown to involve a 'fast cycle' and a 'slow cycle' [McAdam, Fox, Lavelle & Fielden, 1977 (Biochem. J. 165, 71-79)]. Further properties of the enzyme was considered in the present paper. Pulse-radiolysis studies, under conditions of low substrate concentration to (i.e. when the fast cycle predominates), showed that enzyme activity decreases as pH increases (6.5-10.2). Activity was unaffected by the addition of H2O2 or NaN3 but slightly decreased by KCN. Both H2O2 and the reducing radical anion CO2-- caused a decrease in A480 of the native enzyme. The rate of the fast catalytic cycle was independent of temperature (5-55 degrees C), and as temperature increases the slow cycle becomes relatively more important. Arrhenius parameters of the rate contants were estimated. The possible identity of the various forms of the enzyme is considered.

Carbon Dioxide↗
PubMed1977

A pulse-radiolysis study of the catalytic mechanism of the iron-containing superoxide dismutase from Photobacterium leiognathi.

The mechanism of the enzymic reaction of an iron-containing superoxide dismutase purified from the marine bacterium Photobacterium leiognathi was studied by using pulse radiolysis. Measurements of activity were done with two different preparations of enzyme containing either 1.6 or 1.15 g-atom of iron/mol. In both cases, identical values of the second-order rate constant for reaction between superoxide dismutase and the superoxide ion in the pH range 6.2-9.0 (k=5.5 X 10(8) M-1-S-1 at pH 8.0) were found. As with the bovine erythrocuprein, there was no evidence for substrate saturation. The effects of reducing agents (H2O2, sodium ascorbate or CO2 radicals) on the visible and the electron-paramagnetic-resonance spectra of the superoxide dismutase containing 1.6 g-atom of ferric iron/mol indicate that this enzyme contains two different types of iron. Turnover experiments demonstrate that only that fraction of the ferric iron that is reduced by H2O2 is involved in the catalysis, being alternately oxidized and reduced by O2; both the oxidation and the reduction steps have a rate constant equal to that measured under turnover conditions. These results are interpreted by assuming that the superoxide dismutase isolated from the organism contains 1 g-atom of catalytic iron/mol and a variable amount of non-catalytic iron. This interpretation is discused in relation to the stoicheiometry reported for iron-containing superoxide dismutases prepared from several other organisms.

Ascorbic Acid↗