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Biomedical subjects

G W Grigg

Publications and source records attributed to G W Grigg.

At least 37 records · Page 2Linked to original sources
PubMed1977

Sulphydryl-mediated DNA breakage by phlemomycin in Escherichia coli.

Sulphydryl-mediated DNA breakage, which is induced by the antibiotic phleomycin in vitro, has been found to contribute significantly to the DNA damage produced by phleomycin in Escherichia coli. The effect of pleomycin was inhibited in vivo, as in vitro, by chelating agents, sulphydryl blocking agents and antioxidants. An increase in the intracellular concentration of free sulphydryl resulted in an increased response to phleomycin, while mutants containing very low levels of free sulphydryl due to a defect in glutathione synthesis showed greatly reduced DNA breakage, particularly at low phleomycin concentrations. In spheroplasts of these gshA mutants, restoration of the response to phleomycin of dithiothreitol. Sulphydryl-mediated breakage appears to be the principal mechanism for DNA damage in E. coli at libly enzymic, operates at higher phleomycin concentrations.

Anti-Bacterial Agents↗
PubMed1977

Selective breakage of DNA alongside 5-bromodeoxyuridine nucleotide residues by high temperature hydrolysis.

The substitution of thymine mucleotides (pT) in oligodeoxynucleotides by bromouracil nucleotides (pBU) changes the properties of the oligonucleotides in two ways: (1) It alters their mobility during DEAE-Cellulose homochromatography1. (2) It substantially enhances their sensitivity to high temperature hydrolysis under mildly alkaline conditions (pH 8.9). The resultant breaks occur adjacent to pBU residues and leave terminal phosphates on the breakage products. With more extreme conditions some loss of terminal phosphates can occur. Heating at 100 degrees for 16 hr at pH 8.9 produces cleavage at about half of the pBU residues with minimal loss of terminal phosphates. The properties described here may explain the thermal sensitivity of bacteria grown in 5BU2 and may have a use in DNA sequencing technology.

Base Sequence↗
PubMed1977

Amplifications of phleomycin and bleomycin-induced antibiotic activity in Escherichia coli by aromatic cationic compounds.

A wide range of aromatic compounds has been shown to amplify phleomycin-induced cell killing in Escherichia coli. They include acridines, acridinium chlorides, dihydroanthracenes, anthracenes, dianthracenes, phenanthridinium salts, phenazinium chlorides, phenoxazones, triphenyl methane dyes, benzoquinolizinium chloride, diphenylmethane derivatives, stilbene and diphenyl derivatives. Low concentrations of these amplifiers also amplified the DNA breakage and degradation effects of phleomycin. The minimum structural specification for activity as an amplifying agent is suggested. A representative sample of compounds effective as amplifiers of phleomycin also amplified the antibiotic effects of bleomycins B4 and B6. The amplifiers described are known to vary in their ability to penetrate and accumulate in different organisms or tissues. This suggests the possibility of developing a series of antibiotic regimes using these amplifiers (or the large number of derivative compounds also likely to be active) where the therapeutic index is determined by the properties of the amplifier chosen rather than of the phleomycin or the bleomycin.

Anti-Bacterial Agents↗
PubMed1976

The mechanism of sensitivity to phleomycin in growing Escherichia coli cells.

Stationary-phase Escherichia coli B cells transferred to new growth medium are initially resistant to net DNA breakage by low concentrations of phleomycin, and become sensitive as DNA replication commences. From studies with inhibitors of various stages of the DNA replication cycle it is evident that it is not DNA synthesis itself that is required for induction of DNA breakage by phleomycin, but events associated with the initiation of DNA replication. Termination of replication in the absence of further initiaiton results in resistance to phleomycin. The cellular change responsible for changes in sensitivity to phleomycin could be the attachment of the bacterial chromosome to the cell membrane at initiation and detachment on termination of replication, suggesting an alteration in the balance between cellular DNA breakage and repair processes for membrane-associated compared with non-membrane-associated DNA.

Anti-Bacterial Agents↗
PubMed1971

Effects of coumarin, thiopurines, and pyronin Y on amplification of phleomycin-induced death and deoxyribonucleic acid breakdown in Escherichia coli.

Phleomycin (</=2 mug/ml) induces neither deoxyribonucleic acid (DNA) breakdown nor cell death in stationary-phase Escherichia coli B cells, but the addition of 8 mm caffeine immediately initiates these changes in the same way as increasing the phleomycin concentration 10-fold. This phenomenon is termed "amplification" (6). Pyronin Y, a number of nontoxic thio- and mercaptopurines (of which the most active were 6,7- and 6,9-dimethyl-2-methylthiopurine), and coumarin have been found to be considerably more efficient amplifiers of phleomycin activity than caffeine. Thus 2 mm 6,7- and 6,9-dimethyl-2-methylthiopurine, 0.16 mm pyronin, and 4 mm coumarin killed 10 to 100 times more phleomycin-treated bacteria within 2 hr than 8 mm caffeine. As with caffeine, amplification of cell death by these compounds was accompanied by degradation of DNA to acid-soluble fragments. A number of compounds including 2,6-dichloropurine, 6-hydroxy-2-methylthiopurine, alpha-naphthol, beta-naphthol, naphthionic acid, and alpha-naphthol-4,8-disulphonic acid inhibited the action of phleomycin, if they were present in the cell suspension during phleomycin treatment, but some caused amplification if added subsequent to the phleomycin. Although no mutants resistant to >/=10 mug of phleomycin per ml were observed among 10(11)E. coli B cells screened, such mutants occurred with a frequency of 10(-6) to 10(-7) among cultures resistant to 1 to 2 mug of phleomycin per ml. These double mutants were cross-resistant to phleomycin plus caffeine. The amplifying compounds, though structurally dissimilar, shared the common characteristic of binding selectively to denatured DNA as measured by equilibrium dialysis methods. The implications of these observations in supporting a model of phleomycin amplification proposed previously (6) and their utility in providing a logic for developing a new class of antibiotics are discussed.

Antibiotics, Antineoplastic↗