ABSTRACT
A new bioassay method utilizing laser light scattering from suspensions of drug-sensitive bacteria has been developed for the estimation of antitumor drugs in biologic samples. Changes in the light-scattering patterns of antibiotic-treated bacteria have recently been shown to provide a rapid and accurate indication of antibiotic sensitivity. Similar considerations for several antitumor drugs have shown the method capable of assaying 0.1 ml with drug concentrations as low as a few nanograms of drug per milliliter of sample. The first successful application of the methodology is described for the antitumor agent methotrexate. Studies of both drug-treated human serum specimens and dog serum levels and urinary excretion as a function of time indicate that assay results are available within 3 hours of preparing the serum. Time variations of drug serum levels and urinary excretion rates are compared via laser differential light-scattering assay, standard disc-diffusion assay, and previously published radioisotopic assays.
Subject(s)
Biological Assay/methods , Lasers , Methotrexate/analysis , Animals , Dogs , Dose-Response Relationship, Drug , Enterococcus faecalis/drug effects , Female , Humans , Methotrexate/pharmacology , Scattering, RadiationSubject(s)
Plicamycin/metabolism , Animals , Bacteriological Techniques , Biological Assay , Brain/metabolism , Female , Kidney/metabolism , Liver/metabolism , Lung/metabolism , Male , Methods , Mice , Mice, Inbred Strains , Plicamycin/analysis , Plicamycin/blood , Plicamycin/pharmacology , Sarcina/drug effects , Spleen/metabolism , Time FactorsSubject(s)
Impetigo/pathology , Skin/pathology , Streptococcal Infections/pathology , Animals , Biopsy , Connective Tissue/pathology , Guinea Pigs , Impetigo/microbiology , Phagocytosis , Streptococcal Infections/microbiology , Streptococcus/growth & development , Streptococcus/isolation & purificationABSTRACT
Streptonigrin, a quinone antitumor antibiotic, has been reported to be effective in human trials. A sensitive and precise microbiological assay for the determination of distribution and concentrations of streptonigrin in the body fluids and tissues of treated mice has been developed in an attempt to supplement successful clinical application of this drug.
Subject(s)
Body Fluids/analysis , Streptonigrin/analysis , Animals , Bacillus subtilis/drug effects , Biological Assay , Methods , Mice , Mice, Inbred Strains , Streptonigrin/pharmacologySubject(s)
Cytarabine/metabolism , Leukemia L1210/metabolism , Thioguanine/metabolism , Animals , Biological Assay , Brain/metabolism , Cytarabine/pharmacology , Cytarabine/therapeutic use , Drug Therapy, Combination , Enterococcus faecalis/drug effects , Escherichia coli/drug effects , Leukemia L1210/drug therapy , Liver/metabolism , Lung/metabolism , Mice , Mice, Inbred Strains , Spleen/metabolism , Thioguanine/pharmacology , Thioguanine/therapeutic use , Time FactorsABSTRACT
Of 142 purines, purine nucleosides, and analogues tested for inhibition of growth of Escherichia coli B Hill, 45 were active. Of these, 27 were evaluated for inhibition of other E. coli lines, including those resistant to 6-thioguanine, 2-fluoroadenosine, 2,6-diaminopurine, or 6-mercaptopurine. Most toxic to the parent lines were 2-fluoroadenosine, 2-fluoroadenine, 2-fluoro-5'-deoxyadenosine, adenosine, 6-thioguanosine, 6-thioguanine, 6-mercaptopurine, 6-mercaptopurine ribonucleoside, 2-azaadenine, 2'-deoxyinosine, 6-N-aminoadenine, and inosine. Hypoxanthine was strongly inhibitory only to E. coli B Hill. Evidence regarding the substrate specificity of the three purine phosphoribosyltransferases was obtained by assaying for these enzymes in extracts of the various cell lines and by cross-resistance studies. The line selected for resistance to 6-thioguanine had low guanine phosphoribosyltransferase activity (guanosine monophosphate: pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) and was deficient in activity for xanthine and 6-thioguanine. The lines selected for resistance to 2-fluoroadenosine and 2,6-diaminopurine were deficient in adenine phosphoribosyltransferase activity (adenosine monophosphate: pyrophosphate phosphoribosyltransferase, EC 2.4.2.7), and that selected for resistance to 6-mercaptopurine had low hypoxanthine phosphoribosyltransferase activity and undetectable activity with 6-mercaptopurine as a substrate. Purine, 6-methylpurine, 2-fluoroadenine, 2,6-diaminopurine, and 2-azaadenine were classified as adenine analogues; 6-mercaptopurine and 8-aza-2,6-diaminopurine, as hypoxanthine analogues; and 6-thioguanine and 2-amino-6-chloropurine, as analogues of guanine. The inhibition of bacterial growth by hypoxanthine, inosine, 2'-deoxyinosine, or adenosine was prevented by small amounts of thiamine or by relatively high concentrations of either cytidine or uridine. Cytidine also reversed the inhibition by some purine and purine ribonucleoside analogues. Orotate phosphoribosyltransferase (OMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.10), a possible site of action for these compounds, was not inhibited directly by the toxic agents.
Subject(s)
Escherichia coli/drug effects , Purine Nucleosides/pharmacology , Purines/pharmacology , Carbon Radioisotopes , Escherichia coli/enzymology , MutationABSTRACT
A sensitive, precise microbiological assay was developed for the determination of tissue distribution of dl-alanosine, a new antitumor agent.
Subject(s)
Antibiotics, Antineoplastic/analysis , Nitroso Compounds/analysis , Propionates/analysis , Animals , Biological Assay , Escherichia coli/drug effects , Hydroxylamines/analysis , Mice , Mice, Inbred Strains , Time FactorsSubject(s)
Ammonia/biosynthesis , Azaserine/metabolism , Glycolates/biosynthesis , Pyruvates/biosynthesis , Animals , Azo Compounds/metabolism , Chromatography, Ion Exchange , Dogs , Female , Glutamine/analysis , Kidney/analysis , Liver/analysis , Liver/metabolism , Male , Mice , Mice, Inbred Strains , Prostate/metabolism , Rats , Rats, Inbred Strains , Salivary Glands/analysis , Salivary Glands/metabolism , SpectrophotometryABSTRACT
A microbiological assay was developed for bleomycin, an antitumor antibiotic reported to be active in human trials. The assay bacterium was a strain of Escherichia coli which is resistant to ethionine. Studies revealed relatively high concentrations of bleomycin in the blood and urine of mice after a single dose, < 0.33 ld(10), injected intraperitoneally.
Subject(s)
Antibiotics, Antineoplastic/pharmacology , Drug Resistance, Microbial , Escherichia coli/drug effects , Animals , Biological Assay , Bleomycin/analysis , Bleomycin/blood , Bleomycin/pharmacology , Bleomycin/urine , Ethionine/pharmacology , Female , Injections, Intraperitoneal , Male , Methods , Mice , Mice, Inbred StrainsABSTRACT
A microbiological assay has been developed for chromomycin A(3), an antitumor antibiotic showing promise in human trials. The assay bacterium is a derived strain of Streptococcus faecalis resistant to methotrexate. Studies with mice revealed that relatively high concentrations of this antibiotic were maintained in the blood, kidneys, and liver of mice after a single-dose intraperitoneal injection of the drug.
Subject(s)
Antibiotics, Antineoplastic/analysis , Biological Assay , Kidney/analysis , Liver/analysis , Animals , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/blood , Antibiotics, Antineoplastic/pharmacology , Antibiotics, Antineoplastic/urine , Chemical Phenomena , Chemistry , Colorimetry , Drug Resistance, Microbial , Enterococcus faecalis/drug effects , Female , Injections, Intraperitoneal , Male , Methods , Methotrexate/pharmacology , Mice , Mice, Inbred StrainsSubject(s)
Antibiotics, Antineoplastic/pharmacology , Bacteria/drug effects , Fungi/drug effects , Naphthacenes/pharmacology , Antibiotics, Antineoplastic/analysis , Bacillus subtilis/metabolism , Biological Assay , Culture Media , Daunorubicin/pharmacology , Drug Resistance, Microbial , Enterococcus faecalis/drug effects , Enterococcus faecalis/metabolism , Enterococcus faecalis/radiation effects , Escherichia coli/drug effects , Glycosides/analysis , Glycosides/pharmacology , Lactobacillus/drug effects , Mercaptopurine/pharmacology , Microbial Sensitivity Tests , Naphthacenes/analysis , Radiation Effects , Radiation-Sensitizing Agents/pharmacology , Streptomyces/metabolismABSTRACT
A sensitive, precise microbiological assay has been developed for the determination of tissue distribution of 5-diazouracil, a potential antitumor and antimicrobial agent.
Subject(s)
Biological Assay , Uracil/analysis , Uracil/metabolism , Animals , Azo Compounds/analysis , Azo Compounds/blood , Azo Compounds/metabolism , Escherichia coli , Methods , Mice , Spleen/analysis , Uracil/bloodSubject(s)
Bacteria/isolation & purification , Diaper Rash/microbiology , Fungi/isolation & purification , Skin/microbiology , Bacillus/isolation & purification , Candida/isolation & purification , Corynebacterium/isolation & purification , Diaper Rash/etiology , Enterobacter/isolation & purification , Escherichia coli/isolation & purification , Female , Humans , Infant , Infant, Newborn , Klebsiella/isolation & purification , Male , Micrococcus/isolation & purification , Proteus/isolation & purification , Pseudomonas/isolation & purification , Shigella dysenteriae/isolation & purification , Staphylococcus/isolation & purification , Statistics as Topic , Streptococcaceae/isolation & purification , Streptococcus/isolation & purificationSubject(s)
Rifampin/blood , Animals , Chemical Phenomena , Chemistry , Dogs , Female , Half-Life , Hydrogen-Ion Concentration , Kinetics , Microbial Sensitivity Tests , Oxidation-Reduction , Peroxides , Rifampin/administration & dosage , Rifampin/pharmacology , Spectrometry, Fluorescence , Staphylococcus/drug effects , Time FactorsSubject(s)
Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/pharmacology , Enterococcus faecalis/drug effects , Escherichia coli/drug effects , Staphylococcus/drug effects , Antibiotics, Antineoplastic/pharmacology , Bacillus cereus/drug effects , Bacteriolysis , Carbenicillin/pharmacology , Cell Survival/drug effects , Cephalothin/pharmacology , Chlortetracycline/pharmacology , Colistin/pharmacology , Daunorubicin/pharmacology , Fluorouracil/pharmacology , Mechlorethamine/pharmacology , Mercaptopurine/pharmacology , Methotrexate/pharmacology , Microbial Sensitivity Tests , Nucleosides/pharmacology , Penicillins/pharmacology , Pseudomonas/drug effects , Rifampin/pharmacology , Sarcina/drug effects , Sulfonamides/pharmacology , Triazenes/pharmacologySubject(s)
Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/pharmacology , Enterococcus faecalis/drug effects , Escherichia coli/drug effects , Staphylococcus/drug effects , Antibiotics, Antineoplastic/pharmacology , Bacillus cereus/drug effects , Bacteriolysis , Carbenicillin/pharmacology , Cell Survival/drug effects , Cephalothin/pharmacology , Chlortetracycline/pharmacology , Colistin/pharmacology , Daunorubicin/pharmacology , Fluorouracil/pharmacology , Imidazoles/pharmacology , Mechlorethamine/pharmacology , Mercaptopurine/pharmacology , Methotrexate/pharmacology , Microbial Sensitivity Tests , Nucleosides/pharmacology , Penicillins/pharmacology , Pseudomonas/drug effects , Rifampin/pharmacology , Sarcina/drug effects , Sulfonamides/pharmacology , Triazenes/pharmacologyABSTRACT
A sensitive, precise microbiological assay has been developed for the determination of tissue distribution of beta-thioguanine deoxyriboside, a new antitumor agent.
