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S G Amyes

Publications and source records attributed to S G Amyes.

At least 91 records · Page 5Linked to original sources
PubMed1991

The role of the SOS response in bacteria exposed to zidovudine or trimethoprim.

Trimethoprim was more potent than zidovudine as an inducer of the SOS response in Escherichia coli. The level of induction by each compound initially increased with rising drug concentration and then fell; this effect was less marked with zidovudine than with trimethoprim. The SOS response did not appear to be involved in the inhibition of bacterial multiplication as the MICs of trimethoprim or zidovudine for recA430 and lexA3 mutants, which are unable to induce the SOS response, were identical to the MICs for the parent strains. However, the bactericidal activity of each compound against strains deficient in the SOS response was reduced. This suggest that induction of the DNA repair system contributes to the bactericidal activity of the drugs.

Cell Survival↗
PubMed1991

In vitro susceptibility of the fish pathogen Aeromonas salmonicida to flumequine.

The activity of the fluoroquinolone flumequine was investigated against the fish pathogen Aeromonas salmonicida and was compared with that of oxolinic acid. Flumequine was more active than oxolinic acid in terms of its MIC against oxolinic acid-resistant isolates of A. salmonicida and was as active as oxolinic acid against susceptible isolates. In contrast to oxolinic acid, flumequine was bactericidal, with only 1% of the bacteria surviving 6 h of exposure to the drug at concentrations slightly above the MIC. Mutation to resistance to flumequine was found to occur at a lower frequency than that to oxolinic acid. Hence, in vitro, flumequine appears to possess some advantages over oxolinic acid against this fish pathogen.

Aeromonas↗
PubMed1990

Separation of plasmid-mediated extended spectrum beta-lactamases by fast protein liquid chromatography (FPLC system).

We have devised a reliable procedure for the separation of three beta-lactamases of isoelectric focusing points (pI), 5.4, 6.5, and 7.9 by Fast Protein Liquid Chromatography (FPLC System). All of these enzymes were transferable and originated from a ceftazidime and cefotaxime resistant Klebsiella pneumoniae isolated in Bombay, India. The complete separation of the enzymes, achievable by this method, allowed each of the different individual beta-lactamases to be characterized biochemically. This analysis revealed that the enzymes of pI 6.5 and pI 7.9 hydrolysed ceftazidime and cefotaxime, and were responsible for the resistance of K. pneumoniae, and its Escherichia coli J53-2 transconjugant to third generation cephalosporins. The enzyme of pI 5.4 was the TEM-1 beta-lactamase. The beta-lactamase of pI 7.9 appears quite different from any previously reported third generation cephalosporin hydrolysing beta-lactamase, and consequently given the preliminary designation DJP-1. This is also the first example of extended spectrum hydrolysing beta-lactamases found in Asia.

Cefotaxime↗
PubMed1990

Cross resistance between oxytetracycline and oxolinic acid in Aeromonas salmonicida associated with alterations in outer membrane proteins.

Oxytetracycline resistant mutants of Aeromonas salmonicida isolated from mutation frequency experiments showed decreased susceptibility to oxolinic acid. Outer membrane preparations of these resistant mutant strains revealed a major protein, with a molecular mass of approximately 37 kDa, which was not present in significant quantities in the parent strain.

Aeromonas↗
PubMed1990

Novel dihydrofolate reductases isolated from epidemic strains of trimethoprim/sulfamethoxazole-resistant Shigella sonnei.

Two strains of trimethoprim-resistant Shigella sonnei bearing R plasmids pBH600 and pBH700 each elaborated a dihydrofolate reductase (DHFR) and were moderately resistant to trimethoprim (minimum inhibitory concentrations, 128 and 256 micrograms/ml, respectively). Neither plasmid hybridized to probes for DHFR types I, II, or III. The trimethoprim resistance genes from the R plasmids resided on a 1600-base pair (bp) PstI fragment of pBH600 and an 1800-bp PstI fragment of pBH700. Isoelectric focusing showed distinct isoelectric points for the enzymes coded for on pBH600 (5.3) and pBH700 (5.6-5.7). Trimethoprim-resistant S. sonnei from 10 locations in nine states were examined. Isolates from 8 locations hybridized only to a pBH700-derived probe and one isolate hybridized to a pBH600-derived probe. These two trimethoprim resistance genes appear novel. The gene on plasmid pBH700 codes for an enzyme that seems widespread among S. sonnei isolates in the USA.

Ampicillin Resistance↗
PubMed1990

Plasmid-mediated beta-lactam resistance in pathogenic gram-negative bacteria isolated in south India.

A field study was undertaken at the Christian Medical College Hospital in Vellore, South India in 1984. Two hundred and eighty-four clinical isolates of enterobacteria were collected from patients with significant bacteriuria and their sensitivities to a series of beta-lactam antibiotics were determined. There was a very high incidence of beta-lactam resistance (ampicillin resistance greater than 80% and cephaloridine resistance greater than 65%) amongst these isolates. There was significant resistance to the combination of ampicillin and clavulanic acid but few of the isolates were resistant to cefuroxime or ceftazidime. Seventy seven per cent of the Escherichia coli isolates were resistant to ampicillin and 57% were resistant to cephaloridine. Most of the resistant Esch. coli isolates carried resistance genes for both ampicillin and cephaloridine which were transferable, either by direct conjugation or by mobilization with the X+ plasmid. Characterization of the R-plasmids revealed considerable diversity. Although the distribution of plasmid-mediated beta-lactamases was broadly similar to those found in other surveys, three new beta-lactamases were identified and atypically some of the Esch. coli isolates produced PSE beta-lactamases.

Ampicillin↗
PubMed1990

Assessment of the interaction between ciprofloxacin and teicoplanin in vitro and in neutropenic patients.

The interactions of ciprofloxacin and teicoplanin were investigated against methicillin-sensitive and methicillin-resistant Staphylococcus aureus and Staph. epidermidis isolated during a clinical trial of the efficacy of this combination. In-vitro studies of the combination of ciprofloxacin and teicoplanin found no evidence of any antagonism between these two drugs in terms of inhibition of bacterial multiplication, as determined by Fractional Inhibitory Concentration Indices, and lethality. Clinical use of teicoplanin plus ciprofloxacin as empirical therapy in 29 febrile neutropenic patients revealed an overall response rate of 75%. Response rate for staphylococcal infections, which accounted for 53% of isolated pathogens, was 80%. No serious adverse drug reactions were seen. Our results show that both in vitro, and in the treatment of febrile neutropenic patients, teicoplanin plus ciprofloxacin is an effective anti-staphylococcal combination.

Adult↗
PubMed1990

N-terminal amino acid sequence of the novel type IIIb trimethoprim-resistant plasmid-encoded dihydrofolate reductase from Shigella sonnei.

The type IIIb dihydrofolate reductase, a novel plasmid-encoded enzyme recently identified in Shigella sonnei, has been shown to have some similar biochemical properties to the type IIIa dihydrofolate reductase which was first identified in New Zealand in 1979. However, the type IIIb enzyme has a Ki for trimethoprim of 0.4 microM, and a pI of 5.35 (as compared to 19 nM and 6.1 for the type IIIa); both these results suggest that it is a different enzyme from the prototype type IIIa. The type IIIb dihydrofolate reductase was purified by methotrexate agarose affinity chromatography, yielding a pure protein as determined by HPLC. Automatic amino acid analysis of the purified enzyme showed it to be distinct from all other known plasmid-encoded dihydrofolate reductases and quite different from the type IIIa enzyme. The purified enzyme was examined by SDS-PAGE, which revealed that the type IIIb dihydrofolate reductase was a monomeric protein of Mr 17,200.

Amino Acid Sequence↗
PubMed1990

Identification and cloning of the type IIIa plasmid-encoded dihydrofolate reductase gene from trimethoprim-resistant gram-negative bacteria isolated in Britain.

A clinical strain of Escherichia coli isolated in Nottinghamshire in 1980 was shown to harbour the type IIIa trimethoprim-resistant dihydrofolate reductase gene, previously identified on only one occasion, in New Zealand in 1979. The gene was identified by hybridisation with an 855-bp type III gene probe and its classification as a type IIIa dihydrofolate reductase was confirmed by detailed biochemical analysis of the enzyme product. The dihydrofolate reductase was identical in size and isoelectric point with the original type IIIa enzyme and shared similar inhibitory and kinetic profiles. The trimethoprim resistance gene was subsequently cloned and the type IIIa dihydrofolate reductase gene was localised to a 700-bp EcoRI-PstI fragment. This smaller fragment may prove to be a more specific DNA probe for the future identification of type IIIa dihydrofolate reductase genes.

Cloning, Molecular↗
PubMed1990

Characterisation of a unique ceftazidime-hydrolysing beta-lactamase, TEM-E2.

A strain of Klebsiella oxytoca, originally isolated in Liverpool in 1982, has been found to produce a novel transferable beta-lactamase, TEM-E2. This enzyme confers resistance to ceftazidime and focused as a doublet band with an iso-electric point (pI) of 5.3. The strain also produced the TEM-1 beta-lactamase. Both TEM-1 and TEM-E2 beta-lactamases were encoded by a transferable 103 kb plasmid; these two enzymes also had similar molecular weights, were inhibited by clavulanic acid, and hydrolysed ampicillin, carbenicillin and cephaloridine at similar rates. However, unlike the TEM-1 enzyme, the TEM-E2 beta-lactamase hydrolysed ceftazidime and cefotaxime with similar efficiency, although it conferred much greater resistance to ceftazidime in the host strain. This is the earliest documented example of a TEM-like enzyme which confers transferable resistance to ceftazidime and related cephalosporins.

Cefotaxime↗
PubMed1990

Conditions required for the bactericidal activity of fleroxacin and pefloxacin against Escherichia coli KL 16.

Pefloxacin and fleroxacin showed biphasic bactericidal activity against Escherichia coli KL16 in nutrient broth. Bacteriostatic concentrations of chloramphenicol, an inhibitor of protein synthesis, and rifampicin, an inhibitor of RNA synthesis, could not completely abolish the bactericidal activity of either drug. Pefloxacin and fleroxacin were also active against non-dividing E. coli KL16. Therefore, pefloxacin and fleroxacin are able to kill bacteria which are not dividing nor actively synthesising protein or RNA.

Anti-Infective Agents↗
PubMed1990

N-terminal amino-acid sequence and subunit structure of the type IV trimethoprim-resistant plasmid-encoded dihydrofolate reductase.

The type IV plasmid-mediated dihydrofolate reductase (DHFR), from a clinical strain of Escherichia coli isolated in South India, was prepared from a transconjugant containing the original clinical plasmid, E. coli J62-2 (pUK1123), and from E. coli C600 (pUK1150) containing a 2.6-kb HindIII fragment of pUK1123 cloned into plasmid pBR322. Both preparations were purified by methotrexate affinity chromatography. Automatic amino-acid sequencing of the N-terminal of the purified type IV enzyme from both sources gave an identical sequence which was clearly distinct from other plasmid-mediated trimethoprim-resistant DHFRs. The type IV DHFR showed most homology with the endogenous, chromosomally-encoded E. coli enzyme. Amino-acid sequence analysis also showed that the type IV enzyme preparation from E. coli J62-2 harbouring the original clinical plasmid, pUK1123, also contained the E. coli DNA-binding protein NS1. Analysis by polyacrylamide gel electrophoresis suggested that the type IV enzyme, in its native form, consists of a DHFR of Mr 33,000 coupled to a DNA-binding protein.

Amino Acid Sequence↗
PubMed1990

Bactericidal action of PD127,391, an enhanced spectrum quinolone.

The 4-quinolone PD127,391 displays a biphasic effect on Escherichia coli, Staphylococcus aureus and S. epidermidis in nutrient broth. It is as active as ciprofloxacin in terms of its optimum bactericidal concentration against E. coli. However, against staphylococci it is six times as active as ciprofloxacin or any other 4-quinolone previously investigated. Although protein and RNA synthesis are not required for bactericidal activity, cell division is essential.

4-Quinolones↗