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Biomedical subjects

P M Bennett

Publications and source records attributed to P M Bennett.

At least 37 records · Page 2Linked to original sources

Carriage of antibiotic-resistant bacteria by healthy children.

The frequency of carriage of antibiotic-resistant bacteria in healthy 7- and 8-year-old children in Bristol was studied. Children born in Avon between 1 April 1991 and 31 December 1992, attending the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC) 7 year follow-up clinic, formed the study population. Carriage was estimated using mouth and stool samples. None of 105 children on whom information was available had received tetracycline, chloramphenicol, ciprofloxacin or an extended-spectrum cephalosporin in the previous year. Staphylococcus aureus was isolated from mouthwashes from 200 (37.1%) of 539 children sampled. Six (3%) of the isolates were resistant to chloramphenicol or tetracycline and four (2%) were methicillin resistant. Haemophilus spp. were isolated from 369 (72%) of 513 samples and 63 (17%) were ampicillin resistant, 49 (13.3%) were erythromycin resistant and seven (1.9%) were tetracycline resistant. Branhamella catarrhalis was isolated from 333 (74%) of 450 samples. Twenty-eight (8.4%) were erythromycin resistant and 14 (4.2%) strains were tetracycline resistant. Group A beta-haemolytic streptococci were isolated from 17 of 507 children sampled. One (5.9%) was tetracycline resistant. Stool samples were returned from 335 (62%) of 539 children from whom they were requested. Eleven per cent of samples yielded Gram-negative bacilli with high-level resistance to chloramphenicol, which was frequently linked to resistance to ampicillin, spectinomycin and streptomycin. Isolates demonstrating resistance to the third-generation cephalosporin ceftazidime were recovered from 17 subjects (3.2%). Six (35%) of 17 isolates possessed extended-spectrum beta-lactamases. Healthy children carry bacteria resistant to antibiotics to which children are not usually exposed. Resistance to ceftazidime, chloramphenicol and tetracycline may be co-selected by exposure to other antibiotics used in children or may be acquired from family members, pets, other children or food. These results suggest that antibiotic-resistant bacteria are widely disseminated and may be acquired by children before exposure to specific selection pressure.

Bacteria↗

Characterization, cloning and sequence analysis of the inducible Ochrobactrum anthropi AmpC beta-lactamase.

Ochrobactrum anthropi is resistant to most cephalosporins and penicillins due, at least in part, to the inducible expression of a single beta-lactamase. The beta-lactamase gene has been cloned and sequenced. It encodes an AmpC-type class 1 serine active-site enzyme that hydrolyses mainly cephalosporins and is resistant to inhibition by clavulanic acid. Expression of the ampC gene is inducible via a typical AmpR regulator, which is encoded upstream of ampC. Inducible expression is retained following cloning of O. anthropi ampR-ampC into Escherichia coli, confirming that the signal for AmpR activation in O. anthropi is the same as that used in the Enterobacteriaceae. This is the first reported example of an AmpC beta-lactamase outside of the gamma-subdivision of the bacterial kingdom. Genomic searches of other non-gamma-subdivision bacteria revealed a homologous ampR-ampC cluster in the plant symbiont, Sinorhizobium meliloti.

Amino Acid Sequence↗

The activity of vancomycin against heterogeneous vancomycin-intermediate methicillin-resistant Staphylococcus aureus explored using an in vitro pharmacokinetic model.

Heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) may account for treatment failure with vancomycin and act as a precursor of vancomycin-intermediate or -resistant S. aureus. The activity of vancomycin was assessed against vancomycinsusceptible, hVISA and VISA strains in a dilutional pharmacokinetic model. Over a 48 h period, total bacteria and cells with a vancomycin-intermediate phenotype were quantified. Total counts of hVISA were reduced by vancomycin in a similar way to a vancomycin-susceptible control. The vancomycin-intermediate sub-population was eradicated from the model within one dose interval. Exposure to low vancomycin concentrations did not result in an increase in the proportion of cells which were vancomycin intermediate. Short-term exposure of hVISA to vancomycin at gradient concentrations did not increase the proportion of cells with vancomycin-intermediate phenotype.

Anti-Bacterial Agents↗

Plasmid location and molecular heterogeneity of the L1 and L2 beta-lactamase genes of Stenotrophomonas maltophilia.

An approximately 200-kb plasmid has been purified from clinical isolates of Stenotrophomonas maltophilia. This plasmid was found in all of the 10 isolates examined and contains both the L1 and the L2 beta-lactamase genes. The location of L1 and L2 on a plasmid makes it more likely that they could spread to other gram-negative bacteria, potentially causing clinical problems. Sequence analysis of the 10 L1 genes revealed three novel genes, L1c, L1d, and L1e, with 8, 12, and 20% divergence from the published strain IID 1275 L1 (L1a), respectively. The most unusual L1 enzyme (L1e) displayed markedly different kinetic properties, with respect to hydrolysis of nitrocefin and imipenem, compared to those of L1a (250- and 100-fold lower k(cat)/K(m) ratios respectively). L1c and L1d, in contrast, displayed levels of hydrolysis very similar to that of L1a. Several nonconservative amino acid differences with respect to L1a, L1b, L1c, and L1d were observed in the substrate binding-catalytic regions of L1e, and this could explain the kinetic differences. Three novel L2 genes (L2b, L2c, and L2d) were sequenced from the same isolates, and their sequences diverge from the published sequence of strain IID 1275 L2 (L2a) by 4, 9, and 25%, respectively. Differences in L1 and L2 gene sequences were not accompanied by similar divergences in 16S rRNA gene sequences, for which differences of <1% were found. It is therefore apparent that the L1 and L2 genes have evolved relatively quickly, perhaps because of their presence on a plasmid.

Alleles↗

Ecological basis of extinction risk in birds: habitat loss versus human persecution and introduced predators.

Understanding the ecological mechanisms that underlie extinction is fundamental to conservation. It is well established that not all taxa are equally vulnerable to extinction, but the reasons for these differences are poorly understood. This may be, in part, because different taxa are threatened by different mechanisms. Theoretically, sources of extinction risk that perturb the balance between fecundity and longevity, such as human persecution and introduced predators, should be particularly hazardous for taxa that have slow rates of population growth. In contrast, sources of extinction risk that reduce niche availability, such as habitat loss, should represent a particular threat to taxa that are ecologically specialized. Here we test these predictions by using a phylogenetic comparative method and a database on 95 families of birds. As theory predicts, extinction risk incurred through persecution and introduced predators is associated with large body size and long generation time but is not associated with degree of specialization, whereas extinction risk incurred through habitat loss is associated with habitat specialization and small body size but not with generation time. These results demonstrate the importance of considering separately the multiple mechanisms that underlie contemporary patterns of extinction. They also reveal why it has previously proven so difficult to identify simple ecological correlates of overall extinction risk.

Animals↗

Pulmonary pathology of harbour porpoises (Phocoena phocoena) stranded in England and Wales between 1990 and 1996.

The pathological changes observed in the lungs of 197 freshly dead to moderately decomposed harbour porpoises (Phocoenaphocoena) stranded in England and Wales between October 1990 and December 1996 were reviewed. In 135 (69 per cent of the cases) macroscopic nematode infections of the bronchial tract with Pseudalius inflexus and Torynurus convolutus, either singly or in combination, were recorded, and 106 (54 per cent) also had P inflexus within the pulmonary blood vessels. All the macroscopically parasitised porpoises were adults or juveniles although two neonates had histological evidence of nematode infection. There were 62 cases of mild to severe, subacute to chronic bronchitis and bronchiolitis, 113 cases of mild to severe chronic granulomatous interstitial pneumonia, and 34 cases of mild to severe vasculitis or thrombovasculitis of pulmonary blood vessels attributable to these nematode infections. In 35 cases necropurulent or purulent (broncho)pneumonias were attributed either to secondary bacterial infections of the lungs or to septicaemias associated in seven cases with Streptococcus canis, in two cases with group B Salmonella species, in one case with Escherichia coli and in one case with Streptococcus lactis. The pulmonary lesions in 67 animals known or diagnosed to have been entrapped in fishing gear were non-specific and included persistent foam in the airways in 45 cases, diffuse congestion in 53, oedema in 50, and multifocal intra-alveolar haemorrhage in 33 cases. Seven cases of acute fibrinous or chronic fibrous pleuritis, seven cases of chronic necropurulent pneumonia associated with mycotic infections, four porpoises with traumatic lesions of the thorax and other parts of the body consistent with fatal attack from bottlenose dolphins (Tursiops truncatus), and one case of diffuse bronchointerstitial pneumonia associated with generalised morbillivirus infection were also recorded.

Animals↗

beta-lactamase expression in Plesiomonas shigelloides.

We have examined the production of beta-lactamases by 11 clinical and nine environmental isolates of Plesiomonas shigelloides from Czechoslovakia, the Czech Republic and Cuba. Ten isolates (50%) expressed detectable amounts of a chromosomally encoded, non-inducible beta-lactamase, though all isolates showed a broadly similar resistance profile: low-level resistance to ampicillin and higher-level resistance to carbenicillin. All strains were susceptible to cephalosporins and meropenem. Three clinical isolates expressed a beta-lactamase similar to a class 2c carbenicillinase, with a pI of 5.2 and three expressed an enzyme similar to a class 2d oxacillinase, with a pI of 5.3. The environmental isolates produced a variety of penicillinases, indicating that there is a reservoir of heterogeneous beta-lactamase genes in this species.

Anti-Bacterial Agents↗

Aeromonas hydrophila AmpH and CepH beta-lactamases: derepressed expression in mutants of Escherichia coli lacking creB.

The class 1 cephalosporinase (CepH) and class 2d oxacillinase (AmpH) from an Aeromonas hydrophila clinical isolate, strain T429125, have been cloned and sequenced. Both enzymes are typical of their equivalents in other species of Aeromonas Both cloned beta-lactamase genes were expressed at a low level in a standard laboratory Escherichia coli strain, but when cloned into a cre deletion E. coli mutant, they were expressed at significantly higher levels. Specific disruption of the creB gene resulted in similar increased levels of beta-lactamase expression, so it was concluded that CreB represses the transcription of ampH and cepH in a cre(+) E. coli strain. The expression of cepH was four times that of ampH in the deltacre mutant because of an additional factor encoded on the cloned T429125 chromosomal fragment containing cepH. This factor was able to trans-activate expression of co-resident ampH in the deltacre mutant such that expression of the two genes was approximately equal. The entire cepH-containing fragment was sequenced, but it contained no genes that were obviously related to any known class of DNA-binding protein.

Aeromonas hydrophila↗

A TEM-2beta-lactamase encoded on an active Tn1-like transposon in the genome of a clinical isolate of Stenotrophomonas maltophilia.

A constitutively expressed beta-lactamase gene from a clinical isolate of Stenotrophomonas maltophilia, J675Ia, has been cloned. Its DNA sequence is almost identical to that of bla(TEM2) (one nucleotide change) and the expressed enzyme is a Bush type 2a penicillinase with an amino acid sequence identical to that of TEM-2. The bla(TEM) gene was present within a novel Tn1/Tn3-type transposon in the genome of isolate J675Ia and the transposon was able to mobilize bla(TEM) on to the broad host-range conjugative plasmid, R388. When transferred to an Escherichia coli recipient, R388::Tn conferred high-level ampicillin resistance. This represents the first identification of a TEM beta-lactamase in S. maltophilia and the first evidence that this important clinical pathogen is able to act as a reservoir for mobile beta-lactamase genes in the hospital environment.

DNA Transposable Elements↗

Investigating potential associations between chronic exposure to polychlorinated biphenyls and infectious disease mortality in harbour porpoises from England and Wales.

Bioaccumulation of immunosuppressive organochlorines like polychlorinated biphenyls (PCBs) may pose a threat to the health and viability of cetacean populations. To investigate possible associations between chronic exposure to PCBs and infectious disease mortality in harbour porpoises Phocoena phocoena in UK waters, blubber concentrations of 25 individual chlorobiphenyl (CB) congeners in 34 healthy harbour porpoises that died due to physical trauma (mainly by-catch) were compared with CB concentrations in 33 animals that died due to infectious disease. The infectious disease group had significantly greater total 25 CBs (sigma 25CBs) concentrations than the physical trauma group (P < 0.001). The mean sigma 25CBs concentration in animals that died due to physical trauma was 13.6 mg kg-1 extractable lipid whereas the mean concentration in the infectious disease group was 31.1 mg kg-1 extractable lipid. The relationship between higher sigma 25CBs and the infectious disease group was not confounded by age, sex, nutritional status, season, location or year of stranding. In addition, adult females had significantly lower sigma 25CBs levels than adult males (P < 0.05) due to maternal transfer of CBs to offspring. These findings are consistent with the hypothesis that chronic PCB exposure predisposes harbour porpoises in UK waters to infectious disease mortality, although further research is required to test these associations more robustly.

Adipose Tissue↗

Retrograde renoscopic fragmentation of moderate-size (1.5-3.0-cm) renal cystine stones.

PURPOSE: We present a series of cystinuric patients with renal cystine calculi between 1.5 and 3.0 cm treated with retrograde renoscopy and intracorporeal lithotripsy and report our results, complications, and inpatient utilization with this approach. PATIENTS AND METHODS: The hospital and office charts of five consecutive patients with six treated renal units who underwent retrograde renoscopy and electrohydraulic lithotripsy for renal cystine stones between 1.5 and 3.0 cm were reviewed. Data on stone size and location, procedures performed, results, complications, and inpatient hospital days were compiled. RESULTS: Five of the six renal units were either rendered stone free or had fragments totalling 3 mm or less. Three renal units required only a single procedure, one required repeat ureteroscopy for Steinstrasse, and one required SWL and repeat ureteroscopy for Steinstrasse. One renal unit was left with a 6-mm fragment for which the patient refused further treatment. There were no major complications. The mean hospital stay was 1 day, and the mean number of procedures per patient was 1.3. CONCLUSION: Retrograde renoscopy and intracorporeal lithotripsy for renal cystine stones 1.5 to 3.0 cm is safe and effective and should be considered as an alternative to percutaneous nephrolithotomy in these patients.

Cystinuria↗

Expression and detection of hetero-vancomycin resistance in Staphylococcus aureus.

Isolates of Staphylococcus aureus resistant to vancomycin have been reported but appear to be extremely rare. However, isolates displaying hetero-resistance to vancomycin (hVRSA) are reportedly common in parts of Japan (9.3% of MRSA isolated from a group of university hospitals). We have investigated the reliability of the proposed method for detection of hetero-resistant isolates and the ability of clinical S. aureus isolates to express vancomycin resistance. The original method for identification of hVRSA was found to have poor reproducibility and may select for, rather than detect, vancomycin resistance. There appears to be a spectrum of heterogeneity in the expression of resistance to vancomycin among S. aureus. Until there is a clearer understanding of the mechanism and control of vancomycin resistance in S. aureus, and reliable tests are devised, the clinical relevance of different degrees of hetero-resistance cannot be assessed.

Anti-Bacterial Agents↗

Interactions between methicillin and vancomycin in methicillin-resistant Staphylococcus aureus strains displaying different phenotypes of vancomycin susceptibility.

Vancomycin-sensitive, -intermediate, and -heterointermediate methicillin-resistant Staphylococcus aureus isolates were tested by using E-tests to explore the interaction of methicillin and vancomycin. For the vancomycin-intermediate and -heterointermediate strains both drugs showed antagonism at concentrations below their MICs but synergy at methicillin concentrations near the MIC. This property could be used to screen for heterointermediate S. aureus strains.

Anti-Bacterial Agents↗