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Biomedical subjects

M Midgley

Publications and source records attributed to M Midgley.

At least 19 recordsLinked to original sources

Substrate specificity and energetics of antiseptic and disinfectant resistance in Staphylococcus aureus.

Clinical isolates of Staphylococcus aureus carry various antiseptic and disinfectant resistance determinants (qac genes) on a variety of plasmids. The biochemistry and specificity of these resistance genes in S. aureus is the subject of this report. The qac genes were separated into two families on the basis of resistance profiles and DNA homology. Isotopic and fluorimetric assays demonstrated that the qac genes encode efflux systems that rely on proton motive force.

Anti-Infective Agents, Local

An evaluation of two systems of in-patient care in a general hospital psychiatric unit I: staff and patient perceptions and attitudes.

This study reports a comparison of a single ward versus a two-ward system of acute in-patient care in a general hospital psychiatric unit. It focuses on staff and patient perceptions and attitudes towards the two types of ward. A continuous care ward system was preferred by the majority of staff who described an improvement in 'ward atmosphere'. Patients who had experienced both ward systems also preferred the continuous care system. Both patients and staff felt that the continuous care system still needed to place greater emphasis on 'ward atmosphere' dimensions of involvement, support and spontaneity. The implications of these findings for improving the functioning of the preferred ward are discussed.

Adult

Cloning of the ethidium efflux gene from Escherichia coli.

The gene specifying the ethidium efflux system of Escherichia coli has been cloned on a 3.2 kbp HindIII fragment and located on a 1.2 kbp fragment within this. Cross-resistance studies indicate that the system has a broad specificity for monovalent cations and the gene shows no hybridisation with similar genes found in Staphylococci.

Blotting, Southern

An evaluation of two systems of in-patient care in a general hospital psychiatric unit. II: Measures of staff and patient performance.

This study reports on a comparison between a single ward and a two-ward system of acute in-patient care in a general hospital psychiatric unit. It looked at job satisfaction, levels of stress, absenteeism/sickness and work proficiency amongst the staff and also at patients' level of disturbed behaviour, length of stay and re-admission rates. It was found that a continuous-care ward was associated with a lower level of behavioural disturbance and was preferred by the majority of staff, with some increase in job satisfaction and a decrease in stress. The continuous-care ward was also associated with shorter admissions. However, those who progressed through a two-ward system were less likely to be re-admitted within 3 months. The implications of these results are discussed.

Absenteeism

Physical and biochemical characterization of the qacA gene encoding antiseptic and disinfectant resistance in Staphylococcus aureus.

We have previously cloned a 3.5 kb fragment from the Staphylococcus aureus multiresistance plasmid pSK1 which carries the qacA determinant responsible for linked resistance to acriflavine (Acr), ethidium bromide (Ebr), quaternary ammonium compounds (Qar), propamidine isethionate (Pir), and diamidinodiphenylamine dihydrochloride (Ddr). This report presents a biochemical and physical analysis of qacA and shows the widespread carriage of this gene on S. aureus resistance plasmids. Tn5 insertion mutagenesis defined the extent of qacA to within 2.40 kb of pSK1 DNA. Examination of the expression of insertion and deletion mutants of the cloned qacA sequences in both maxicells and minicells led to the association of a 50 kDa protein, designated QacA, with the AcrEbrQarPirDdr phenotype. Based on fluorimetric and isotopic assays used to determine the extent of accumulation of ethidium bromide by S. aureus strains harbouring pSK1, we propose that the basis of AcrEbrQarPirDdr in S. aureus is a qacA-mediated efflux system.

Anti-Infective Agents, Local

Experiments with the Cornell intermittent lighting system for laying hens.

1. Two experiments were conducted to provide further evidence about rate of lay under the Cornell lighting system (2L:4D:8L:10D). Each used 1728 hens of each of 2 brown-egg stocks in 12 light-proof rooms. 2. In the first the Cornell system was compared at 2 light intensities (average values 2 and 10 lux) with a conventional step up lighting programme. In the second, Cornell lighting was introduced at 18, 21 or 24 weeks of age and compared with a step up programme. 3. Total egg output was essentially the same from the Cornell lighting system, using 10 h light/d, as from the step up programme using 16 h/d. When the Cornell system was applied abruptly at 18 weeks to pullets which had been reared on short days (8L:16D) sexual maturity was advanced, resulting in an increase in mean rate of lay to 72 weeks of age and a reduction in mean egg size. Application of the Cornell system from 21 or 24 weeks gave the same egg numbers and the same egg size as the step up programme. 4. Food intake was about 2% lower with the Cornell treatment in both experiments. Although this difference was not quite significant in either, it probably reflects a real effect of the reduced hours of light. It represents a greater potential cost saving than the reduced electricity consumption. 5. Birds in rooms with an average light intensity of 2 lux laid slightly fewer eggs but their eggs were 0.5 g heavier than those laid in rooms maintained at 10 lux. There were no interactions between light intensity and light pattern or between stocks and light pattern.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Experiments with the Bio-mittent lighting system for laying hens.

1. Two experiments are described in which a system of intermittent lighting (15 min light followed by 45 min dark for 15 h, then 15 min light, 30 min dark, 15 min light and 8 h dark) was applied to laying pullets from 37 to 72 weeks of age. A step-up lighting programme was used as a control treatment (8L:16D from 0 to 18 weeks, photoperiod increased by 20 min each week from 18 to 41 weeks, 16L:8D from 41 to 72 weeks of age). 2. Food consumption was reduced by about 5% when intermittent lighting was in use and by 3.8% for the period from 18 to 72 weeks. 3. Rate of lay and egg weight were similar for intermittent lighting and the control treatment, provided that protein content of the diet was adjusted to maintain an adequate amino acid intake. 4. In the second trial 2 stocks, 2 stocking densities confounded with 2 temperatures and 2 types of food trough were used. Each of these factors affected food intake and it was found that more food was saved by intermittent lighting when intake was high and less when it was low. The proportion saved was approximately 5%. 5. Mortality was slightly but not significantly lower in both experiments where intermittent lighting was used. This may indicate that caged pullets are under less stress when intermittent lighting is used.

Animals

The phosphonium ion efflux system of Escherichia coli: relationship to the ethidium efflux system and energetic studies.

The extent of accumulation of methyltriphenylphosphonium ion by Escherichia coli was shown to be dependent on the permeability of the outer membrane and the activity of an efflux system for this compound. Evidence consistent with the operation of a single efflux system for compounds such as phosphonium ions, phenanthridiniums and flavines is presented. Studies on the energy coupling mechanism for this efflux system indicated that it was driven by the transmembrane proton electrochemical gradient.

Energy Metabolism

Characteristics of ethidium uptake by the trypanosomatid flagellates Crithidia fasciculata and Leptomonas seymouri.

The uptake of the phenanthridinium ethidium by the trypanosomatids Crithidia fasciculata and Leptomonas seymouri was studied. The time course of uptake of ethidium was biphasic for both organisms, consisting of an initial rapid phase and a protracted slow phase. The characteristics of these phases were consistent with the hypothesis that the initial phase represented specific external binding, while the second phase represented transport into the cells. In L. seymouri the transport phase was inhibited by inhibitors of energy transduction and putative inhibitors of a transport ATPase. Ethidium transport could not be saturated over a large concentration range of ethidium. Phenanthridiniums and related compounds displayed both inhibitory and stimulatory effects on ethidium transport.

Adenosine Triphosphatases

The transport of alpha-aminoisobutyrate into Crithidia fasciculata.

The transport of alpha-aminoisobutyrate into Crithidia fasciculata was characterized under aerobic and anaerobic conditions. Kinetic data for alpha-aminoisobutyrate transport were consistent with the operation of a single system of broad specificity that showed no marked dependence on Na+. Under anaerobic conditions alpha-aminoisobutyrate transport was inhibited by uncouplers such as 2,4-dinitrophenol, lipophilic cations such as methyltriphenylphosphonium ion and adenosine triphosphatase inhibitors such as dicyclohexylcarbodi-imide and NaN3. A working model in which alpha-aminoisobutyrate enters this organism by an H+-symport mechanism, the electrochemical gradient of protons being maintained by an H+-translocating adenosine triphosphatase on the cytoplasmic membrane, is proposed.

Aerobiosis

Transport of alpha-aminoisobutyrate by cells and membrane vesicles of Pseudomonas fluorescens.

The transport of alpha-aminoisobutyrate into Pseudomonas fluorescens NCIB 8865 and membrane vesicles prepared from this organism has been studied. Uptake by cells was mediated by two active transport systems with different apparent Km values, while transport into membrane vesicles was mediated by a single component. The effect of inhibitors on the energy-coupling mechanism for alpha-aminoisobutyrate transport in these systems suggests that a membrane potential may play a significant role in supporting alpha-aminoisobutyrate transport. The magnitude of the membrane potential in the vesicle system, and the sensitivity of its generation to inhibitors, has been measured using 137Cs in the presence of valinomycin. Direct attempts to demonstrate a protonsymport mechanism for alpha-aminoisobutyrate transport were negative.

Aminoisobutyric Acids

Anaerobic transport of serine and 2-aminoisobutyric acid by Staphylococcus epidermidis.

A membrane-bound ATPase detected in extracts of anaerobically grown Staphylococcus epidermidis was inhibited by a variety of compounds which inhibit ATPases in other organisms. Serine and 2-aminoisobutyric acid (AIB) were shown to enter the organism via the same transport system. The transport of AIB, the membrane potential and the transmembrane pH gradient were partially or completely abolished by the same inhibitors and also by uncoupling agents and lipid-soluble ions. It is proposed therefore that this ATPase generates and maintains an electrochemical gradient of protons across the cytoplasmic membrane of S. epidermidis capable of driving AIB uptake. Studies of AIB-induced proton movements suggested that AIB enters via a proton symport mechanism.

Adenosine Triphosphatases

Localization and possible role of an adenosine triphosphatase in Chlorobium thiosulfatophilum.

1. Evidence is presented that the ATPase activity detected in cell extracts of Chlorobium thiosulfatophilum is bound to the cytoplasmic membrane rather than to the chlorobium vesicles. 2. The activity of this ATPase is inhibited in vitro by various carbodiimides, phloridzin and sodium azide. 3. The apparent Km for ATP is approximately 0.2 mM and the enzyme shows product inhibition by ADP. 4. Photophosphorylation, characterized in vivo, is inhibited by many of the compounds that inhibit the ATPase.

Adenosine Diphosphate

The regulation of transport of glucose, gluconate and 2-oxogluconate and of glucose catabolism in Pseudomonas aeruginosa.

1. The induction by glucose and gluconate of the transport systems and catabolic enzymes for glucose, gluconate and 2-oxogluconate was studied with Pseudomonas aeruginosa PAO1 growing in a chemostat under conditions of nitrogen limitation with citrate as the major carbon source. 2. In the presence of a residual concentration of 30mM-citrate an inflowing glucose concentration of 6-8 mM was required to induce the glucose-transport system and associated catabolic enzymes. When the glucose concentration was raised to 20mM the glucose-transport system was repressed, but the transport system for gluconate, and at higher glucose concentrations, that for 2-oxogluconate, were induced. No repression of the glucose-catabolizing enzymes occurred at the higher inflowing glucose concentrations. 3. In the presence of 30mM-citrate no marked threshold concentration was required for the induction of the gluconate-transport system by added gluconate. 4. In the presence of 30mM-citrate and various concentrations of added glucose and gluconate, the activity of the glucose-transport system accorded with the proposal that a major factor concerned in the repression of this system was the concentration of gluconate, produced extracellularly by glucose dehydrogenase. 5. This proposal was supported by chemostat experiments with mutants defective in glucose dehydrogenase. Such mutants showed no repression of the glucose-transport system by high inflowing concentrations, but with a mutant apparently defective only in glucose dehydrogenase, the addition of gluconate caused repression of the glucose-transport system. 6. Studies with the mutants showed that both glucose and gluconate can induce the enzymes of the Entner-Doudoroff system, whereas for the induction of the gluconate-transport system glucose must be converted into gluconate.

Citrates

The role of glucose limitation in the regulation of the transport of glucose, gluconate and 2-oxogluconate, and of glucose metabolism in Pseudomonas aeruginosa.

The pathway of glucose metabolism in Pseudomonas aeruginosa was regulated by the availability of glucose and related compounds. On changing from an ammonium limitation to a glucose limitation, the organism responded by adjusting its metabolism substantially from the extracellular direct oxidative pathway to the intracellular phosphorylative route. This change was achieved by repression of the transport systems for gluconate and 2-oxogluconate and of the associated enzymes for 2-oxogluconate metabolism and gluconate kinase, while increasing the levels of glucose transport, hexokinase and glucose 6-phosphate dehydrogenase. The role of gluconate, produced by the action of glucose dehydrogenase, as a major inhibitory factor for glucose transport, and the possible significance of these regulatory mechanisms to the organism in its natural environment, are discussed.

Alcohol Oxidoreductases

The regulation of transport of glucose and methyl alpha-glucoside in Pseudomonas aeruginosa.

The methyl alpha-glucoside-transport system of Pseudomonas aeruginosa has been characterized with respect to its specificity, energy-dependence, kinetics and regulation. The uptake of glucose involved two components, one of which transported glucose (K(m)=8mum) and methyl alpha-glucoside (K(m)=2.8mm) whereas the other was more complex, involving the extracellular activity of glucose dehydrogenase. Mutants defective in this enzyme have been isolated and characterized. The methyl alpha-glucoside-glucose-transport system was repressed when the organism was grown in the absence of glucose; the induction of this transport system by glucose, and its activity once induced, were inhibited by products of citrate metabolism.

Azides