Biomedical subjects

R J Miles

Publications and source records attributed to R J Miles.

At least 55 records · Page 3Linked to original sources

PubMed1992

Refinement of the transpalatine exposure of the nasopharynx.

The intrasulcular incision provides excellent transpalatine exposure of the nasopharynx with a viable palatal flap. There is less chance for naso-oral fistula formation since the resulting suture line is always over underlying bone. This safely allows extensive removal of the hard palate. Additionally, the continuous sling suture allows excellent flap reapproximation using the teeth as anchors for the replaced palatal flap. Patients tolerate oral feedings within 24 hours of their operations and require minimal postoperative analgesics. The extended intrasulcular incision offers many advantages over other methods for transpalatine exposures to the nasopharynx.

Gingivectomy↗

PubMed1991

Oxygen uptake and H2O2 production by fermentative Mycoplasma spp.

Oxygen uptake and H2O2 accumulation during the metabolism of glucose and glycerol by whole cells, and of L-alpha-glycerophosphate (GP) and NADH by cells lysed with Triton, was determined for the type strains of six fermentative Mycoplasma species. Oxidation of glucose and of NADH by M. mycoides, M. pneumoniae and M. putrefaciens was accompanied by the accumulation of relatively small quantities of H2O2 (less than 0.05 mol/mol O2), though larger quantities (0.17-0.24 mol/mol O2) were produced by M. dispar. M. fermentans and M. canis were distinguished from the other strains used in that O2 uptake in the presence of glucose could not be demonstrated. However, metabolism of glucose was indicated by a reduction in the pH of the suspending medium and lysed cells oxidised NADH with the production of approximately 1.0 mol H2O2/mol O2 taken up. Glycerol was oxidised by all the strains studied except M. fermentans, and large quantities of H2O2 (0.48-1.07 mol/mol O2) accumulated. Cells of the glycerol-oxidising strains, lysed with Triton, oxidised GP with the production of approximately 1.0 mol H2O2/mol O2 utilised, which indicated the presence of a GP oxidase. The importance of H2O2 production as a factor in the pathogenicity of some mycoplasmas might depend upon the availability of glycerol in vivo.

Glucose↗

PubMed1990

A mutant of Mycoplasma mycoides subsp. mycoides lacking the H2O2- producing enzyme L-alpha-glycerophosphate oxidase.

Cells of Mycoplasma mycoides subsp. mycoides grown without stirring or aeration in batch culture, and resuspended in a salts solution, oxidised a range of carbohydrates including glycerol. The rate of glycerol oxidation was not reduced when cells were passaged more than 20 times in batch culture. However, in cells grown in stirred and aerated chemostat culture for 100 generations the ability to oxidise glycerol, but not other carbohydrates, was lost or greatly reduced. A mutant strain isolated from chemostat even after several passages in batch culture. The growth rate and growth-yield of the mutant strain in batch culture were similar to those of the parent strain. The mutant possessed activity for glycerol kinase but had lost that for the hydrogen peroxide-producing enzyme, L-alpha-glycerophosphate oxidase. The selection pressure in favour of the mutant strain in chemostat culture may be a decreased production of hydrogen peroxide.

Culture Media↗

PubMed1990

Expression of carbonic anhydrase in neisseriae and other heterotrophic bacteria.

A diverse range of heterotrophic bacteria was screened for the presence of carbonic anhydrase (CA) activity, sensitivity to inhibition of growth by acetazolamide (CA inhibitor), and the presence of protein binding monospecific antibody prepared against purified Neisseria sicca CA. CA activity was demonstrated only in strains of N. sicca and N. gonorrhoeae. However, all Neisseria strains, including various isolates of N. meningitidis and N. lactamica, were sensitive to acetazolamide, when grown in air, and showed serological cross-reaction with N. sicca CA. Strains of other genera were resistant to acetazolamide. A number of strains including members of the genera Pseudomonas, Staphylococcus, Streptococcus, Serratia and Proteus also strongly expressed a gene product(s) immunologically related to CA. The presence of CA cross-reacting proteins, which lack hydrase activity, is discussed in relation to the function of the various mammalian CA isoenzymes.

Acetazolamide↗

PubMed1989

Flow microcalorimetric assay of antibiotics--I. Polymyxin B sulphate and its combinations with neomycin sulphate and zinc bacitracin on interaction with Bordetella bronchiseptica (NCTC 8344).

A flow microcalorimetric assay for polymyxin B sulphate has been developed which has a better reproducibility (relative standard deviation less than 3%) and sensitivity (0.35 micrograms ml-1) than conventional microbiological assays, and requires an assay time of ca. 4.5 h. The combinations with zinc bacitracin, with neomycin sulphate, and with both zinc bacitracin and neomycin sulphate indicate antagonism between these antibiotics upon interaction with Bordetella bronchiseptica (NCTC 8344). The combinations of all three antibiotics assayed were: (1) equimolar proportions; and (2) those proportions present in the commercial preparation TrisepR (ICI, Macclesfield, UK).

Bacitracin↗

PubMed1989

Flow microcalorimetric assay of antibiotics--II. Neomycin sulphate and its combinations with polymyxin B sulphate and zinc bacitracin on interaction with Bacillus pumilus (NCTC 8241).

A flow microcalorimetric assay for Neomycin has been developed which is monitored through interaction of the antibiotic with Bacillus pumilus as the test organism. The assay has better reproducibility (relative standard deviation 2.3%) and is more sensitive than conventional microbiological bioassay (0.5-2 micrograms ml-1). The effects of combinations with zinc bacitracin, with polymyxin B sulphate, and with both zinc bacitracin and polymyxin B sulphate (both in equimolar proportions), and in those proportions present in the commercial preparation TrisepR (ICI, Macclesfield, UK) have also been investigated. Synergy was observed for the combinations of Neomycin with the other two antibiotics in binary mixtures at the relative proportions found in TrisepR. The addition of all three antibiotics at the levels used in TrisepR did not show synergy. However, addition of all three antibiotics at equimolar concentrations did show synergy. It is suggested that microcalorimetry may be useful in in vitro experiments for exploring the relative proportions required for maximal effect in antibiotic combinations.

Bacillus↗

PubMed1989

Flow microcalorimetric assay of antibiotics--III. Zinc bacitracin and its combinations with polymyxin B sulphate and neomycin sulphate on interaction with Micrococcus luteus.

A flow microcalorimetric assay for zinc bacitracin has been developed which has better reproducibility (relative standard deviation less than 2%) and sensitivity (0.02 micrograms ml-1) than conventional microbiological assays, and requires an assay time of between 7.5-9 h. The assay is not suitable for zinc bacitracin determinations in the presence of equimolar concentrations of polymyxin B sulphate or neomycin sulphate, or of these antibiotics in the proportions in which they occur in the commercial preparation Trisep (ICI, Macclesfield, UK).

Bacitracin↗

PubMed1989

Flow microcalorimetric assay of antibiotics--IV. Polymyxin B sulphate, neomycin sulphate, zinc bacitracin and their combinations with Escherichia coli suspended in buffer plus glucose medium.

Flow microcalorimetric assays for polymyxin B sulphate and neomycin sulphate have been developed using Escherichia coli as the test organism, suspended in a glucose plus buffer medium. These assays have a better reproducibility (relative standard deviations 3.2 and 2.0%, respectively), and require a shorter time (1 h including time required for preparation of the calorimeter) than do conventional microbiological assays, but are not as sensitive. It is suggested that a screening programme might produce a small group of more suitable (i.e. more sensitive) test organisms, which could be used to develop rapid and reproducible flow microcalorimetric assays for a wide range of antibiotics by the procedure described. The effect of combinations of polymyxin B sulphate, neomycin sulphate and zinc bacitracin (the antibiotic components of the commercial preparation Trisep, ICI, Macclesfield, UK) on the power output of cells suspended in glucose plus buffer medium is also reported. In defined combinations, the effects of neomycin sulphate appeared to be exerted before those of polymyxin B sulphate.

Bacitracin↗

PubMed1988

Microcalorimetric determination of the kinetics of substrate utilisation by non-growing suspensions of Neisseria sicca.

The metabolism of various substrates by non-growing suspensions of Neisseria sicca was investigated by a flow-microcalorimetric technique. Substrate utilisation showed Michaelis kinetics allowing determination of saturation constants (Km) and maximum specific rates of substrate utilisation (Vmax). Pyruvate, lactate, a number of tricarboxylic acid cycle intermediates, and amino acids (aspartate, glutamate and proline) were rapidly metabolised [Vmax 5-35 mumol (g dry wt cells)-1 min-1]; Km values were between 4 and 20 microM. Glucose, glycerol, acetate and the other amino acids investigated gave only a slight or no increase in power. The pattern of substrate utilisation is discussed in relation to the role of carbonic anhydrase in N. sicca.

Acetates↗

PubMed1988

Factors affecting the survival of Neisseria sicca.

Cultures of Neisseria sicca incubated at 37 degrees C died rapidly (within 36 h) after growth ceased. Re-suspending cells in a brain heart infusion broth and storing at 4 degrees C greatly reduced the rate of decline in viability (decimal reduction time 6 days). An important factor in maintaining viability was apparently the presence of external energy source(s). Survival comparable to that in broth was obtained by incubation in Ringer's solution with pyruvate plus glucose (but not with pyruvate or glucose alone). Medium pH had little effect on survival in the range pH 7.0 to 8.5. Energy sources also promoted survival of cells in Ringer's solution or a buffered salts solution at 37 degrees C. Highest levels of survival (up to 30% at 24 h) were obtained with pyruvate, lactate, proline and glutamate. A number of other amino acids and the tricarboxylic acid cycle intermediates, isocitrate, oxoglutarate, succinate, fumarate, malate and oxaloacetate, enhanced survival to a lesser extent.

Brain↗

PubMed1987

Antibiotic sensitivity and mutation rates to antibiotic resistance in Mycoplasma mycoides ssp. mycoides.

The antibiotic resistance of Mycoplasma mycoides ssp. mycoides strain T1 was investigated. This strain was resistant to high levels (greater than 100 micrograms ml-1) of rifampicin and nalidixic acid. It was sensitive to streptomycin, spectinomycin and novobiocin; however, single step mutants with high levels of resistance (greater than 100 micrograms ml-1) were readily isolated. With erythromycin and tylosin for which the minimum inhibitory concentration (MIC) for the parent strain was less than 0.1 microgram ml-1, mutants resistant to greater than 100 micrograms ml-1 were obtained in two and three steps respectively. The MIC of tetracycline in single step resistant mutants (0.6 microgram ml-1) was tenfold higher than the parent strain, but could not be increased further. There was only a twofold increase in resistance to chloramphenicol in single step mutants. The frequency of resistant mutants varied with the antibiotic and was between 4 X 10(-6) and 2 X 10(-8). The mutation rate to antibiotic resistance to streptomycin, spectinomycin, novobiocin, erythromycin and tylosin was between 3 X 10(-8) and 5 X 10(-9) per cell per generation. There was a fivefold decrease in mutation rate to resistance to 60 micrograms ml-1 streptomycin compared to that to 20 micrograms ml-1.

Anti-Bacterial Agents↗

PubMed1986

Growth of Mycoplasma mycoides subspecies mycoides on media containing various sugars and amino sugars: an ampoule microcalorimetric study.

The growth of Mycoplasma mycoides subspecies mycoides strain T1 on media containing various sugars, tryptose, yeast extract, salts and either pig or calf-serum or a mixture of bovine serum albumin (BSA) plus lipid was followed by ampoule microcalorimetry. Power-time (p-t) curves were reproducible and showed details of growth not observable by conventional microbiological techniques. In media with metabolisable sugars p-t curves typically showed three periods of exponential increase in power separated by transient declines or plateaux. Maximum power (Pmax) was dependent upon the nature and concentration of sugar, whether ampoules were capped in air or nitrogen, and whether the medium contained pig or calf-serum or BSA plus lipid. The highest Pmax was observed in pig-serum medium with glucose, in ampoules capped in air. Decline in power from Pmax was essentially exponential.

Acetylglucosamine↗

PubMed1986

A nystatin-resistant mutant of Saccharomyces cerevisiae: isolation and characterization by electron microscopy and chemical analysis of whole cells, cell-walls and protoplasts.

A mutant of Saccharomyces cerevisiae NCYC 239 with a high minimum inhibitory concentration (35 micrograms ml-1) for nystatin, compared to that of the parent strain (2 micrograms ml-1), was derived by a series of subcultures in media containing increasing antibiotic concentrations. In the absence of nystatin, the growth rate of the mutant was significantly lower than the parent strain, although mean cell-size and size-distribution were similar. No differences between strains were detectable by electron microscopy. Analysis of whole cells showed the total sterol present and the ratio of ergosterol:24(28)dehydroergosterol was similar. However, there were marked differences in amino acid content and chain-length of fatty acids in the cell wall, and protoplasts from resistant cells had decreased amounts of unsaturated fatty acids. It is suggested that alterations in cell wall components in the mutant may be directly linked to the mechanism of nystatin resistance.

Amino Acids↗

PubMed1986

Uptake of nystatin by sensitive and resistant cells of Saccharomyces cerevisiae.

The uptake of nystatin by sensitive and by resistant cells of Saccharomyces cerevisiae was studied as a function of nystatin concentration, temperature and pH. The presence or absence of glucose in nystatin uptake experiments was also studied. The rate data, effects of glucose and the derived activation energies for nystatin uptake revealed significant differences in response between sensitive and resistant cells. The role of the cell wall in the uptake process is discussed.

Drug Resistance, Microbial↗

PubMed1986

Uptake of nystatin by protoplasts of sensitive and resistant cells of Saccharomyces cerevisiae.

The uptake of nystatin by protoplasts derived from sensitive and resistant cells of Saccharomyces cerevisiae has been studied as a function of nystatin concentration, temperature and pH. The presence or absence of glucose in the uptake experiments was also studied. Activation energies (Ea) for nystatin uptake revealed profound differences between protoplasts derived from sensitive and resistant cells. Those for the latter closely resembled their whole cell counterparts. The values of Ea for the uptake of nystatin under all the conditions studied indicate the importance of the cell wall in the uptake process.

Biological Transport↗

PubMed1985

The effect of chemical modification of Saccharomyces cerevisiae on electrophoretic mobility, cell-wall structure and amphotericin B uptake.

Saccharomyces cerevisiae NCYC 239 suspended in solutions of NaCl showed two distinct plateaus in plots of electrophoretic mobility vs. pH, corresponding to pKa values of approx. 2 and 5. This is in contrast to cells suspended in buffer where only a single pKa (4) can be determined. Modification of cells with KI/I2 or nitrous acid led to altered electrophoretic mobility, indicating the presence of sulphydryl and amino groups, respectively, in the yeast cell surface, whereas uranyl nitrate modification had little effect, suggesting phosphate groups to be absent. Electron micrographs showed visible effects of KI/I2 and nitrous acid modification on cell membrane structure, and in these modified cells amphotericin B uptake was rapid. It is suggested that diffusion through the cell wall is the rate-limiting step for amphotericin B uptake. An activation energy of 20 kJ X mol-1 was determined for uptake of amphotericin B by unmodified cells.

Amphotericin B↗

PubMed1985

The mycoplasmacidal properties of sodium hypochlorite.

The effect of hypochlorite concentration on Mycoplasma mycoides ssp. mycoides viability was tested under a variety of conditions. The experimental variables employed included chlorine-cell contact time, chlorine concentration, carrier system and organic loading. Initial populations of 10(6) c.f.u./ml were killed (no survivors in 1 ml) by hypochlorite solution containing 25 p.p.m. available chlorine in 15 s in the absence of organic load and 50 p.p.m. available chlorine in 5 min in the presence of 1% protein. Higher concentrations of hypochlorite were required to disinfect a porous carrier system in the absence or presence of protein. The results are in contrast to previous reports that M. bovis is killed only by high hypochlorite concentrations.

Animals↗

PubMed1985

Kinetics of utilization of organic substrates by Mycoplasma mycoides subsp. mycoides in a salts solution: a flow-microcalorimetric study.

The metabolism of various organic substrates by suspensions of Mycoplasma mycoides subsp. mycoides in a salts solution was followed by microcalorimetry. Enthalpy changes associated with metabolism were in good agreement with theoretical values. Substrate utilization showed Michaelis kinetics, allowing saturation constants (Km) and maximum specific rates of substrate utilization (Vmax) to be determined. In cells grown on a complex medium containing glucose, Km values were: glucose, fructose, N-acetylglucosamine, glycerol and pyruvate, less than 5 microM; lactate, 20 microM; glucosamine, 130 microns, and mannose, 1 mM. Values of Vmax for glycerol, pyruvate and lactate were similar and approximately twice those for glucose, mannose, glucosamine and N-acetylglucosamine; Vmax for fructose was one-quarter of that for glucose. In cells grown on complex medium in which glucose was replaced by mannose, glucosamine or N-acetylglucosamine, Vmax and Km for the respective growth sugars and for glucose were not significantly affected. However, in cells grown in the presence of fructose, Vmax for fructose increased to the value observed for glucose. It is suggested that M. mycoides is adapted to, and is constitutive for, the utilization of a single sugar (glucose), and a single amino sugar (N-acetylglucosamine), but that in the presence of fructose a fructose-utilizing pathway is induced.

Acetylglucosamine↗