Biomedical subjects

R J Lambert

Publications and source records attributed to R J Lambert.

At least 19 recordsLinked to original sources

PubMed2003

QTL and candidate genes phytoene synthase and zeta-carotene desaturase associated with the accumulation of carotenoids in maize.

Carotenoids are a class of fat-soluble antioxidant vitamin compounds present in maize ( Zea mays L.) that may provide health benefits to animals or humans. Four carotenoid compounds are predominant in maize grain: beta-carotene, beta-cryptoxanthin, zeaxanthin, and lutein. Although beta-carotene has the highest pro-vitamin A activity, it is present in a relatively low concentration in maize kernels. We set out to identify quantitative trait loci (QTL) affecting carotenoid accumulation in maize kernels. Two sets of segregating families were evaluated-a set of F2:3 lines derived from a cross of W64a x A632, and their testcross progeny with AE335. Molecular markers were evaluated on the F2:3 lines and a genetic linkage map created. High-performance liquid chromatography was performed to measure beta-carotene, beta-cryptoxanthin, zeaxanthin, and lutein on both sets of materials. Composite interval mapping identified chromosome regions with QTL for one or more individual carotenoids in the per se and testcross progenies. Notably QTL in the per se population map to regions with candidate genes, yellow 1 and viviparous 9, which may be responsible for quantitative variation in carotenoids. The yellow 1 gene maps to chromosome six and is associated with phytoene synthase, the enzyme catalyzing the first dedicated step in the carotenoid biosynthetic pathway. The viviparous 9 gene maps to chromosome seven and is associated with zeta-carotene desaturase, an enzyme catalyzing an early step in the carotenoid biosynthetic pathway. If the QTL identified in this study are confirmed, particularly those associated with candidates genes, they could be used in an efficient marker-assisted selection program to facilitate increasing levels of carotenoids in maize grain.

Alkyl and Aryl Transferases↗

PubMed2001

Advances in disinfection testing and modelling.

AIMS: To develop a set of kinetic equations which more ably describe the disinfection process. METHODS AND RESULTS: A group of functions, the fat equations, based on the model used for the quantification of microbial inhibition, was produced. These functions introduce a limit to the numbers of micro-organisms capable of being disinfected. These new expressions were shown to be more general forms of currently-used (e.g. log-linear) disinfection models, and accommodate the lags and/or tails of non-linear log-survivor--time plots. An advance in the experimental procedures used to obtain disinfection data, using an optical density technique, was developed concomitantly. CONCLUSION: The methods of analyses (experimental and modelling) allow the researcher to examine, more ably, five-minute disinfection (or specific time disinfection tests) as well as the more important disinfection rate analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: The fat equations are an improvement over commonly-used rate models of disinfection, which are shown to be special cases of these equations. This raises the question as to whether our current understanding of the kinetic basis of disinfection requires revision.

Chloramphenicol↗

PubMed2001

The effect of interfering substances on the disinfection process: a mathematical model.

AIMS: To gain a greater understanding of the effect of interfering substances on the efficacy of disinfection. METHODS AND RESULTS: Current kinetic disinfection models were augmented by a term designed to quantify the deleterious effect of soils such as milk on the disinfection process of suspended organisms. The model was based on the assumption that inactivation by added soil occurred at a much faster rate than microbial inactivation. The new model, the fat-soil model, was also able to quantify the effect of changing the initial inoculum size (1 x 10(7)-5 x 10(7) ml(-1) of Staphylococcus aureus) on the outcome of the suspension tests. Addition of catalase to the disinfection of Escherichia coli by hydrogen peroxide, resulted in changes to the shape of the log survivor/time plots. These changes were modelled on the basis of changing biocide concentration commensurate with microbial inactivation. CONCLUSIONS: The reduction in efficacy of a disinfectant in the presence of an interfering substance can be quantified through the use of adaptations to current disinfection models. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the effect of soil on disinfection efficacy allows us to understand the limitations of disinfectants and disinfection procedures. It also gives us a mechanism with which to investigate the soil tolerance of new biocides and formulations.

Animals↗

PubMed2001

A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol.

AIMS: The minimum inhibitory concentration (MIC) of oregano essential oil (OEO) and two of its principle components, i.e. thymol and carvacrol, against Pseudomonas aeruginosa and Staphylococcus aureus was assessed by using an innovative technique. The mechanism of action of the above substances was also investigated. METHODS AND RESULTS: The applied technique uses 100-well microtitre plate and collects turbidimetric growth data. To produce the inhibition profiles, a wide range of concentrations were tested for each of the three compounds, as well as for carvacrol-thymol mixtures. Following a specific mathematical analysis of the observed inhibition profiles from all compounds, it was suggested that mixtures of carvacrol and thymol gave an additive effect and that the overall inhibition by OEO can be attributed mainly to the additive antimicrobial action of these two compounds. Addition of low amounts of each additive: (a) increased permeability of cells to the nuclear stain EB, (b) dissipated pH gradients as indicated by the CFDA-SE fluorescent probe irrespective of glucose availability and (c) caused leakage of inorganic ions. CONCLUSION: Mixing carvacrol and thymol at proper amounts may exert the total inhibition that is evident by oregano essential oil. Such inhibition is due to damage in membrane integrity, which further affects pH homeostasis and equilibrium of inorganic ions. SIGNIFICANCE AND IMPACT OF THE STUDY: The knowledge of extent and mode of inhibition of specific compounds, which are present in plant extracts, may contribute to the successful application of such natural preservatives in foods, since certain combinations of carvacrol-thymol provide as high inhibition as oregano essential oil with a smaller flavour impact.

Cell Membrane↗

PubMed2001

The relationships and susceptibilities of some industrial, laboratory and clinical isolates of Pseudomonas aeruginosa to some antibiotics and biocides.

AIMS: To provide evidence to support or refute the hypothesis that cross-resistance between antibiotics and biocides can occur. METHODS AND RESULTS: Fifty-five strains of Pseudomonas aeruginosa were tested for their resistance to anti-pseudomonal antibacterials. Twenty clinical, 19 industrial and 16 culture collection isolates were used. The MIC was found for the antibiotics amikacin, ceftazidime, ciprofloxacin, gentamycin, ticarcillin, tobramycin, imipenem and polymyxin B. The MIC was also found for the biocides benzalkonium chloride and chlorhexidine. The analysis of the data was based on the production of a normal distribution of the log (MIC) plots for each antimicrobial. Strains were then labelled as resistant, intermediate or sensitive based on the mean and standard deviation of the distributions. CONCLUSIONS: In general the clinical isolates were the most recalcitrant organisms, with the industrial isolates being the most sensitive. SIGNIFICANCE AND IMPACT OF THE STUDY: The work shows that antibiotic/biocide correlations do occur, especially with clinical strains. That such correlations were not found with industrial isolates suggests that the clinical environment is responsible for the correlation. We could infer that it is the selective pressure of antibiotic usage that differentiates the clinical environment from the industrial.

Animals↗

PubMed2000

One explanation for the variability of the bacterial suspension test.

Disinfection kinetic studies of sodium dodecyl sulphate, benzalkonium chloride and sodium hypochlorite against Staphylococcus aureus revealed that when a higher inoculum level of Staph. aureus than normal was used (approximately 1 log higher), the efficacy of disinfection was severely attenuated. Kinetic analysis using the Hom model for experiments carried out on tests using 3 x 108 organisms ml-1 were unable to account for the large increase in disinfection power observed when smaller inoculum levels were used. Since the inoculum was the same in every way except for the numbers used, the large variations in the log reduction/time curves could not be explained by a variation in the resistance of the population to the biocide, as identical log reduction-time curves should have resulted. The level of disinfection achieved for a given concentration of biocide was found to be approximately linearly related to the cell number ml-1 of test solution and not to the log number. The variation observed is believed to occur due to intrinsic self-quenching of the biocide by the microbes during the course of the disinfection test. As the level of free biocide decreases, the rate of reaction decreases, giving the tails of the log reduction/time curves. Such intrinsic self-quenching could explain the large variations known to occur in the legally required disinfection suspension tests.

Bacteriological Techniques↗

PubMed2000

Susceptibility testing: accurate and reproducible minimum inhibitory concentration (MIC) and non-inhibitory concentration (NIC) values.

Measuring the minimum inhibitory concentration (MIC) of a substance by current methods is straightforward, whereas obtaining useful comparative information from the tests can be more difficult. A simple technique and a method of data analysis are reported which give the experimentalist more useful information from susceptibility testing. This method makes use of a 100-well microtitre plate and the analysis uses all the growth information, obtained by turbidometry, from each and every well of the microtitre plate. A modified Gompertz function is used to fit the data, from which a more exact value can be obtained for the MIC. The technique also showed that at certain concentrations of inhibitor, there was no effect on growth relative to a control well (zero inhibitor). Above a threshold value, which has been termed the non-inhibitory concentration or NIC, growth becomes limiting until it reaches the MIC, where no growth relative to the control is observed.

Anti-Bacterial Agents↗

PubMed2000

Disinfection kinetics: a new hypothesis and model for the tailing of log-survivor/time curves.

A new hypothesis for the understanding of chemical disinfection, which we have termed the Intrinsic Quenching hypothesis, is presented. This mechanistic treatment of disinfection kinetics is based on the hypothesis that the biocide concentration may not be in vast excess over the microbes, as is normally assumed. A mathematical model was developed and found to be useful in describing the observed kinetics of several disinfectants. The model suggested that the reason for the observation of non-linear, log-survivor curves was due to the ability of the microbes, in clean, soil-free conditions, to intrinsically quench the bulk concentration of biocide.

Disinfectants↗

PubMed2000

Susceptibility testing: inoculum size dependency of inhibition using the Colworth MIC technique.

The minimum inhibitory concentration, MIC, is an accepted and well used criterion for measuring the susceptibility of organisms to inhibitors. Many factors influence the MIC value obtained, including temperature, inoculum size and type of organism. A modification of the method developed in this laboratory to obtain inhibition profiles of antimicrobials was used to examine the effect of inoculum size on the degree of inhibition observed with respect to inhibitor concentration. The data obtained enabled the production of an empirical model of inhibition, based on a Gompertz function, relating the level of growth observed to both the inoculum size and concentration of the inhibitor. The inoculum size dependencies of phenethyl alcohol, phenoxyethanol, p-chloro-m-cresol, trichloro-phenol, thymol and dodecyltrimethylammmonium bromide against Staphylococcus aureus were obtained.

Anti-Bacterial Agents↗

PubMed2000

Development of resistance to chlorhexidine diacetate in Pseudomonas aeruginosa and the effect of a "residual" concentration.

Stable resistance in Pseudomonas aeruginosa NCIMB 10421 was obtained by step-wise exposure to gradually increasing concentrations of chlorhexidine diacetate (CHX). Repeated exposure to a proposed "residual" (sub-MIC) concentration of CHX also created stable resistance. Resistance was also developed by a single exposure to the "residual" concentration of CHX, but this was unstable. Similar experiments with Escherichia coli and CHX or cetylpyridinium chloride resulted in no significant increase in resistance. Antibiotic susceptibility profiles of the CHX-resistant P. aeruginosa cultures showed no cross-resistance, although some of the cultures were resistant to benzalkonium chloride.

Anti-Infective Agents, Local↗

PubMed1999

Experimental neodymium:YAG laser damage to acrylic, poly(methyl methacrylate), and silicone intraocular lens materials.

PURPOSE: To compare neodymium:YAG (Nd:YAG) laser effects on acrylic, silicone, and poly(methyl methacrylate) (PMMA) intraocular lens (IOL) polymers. METHODS: Ten Nd:YAG laser exposures were produced in each of 6 implantation-quality acrylic (Alcon MA60BM), silicone (Staar AQ1016), and PMMA (Alcon MC60BM) IOLs under identical conditions. Each polymer type was irradiated at 6 power settings (0.3, 0.5, 1.0, 1.5, 2.0, and 3.0 mJ) and at 2 focal points (midpoint of lens optic and on the posterior surface to which a cellophane membrane was affixed). The linear extent of the damage was measured using light microscopy. Specimens exposed to 1.0 mJ were processed for scanning electron microscopy. RESULTS: The damage threshold (> or = 5 microns depth) was 0.3 mJ for silicone and 1.0 mJ for acrylic and PMMA IOLs. At the clinically relevant power levels, 1.0 to 2.0 mJ, the depth of damage in the acrylic polymer was 11.9 to 30.5 times less than the depth in the silicone polymer. Similarly, the depth of damage in the PMMA polymer was 5.4 to 52.6 times less than the depth in the silicone polymer. The morphologic pattern of damage in the silicone IOL showed a deep, irregularly configured trough with meandering tendrils. Acrylic IOL damage morphology consisted of an ameboid-shaped entry site without radiating fractures and mild posterior penetration. Poly(methyl methacrylate) IOL damage consisted of a shallow focal trough with radiating fractures. CONCLUSIONS: The silicone IOL polymer had the lowest threshold for laser-induced damage and greater linear extension of damage than the PMMA and acrylic IOL polymers. Poly(methyl methacrylate) and silicone polymers exhibited collateral damage or ejected particulates adjacent to the entry site, whereas the acrylic polymer showed a discrete locus of damage.

Acrylic Resins↗

PubMed1999

Weak-acid preservatives: modelling microbial inhibition and response.

Weak-acid preservatives are widely used to prevent microbial spoilage of acidic foods and beverages. Characteristically, weak-acid preservatives do not kill micro-organisms but inhibit growth, causing very extended lag phases. Preservatives are more effective at low pH values where solutions contain increased concentrations of undissociated acids. Inhibition by weak-acids involves rapid diffusion of undissociated molecules through the plasma membrane; dissociation of these molecules within cells liberates protons, thus acidifying the cytoplasm and preventing growth. By modelling preservative action in yeast, using a thermodynamic and kinetic approach, it was possible to demonstrate that: (i) inhibition depends more on the degree to which individual preservatives are concentrated within cells, rather than on undissociated acid concentration per se; (ii) it is entirely feasible for microbes to pump protons out of the cell during extended lag phase and raise internal pH (pHi), despite further influx of preservatives; (iii) the duration of the lag phase can be predicted from the model, using a Gaussian fit of proton-pumping H(+)-ATPase activity against pHi; (iv) theoretical ATP consumption for proton pumping can be directly correlated with the reduction in cell yield observed in glucose-limited cultures.

Acids, Acyclic↗

PubMed1999

An investigation into the differences between the Bioscreen and the traditional plate count disinfectant test methods.

Investigations of biocide efficacy by automated methods involving optical density measurements, e.g. using the recently published 'Bioscreen' method, are complicated by the fact that a poor correlation often exists between the log reductions obtained using the automated method vs those obtained by the traditional plate count methods. It was hypothesized that the differences observed between the two methods were due to the level of cell injury, which was masked by the optical density methods but which was recognized by the plate counts. Comparisons of log reductions following a disinfection test always showed the Bioscreen method to be overestimating the log reductions with respect to the plate counts. A correlation between colony size on the plates and the 'Bioscreen' results for a fixed disinfectant concentration and contact time was found using Global Imaging software. The results obtained also suggested that the observed colony area was dependent on the amount of injury incurred by a microbe during the disinfection process. A mathematical model of injury was developed which predicted the observed differences between the Bioscreen and the traditional plate method. The model further suggested a possible reason for biocidal lags.

Colony Count, Microbial↗

PubMed1999

A kinetic study of the effect of hydrogen peroxide and peracetic acid against Staphylococcus aureus and Pseudomonas aeruginosa using the bioscreen disinfection method.

Hydrogen peroxide and peracetic acid at pH 4 were examined against Staphylococcus aureus and Pseudomonas aeruginosa using the published 'Bioscreen' technique of biocide analysis. The data were examined using either classical Chick-Watson (CW) log-linear disinfection kinetics or the empirical, non-linear time Hom model. In some cases, modelling the data with the classical CW method gave good linear correlations, in others, however, deviations from this model were observed. In such cases the Hom model proved an adequate descriptor of the data. The Bioscreen technique therefore gives data which can be analysed using the normal mechanistic and empirical models currently available.

Disinfection↗

PubMed1998

Intraocular lens changes after short- and long-term exposure to intraocular silicone oil. An in vivo study.

OBJECTIVE: Silicone oil frequently is used as a vitreous substitute after complex vitreoretinal procedures. The authors sought to study the effect of short- and long-term exposure to silicone oil on polymethyl methacrylate (PMMA, MC60BM; Alcon, Ft. Worth, TX), silicone (SI-30NB; AMO, Irvine, CA), and soft acrylic (MA60BM; Alcon) intraocular lenses (IOLs). DESIGN: An experimental animal study. INTERVENTION: Forty-one New Zealand white rabbits underwent lensectomy, vitrectomy, capsulotomy, and placement of one of the three types of IOLs into the ciliary sulcus. All lenses were weighed before implantation and 24 hours after explanation. In the short-term study, an fluid-air exchange was performed followed by the use of silicone oil (1000 centistokes) to coat the posterior lens surface. Immediately thereafter, an air-fluid exchange was performed and the remaining silicone on the posterior lens surface was aspirated or wiped or both for 1 minute using a soft-tipped extrusion cannula for 1 minute. In the long-term study, the posterior segment was filled with 1000 centistokes silicone oil after fluid-air exchange. Animals were observed by slit-lamp biomicroscopy and photographed at 1 week, 1 month, and 3 months after surgery. At 3 months, all animals underwent silicone-fluid exchange, an attempt to manually remove any remaining silicone oil, and lens explanation. RESULTS: In the short-term study, no silicone oil remained after manual wiping and/or aspiration in any of the four rabbits implanted with PMMA or acrylic IOLs. In the animals with silicone IOLs, a significant amount of silicone oil remained on the posterior lens surface of all lenses (P < 0.01 for silicone vs. acrylic and silicone vs. PMMA). No statistically significant difference was found when comparing the lens weights in each group before and after implantation. In the long-term study, aqueous droplet formation was found on the posterior lens surface of six of nine PMMA IOLs and ten of ten silicone IOLs at 3 months. No opacities were observed in the group with acrylic IOLs (P < 0.001 for acrylic vs. silicone, P = 0.0018 for acrylic vs. PMMA, and P = 0.047 for PMMA vs. silicone). Adherent silicone oil remained on two of nine PMMA IOLs and on none of ten acrylic IOLs. In contrast, a significant amount of silicone oil remained on the posterior lens surface of ten of ten silicone IOLs (P < 0.001 for silicone vs. acrylic and silicone vs. PMMA). Furthermore, there was a statistically significant increase in lens weights before and after implantation in the silicone IOL group but not in the PMMA or acrylic group (P < 0.01). CONCLUSIONS: It is extremely difficult or impossible to remove remaining silicone oil from the posterior surface of a silicone IOL after short- or long-term exposure to silicone oil. This oil may interfere with the surgeon's view of the retina and may diminish the patient's visual acuity. In contrast, oil is readily removed from the posterior surface of an acrylic IOL. The authors therefore recommend the use of a soft acrylic or PMMA IOL over a silicone IOL when choosing a lens for implantation in patients who may require vitreoretinal procedures with silicone oil tamponade.

Acrylic Resins↗

PubMed1998

Viscoelastic adherence to corneal endothelium following phacoemulsification.

PURPOSE: To compare the residual adherence of viscoelastics to the corneal endothelium following phacoemulsification in an in vitro rabbit model. SETTING: Departments of Ophthalmology and Anatomy, Wayne State University, Detroit, Michigan, USA. METHODS: Three groups of 10 rabbit eyes each had a lensectomy via phacoemulsification using sodium hyaluronate (Amvisc Plus, Healon GV) or sodium chondroitin sulfate-sodium hyaluronate (Viscoat) as the viscoelastic agent. After phacoemulsification and cortex removal, a central corneal block was excised, cryofixed, and processed for light and electron microscopy. Viscoelastic thickness was determined by a calibrated reticule on the light microscope or a calibrated measuring program in the electron microscope. The nonparametric statistical test, Kruskal-Wallis, was used to compare viscoelastic groups. RESULTS: Median phacoemulsification time between viscoelastic agents was not significantly different. Median viscoelastic thicknesses were 13.0 microns for Amvisc Plus, 0.4 micron for Healon GV, and 375.0 microns for Viscoat. Each was significantly different from the others (Kruskal-Wallis, P < .001). CONCLUSIONS: Median thickness of Amvisc Plus, Healon GV, and Viscoat remaining adherent to the corneal endothelium after phacoemulsification was markedly different. Viscoat provided the greatest amount of viscoelastic material adjacent to the corneal endothelium.

Animals↗

PubMed1998

Disinfectant testing: use of the Bioscreen Microbiological Growth Analyser for laboratory biocide screening.

A new method is described for screening potential biocides based on the traditional suspension test using the Bioscreen optical plate reader. This new method is rapid, reproducible, quantitative and cost effective. Data obtained by this new method are not directly equivalent to the log reduction normally quoted, but give a measurement of the total effect of the biocide on the microbe population, measuring the effect of injury as well as death (non-viability). The method allows for the routine examination of disinfection kinetics, the study of which leads to greater scientific insight into disinfection than that achieved by the standard 5 min, one-point, disinfection tests currently employed.

Colony Count, Microbial↗

PubMed1997

Incision sizes for foldable intraocular lenses.

OBJECTIVE: The purpose of the study is to determine the minimal incision sizes required for implantation of a variety of different foldable intraocular lenses (IOLs) and to evaluate the effect of incision size on tissue damage. DESIGN: Randomized experimental study. PARTICIPANTS: Sixty-nine fresh human cadaveric eyes: 15 (pilot study), 48 (main study), and 6 (scanning electron microscopy study). INTERVENTION: Implantation of foldable IOLs into cadaveric eyes. MAIN OUTCOME MEASURES: In 48 fresh human cadaveric eyes, limbal corneal tunnel incisions were made, and in a randomized fashion, 8 different foldable IOLs of 20.5 diopters (D) were inserted: 4 silicone (SI-30NB, Allergan Medical Optics, Irvine, CA; C10UB, Chiron Ophthalmics, Inc, Irvine, CA; LI41U, IOLAB, Chiron Ophthalmics, Inc, Irvine, CA; AA-4203, Staar Surgical Company, Monrovia, CA); two soft acrylic (MA60BM and MA30BA, both Alcon Laboratories, Inc, Ft. Worth, TX); and two hydrogel models (SH30BC, Alcon; H60M, Storz Ophthalmics, Inc, St. Louis, MO). For each IOL model, six insertions were performed with a recommended implantation device. Using calipers, the authors measured internal and external incision sizes before and after insertion. Scanning electron microscopy was performed on selected incisions in six additional human cadaveric eyes. RESULTS: Incision sizes after insertion ranged from 3.2 to 3.8 mm. The smallest incisions permitting IOL insertion were associated with the injectors. However, these incisions enlarged after insertion by approximately 11% and then were similar to the incision sizes after forceps insertion of the high-refractive-index silicone, the 5.5-mm optic acrylic, and the one-piece hydrogel IOL. The largest incisions were associated with the 6-mm acrylic IOL and the three-piece silicone IOL with a lower refractive index. The scanning electron microscopy showed tearing of corneal tissue after implantation through the smallest incision; this was more pronounced with injectors than with forceps. CONCLUSIONS: Corneal tunnel incisions enlarged up to 11% after insertion of foldable IOLs through the smallest possible incision. With current technology, the smallest postinsertion incision size of a 20.5-D foldable IOL is 3.2 mm.

Cadaver↗