Biomedical subjects

K A Dawson

Publications and source records attributed to K A Dawson.

At least 19 recordsLinked to original sources

PubMed2006

Novel "plum pudding" gels as potential drug-eluting stent coatings: controlled release of fluvastatin.

This study evaluated novel structural motifs known as "plum pudding" gels as potential drug-eluting stent coatings. Controlled delivery of a HMG-CoA reductase inhibitor (statin) from the intravascular stent surface represents a potential therapeutic modality for prevention of in-stent restenosis (ISR). In this study, gels were comprised of fluvastatin-loaded thermoresponsive microgel particles containing the relatively hydrophilic N-isopropylacrylamide (NiPAAm), mixed with the more hydrophobic N-tert-butylacrylamide (NtBAAm) in different wt/wt ratios: 85/15, 65/35, and 50/50, randomly dispersed in a 65/35 or 85/15 NiPAAm/NtBAAm copolymer matrix. Fluvastatin release from 5 microm copolymer films was greatest from the most hydrophilic systems and least from the more hydrophobic systems. Release from hydrophobic matrices appeared to be via Fickian diffusion, enabling use of the Stokes-Einstein equation to determine diffusion coefficients. Release from hydrophilic matrices was non-Fickian. Eluted drug retained its bioactivity, assessed as selective inhibition of human coronary artery smooth muscle cell proliferation. When stainless steel stent wires were coated (25 microm thickness) with fluvastatin-loaded 65/35 microgels in an 85/15 copolymer matrix, drug elution into static and perfused flow environments followed similar elution profiles. In contrast to elution from copolymer films cast on flat surfaces, diffusion from stent wires coated with hydrophilic and hydrophobic systems both followed Fickian patterns, with slightly larger diffusion coefficients for elution from the flow system. We conclude that manipulation of the relative hydrophobicities of both microgel and matrix components of "plum pudding" gels results in tightly regulated release of fluvastatin over an extended time period relevant to initiation and propagation of ISR.

Anticholesteremic Agents↗

PubMed2005

Simultaneous release of hydrophobic and cationic solutes from thin-film "plum-pudding" gels: a multifunctional platform for surface drug delivery?

The release of two compositionally different solutes from a composite gel composed of two different populations of microgel particles embedded in a single bulk gel matrix is described, showing the potential of the "plum-pudding gel" as a multifunctional platform for controlled surface release. One hydrophobic solute (pyrene) and one hydrophobic and charged solute (rhodamine 123) were chosen as the solutes to be released. Hydrophobic microgels composed of 50% N-isopropylacrylamide (NIPAM) and 50% N-tert-butylacrylamide (BAM) were loaded with pyrene, and anionic microgels composed of 30% acrylic acid (AAc), 20% NIPAM, and 50% BAM were loaded with rhodamine 123. The two solute-loaded microgel populations were incorporated into a single bulk gel network, from which the two solutes were released simultaneously and independently. Using this structural motif, solutes that are mutually incompatible can be incorporated into a single matrix with which they may also be incompatible. The electrostatically incorporated solute was released much more slowly than the hydrophobically attracted solute, indicating that the microgel composition can be tailored to the specific solute, and thus control its release rate. The choice of bulk matrix was also found to influence the release rate much more than expected, offering a further control element to the system.

Drug Delivery Systems↗

PubMed2005

Poly(N-isopropylacrylamide) copolymer films as vehicles for the sustained delivery of proteins to vascular endothelial cells.

The aim of this study was to establish the capacity of thermoresponsive poly(N-isopropylacrylamide) copolymer films to deliver bioactive concentrations of vascular endothelial growth factor (VEGF165) to human aortic endothelial cells (HAEC) over an extended time period. Films were prepared using a 50:50 (w/w) mixture of non-crosslinkable and crosslinkable copolymers of the following monomer compositions (w/w): 85:15, N-isopropylacrylamide (NiPAAm):N-tert-butylacrylamide (NtBAAm); and 85:13:2 NiPAAm:NtBAAm:acrylamidobenzophenone (ABzPh, crosslinking agent), respectively. After crosslinking by UV irradiation, the ability of films to incorporate a fluorescently labeled carrier protein (FITC-labeled BSA, 1 mg loaded per film), at 4 degrees C, was first established. Incorporation into the matrix was confirmed by the observation that increasing film thickness from 5 to 10 microm increased release from collapsed films at 37 degrees C (1.76 +/- 0.15 and 10.98 +/- 3.38 microg/mL, respectively, at 24 h postloading) and that this difference was maintained at 5 days postloading (1.81 +/- 0.25 and 13.8 +/- 2.3 microg/mL, respectively). Incorporation was also confirmed by visualization using confocal microscopy. When 10-microm films were loaded with a BSA solution (1 mg/mL) containing VEGF165 (3 microg/mL), sustained release of VEGF165 was observed (10.75 +/- 3.11 ng at 24 h; a total of 31.32 +/- 8.50 ng over 7 days). Furthermore, eluted VEGF165 increased HAEC proliferation by 18.2% over control. The absence of cytotoxic species in medium released from the copolymer films was confirmed by the lack of effect of medium (incubated with copolymer films for 3 days) on HAEC viability. In conclusion this study has shown that NiPAAm:NtBAAm copolymers can be loaded with a therapeutic protein and can deliver bioactive concentrations to human vascular endothelial cells over an extended time period.

Acrylic Resins↗

PubMed2004

Star polymers: a study of the structural arrest in the presence of attractive interactions.

Simulations and mode-coupling theory calculations, for a large range of the arm number f and packing fraction eta have shown that the structural arrest and the dynamics of star polymers in a good solvent are extremely rich: the systems show a reentrant melting of the disordered glass nested between two stable fluid phases that strongly resemble the equilibrium phase diagram. Starting from a simple model potential we investigate the effect of the interplay between attractive interactions of different range and ultrasoft core repulsion, on the dynamics and on the occurrence of the ideal glass transition line. In the two cases considered so far, we observed some significant differences with respect to the purely repulsive pair interaction. We also discuss the interplay between equilibrium and nonequilibrium phase behavior. The accuracy of the theoretical tools we utilized in our investigation has been checked by comparing the results with molecular dynamics simulations.

Journal Article↗

PubMed2004

Effects of dietary mannan oligosaccharide, bacitracin methylene disalicylate, or both on the live performance and intestinal microbiology of turkeys.

Hybrid male turkeys were fed to 18 wk of age in a completely randomized design with 10 replicate pens (18 birds each) per treatment to compare growth promoters. Four dietary treatments were used: negative control (CON), bacitracin methylene disalicyate (BMD) at 55 mg/kg to 6 wk and 27.5 mg/kg thereafter, mannan oligosaccharide (MOS) at 0.1% to 6 wk and 0.05% thereafter, and BMD and MOS at concentrations listed above. There were 3 toms/m2 (3.59 ft2/tom) on fresh pine shavings inoculated with used litter. A 6-phase feeding program was used, with crumbles the first 3 wk and pellets thereafter. At wk 6 and 18, one bird per pen was killed, and the large intestinal microbial populations, after being frozen, were enumerated (i.e., bifidobacteria, Clostridium perfringens, coliforms, enterococci, Escherichia coli, lactobacilli, and total anerobes). Body weights at wk 18 were as follows: CON, 11.87 kg; BMD, 12.46 kg; MOS: 12.56 kg; and BMD + MOS, 12.79 kg. The BMD and MOS turkeys were (P < 0.05) heavier than CON birds, and those fed the combination were significantly heavier than all other treatments. At wk 18, BMD + MOS feed conversion ratio of 2.66 was significantly lower than CON at 3.00 with BMD and MOS being intermediate (2.83 and 2.79, respectively). Mortality was not affected by treatment. The BMD and MOS each reduced large intestinal concentrations of Clostridium perfringens (log transformed data analysis) at wk 6 but not at wk 18. The BMD or MOS each improved turkey performance, and when used together, exhibited further beneficial effects.

Animal Nutritional Physiological Phenomena↗

PubMed2003

Extended delivery of the antimitotic agent colchicine from thermoresponsive N-isopropylacrylamide-based copolymer films to human vascular smooth muscle cells.

The aim of this study was to establish the capacity of thermoresponsive poly(N-isopropylacrylamide) copolymer films to deliver bioactive concentrations of an antimitotic agent to human vascular smooth muscle cells (HASMC) over an extended period of time. Copolymer films were prepared using a 50:50 (w/w) ratio of N-isopropylacrylamide (NiPAAm) monomer to the more hydrophobic N-tert-butylacrylamide (NtBAAm) and loaded with the antimitotic agent colchicine (0.1 micromol per film) at room temperature. Colchicine release from films was sustained over a 14-day period. At 24 h postloading, the concentration of colchicine in the medium overlying films was 2.12 +/- 0.16 microM; this fell to 0.20 +/- 0.01 microM at 7 days and decreased further to 0.12 +/- 0.01 microM after 14 days. Colchicine released from copolymer films inhibited proliferation when subsequently placed on HASMC: at 0.1 microM, released colchicine reduced proliferation to 18.5 +/- 0.8% of control cells (p < 0.001, n = 9). The antiproliferative effect of released colchicine was comparable to that of native colchicine, as observed in separate experiments. Furthermore, colchicine released from 50:50 polymer films inhibited the proliferation of cells grown in the same environment as the copolymer. Inhibition of cell proliferation was not due to the release of cytotoxic particles from the copolymer because medium incubated with copolymer for 5 days and then applied to HASMC did not alter cell viability. In conclusion, this study demonstrates that 50:50 NiPAAm:NtBAAm copolymers can deliver bioactive concentrations of the antimitotic agent colchicine to human vascular cells over an extended period of time.

Acrylamides↗

PubMed2003

Structural arrest in dense star-polymer solutions.

The dynamics of star polymers is investigated via extensive molecular and Brownian dynamics simulations for a large range of functionality f and packing fraction eta. The calculated isodiffusivity curves display both minima and maxima as a function of eta and minima as a function of f. Simulation results are compared with theoretical predictions based on different approximations for the structure factor. In particular, the ideal glass transition line predicted by mode-coupling theory is shown to exactly track the isodiffusivity curves, offering a theoretical understanding for the observation of disordered arrested states in star-polymer solutions.

Journal Article↗

PubMed2002

Confirmation of anomalous dynamical arrest in attractive colloids: a molecular dynamics study.

Previous theoretical simulation and experimental studies have indicated that particles with a short-ranged attraction exhibit a range of dynamical arrest phenomena. These include very pronounced reentrance in the dynamical arrest curve, a logarithmic singularity in the density correlation functions, and the existence of "attractive" and "repulsive" glasses. Here we carry out extensive molecular dynamics calculations on dense systems interacting via a square-well potential. This is one of the simplest systems with the required properties, and may be regarded as canonical for interpreting the phase diagram, and now also the dynamical arrest. We confirm the theoretical predictions for reentrance, logarithmic singularity, and give a direct evidence of the existence, independent of theory, of two distinct glasses. We now regard the previous predictions of these phenomena as having been established.

Journal Article↗

PubMed2002

Evidence for an unusual dynamical-arrest scenario in short-ranged colloidal systems.

Extensive molecular dynamics simulation studies of particles interacting via a short-ranged attractive square-well potential are reported. The calculated loci of constant diffusion coefficient D in the temperature-packing fraction plane show a reentrant behavior, i.e., an increase of diffusivity on cooling, confirming an important part of the high volume-fraction dynamical-arrest scenario earlier predicted by theory for particles with short-ranged potentials. The more efficient localization mechanism induced by the short-range bonding provides, on average, additional free volume as compared to the hard-sphere case and results in faster dynamics.

Journal Article↗

PubMed2002

Effects of supplemental fructooligosaccharides plus mannanoligosaccharides on immune function and ileal and fecal microbial populations in adult dogs.

The goal of this study was to examine whether supplemental fructooligosaccharides (FOS) plus mannanoligosaccharides (MOS) influenced immune function and ileal and fecal microbial populations of adult dogs. Eight adult dogs surgically fitted with ileal cannulas were used in a crossover design. Dogs were fed 200 g of a dry, extruded, kibble diet twice daily. At each feeding, dogs were dosed with either 1 g sucrose (placebo) or 2 g FOS plus 1 g MOS orally via gelatin capsule. Fecal, ileal, and blood samples were collected at the end of each 14-d period to measure microbial populations and immune characteristics. Treatment least squares means were compared using the GLM procedure of SAS. Supplementation of FOS plus MOS increased fecal bifidobacteria and fecal and ileal lactobacilli concentrations. Dogs fed FOS plus MOS also tended to have lower blood neutrophils and greater blood lymphocytes vs placebo. Serum, fecal, and ileal immunoglobulin concentrations were unchanged by treatment. Supplementation of FOS plus MOS beneficially altered indices of gut health by improving ileal and fecal microbial ecology. Supplementation of FOS plus MOS also altered immune function by causing a shift in blood immune cells.

Animals↗

PubMed2001

Mechanical properties of a model of attractive colloidal solutions.

We review the nature of glass transitions and the glasses arising from a square-well potential with a narrow and deep well. Our discussion is based on the mode coupling theory (MCT), and the predictions of glasses that we make refer to the "ideal" glasses predicted by this theory. We believe that the square-well system well represents colloidal particles with attractive interactions produced by grafted polymers, or depletion interactions. It has been recently shown that two types of glasses, an attractive and a repulsive one, are predicted by MCT for this model. The former can form at quite low densities. Most of our attention is directed at the mechanical properties of the glasses predicted by this theory. In particular we calculate the elastic shear modulus at zero frequency and the longitudinal stress modulus in the long wavelength limit. Results for both are presented along the glass-liquid transition curves and their interesting behavior is explained in terms of the underlying physics of the system.

Journal Article↗

PubMed2001

Size-dependent sex allocation within flowers of the annual herb Clarkia unguiculata (Onagraceae): ontogenetic and among-plant variation.

The relative allocation of resources to male and female functions may vary among flowers within and among individual plants for many reasons. Several theoretical models of sex allocation in plants predict a positive correlation between the resource status of a flower or individual and the proportion of reproductive resources allocated to female function. These models assume that, independent of resource status, a negative correlation exists between male and female investment. Focusing on the allocation of resources within flowers, we tested these theoretical predictions and this assumption using the annual Clarkia unguiculata (Onagraceae). We also sought preliminary evidence for a genetic component to these relationships. From 116 greenhouse-cultivated plants representing 30 field-collected maternal families, multiple flowers and fruits per plant were sampled for gamete production, pollen : ovule ratio, seed number, ovule abortion, seed biomass/fruit, mean individual seed mass, and petal area. If sex allocation changes as predicted, then (1) assuming that flowers produced early have access to more resources than those produced later, basal flowers should exhibit a higher absolute and proportional investment in female function than distal flowers and (2) plants of high resource status (large plants) should produce flowers with a higher proportional investment in female function than those of low resource status. Within plants, variation in floral traits conformed to the first prediction. Among plants and families, no significant effects of plant size (dry stem biomass) on intrafloral proportional sex allocation were observed. We detected no evidence for a negative genetic correlation between male and female investment per flower, even when controlling for plant size.

Journal Article↗

PubMed2000

Toxicity effects of compressed and supercritical solvents on thermophilic microbial metabolism.

Selection of biocompatible solvents is critical when designing bioprocessing applications for the in situ biphasic extraction of metabolic end-products. The prediction of the biocompatibility of supercritical and compressed solvents is more complicated than for liquid solvents, because their properties can change significantly with pressure and temperature. The activity of the anaerobic thermophilic bacterium, Clostridium thermocellum, was studied when the organism was incubated in the presence of compressed nitrogen, ethane, and propane at 333 K and multiple pressures. The metabolic activity of the organisms in contact with compressed solvents was analyzed using traditional indicators of solvent biocompatibility, such as log P, interfacial tension, and solvent density. The toxicity of the compressed solvents was compared with the phase and molecular toxicity effects measured in liquid alkanes at atmospheric pressure. Inactivation increased with time in the presence of the compressed solvents, but was constant in the presence of atmospheric liquid solvents. Knowledge of molecular and phase toxicity provides a framework for the interpretation of C. thermocellum metabolism in contact with atmospheric and compressed solvents.

Clostridium↗

PubMed2000

Organization and sequence of histidine biosynthesis genes hisH, -A, -F, and -IE in Thermoanaerobacter ethanolicus.

Nucleotide sequence analysis of a 3.5-kb chromosomal fragment from the low G + C Gram-positive bacterium Thermoanaerobacter ethanolicus revealed a cluster of five contiguous open reading frames (ORFs) designated hisH, hisA, hisF, hisIE, and ORF5. The first four ORFs showed homology to genes of the histidine biosynthesis pathway, and ORF5 encoded a product with no significant similarities to polypeptides presently known. The hisH ORF was partial (truncated by cloning) and ORF5 was adjacent to xylF, which codes for a xylose-binding periplasmic protein. The five genes encoded putative proteins of >104, 237, 254, 216, and 169 amino acids, respectively. Amino acid sequence comparison of the four his gene products indicated closely related homologs in prokaryotes, varying from low G + C Gram-positive bacteria to archaea. This is the first report of his anabolic genes in a thermophilic anaerobic bacterium.

Bacteria, Anaerobic↗

PubMed2000

A simple and effective separation and purification procedure for DNA fragments using dodecyltrimethylammonium bromide.

In this work we describe a simple two step separation procedure for the separation and purification of short DNA fragments. The first step involves precipitating the DNA using the cationic surfactant dodecyltrimethylammonium bromide. Dodecyltrimethylammonium bromide, unlike cetyltrimethylammonium bromide will not precipitate DNA before complexation is complete thus providing a high purity DNA. The second step involves dissolution of the DNA-dodecyltrimethylammonium complex in 75% ethanol, followed by precipitation of the Sodium-DNA salt, by titrating in a salt solution. This method is particularly suited to purification of short fragments as it does not require high salt concentrations in the ethanol precipitation step, which can be damaging for short DNA. The ability of dodecyltrimethylammonium bromide to remove ethidium bromide from intercalation sites on the DNA is also discussed

Chemical Precipitation↗

PubMed2000

The effects of dietary mannaoligosaccharides on cecal parameters and the concentrations of enteric bacteria in the ceca of salmonella-challenged broiler chicks.

The ability of different enteric pathogens and coliforms to trigger agglutination of yeast cells (Saccharomyces cerevisiae, NCYC 1026) and a yeast cell wall preparation (MOS) was examined. Five of seven strains of Escherichia coli and 7 of 10 strains of Salmonella typhimurium and Salmonella enteritidis agglutinated MOS and Sac. cerevisiae cells. Strains of Salmonella choleraesuis, Salmonella pullorum, and Campylobacter did not lead to agglutination. Two strains that agglutinated MOS (S. typhimurium 29E and Salmonella dublin) and one nonagglutinating strain (S. typhimurium 27A) were selected as challenge organisms for in vivo studies in chicks under controlled conditions. In a series of three trials in which 3-d-old chicks were orally challenged with 10(4) cfu of S. typhimurium 29E, birds receiving 4,000 ppm of dietary MOS had reduced cecal S. typhimurium 29E concentrations (5.40 vs 4.01 log cfu/ g; P < 0.05) at Day 10. In a second series of three trials with S. dublin as challenge organism, the number of birds that tested salmonella positive in the ceca at Day 10 was less when MOS was part of the diet (90 vs 56%; P < 0.05). To test the effect of MOS on concentrations of bacteria that do not express Type 1 fimbriae, a challenge trial was conducted with S. typhimurium 27A. However, strain 27A did not colonize the birds sufficiently to evaluate whether MOS affected its cecal concentration. Mannanoligosaccharide did not significantly reduce the concentrations of cecal coliforms (P < 0.10) although they were numerically lower. It had no effect on cecal concentrations of lactobacilli, enterococci, anaerobic bacteria, lactate, volatile fatty acid, or cecal pH.

Agglutination Tests↗

PubMed2000

High-affinity maltose binding and transport by the thermophilic anaerobe Thermoanaerobacter ethanolicus 39E.

Thermoanaerobacter ethanolicus is a gram-positive thermophile that produces considerable amounts of ethanol from soluble sugars and polymeric substrates, including starch. Growth on maltose, a product of starch hydrolysis, was associated with the production of a prominent membrane-associated protein that had an apparent molecular weight of 43,800 and was not detected in cells grown on xylose or glucose. Filter-binding assays revealed that cell membranes bound maltose with high affinity. Metabolic labeling of T. ethanolicus maltose-grown cells with [(14)C]palmitic acid showed that this protein was posttranslationally acylated. A maltose-binding protein was purified by using an amylose resin affinity column, and the binding constant was 270 nM. Since maltase activity was found only in the cytosol of fractionated cells and unlabeled glucose did not compete with radiolabeled maltose for uptake in whole cells, it appeared that maltose was transported intact. In whole-cell transport assays, the affinity for maltose was approximately 40 nM. Maltotriose and alpha-trehalose competitively inhibited maltose uptake in transport assays, whereas glucose, cellobiose, and a range of disaccharides had little effect. Based on these results, it appears that T. ethanolicus possesses a high-affinity, ABC type transport system that is specific for maltose, maltotriose, and alpha-trehalose.

Amino Acid Sequence↗

PubMed2000

Efficacy of laidlomycin propionate in low-protein diets fed to growing beef steers: effects on steer performance and ruminal nitrogen metabolism.

We conducted two experiments to evaluate the effect of the ionophore laidlomycin propionate (LP) on steer performance and ruminal N metabolism. Experiment 1 was a 91-d growth study evaluating the growth and ruminal characteristics of steer calves consuming supplemental LP. Steers (n = 96; 255 +/- 3 kg; four steers/pen; six pens/treatment) were used in a randomized complete block design with a 2 x 2 factorial arrangement of treatments consisting of two levels of dietary CP (formulated to be 10.5 and 12.5% of DM) with and without LP (11 mg/kg diet DM). Ruminal fluid was collected via stomach tube on d 91 from one steer randomly selected from each pen. No CP x LP interactions were observed with performance data (P > .64). Final weight and total gain were greater (P < .07) for 12.5% CP and LP compared with 10.5% CP and control steers, respectively. Also, DMI was increased (P = .08) with 12.5% CP but not with LP supplementation (P = .36). In addition, ADG and gain:feed ratio were greater (P < .03) for both 12.5% CP and supplemental LP. Ruminal NH3 N concentration was greater (P < .09) with 12.5% CP and LP. Total VFA concentration and molar proportion of acetate were not affected by treatment (P > .11). However, propionate concentration was increased (P < .09) with 12.5% CP and LP, and acetate:propionate was lower (P = .02) with LP supplementation. In Exp. 2, six steers were used in a replicated 3 x 3 Latin square design to compare ruminal fermentation and protein degradation in steers without ionophore feeding or adapted to LP or monensin. In vitro deamination of amino acids by adapted ruminal microbes was also assessed. Ionophore supplementation decreased (P = .07) ruminal NH3 N concentration compared with control steers, and LP increased (P = .02) ruminal NH3 N compared with monensin. Molar proportion of acetate was decreased (P = .02) and propionate increased (P = .01) with ionophore treatment. Consequently, ionophore supplementation depressed the acetate:propionate ratio (P = .01). In situ degradation rate of soybean meal (SBM) CP was greater (P = .09) with ionophore treatment, but estimates of SBM undegradable intake protein were not altered by treatment (P > .25). Microbial specific activity of net NH3 N release and alpha-amino N degradation were decreased (P < .04) with ionophores. Based on this study, LP and monensin did not affect the extent of ruminal degradation of SBM CP but decreased amino acid deamination.

Animal Feed↗