Pharmacokinetic profile and effect on the faecal microbiome of a single dose of pradofloxacin oral suspension in the rabbit (Oryctolagus cuniculus).

Fluoroquinolones are often administered to pet rabbits given their perceived safety and limited effects on anaerobic gut microbiota. However, the pharmacokinetics and relative safety of pradofloxacin, a third-generation veterinary fluoroquinolone with a much broader spectrum of activity, have not been reported in this species. Here, we determined the pharmacokinetic profile of a single dose of oral pradofloxacin in rabbits and evaluated effects on the faecal microbiome. Four mature female rabbits were administered pradofloxacin (25 mg/ml oral suspension), at a dose of 7.5 mg/kg. The pradofloxacin median (range) Tmax was 4.50 (2.00-5.00) h, Cmax 600.66 (395.85-886.72) ng/ml and t½ was 1.27 (0.12-1.39) h. These results indicated that oral absorption of pradofloxacin was slower, and elimination faster compared with other fluoroquinolones in healthy rabbits, as well as relative to cats and dogs. Following treatment with pradofloxacin, faecal microbiota profiling showed some compositional differences between treated and control animals. This was the result of a significant decrease in the abundance of Proteobacteria, in particular bacteria belonging to the Pseudomonas, Atopostipes and Parabacteroides genera. The pharmacokinetic profile of pradofloxacin in rabbits should be further studied by increasing the sample size and using multiple-dose protocols (i.e. 7 days) to confirm safety. Further information on the effects of protein binding, higher dosages and disease on pradofloxacin pharmacokinetics in rabbits are needed before an accurate dosing regimen can be recommended.

Atomic force microscopy as a nanoscience tool in rational food design.

Atomic force microscopy (AFM) is a nanoscience tool that has been used to provide new information on the molecular structure of food materials. As an imaging tool it has led to solutions to previously intractable problems in food science. This type of information can provide a basis for tailoring food structures to optimise functional behaviour. Such an approach will be illustrated by indicating how a basic understanding of the role of interfacial stability in complex foods systems can be extended to understand how such interfacial structures behave on digestion, and how this in turn suggests routes for the rational design of processed food structures to modify lipolysis and control fat intake. As a force transducer AFM can be used to probe interactions between food structures such as emulsion droplets at the colloidal level. This use of force spectroscopy will be illustrated through showing how it allows the effect of the structural modification of interfacial structures on colloidal interactions to be probed in model emulsion systems. Direct studies on interactions between colliding soft, deformable droplets reveal new types of interactions unique to deformable particles that can be exploited to manipulate the behaviour of processed or natural emulsion structures involved in digestion processes. Force spectroscopy can be adapted to probe specific intermolecular interactions, and this application of the technique will be illustrated through its use to test molecular hypotheses for the bioactivity of modified pectin molecules.

Probing the in situ competitive displacement of protein by nonionic surfactant using atomic force microscopy.

Force-distance data obtained from an atomic force microscope have been used to follow the in situ displacement of beta-lactoglobulin from tetradecane droplets by Tween 20 (polyoxyethylenesorbitan monolaurate). Interpretation of the force-distance curves has shown that the slope of the region, traditionally termed the constant compliance region, is a useful indicator of droplet deformation within a given experiment. The magnitude of this slope can be used to monitor how the deformability of the droplet changes upon addition of surfactant. It has been found that, immediately after initial addition of surfactant, there is an increase in magnitude of this slope, indicating a stiffening of the droplet, attributed to a stiffening of the protein network formed at the surface of the droplet. Subsequent additions of Tween 20 reduce the magnitude of the slope until an equilibrium value is reached, where the interface becomes surfactant-dominated. These observations suggest that it is possible to monitor in situ the displacement of protein from individual oil droplets. The data have been interpreted in terms of the "orogenic" model of displacement, which is based on studies made on model interfaces. These data have been compared to those obtained using the more traditional techniques of dilatational rheology, surface loading, and surface potential measurements for analogous beta-lactoglobulin-stabilized droplets or emulsions.

Effect of substituent pattern and molecular weight of cellulose ethers on interactions with different bile salts.

Some known mechanisms proposed for the reduction of blood cholesterol by dietary fibre are: binding with bile salts in the duodenum and prevention of lipid absorption, which can be partially related with the bile salt binding. In order to gain new insights into the mechanisms of the binding of dietary fibre to bile salts, the goal of this work is to study the main interactions between cellulose derivatives and two types of bile salts. Commercial cellulose ethers: methyl (MC), hydroxypropyl (HPC) and hydroxypropylmethyl cellulose (HPMC), have been chosen as dietary fibre due to their highly functional properties important in manufactured food products. Two types of bile salts: sodium taurocholate (NaTC) and sodium taurodeoxycholate (NaTDC), have been chosen to understand the effect of the bile salt type. Interactions in the bulk have been investigated by means of differential scanning calorimetry (DSC) and linear mechanical spectroscopy. Results show that both bile salts have inhibitory effects on the thermal structuring of cellulose ethers and this depends on the number and type of substitution in the derivatised celluloses, and is not dependent upon molecular weight. Concerning the bile salt type, the more hydrophobic bile salt (NaTDC) has greater effect on these interactions, suggesting more efficient adsorption onto cellulose ethers. These findings may have implications in the digestion of cellulose-stabilised food matrices, providing a springboard to develop new healthy cellulose-based food products with improved functional properties.

Effect of processing on the displacement of whey proteins: applying the orogenic model to a real system.

Atomic force microscopy (AFM) has been used to investigate the displacement of a commercial whey protein system and the behavior as compared to that of beta-lactoglobulin (Mackie, A. R.; Gunning, A. P.; Wilde, P. J.; Morris, V. J. Orogenic displacement of protein from the air-water interface by competitive adsorption. J. Colloid Interface Sci. 1999, 210, 157-166). The whey protein isolate (WPI) was displaced from an air-water interface by the surfactants Tween 20 and Tween 60. Displacement data obtained were compared with data obtained for pure beta-lactoglobulin and have shown that WPI was more resistant to displacement from the air-water interface than native beta-lactoglobulin. This was related to the greater surface elasticity of WPI at higher surface stresses. In the presence of Tween 20, WPI was observed to remain on the interface at surface pressures up to 8 mN/m greater than the surface pressure at which complete displacement of beta-lactoglobulin was observed. Displacement of WPI and beta-lactoglobulin films by the surfactant Tween 60 showed similar results. However, because of the lower surface activity of Tween 60, it was not possible to reach surface tension values similar to those obtained for Tween 20. Despite the lower surface activity of Tween 60, WPI was still observed to be present at the interface at surface pressure values greater than those by which beta-lactoglobulin had been completely displaced.

Interfacial characterization of beta-lactoglobulin networks: displacement by bile salts.

The competitive displacement of a model protein (beta-lactoglobulin) by bile salts from air-water and oil-water interfaces is investigated in vitro under model duodenal digestion conditions. The aim is to understand this process so that interfaces can be designed to control lipid digestion thus improving the nutritional impact of foods. Duodenal digestion has been simulated using a simplified biological system and the protein displacement process monitored by interfacial measurements and atomic force microscopy (AFM). First, the properties of beta-lactoglobulin adsorbed layers at the air-water and the olive oil-water interfaces were analyzed by interfacial tension techniques under physiological conditions (pH 7, 0.15 M NaCl, 10 mM CaCl2, 37 degrees C). The protein film had a lower dilatational modulus (hence formed a weaker network) at the olive oil-water interface compared to the air-water interface. Addition of bile salt (BS) severely decreased the dilatational modulus of the adsorbed beta-lactoglobulin film at both the air-water and olive oil-water interfaces. The data suggest that the bile salts penetrate into, weaken, and break up the interfacial beta-lactoglobulin networks. AFM images of the displacement of spread beta-lactoglobulin at the air-water and the olive oil-water interfaces suggest that displacement occurs via an orogenic mechanism and that the bile salts can almost completely displace the intact protein network under duodenal conditions. Although the bile salts are ionic, the ionic strength is sufficiently high to screen the charge allowing surfactant domain nucleation and growth to occur resulting in displacement. The morphology of the protein networks during displacement is different from those found when conventional surfactants were used, suggesting that the molecular structure of the surfactant is important for the displacement process. The studies also suggest that the nature of the oil phase is important in controlling protein unfolding and interaction at the interface. This in turn affects the strength of the protein network and the ability to resist displacement by surfactants.

Effect of surfactant type on surfactant--protein interactions at the air-water interface.

The displacement of the proteins (beta-lactoglobulin and beta-casein) from an air-water interface by the nonionic (Tween 20 and Tween 60) and ionic (sodium dodecyl sulfate, cetyltrimethylammonium bromide, and lyso-phosphatidylcholine-lauroyl) surfactants has been visualized by atomic force microscopy (AFM). The surface structure has been sampled by the use of Langmuir-Blodgett deposition onto mica substrates to allow imaging in the AFM. In all cases, the displacement process was found to occur through the recently proposed orogenic mechanism (Mackie et al. J. Colloid Interface Sci. 1999, 210, 157-166). In the case of the nonionic surfactants, the displacement involved nucleation and growth of surfactant domains leading to failure of the protein network and subsequent loss of protein into the bulk phase. The surface pressure dependence of the growth of surfactant domains and the failure of the network were found to be the same for both Tween 20 and Tween 60, demonstrating that the breakdown of the protein film was dominated by the mechanical properties of the network. The displacement of protein by ionic surfactants was found to be characterized by nucleation of surfactant domains with little domain growth prior to failure of the network. The size of the domains formed by ionic surfactants was found to be limited by the strong intersurfactant repulsive forces between the charged headgroups. Screening of these charges led to an increase in the size of the domains. The surface pressure at which the network continuity was lost was found to be dependent on the type of surfactant and, in all cases, to occur at higher surface pressures than that required for nonionic surfactants. This has been attributed to surfactant-protein binding that initially strengthens the protein network at low surfactant concentrations. Evidence obtained from surface shear rheology supports this assertion.