Rhett W. Stout

A Novel Solubility-Enhanced Curcumin Formulation Showing Stability and Maintenance of Anticancer Activity

Curcumin (CUR) is an active food compound, but its insolubility and instability in water contributes to low bioavailability. In this study, the solubility of CUR was enhanced by utilizing the solubilizing properties of rubusoside (RUB). The solubility of CUR in water increased linearly from 61 μg/mL to 2.318 mg/mL in the presence of RUB ranging from 1% to 10% (w/v). Dynamic light scattering and transmission electron microscopy studies found that CUR and RUB formed CUR-RUB nanoparticle (∼8 nm) complexes. The RUB-solubilized CUR was stable in physiological conditions and did not precipitate when diluted or degrade when spray-dried to a completely reconstitutable powder. Furthermore, cell viability assays demonstrated the efficacy of RUB-solubilized CUR against human colon, breast, and pancreatic cancer cell lines. The development of this new solubilized, stable, and biologically active CUR formulation lays the foundation for future bioavailability improvement.

Variability associated with repeated measurements of gastrointestinal tract motility in dogs obtained by use of a wireless motility capsule system and scintigraphy

OBJECTIVE To compare repeatability of measurements of gastrointestinal tract motility in healthy dogs obtained by use of a wireless motility capsule (WMC) and scintigraphy. ANIMALS 6 healthy adult dogs (mean +/- SD body weight, 21.5 +/- 1.8 kg). PROCEDURES A radiolabeled test meal was offered immediately after oral administration of a WMC. Serial static scintigraphic abdominal images were acquired for 270 minutes. A dedicated remote receiver was used for data collection from the WMC until the WMC was expelled in the feces. Each dog was evaluated 3 times at intervals of 1 to 2 weeks. RESULTS Mean gastric emptying half-time measured by use of scintigraphy (T(1/2)-GES) for each dog ranged from 99.9 to 181.2 minutes. Mean gastric emptying time (GET) measured by use of the WMC (GET-WMC) in each dog ranged from 385.3 to 669.7 minutes. Mean coefficient of variation was 11.8% for T(1/2)-GES and 7.8% for GET-WMC. The intraclass correlation coefficient was 69% for T(1/2)-GES and 71% for GET-WMC. Results for a nested analysis of covariance suggested that both methods were comparable for the evaluation of gastric emptying. CONCLUSIONS AND CLINICAL RELEVANCE Scintigraphy and a WMC system had similar variation for assessment of gastric emptying. Moderate intraindividual variability was detected for both methods and must be considered when interpreting test results for individual dogs. Repeatability of measurements obtained by use of the WMC was equivalent to that obtained by use of scintigraphy. The WMC system offers a nonradioactive, user-friendly method for assessment of gastric emptying in dogs.

Prevalence and Infection Load Dynamics of Rickettsia felis in Actively Feeding Cat Fleas

Background Rickettsia felis is a flea-associated rickettsial pathogen recurrently identified in both colonized and wild-caught cat fleas, Ctenocephalides felis. We hypothesized that within colonized fleas, the intimate relationship between R. felis and C. felis allows for the coordination of rickettsial replication and metabolically active periods during flea bloodmeal acquisition and oogenesis. Methodology/Principal Findings A quantitative real-time PCR assay was developed to quantify R. felis in actively feeding R. felis-infected fleas. In three separate trials, fleas were allowed to feed on cats, and a mean of 3.9×106 R. felis 17-kDa gene copies was detected for each flea. A distinct R. felis infection pattern was not observed in fleas during nine consecutive days of bloodfeeding. However, an inverse correlation between the prevalence of R. felis-infection, which ranged from 96% in Trial 1 to 35% in Trial 3, and the R. felis-infection load in individual fleas was identified. Expression of R. felis-infection load as a ratio of R. felis/C. felis genes confirmed that fleas in Trial 3 had significantly greater rickettsial loads than those in Trial 1. Conclusion/Significance Examining rickettsial infection dynamics in the flea vector will further elucidate the intimate relationship between R. felis and C. felis, and facilitate a more accurate understanding of the ecology and epidemiology of R. felis transmission in nature.

Hepatitis E antibodies in laboratory rabbits from 2 US vendors.

We tested laboratory rabbits from 2 US vendors for antibodies against hepatitis E virus (HEV); Seroprevalences were 40% and 50%. Retrospective analysis of an ocular herpes simplex 1 experiment demonstrated that HEV seropositivity had no effect on experiment outcome. HEV probably is widespread in research rabbits, but effects on research remain unknown.

Laboratory evaluation of oral treatment of rodents with systemic insecticides for control of bloodfeeding sand flies (Diptera: Psychodidae).

The purpose of this study was to evaluate the efficacy of oral treatment of rodents with diets containing the systemic insecticides ivermectin, abamectin, imidacloprid, or spinosad, to control bloodfeeding sand flies. We found that diets containing concentrations higher than 10 mg/kg abamectin were not palatable to rodents, and that a diet containing 10 mg/kg abamectin (a palatable concentration) did not cause 100% mortality of bloodfeeding sand flies. Treatment of rodents with imidacloprid was effective for less than 3 days post-treatment. Treatment of rodents with diets containing 20 mg/kg ivermectin or 5000 mg/kg spinosad caused 100% mortality of bloodfeeding sand flies for at least 1 week. The efficacy of ivermectin and spinosad also were not reduced when combined with the fluorescent tracer dye rhodamine B in a single diet. We also did not observe significant benefits by increasing the feeding period of the rodents from 3 to 6 or 9 days. We conclude that ivermectin and spinosad are effective as rodent systemic insecticides against bloodfeeding sand flies, and suggest that weekly treatment of wild rodent reservoirs of Leishmania major with bait containing one of these systemic insecticides could be a useful tool as part of a sand fly control program.

Oral treatment of rodents with insecticides for control of sand flies (Diptera: Psychodidae) and the fluorescent tracer technique (FTT) as a tool to evaluate potential sand fly control methods

ABSTRACT: In laboratory studies, insecticides (diflubenzuron, novaluron, methoprene and, pyriproxyfen) that have been incorporated into rodent diets were effective as feed-throughs against sand fly larvae. Novaluron also was effective against sand fly larvae at low concentrations and under simulated field conditions. Ivermectin has been shown to be effective as a systemic insecticide, killing 100% of blood-feeding sand flies for up to seven d after rodents were treated. The fluorescent tracer technique (FTT) is the use of certain fluorescent dyes (rhodamine B or uranine O) as feed-through transtadial biomarkers for phlebotomine sand flies, systemic biomarkers for blood-feeding sand flies, and permanent markers for nectar-feeding sand flies. The results of these laboratory studies provide proof of concept for the FTT and indicate that the FTT could be used to delineate specific foci with rodent/sand fly associations that would be susceptible to control by using feed-through or systemic insecticides, or foci where insecticide-treated sugar baits could be used against sand flies.

An Indirect Indentation Method for Evaluating the Linear Viscoelastic Properties of the Brain Tissue

Indentation experiments offer a robust, fast, and repeatable testing method for evaluating the mechanical properties of the solid-state materials in a wide stiffness range. With the advantage of requiring a minimal sample preparation and multiple tests on a small piece of specimen, this method has recently become a popular technique for measuring the elastic properties of the biological materials, especially the brain tissue whose ultrasoft nature makes its mechanical characterization very challenging. Nevertheless, some limitations are associated with the indentation of the brain tissue, such as improper surface detection, negative initial contact force due to tip-tissue moisture interaction, and partial contact between the tip and the sample. In this study, an indirect indentation scheme is proposed to overcome the aforementioned difficulties. In this way, the indentation force is transferred from a sharp tip to the surface of the tissue slices via a rigid coverslip. To demonstrate the accuracy of this method, the linear viscoelastic properties of the white and gray matters of the bovine brain samples are measured by imposing small cyclic loads at different frequencies. The rate, regional, directional, and postmortem time dependence of the viscoelastic moduli are investigated and compared with the previous results from cyclic shear and monotonic experiments on the brain tissue. While findings of this research present a comprehensive set of information for the viscoelastic properties of the brain at a wide frequency range, the central goal of this paper is to introduce a novel experimentation technique with noticeable advantages for biomechanical characterization of the soft tissue.

Cytotoxic and antiangiogenic paclitaxel solubilized and permeation-enhanced by natural product nanoparticles

Paclitaxel (PTX) is one of the most potent intravenous chemotherapeutic agents to date, yet an oral formulation has been problematic because of its low solubility and permeability. Using the recently discovered solubilizing properties of rubusoside (RUB), we investigated the unique PTX–RUB formulation. PTX was solubilized by RUB in water to levels of 1.6–6.3 mg/ml at 10–40% weight/volume. These nanomicellar PTX–RUB complexes were dried to a powder, which was subsequently reconstituted in physiologic solutions. After 2.5 h, 85–99% of PTX–RUB remained soluble in gastric fluid, whereas 79–96% remained soluble in intestinal fluid. The solubilization of PTX was mechanized by the formation of water-soluble spherical nanomicelles between PTX and RUB, with an average diameter of 6.6 nm. Compared with Taxol, PTX–RUB nanoparticles were nearly four times more permeable in Caco-2 cell monocultures. In a side-by-side comparison with dimethyl sulfoxide-solubilized PTX, PTX–RUB maintained the same level of cytotoxicity against three human cancer cell lines with IC50 values ranging from 4 to 20 nmol/l. In addition, tubule formation and migration of human umbilical vein endothelial cells were inhibited at levels as low as 5 nmol/l. These chemical and biological properties demonstrated by the PTX–RUB nanoparticles may improve oral bioavailability and enable further pharmacokinetic, toxicologic, and efficacy investigations.

Cardiovascular tolerance of intravenous lidocaine in broiler chickens (Gallus gallus domesticus) anesthetized with isoflurane

OBJECTIVE To determine the cardiovascular effects of lidocaine infused intravenously (IV) in broiler chickens. STUDY DESIGN Two phase study: Phase 1, randomized up-and-down study to determine effective dose 50 (ED50) for lidocaine; Phase 2, prospective randomized study to determine the cardiovascular effects of lidocaine. ANIMALS Seventeen Ross-708 broiler chickens (Gallus gallus domesticus) [11 chickens (Phase 1) and 6 chickens (Phase 2)], weighing 2.6-4.3 kg. METHODS After induction of anesthesia with isoflurane and placement of monitoring equipment including invasive blood pressure, chickens were administered lidocaine IV. During Phase 1, using an up-and-down design, each animal received a variable dose selected based on the response of the previous animal. During Phase 2, each animal was administered 6 mg kg(-1) of lidocaine IV over 2 minutes. Clinically irrelevant cardiovascular effects were defined as a relative decrease of heart rate (HR) and mean blood pressure (MAP) <30% subsequent to IV lidocaine administration. The ED50 was defined as the dose rate that would cause clinically irrelevant cardiovascular depression in 50% of the population. RESULTS During Phase 1, using an up-and-down study design (n = 11), the ED50 of lidocaine was determined to be 6.30 mg kg(-1) and 6.22 mg kg(-1) (95% confidence interval, 5.30-7.13 mg kg(-1)), when calculated by Dixons up-and-down method, and logistic regression, respectively. During Phase 2, following infusion of lidocaine (6 mg kg(-1)), no clinically relevant effects on HR or MAP were detected in any animal. CONCLUSIONS AND CLINICAL RELEVANCE Previous reports state that the dose of lidocaine used in birds should be ≤4 mg kg(-1). In this study, 6 mg kg(-1) of lidocaine injected IV was not associated with adverse cardiovascular effects. These results suggest that the dose of 4 mg kg(-1) can be exceeded, at least in chickens, and opens the possibility of other therapeutic uses for lidocaine in birds.

Oral Treatment of Rodents With Fipronil for Feed-Through and Systemic Control of Sand Flies (Diptera: Psychodidae)

ABSTRACT The sand fly Phlebotomus papatasi Scopoli is the vector of Leishmania major (Yakimoff & Schokhor), which is maintained in populations of burrowing rodents. The purpose of this study was to conduct a laboratory study to determine the efficacy of oral treatment of rodents with fipronil for control of sand flies that feed on rodent feces as larvae or on rodent blood as adults. We determined through larval bioassays that fipronil was eliminated in feces of orally-treated hamsters at a level that was significantly toxic to sand fly larvae for 21 d after the hamsters had been withdrawn from a fipronil-treated diet. Through bloodfeeding bioassays, we also found that fipronil was present in the peripheral blood of hamsters at a concentration that was significantly toxic to bloodfeeding adult female sand flies for 49 d after the hamsters had been withdrawn from their treated diet. The results of this study suggest that fipronil acts as well as or better than feed-through or systemic insecticides that previously have been measured against sand flies, and is particularly promising because this single compound acts against both larvae and bloodfeeding adults. An area-wide approach using rodent baits containing a fipronil could suppress vector populations that originate in the vicinity of rodent reservoirs, and could be used to eliminate the most epidemiologically important part of the vector population: female sand flies that take bloodmeals on rodent reservoirs.