Autoradiographic and scintillation analysis of 5-aminolevulinic acid permeation through epithelialised tissue: Implications for topical photodynamic therapy of superficial gynaecological neoplasias

Purpose. Aminolevulinic acid (5-ALA) diffusion through both keratinised and non-keratinised tissue, used as a model tissue substrates, was evaluated, together with the depth of permeation and the concentration achieved following delivery from bioadhesive patch and proprietary cream formulations. Materials and Methods. Moisture-activated, bioadhesive patches loaded with 5-ALA at concentrations of 19.0, 38.0 and 50.0 mg cm(-2) and an o/w cream (20% w/w 5-ALA) were radiolabelled with C14 5-ALA and applied to excised human vaginal tissue and porcine skin. After 1, 2 and 4 h, tissue was sectioned in two orientations and the 5-ALA concentration at specific depths determined using autoradiography and liquid scintillation counting (LSC). Results. The stratum corneum was a significant barrier to 5-ALA permeation, with concentrations in tissue dependent on application time and drug loading. 5-ALA was detected at 6 mm using autoradiography after 2 h, with LSC showing phototoxic concentrations at 2.375 mm after 4 h of application. Inclusion of oleic acid and dimethyl sulphoxide in bioadhesive patches increased 5-ALA significantly in neonate porcine tissue, but only for patches cast from blends containing 5% w/w oleic acid. Conclusions. The bioadhesive patch described delivered 5-ALA to depths of at least 2.5 mm in tissue types indicative of vulval skin, suggesting that photodynamic therapy of deep vulval intraepithelial neoplasia is feasible using this means of bioadhesive 5-ALA delivery.

The effect of triclabendazole ("Fasinex") on protein synthesis by the liver fluke, Fasciola hepatica

The effect of the active sulphoxide metabolite of the anthelmintic triclabendazole (TCBZ-SX, 15-50 mu g ml(-1)) on the incorporation of radioactively labelled [C-14] leucine by adult Fasciola hepatica tissue slices was measured by liquid scintillation counting. In addition, the ability of the microfilament-disrupting drug, cytochalasin B, and the microtubule-disrupting drug, tubulozole-C, to inhibit protein synthesis, was assessed by similar methods and compared with TCBZ-SX. The established protein synthesis inhibitors, cycloheximide and actinomycin D were used as positive controls. All the drugs showed a significant inhibition of protein synthesis, albeit to different extents; however, TCBZ-SX was the most potent, with no significant difference between its effect and that of cycloheximide or actinomycin D. Moreover, the concentration of TCBZ-SX, above 15 mu g ml(-1), had little further influence on incorporation of [C-14] leucine. This investigation demonstrates the inhibitory effect of TCBZ-SX, cytochalasin B and tubulozole-C on protein synthesis in F. hepatica and confirms the qualitative observations made in several previous ultrastructural studies.

THE EFFECT OF DIAMPHENETHIDE ON PROTEIN-SYNTHESIS BY THE LIVER FLUKE, FASCIOLA-HEPATICA

The effect of the deacetylated (amine) metabolite of diamphenethide (DAMD, 10 mug ml-1) on the uptake and incorporation by adult Fasciola hepatica of radioactively labelled precursors of DNA, RNA and protein synthesis ([H-3]thymidine, [H-3]uridine and [H-3]leucine, respectively) was measured by liquid scintillation counting. Comparison was made between the effects of DAMD and those of specific inhibitors of DNA, RNA and protein synthesis, namely, 5-fluorouracil, cordycepin and cycloheximide, respectively. DAMD caused a significant decrease in the overall uptake and incorporation of [H-3]uridine by F. hepatica, decreased the incorporation of [H-3]leucine and also caused a significant decrease in the overall protein content of the flukes. The effect of DAMD was similar to that of cycloheximide (I x 10(-3) M), a potent inhibitor of protein synthesis, which also caused a significant decrease in the incorporation of [H-3]leucine by the fluke and a decrease in the overall protein content of the fluke. Cordycepin(100 mug ml-1) caused a significant decrease in the protein content of the fluke, but had no effect on the uptake or incorporation of [H-3]uridine. 5-Fluorouracil (I x 10(-4) m) did not affect the uptake or incorporation of VH]thymidine, nor did it decrease the protein content of the fluke. The results indicate that DAMD inhibits protein synthesis by F. hepatica, possibly by inhibiting RNA synthesis. The results are also consistent with previous morphological investigations involving DAMD.

Time of Flight Measurements for Neutrons Produced in Reactions Driven by Laser-Target Interactions at Petawatt level

Abstract Short intense pulses of fast neutrons were produced in a two stage laser-driven experiment. Protons were accelerated by means of the Target Normal Sheath Acceleration (TNSA) method using the TITAN facility at the Lawrence Livermore National Laboratory. Neutrons were obtained following interactions of the protons with a secondary lithium fluoride (LiF) target. The properties of the neutron flux were studied using BC-400 plastic scintillation detectors and the neutron time of flight (nTOF) technique. The detector setup and the experimental conditions were simulated with the Geant4 toolkit. The effects of different components of the experimental setup on the nTOF were studied. Preliminary results from a comparison between experimental and simulated nTOF distributions are presented.

Chromatography- High Performance Liquid Chromatography

High-performance liquid chromatography (HPLC) is a major analytic tool in contemporary science, with possibly the highest number of systems installed and running globally. Modern HPLC offers high resolutions allowing the quantitative determination of target analytes within complex matrices by its compatibility with a number of detectors. The article describes the major technological characteristics of HPLC, reviewing separation mechanisms and their application in health and food science. Separation modes and media, key instrumental parameters, compatibility with detection modes, and applications are briefly discussed, aiming to provide helpful hints to the reader in the search for appropriate analytic techniques for a given task.