A two compartment in vitro release model for the testing of HIV microbicide formulations

BACKGROUND: In vitro release testing of vaginal formulations is usually performed in a one-compartment model (OCM) where the release medium, usually comprising pH-adjusted water, an aqueous surfactant solution or a solvent-water solution, provides sink conditions throughout the release experiment. Although this model is useful in evaluating the effect of formulation parameters upon release, it rarely reflects in vivo conditions. Here we report use of a two-compartment diffusion cell model (TCM, comprising a small volume donor, a large volume receptor, and separated by a model epithelial membrane) to more closely mimic in vivo vaginal release and tissue absorption following administration of a UC781 vaginal ring.

METHODS: Macaque-sized matrix silicone elastomer vaginal rings containing 100mg UC781 were prepared by injection molding, and in vitro release testing performed using both OCM (20mL simulated vaginal fluid, SVF) and TCM (5mL SVF in donor cell and variable quantities of Tween 80; silicone elastomer membrane; 100mL 3:2 ethanol/water in receptor cell). In the TCM, drug levels were measured by HPLC in both donor and receptor cells, representing fluid and tissue levels respectively. Rings containing 100mg UC781 and 10% w/w Tween 80 were also manufactured and tested.

RESULTS: The amount of UC781 released from rings was significantly influenced by the choice of release model. Greatest release (56mg/14 days) was measured in the ethanol/water OCM, compared with no measurable release into SVF only. Increasing the concentration of Tween 80 in the SVF medium (1, 3 and 5% w/w) led to increased UC781 release (11, 16 and 18mg, respectively), demonstrating that vaginal fluid solubility of UC781 may be rate-determining in vivo. In the TCM, UC781 accumulates in the receptor cell medium over time, despite not being measured in the donor medium containing the ring device. Incorporation of Tween 80 directly into the ring provided enhanced release in both donor and receptor cells.

CONCLUSIONS: Release of UC781 was influenced by the choice of release medium and the inclusion of Tween 80 in the ring. Although use of SVF-only in the OCM indicated no measurable UC781 release from rings, data from the TCM confirms that UC781 is not only released but is also capable of penetrating across the model epithelial membrane. The TCM may therefore provide a more representative in vitro release model for mimicking in vivo absorption.

Testing nonlocal realism with entangled coherent states

We investigate the violation of nonlocal realism using entangled coherent states (ECSs) under nonlinear operations and homodyne measurements. We address recently proposed Leggett-type inequalities, including a class of optimized incompatibility inequalities proposed by Branciard et al. [Nature Phys. 4, 681 (2008)], and thoroughly assess the effects of detection inefficiency.

Building and testing age models for radiocarbon dates in Lateglacial and Early Holocene sediments

The growing importance of understanding past abrupt climate variability at a regional and global scale has led to the realisation that independent chronologies of past environmental change need to be compared between various archives. This has in turn led to attempts at significant improvements in the required precision at which records can be dated. Radiocarbon dating is still the most prominent method for dating organic material from terrestrial and marine archives, and as such many of the recent developments in improving precision have been aimed at this technique. These include: (1) selection of the most suitable datable fractions within a record, (2) the development of better calibration curves, and (3) more precise age modelling techniques. While much attention has been focussed oil the first two items, testing the possibilities of the relatively new age modelling approaches has not received much attention. Here, we test the potential for methods designed to significantly improve precision in radiocarbon-based age models, wiggle match dating and various forms of Bayesian analyses. We demonstrate that while all of the methods can perform very well, in some scenarios, caution must be taken when applying them. It appears that an integrated approach is required in real life dating situations where more than one model is applied, with strict error calculation, and with the integration of radiocarbon data with sedimentological analyses of site formation processes. (C) 2007 Elsevier Ltd. All rights reserved.

Hylambatin and (Thr)11-hylambatin from the skin secretion of the African hyperoliid frog, Kassina maculata: Structural characterisation, precursor cDNA cloning and preliminary pharmacological testing

The tachykinins hylambatin and (Thr)11-hylambatin have been isolated from the defensive skin secretion of the African hyperoliid frog, Kassina maculata,. Hylambatin (DPPDPNRFYGMMamide) is revised in structure from the original sequence by a single site substitution (Asn/Asp at position 6), and (Thr)11-hylambatin, a novel tachykinin, differs in structure from hylambatin by a single Thr/Met substitution. (Thr)11-hylambatin is five- to ten-fold more abundant than hylambatin in secretions. Synthetic replicates of both peptides were active in smooth muscle preparations including the rat tail artery, rat ileum and bovine trachea. While hylambatin displayed activity consistent with an NK1-receptor ligand, (Thr)11-hylambatin was more active than either substance P or neurokinin A in both NK1- and NK-2 receptor rich preparations. Incorporation of a threoninyl residue rather than the canonical leucyl residue at the penultimate position in both substance P and neurokinin A, generated active ligands in both arterial and intestinal smooth muscle preparations. Hylambatin precursor cDNAs, designated HYBN-1 and HYBN-2, respectively, were cloned from a skin library by 3'- and 5'-RACE reactions. Both were highly-homologous containing open-reading frames of 66 amino acids encoding single copies of either hylambatin or (Thr)11-hylambatin. These data reveal a hitherto unrecognized structure/activity attribute of mammalian tachykinin receptors revealed though discovery of a novel amphibian skin-derived, site-substituted peptide ligand.

Use of breakpoint combination sensitivity testing as a simple and convenient method to evaluate the combined effects of ceftazidime and tobramycin on Pseudomonas aeruginosa and Burkholderia cepacia complex isolates in vitro

An in vitro method of determining the activity of antibiotics in combination which is simple and convenient to perform and which could be used routinely in clinical microbiology laboratories is desirable. We investigated the activity, against Pseudomonas aeruginosa and Burkholderia cepacia complex clinical isolates, of ceftazidime and tobramycin in combination using a broth macrodilution sensitivity method based on breakpoint minimum inhibitory concentrations and compared the results obtained using this method with those obtained using the microtitre checkerboard method. There was good agreement in interpretation of results between the two methods for both P. aeruginosa (90%) and B. cepacia complex isolates (70%) with tobramycin and for P. aeruginosa isolates (70%) with ceftazidime. As the breakpoint combination sensitivity testing method employs only four tubes and does not require initial determination of individual antibiotic minimum inhibitory concentrations, it is simpler and more convenient for determining the activity of antibiotics in combination than the microtitre checkerboard method. The use of this method in routine microbiology laboratories to determine the activity of antibiotic combinations against clinical isolates should optimise treatment of infection by ensuring that appropriate antibiotic combinations are prescribed. (C) 2004 Elsevier B.V. All rights reserved.

Rapid colorimetric assay for antimicrobial susceptibility testing of Pseudomonas aeruginosa

A colorimetric assay based on the reduction of a tetrazolium salt {2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT)} for rapidly determining the susceptibility of Pseudomonas aeruginosa isolates to bactericidal antibiotics is described. There was excellent agreement between the tobramycin and ofloxacin MICs determined after 5 h using the XTT assay and after 18 h using conventional methods. The data suggests that an XTT-based assay could provide a useful method for rapidly determining the susceptibility of P. aeruginosa to bactericidal antibiotics.

Marketing secondhand goods in late medieval England

Purpose – The purpose of this paper is to identify the main practitioners, goods, customers and locations of secondhand marketing activities in late medieval England. It questions how important was the economic role played by such markets and what was the interaction with more formal market structures?

Design/methodology/approach – A broad range of evidence was examined, covering the period from 1200 to 1500: regulations, court rolls, wills, manorial accounts, literature, and even archaeology. Such material often provided mere scraps of information about marginal marketing activity and it was important to recognise the severe limitations of the evidence. Nevertheless, a wide survey of the available sources can give us an insight into medieval attitudes towards such trade, as well as reminding us that much marketing activity occurred beyond the reach of the surviving documentation.

Findings – Late medieval England had numerous outlets for secondhand items, from sellers of used clothes and furs who wandered the marketplaces to craftsmen who recycled and mended old materials. Secondhand marketing was an important part of the medieval makeshift economy, serving not only the needs of the lower sectors of society but also those aspiring to a higher status. However, it is unlikely that such trade generated much profit and the traders were often viewed as marginal, suspicious and even fraudulent.

Originality/value – There is a distinct lack of research into the extent of and significance of medieval secondhand marketing, which existed in the shadowy margins of formal markets and is thus poorly represented in the primary sources. A broad-based approach to the evidence can highlight a variety of important issues, which impact upon the understanding of the medieval English economy.