Quantification of corticosteroid-induced skin vasoconstriction: visual ranking, chromameter measurement or digital image analysis

Topical corticosteroid formulations have been evaluated by visual grading protocols for many years. Toward a more objective methodology, several instrumental methods have been evaluated for applicability in quantifying the vasoconstriction side-effect that follows corticosteroid application to the skin. Although the chromameter has been adopted by regulatory bodies throughout the world as the current standard for topical bioequivalence determinations, there is considerable criticism of this instrument from several quarters. A preliminary comparison reported here indicates that digital image analysis provides statistically significant results that are similar to those obtained by visual assessment techniques, and shows considerably greater precision than that obtained by the chromameter. Continued evaluation of objective assessment techniques, such as digital imaging, and continued modernisation of regulatory bioequivalence requirements will assist in protecting patients and optimising clinical results

The use of supersaturated solutions for the percutaneous delivery of rooperol tetra-acetate

A major problem encountered in the transdermal delivery of drugs is the effectiveness of the barrier system imposed by the stratum corneum.To overcome tbe resistance of the skin to the ingress of exogenous chemicals, numerous innovative techniques requiring complex delivery systems have been studied. Many of these systems attempt to alter the barrier potential by the use of enhancer technology. Supersaturation, on the other hand, is a simple and economical technique which is not intended to modify the physical structure or the chemical composition of the stratum corneum, yet may effectively deliver a markedly greater mass of drug to the skin than that achieved by the use of conventional, saturated solutions. Supersaturated systems make use of the elevated thermodynamic activity of the permeant in the delivery vehicle, which results in higher flux rates across the contacting membrane by increasing the concentration gradient. The present study investigated the potential for using supersaturation techniques to transdermally deliver rooperol tetra-acetate (RTA), a lipophilic, cytotoxic agent with potential for use in the treatment of solar keratosis. The diffusion characteristics of the drug from a 60% propylene glycol/water supersaturated solution across silicone membrane and full thickness rat skin were studied using Franz diffusion cells. A comparison was made of the drug diffusion rates from a saturated system and from supersaturated systems prepared with and without an antinucleating agent

Bioequivalence testing of topical dermatological formulations, the gap between science and legislation

Bioavailability concerns for topical dermatological products are complex and it is especially difficult to determine the bioequivalence of similar topical formulations. Since only small amounts of drug dispersed in an appropriate vehicle are applied to the skin, the amount of drug that actually reaches the systemic circulation is often too small to be easily quantified. Additionally, it can be argued that the relevance of any serum/plasma concentration-time curve of a topical agent is questionable, since the curve reflects the amount of drug after the active moiety has left the site of action. For some topical drugs e.g., topical corticosteroids, it is possible to perform a pharmacodynamic bioassay to obtain acceptable bioequivalence data. In this case, the intensity of the side effect of blanching (vasoconstriction) in the skin caused by topical corticosteroids can be measured. The response is directly proportional to the clinical efficacy, and the skin blanching assay has proved to be a reliable procedure for the determination of topical corticosteroid bioavailability. Recently, we had sight of the results of a topical bioequivalence study, which was conducted for the registration of a new generic corticosteroid cream formulation. In this trial the new formulation was compared to two equivalent product from the local market and bioequivalence was demonstrated by the investigators for all three products. These results were examined with interest as the respective reference products have been used repeatedly as standard formulations in our laboratory. However, one of these reference formulations has consistently shown superior bioavailability in our trials, but was not demonstrated to be superior in the study results examined. In the present publication an overview of topical bioequivalence testing in general is given and the difficulties occurring in practice, for topical corticosteroid formulations in particular, are demonstrated

Chromametry: measuring precision of diurnal and local variation of human forearm skin colour

Chromameters are compact portable instruments used for the assessment of surface colour based on the tristimulus analysis of a reflected xenon light pulse, and have been used for the quantification of erythema in the study of irritant dermatitis, and corticosteroid-induced skin blanching in the vasoconstriction assay. The variability and the reproducibility of chromameter results were investigated since it is known that the location and application force of the measuring head on the skin and the orthostatic maneuver of the arms influence the colour measurement. Furthermore the diurnal variation and the homogeneity of forearm skin colour were investigated

Analysis of chromameter results obtained from corticosteroid-induced skin blanching assay: comparison of visual and chromameter data

In a Guidance document, the American FDA recommends the use of a Minolta chromameter rather than the human eye for the quantitative assessment of the pharmacodynamic blanching response produced by topical application of corticosteroids. The purpose of this study was to compare the appropriateness of the human eye and two models of chromameter for the estimation of skin blanching, in terms of the quality of the data generated by each method. The corticosteroid-induced skin blanching from four different betamethasone 17-valerate cream formulations was compared in a typical human skin blanching trial. The optimized assay methodology routinely practised in our laboratories was utilized. The blanching responses were assessed visually by three trained, independent observers and recorded by two chromameters (Minolta model CR-200 and model CR-300). The topical availability of the four creams was determined using visual scoring and chromameter measurements. All data were manipulated in such a manner as to produce a blanching response versus time profile from which AUBC analysis could be performed. Good correlation was observed between the visual assessments made by three independent observers. In contrast, moderate correlation was determined between visual, CR-200 and CR-300 measurements. Surprisingly, no direct linear relationship between the AUBCs produced by the two chromameters was observed indicating that the quality of the data obtained from the two instruments may not be equal. This investigation also indicated that the use of the chromameter is not completely objective. Visual scoring and chromameter measurement produce data sets that differ in quality. Each procedure needs to be validated and investigators have to be trained for both visual assessment and the operation of the chromameter, particularly with regard to the manipulation of the measuring head of the instrument

Rationalisation of Relative »Ring-Current« Sizes in Polycyclic, Conjugated Hydrocarbons

A theoretical basis is established for relating the incidence of relatively \u27high\u27 and \u27low\u27 »ring-current« intensities (as encountered, for example, in the central rings of peropyrene (I) and perylene (II), respectively) to intuitive Valence-Bond Resonance- Theory (VB-RT) ideas about \u27bond fixation\u27. In the present treatment, this aim is not achieved by devising a »ring-current« formalism that is itself actually based on a VB-RT wave-function: the philosophy adopted here is (a) to identify, and classify, those occasions on which simple Molecular-Orbital (MO) theory predicts specific rings in conjugated hydrocarbons to have exceptionally high and low »ring-currents«, and then (b) to use topological arguments that rely simply on the carbon-atom connectivity of the conjugated system in question to show that the situations identified in (a), above, are just those in which the absence, or presence, of VB-RT \u27bond-fixation\u27 is to be expected

CD4+ T-cell responses to Epstein-Barr virus (EBV) latent-cycle antigens and the recognition of EBV-transformed lymphoblastoid cell lines

There is considerable interest in the potential of Epstein-Barr virus (EBV) latent antigen-specific CD4$^+$ T cells to act as direct effectors controlling EBV-induced B lymphoproliferations. Such activity would require direct CD4$^+$ T-cell recognition of latently infected cells through epitopes derived from endogenously expressed viral proteins and presented on the target cell surface in association with HLA class II molecules. It is therefore important to know how often these conditions are met. Here we provide CD4$^+$ epitope maps for four EBV nuclear antigens, EBNA1, -2, -3A, and -3C, and establish CD4$^+$ T-cell clones against 12 representative epitopes. For each epitope we identify the relevant HLA class II restricting allele and determine the efficiency with which epitope-specific effectors recognize the autologous EBV-transformed B-lymphoblastoid cell line (LCL). The level of recognition measured by gamma interferon release was consistent among clones to the same epitope but varied between epitopes, with values ranging from 0 to 35% of the maximum seen against the epitope peptide-loaded LCL. These epitope-specific differences, also apparent in short-term cytotoxicity and longer-term outgrowth assays on LCL targets, did not relate to the identity of the source antigen and could not be explained by the different functional avidities of the CD4$^+$ clones; rather, they appeared to reflect different levels of epitope display at the LCL surface. Thus, while CD4$^+$ T-cell responses are detectable against many epitopes in EBV latent proteins, only a minority of these responses are likely to have therapeutic potential as effectors directly recognizing latently infected target cells

XTE J0111.2-7317 : a nebula-embedded X-ray binary in the SMC

The observed characteristics of the nebulosity surrounding the SMC High Mass X-ray Binary XTE J0111.2-7317 are examined in the context of three possible nebular types: SNR, bowshock and HII region. Observational evidence is presented which appears to support the interpretation that the nebulosity surrounding XTE J0111.2-7317 is an HII region. The source therefore appears to be a normal SMC Be X-ray binary (BeXRB) embedded in a locally enhanced ISM which it has photoionised to create an HII region. This is supported by observations of the X-ray outburst seen with BATSE and RXTE in 1998-1999. It exhibited characteristics typical of a giant or type II outburst in a BeXRB including large spin-up rates, Lx~10E38 erg/sq.cm-s, and a correlation between spin-up rate and pulsed flux. However, the temporal profile of the outburst was unusual, consisting of two similar intensity peaks, with the first peak of shorter duration than the second.Comment: Accepted for publication by MNRA