The effect of an amphiphilic self-assembled lipid lamellar phase on the relief of dry skin.

Humectant and occlusive technologies have traditionally been used for the treatment of dry skin. Originally, non-lamellar-forming ingredients were used such as petrolatum but recent research has shown the advantage of using lamellar-forming ingredients such as ceramides, pseudoceramides and phospholipids in the relief of dry skin. Nevertheless, the importance of using lipid-phase transition inducers, such as long-chain fatty acids, has not been studied clinically. The evaluation of a novel complex of lipophilic ingredients was of interest: cetyl alcohol, isostearyl isostearate, potassium cetyl phosphate, cetyl behenate and behenic acid. The combination of all these ingredients was shown to be more effective than any single component in water vapour transmission rate studies. This was thought to be owing to the formation of a unique structural organization of the lipids upon dry-down from an O/W emulsion as was examined by X-ray diffraction and optical microscopy. When evaluated clinically in a randomized double-blind and vehicle-controlled moisturization efficacy trial, this novel blend of ingredients was shown to not only improve the visible signs of skin dryness to a significantly greater extent than a comparable mineral oil-containing vehicle but also then maintain a better skin condition during the regression no-treatment phase of the study. This combination of ingredients offers a new technology option for the treatment of dry skin.

Superior effect of isostearyl isostearate on improvement in stratum corneum water permeability barrier function as examined by the plastic occlusion stress test.

SYNOPSIS: Dry skin is a major dermatological problem and consumer research indicates that although current moisturizers are effective they are not completely meeting consumer expectation. Several technological approaches have been taken but influencing stratum corneum (SC) lipid phase behaviour as a novel water permeability barrier-enhancing and moisturizing mechanism has only been started to be investigated recently. Both the long periodicity SC lipid lamellar phase and the orthorhombic lipid packing state have been proposed to define optimal SC water permeability barrier properties. Several lipophillic moisturizers have been tested for their ability to modify SC lipid lateral packing namely glyceryl monoisostearate (GMIS), isopropyl isostearate (IPIS) and isostearyl isostearate (ISIS) of which IPIS and ISIS are reported to induce the orthorhombic phase. Despite the improvements in the lateral packing of SC lipids, these ingredients have been shown not to improve transepidermal water loss. However, using a novel skin surface water loss method we have observed for the first time significant improvements in SC water permeability barrier function for ISIS compared with IPIS, GMIS and petrolatum. However, using synthetic membranes and measuring water vapour transport rates we showed that the isostearyl esters were not occlusive like petrolatum. As the effects of ISIS were not because of what would be considered as true occlusion, we propose that the differences in the SC water permeability barrier properties from use of ISIS to the other ingredients tested are because of its reported effects on SC lipid phase behaviour. Further studies probably using spectroscopic approaches, however, will be needed to specifically test this hypothesis in vivo.

Voriconazole therapeutic drug monitoring in allogeneic hematopoietic stem cell transplant recipients.

Voriconazole, a new antifungal agent, is increasingly being used after HSCT. The hepatic cytochrome P450 isoenzyme 2C19 plays a significant role in voriconazole metabolism. As CYP2C19 exhibits significant genetic polymorphism, some patients metabolize voriconazole poorly resulting in increased plasma drug levels. The clinical significance of this is unknown, and the utility of monitoring voriconazole levels is unclear. Steady-state trough plasma voriconazole levels were obtained in 25 allogeneic HSCT recipients using an HPLC assay. Patients had drug levels checked once (n=13), twice (n=10), or > or =3 times (n=2) 5-18 days (median 10) after starting voriconazole or dose modification. The 41 voriconazole levels were 0.2-6.8 microg/ml (median 1.6); 6 (15%) were <0.5 (possibly below the in vitro MIC90 for Aspergillus spp.). Voriconazole concentrations correlated with aspartate aminotranferase (AST) (r=0.5; P=0.0009) and alkaline phosphatase (r=0.34; P=0.03), but not with creatinine, bilirubin and alanine aminotransferase (ALT). Since liver dysfunction is common after HSCT, it was not possible to determine if elevated AST and alkaline phosphatase levels were the cause or the consequence of higher voriconazole levels. We conclude that trough voriconazole levels vary considerably between patients, and suggest monitoring levels in patients receiving voriconazole for confirmed fungal infections, and in those with elevated AST or alkaline phosphatase levels.

Candida krusei renal cyst infection and measurement of amphotericin B levels in cystic fluid in a patient receiving AmBisome.

Candida krusei is an opportunistic pathogen commonly implicated in urinary tract infections in immunocompromised patients. We present the first case of C. krusei renal cyst infection, occurring in a post-liver and kidney transplant patient with autosomal dominant polycystic kidney disease. Her persistent candiduria and fevers were refractory to prolonged therapy with AmBisome (Fujisawa Pharmaceuticals Co. Ltd., Osaka, Japan). She eventually required bilateral nephrectomies of her native kidneys. Cystic fluid was aspirated from six hemorrhagic and six nonhemorrhagic cysts. Cystic fluid cultures yielded C. krusei. Fluid from the nonhemorrhagic cysts was also analyzed for amphotericin B levels, measured using a bioassay. Free amphotericin B levels in the cysts were lower than the minimal inhibitory concentration for amphotericin B for this organism. We provide the first description of amphotericin B levels in cystic fluid obtained during bilateral nephrectomies.

Comparison of high-performance liquid chromatographic and microbiological methods for determination of voriconazole levels in plasma.

A new selective high-performance liquid chromatography (HPLC) method with UV detection for the determination of the investigational triazole voriconazole in human plasma by using acetonitrile precipitation followed by reverse-phase HPLC on a C(18) column was compared with a simple agar well diffusion bioassay method with Candida kefyr ATCC 46764 as the assay organism. Pooled plasma was used to prepare standard and control samples for both methods. The results of analyses with spiked serum samples (run as unknowns) were concordant by the bioassay and HPLC methods, with expected values being obtained. HPLC demonstrated an improved precision (3.47 versus 12.12%) and accuracy (0.81 versus 1.28%) compared to those of the bioassay method. The range of linearity obtained by both methods (from 0.2 to 10 microg/ml for HPLC and from 0.25 to 20 microg/ml for the bioassay) includes the range of concentrations of voriconazole (from 1.2 to 4.7 microg/ml) which are considered clinically relevant. Although either methodology could be used for the monitoring of patient therapy, the smaller variability observed with HPLC compared to that observed with the bioassay favors the use of HPLC for pharmacokinetic studies.

Amphotericin B enzyme-linked immunoassay for clinical use: comparison with bioassay and HPLC.

OBJECTIVE: To evaluate a new enzyme-linked immunosorbent assay (ELISA) for amphotericin B in serum samples. Results are compared with those obtained by HPLC and bioassay. DESIGN: Comparison of results obtained by ELISA, HPLC, and bioassay. METHODS: We developed a new ELISA using a polyclonal rabbit antibody to measure serum amphotericin B concentrations. Blinded samples of amphotericin B in concentrations of 0.15-78 micrograms/mL were prepared in human serum and assayed simultaneously by the ELISA, HPLC, and bioassay. The results of each assay were derived from standard curves and evaluated by using the Table Curve 2D computer program. These data were compared by using correlation analysis with evaluation of Pearson's correlation coefficient by Student's t-test. RESULTS: ELISA and bioassay compared favorably at amphotericin B concentrations of 0.3-20 micrograms/mL with a correlation coefficient of r = 0.993, while ELISA and HPLC compared with a correlation coefficient of r = 0.944. The average coefficient of variation over the range 0.3-20.0 micrograms/mL was 28% +/- 7% for HPLC, 26% +/- 9% for ELISA, and 13% +/- 4% for bioassay. Comparison of all three assays revealed the highest correlation with the ELISA assay (r = 0.998) for the range of concentrations (0.3-20 micrograms/mL) routinely achieved. Samples containing concentrations in excess of 20 micrograms/mL could be diluted. Desiccation for concentrations less than 0.3 microgram/mL was not tested. CONCLUSION: The determination of serum amphotericin B concentrations by ELISA gave results similar to those obtained by a bioassay and HPLC technique. Although variability appears greater with ELISA, the ease of performing yjis assay expedites the evaluation of amphotericin B concentrations from lipid formulations without interference from coadministered antibacterials of azole antifungals.