Review of data on the dermal penetration of mineral oils and waxes used in cosmetic applications.

Mineral oils and waxes used in cosmetic products, also referred to as "personal care products" outside the European Union, are mixtures of predominantly saturated hydrocarbons consisting of straight-chain, branched and ring structures with carbon chain lengths greater than C16. They are used in skin and lip care cosmetic products due to their excellent skin tolerance as well as their high protecting and cleansing performance and broad viscosity options. Recently, concerns have been raised regarding potential adverse health effects of mineral oils and waxes from dermal application of cosmetics. In order to be able to assess the risk for the consumer the dermal penetration potential of these ingredients has to be evaluated. The scope and objective of this review are to identify and summarize publicly available literature on the dermal penetration of mineral oils and waxes as used in cosmetic products. For this purpose, a comprehensive literature search was conducted. A total of 13 in vivo (human, animal) and in vitro studies investigating the dermal penetration of mineral oils and waxes has been identified and analysed. The majority of the substances were dermally adsorbed to the stratum corneum and only a minor fraction reached deeper skin layers. Overall, there is no evidence from the various studies that mineral oils and waxes are percutaneously absorbed and become systemically available. Thus, given the absence of dermal uptake, mineral oils and waxes as used in cosmetic products do not present a risk to the health of the consumer.

Impurity profiling of liothyronine sodium by means of reversed phase HPLC, high resolution mass spectrometry, on-line H/D exchange and UV/Vis absorption.

For the first time, a comprehensive investigation of the impurity profile of the synthetic thyroid API (active pharmaceutical ingredient) liothyronine sodium (LT3Na) was performed by using reversed phase HPLC and advanced structural elucidation techniques including high resolution tandem mass spectrometry (HRMS/MS) and on-line hydrogen-deuterium (H/D) exchange. Overall, 39 compounds were characterized and 25 of these related substances were previously unknown to literature. The impurity classification system recently developed for the closely related API levothyroxine sodium (LT4Na) could be applied to the newly characterized liothyronine sodium impurities resulting in a wholistic thyroid API impurity classification system. Furthermore, the mass-spectrometric CID-fragmentation of specific related substances was discussed and rationalized by detailed fragmentation pathways. Moreover, the UV/Vis absorption characteristics of the API and selected impurities were investigated to corroborate chemical structure assignments derived from MS data.

Levothyroxine sodium revisited: A wholistic structural elucidation approach of new impurities via HPLC-HRMS/MS, on-line H/D exchange, NMR spectroscopy and chemical synthesis.

The structural elucidation of unknown pharmaceutical impurities plays an important role in the quality control of newly developed and well-established active pharmaceutical ingredients (APIs). The United States Pharmacopeia (USP) monograph for the API Levothyroxine Sodium, a synthetic thyroid hormone, features two high pressure liquid chromatography (HPLC) methods using UV-VIS absorption detection to determine organic impurities in the drug substance. The impurity profile of the first USP method ("Procedure 1") has already been extensively studied, however for the second method ("Procedure 2"), which exhibits a significantly different impurity profile, no wholistic structural elucidation of impurities has been performed yet. Applying minor modifications to the chromatographic parameters of USP "Procedure 2" and using various comprehensive structural elucidation methods such as high resolution tandem mass spectrometry with on-line hydrogen-deuterium (H/D) exchange or two-dimensional nuclear magnetic resonance spectroscopy (NMR) we gained new insights about the complex impurity profile of the synthetic thyroid hormone. This resulted in the characterization of 24 compounds previously unknown to literature and the introduction of two new classes of Levothyroxine Sodium impurities. Five novel compounds were unambiguously identified via isolation or synthesis of reference substances and subsequent NMR spectroscopic investigation. Additionally, Collision-Induced Dissociation (CID)-type fragmentation of identified major impurities as well as neutral loss fragmentation patterns of many characterized impurities were discussed.

Rationale for the update of the European Pharmacopoeia general chapter 2.2.48. Raman spectroscopy.

Raman spectroscopy technique is widely used for the identification and characterisation of materials in the laboratory environment and, increasingly, for off-line, at-line, in-line and online Process Analytical Technologies (PAT) applications. The wide range of currently available Raman technologies had led to the requirement to update the existing General Chapter 2.2.48. The purpose of this reflection paper is to explain the rationale behind the establishment of the new wavenumber scale standards and peak position tolerances resulting from a multi-site, multi-instrument, inter-laboratory study of standards and, in particular, to discuss the results obtained with microscopes (including multi-point calibration) by the PAT Working Party.

MEK/Erk-based negative feedback mechanism involved in control of Steel Factor-triggered production of Krüppel-like factor 2 in mast cells.

The receptor tyrosine kinase, c-kit (Steel Factor (SF) receptor) controls survival, proliferation, chemotaxis, and secretion of proinflammatory cytokines in mast cells (MCs). Activation of c-kit results, amongst others, in induction of the PI3K and MEK/Erk pathways. Comparison of two MEK inhibitors, the specific, widely used U0126 and the more selective PD0325901, in different MC models revealed severe differences on SF-induced expression of proinflammatory cytokines IL-6 and TNF-α as well as the transcription factor Krüppel-like factor 2 (KLF2). Expression of the latter in MCs was not investigated so far. Whereas SF-induced expression of IL-6, TNF-α, and KLF2 was unaltered by U0126, it was significantly augmented by PD0325901. The effect of PD0325901 was corroborated by a second selective MEK inhibitor, PD184352 (Cl-1040), indicating the presence of MEK/Erk-based negative feedback mechanism(s) downstream of c-kit activation. Further analysis of KLF2 production revealed a positive function of PI3K. Depending on additional stimuli (e.g. antigen, IGF-1, LPS, thapsigargin), SF-triggered KLF2 expression was differentially modified, most likely controlled by the respective ratio between MEK/Erk and PI3K pathway activation. Moreover, the statin, simvastatin, was demonstrated to upregulate expression of KLF2 in MCs. In conclusion, data obtained by solely using the MEK inhibitor U0126 have to be carefully corroborated by using more selective inhibitors, such as PD0325901 or PD184352. SF-induced expression of the transcription factor KLF2 and its regulation by the MEK/Erk and PI3K pathways could impact on physiological as well as pathophysiological MC functions.

Special aspects of cosmetic spray safety evaluations: principles on inhalation risk assessment.

The consumer exposure to the vast majority of cosmetic products is limited to dermal contact. Even spray applications tend to be topically exposed to skin or hair. Besides this skin contact, spray products require additional considerations in regard to potential inhalation for building a robust and reliable safety assessment. Over the years, cosmetic industry developed prediction models for the best estimate of inhalation exposure combining data from computer simulation programs available in the market, individual real measured data and last but not least the experience from the market. Such attempt is driven by the toxicological profile of individual used ingredients. The focus of this review is on the determination of inhalation exposure, and the derivation of safe exposure levels for cosmetic spray products. Many of the methods employed to ensure product safety of cosmetic sprays in accordance with the general requirements of the EC Cosmetics Directive are based on industry experience which are not necessarily consistent across companies. This paper presents an approach to compile common principles for risk assessment and thus contribute to standardisation of safety assessment methodologies utilized for spray product evaluation without interfering with the flexibility of the individual safety assessor. It is based on the experience within the author's companies and may be useful as a support document as well for SME (Small and Medium Enterprises) companies safety assessors. In this respect it can be seen as one fundamental step in a tiered approach of cosmetic spray safety evaluation.

Bayesian prediction of breeding values by accounting for genotype-by-environment interaction in self-pollinating crops.

In self-pollinating populations, individuals are characterized by a high degree of inbreeding. Additionally, phenotypic observations are highly influenced by genotype-by-environment interaction effects. Usually, Bayesian approaches to predict breeding values (in self-pollinating crops) omit genotype-by-environment interactions in the statistical model, which may result in biased estimates. In our study, a Bayesian Gibbs sampling algorithm was developed that is adapted to the high degree of inbreeding in self-pollinated crops and accounts for interaction effects between genotype and environment. As related lines are supposed to show similar genotype-by-environment interaction effects, an extended genetic relationship matrix is included in the Bayesian model. Additionally, since the coefficient matrix C in the mixed model equations can be characterized by rank deficiencies, the pseudoinverse of C was calculated by using the nullspace, which resulted in a faster computation time. In this study, field data of spring barley lines and data of a 'virtual' parental population of self-pollinating crops, generated by computer simulation, were used. For comparison, additional breeding values were predicted by a frequentist approach. In general, standard Bayesian Gibbs sampling and a frequentist approach resulted in similar estimates if heritability of the regarded trait was high. For low heritable traits, the modified Bayesian model, accounting for relatedness between lines in genotype-by-environment interaction, was superior to the standard model.

Development and validation of a food frequency index using nutritional biomarkers in a sample of middle-aged and older adults.

BACKGROUND: An index of diet quality, which examines different aspects of a diet concurrently, may facilitate the identification of poor dietary habits in population sub-groups. The objectives of the present study were to develop a food frequency index (FFI) and to test its associations with nutritional biomarkers and nutrient intake. METHODS: The study comprised a cross-sectional survey among 444 adults aged 55 years and older in Vienna, Austria, and a sub-sample of 226 subjects who provided fasting blood specimen. Data from a qualitative 28-item food-frequency questionnaire were used to develop the FFI. RESULTS: FFI scores were positively correlated with plasma concentrations of beta-carotene (r = 0.26), beta-cryptoxanthin (r = 0.31), zeaxanthin (r = 0.19), lutein (r = 0.21), phylloquinone (r = 0.19), 25-hydroxyvitamin D (r = 0.20), and serum high-density lipoprotein (HDL)-cholesterol (r = 0.24) and were negatively correlated with the ratio of total : HDL-cholesterol (r = -0.23). Sub-groups with higher FFI scores had, on average, lower intakes of total fat, saturated fat, and dietary cholesterol and higher intakes of total carbohydrates, dietary fibre, and most of the examined micronutrients. CONCLUSION: The FFI as a measure of diet quality has the ability to discern population sub-groups, with reasonable validity, into low- or high-risk dietary habits.

The novel adaptor protein Swiprosin-1 enhances BCR signals and contributes to BCR-induced apoptosis.

B-cell receptor (BCR) signals are essential for B-cell differentiation, homeostasis and negative selection, which are regulated by the strength and quality of BCR signals. Recently, we identified a new adaptor protein, Swiprosin-1, in lipid rafts of B-cell lines that undergo apoptosis after BCR stimulation. During murine B-cell development, Swiprosin-1 exhibited highest expression in immature B cells of the bone marrow, but was also expressed in resting and activated splenic B cells and in non-lymphoid tissue, especially in the brain. Ectopic expression of Swiprosin-1 in the immature murine B-cell line WEHI231 enhanced spontaneous and BCR-induced apoptosis. In contrast, short hairpin RNA (shRNA)-mediated downregulation of Swiprosin-1 impaired specifically spontaneous and BCR-elicited apoptosis, but not BCR-induced G1 cell cycle arrest and upregulation of the cell cycle inhibitor p27(Kip1). In accordance, Swiprosin-1 abundance regulated net cell growth of WEHI231 cell populations through reciprocal regulation of Bcl-xL, but not Bim, thereby controlling spontaneous apoptosis. Swiprosin-1-enhanced apoptosis was blocked through nuclear factor kappaB-activating stimuli, namely B-cell-activating factor of the TNF family, anti-CD40 and lipopolysaccharide (LPS). This correlated with enhanced BCR-induced IkappaB-alpha phosphorylation and degradation in cells expressing a Swiprosin-1-specific shRNA. Finally, ectopic Swiprosin-1 expression enhanced BCR-induced cell death in primary, LPS-stimulated splenic B cells. Hence, Swiprosin-1 may regulate lifespan and BCR signaling thresholds in immature B cells.

Infrainguinal vein graft surveillance: the criteria for intervention.

In most patients, infrainguinal vein grafts are commonly performed for limb-threatening ischemia. Once limb salvage has been achieved, most clinicians agree that regular appointments with the vascular team are important to monitor graft patency. However, the timing of these appointments and what defines an adequate examination remains debatable. The following cases illustrate the natural history and interventions performed in 2 such patients. A review of the literature and a commentary on surveillance strategies is provided to elucidate the advantages and disadvantages of currently available vascular laboratory testing. Recommendations also are made for the appropriate timing of intervention to revise these grafts.

Observation of a P/M interconversion of a gold-phosphine helicate via 31P NMR.

Ligands containing P-CH2-CH2-P elements have been shown to form double-stranded helicates whose axis consists of gold atoms with Au...Au contacts; the interconversion of the P and M forms of the helicate [Au3(mu-pp2)2](OTf)3 (pp2 = PhP(CH2CH2PPh2)2) is monitored via 31P NMR.

Regulation of CD95 expression and CD95-mediated cell death by interferon-gamma in acute lymphoblastic leukemia with chromosomal translocation t(4;11).

The regulatory effects of IFNgamma on CD95 expression and CD95-mediated cell death were investigated in three high-risk pro-B acute lymphoblastic leukemia (ALL) lines that carry the chromosomal translocation t(4;11)(q21;q23). These leukemias are characteristically refractory to conventional chemotherapeutic treatments operating through the induction of apoptosis. However, the mechanisms leading to increased cell survival and resistance to cell death in these leukemias are largely unknown. Interferon-gamma (IFNgamma), a potent inhibitor of hematopoiesis, acts in part by upregulating CD95 and sensitizing cells to CD95-induced apoptosis. The t(4;11) lines SEM, RS4;11, and MV4;11 expressed low levels of CD95, but were completely resistant to CD95-mediated death. Addition of IFNgamma markedly upregulated CD95 expression in SEM (8-9-fold), RS4;11 (2-3-fold), and MV4;11 (2-3-fold) lines. However, after treatment with IFNgamma, only an 11% increase in sensitivity to CD95-mediated cell death was observed in SEM cells, whereas RS4;11 and MV4;11 cells remained resistant. Cycloheximide, but not actinomycin D or brefeldin A, increased CD95-specific cell death only in IFNgamma-treated RS4;11 cells by approximately 12%. Abundant levels of Bcl-2 and Bcl-XL, known to inhibit CD95-signaling in some cells, were present suggesting a possible role for both molecules in the resistance to CD95-mediated cell death. Resistance of the leukemic blasts to CD95-mediated cell death and the failure of IFNgamma to substantially sensitize the CD95-signaling pathway may contribute to the highly malignant phenotype of pro-B ALL with translocation t(4;11).

Cultured composite autografts as coverage for an extensive body surface area burn: case report and review of the technology.

Cultured epithelial autografts (CEA) have been used as an adjunct in the surgical management of extensive thermal burns. Unfortunately, the lack of a dermal matrix makes CEA susceptible to infection, shearing forces and limits their incorporation into the burn wound. A cultured composite autograft (CCA) has been developed in which autologous keratinocytes and fibroblasts are surgically harvested from the burn patient's normal skin. These components are proliferated and then combined to form an epidermal and dermal matrix, grown to confluence then applied. Standard wound coverage techniques as well as CCA technology were utilized for successful wound closure in a 12 yr-old female with an 81% third degree burn. After fascial excision and allograft coverage, autografts were placed on her posterior burns and then 7500 cm2 of CCA was placed onto her anterior thorax, abdomen and lower extremities. Sixty percent of the burn was covered with CCA resulting in a success rate of 40%. No evidence of infection was noted, even in areas where CCA failed, although in those areas random epithelialization appeared to occur which then seemed to facilitate autograft placement. Early debridement and allografting followed by conventional autografts and CCA placement may provide an effective skin coverage strategy in patients with extensive deep burns.

Studies on reproductive toxicity of iloprost in rats, rabbits and monkeys.

A reproduction toxicological test program was performed with the carbaprostacyclin derivative iloprost, an analogue to the endogenous prostacyclin PGI2, in order to detect possible effects on fertility and reproductive performance, on preimplantational, embryonal and fetal development, on delivery as well as on lactation and postpartum development. While in humans iloprost is administered as an i.v. infusion for 6 h/day, it was administered i.v. to rats, rabbits and monkeys by continuous infusion with a subcutaneously implanted pump. No influence on mating or reproductive parameters was found after treatment of male or female rats during the premating phase up to day 7 post coitum (p.c.). Embryonal and fetal development were not remarkably impaired in rabbits or monkeys after treatment throughout the period of organogenesis. The only remarkable observations in the embryotoxicity and peri-/postnatal studies in the rat were defects on the digits (reductions of phalangeal structures) in single individuals. These malformations were interpreted as resulting from a compound-related hypotonia with subsequent change in the regional blood flow and the consequence of temporary impairments of placental blood supply leading to hypoxia in the affected structures.

Evaluation of the neurotoxic potential of chemicals in animals.

This review provides a scientific view on how to evaluate effectively the neurotoxic potential of chemicals in order to provide adequate safeguards for human health. Detection of compounds that may cause direct, persistent, adverse effects on the nervous system should be given the most critical attention. Evaluation of the neurotoxic potential of a chemical should include descriptions of functional and morphological effects as well as the determination of the dose response, no-observed-effect level, time course and reversibility of effects. Evaluation of the nervous system in the context of standard toxicity studies that use a variety of species and study durations are appropriate screening tests (Tier 1) for the detection of potential neurotoxicity. Studies specifically designed to assess neurotoxicity (Tier 2) should be performed with chemicals for which there is an indication of neurotoxic potential and where the available data are inadequate for risk assessment. Tier 2 studies should evaluate the function and structure of the nervous system by comprehensive clinical examinations and neuropathological assessment. These studies may be conducted in conjunction with standard toxicity studies so that any potential neurotoxicity can be interpreted in the context of other systemic toxicity. More specific neurotoxicity tests (Tier 3) may be necessary for advanced characterization of discovered neurotoxicants.