Predictivity of an in vitro model for acute and chronic skin irritation (SkinEthic) applied to the testing of topical vehicles

An in vitro human reconstructed epidermis model (SkinEthic) used for screening acute and chronic skin irritation potential was validated against in vivo data from skin tolerability studies. The irritation potential of sodium lauryl sulfate (SLS), calcipotriol and trans-retinoic acid was investigated. The in vitro epidermis-like model consists of cultures of keratinocytes from human foreskin on a polycarbonate filter. The modulation of cell viability, the release and gene expression of proinflammatory cytokines, interleukins 1α and 8, and morphological changes were evaluated during 3 days as endpoints representative for an inflammatory reaction. The cumulative irritation potential of the topical products was evaluated in a human clinical study by visual scoring and biophysical measurement of inflammatory skin reaction after repeated 24 h applications over 3 weeks under Finn chamber patches. All topical products that were nonirritating in the human study were noncytotoxic and did not induce cytokine expression in the in vitro acute model (day 1 exposure). All irritating controls exhibited specific cell viability and cytokine patterns, which were predictive of the in vivo human data. The ranking of mild to moderate skin irritation potential was based on the lack of cytotoxicity and the presence of cytokine patterns including gene expression specific for each irritant, using the chronic in vitro model (up to 3 days exposure). The human reconstructed epidermis model SkinEthic was shown to be a reliable preclinical tool predicting the irritation potential of topical products. Moreover, it is a useful model in a two-step tiered strategy for screening acute and chronic irritation potential for the selection of vehicles for new topical drugs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Purification and immunohistochemical localization of the auxin binding site i protein from maize coleoptiles

An auxin binding protein fraction prepared by means of affinity chromatography on 2-OH-3,5-diiodobenzoic acid-Sepharose and gel filtration was used as antigen. The obtained rabbit antisera contained antibodies against the auxin, binding protein (ABP) and several contaminating proteins (nonABP). The nonABP could be separated on an appropriate affinity matrix omitting the TIBA analogue. After their immobilization on Sepharose antibodies directed towards contaminating, the proteins were isolated and immobilized, too. This IgGanti nonABP-Sepharose retains almost all contaminating proteins present in the specific eluates of the auxin affinity matrix. In a final affinity chromatography step on IgG-Sepharose a highly purified ABP could be eluted. This ABP was immobilized on Sepharose for the separation of monospecific antibodies against ABP (IgGanti abp). Using these antibodies the ABP could be localized within the outer epidermal cells of the coleoptile by immunofluorescence microscopy. From the inhibition of auxin induced elongation of coleoptile tissue by IgGanti abp it is concluded that the ABP is localized at the plasmalemma of the epidermal cells and that the ABP is involved in auxin action as a true hormone receptor. Presented at the International Symposium “Plant Growth Regulators” held on June 18–22, 1984 at Liblice, Czechoslovakia.     

Dynamic stem cell selection safeguards the genomic integrity of the epidermis

1. Download : Download high-res image (230KB)
2. Download : Download full-size image

     

Comparison of microridges in juvenile and adult sunfish,Lepomis gibbosus

Microridges are F-actin-based surface protrusions of the superficial layer cells of fish epidermis. Microridge patterns progress in complexity during fish embryogenesis, often transitioning from abundant surface microvilli to the classical fingerprint arrangement. This progression suggests pattern changes may also occur during later stages of fish development. Fluorescent labelling of F-actin and morphometric analysis were therefore used to assess changes in epidermal microridge patterns in juvenile and adult sunfish (Lepomis gibbosus). The microridge patterns found in adult pumpkinseed were similar to that described for many fishes, consisting of whorls or complex multi-branched ridges. The microridge patterns of the scales from three different-sized groups of juvenile pumpkinseed were distinctly different from that of adult, however, and were present mainly as unbranched concentric or nearly concentric rings in the two larger juvenile groups. In the smallest juveniles, microridges were often apparent as fragmented ridges with abundant actin puncta. Larger juveniles sometimes displayed mixed patterns, with some microridges similar to that of both adult and juvenile patterns. The results show a transition from simple microridge patterns in juvenile pumpkinseeds to distinctly different, diverse and more complex patterns in adults. The different pattern types may reflect particular microridge functions relevant to fish size and age.     

瘤足蕨科及其近缘类群植物叶表皮微形态扫描电镜观察

瘤足蕨科是一个自然的群,但由于各类群之间的形态差异及种内变异比较大,其分类和系统位置一直有争议。该研究利用扫描电子显微镜对瘤足蕨科10种及其近缘类群桫椤科2种、蚌壳蕨科1种共13种植物,即瘤足蕨(Plagiogyria adnata)、耳形瘤足蕨(P.stenoptera)、镰羽瘤足蕨(P.falcata)、峨嵋瘤足蕨(P.assurgens)、密羽瘤足蕨(P.pycnophylla)、灰背瘤足蕨(P.glauca)、华东瘤足蕨(P.japonica)、华中瘤足蕨(P.euphlebia)、日本瘤足蕨(P.matsumureana)、美洲瘤足蕨(P.pectinata)、桫椤(Alsophila spinulosa)、西亚黑桫椤(A.khasyana)和金毛狗(Cibotium barometz)的叶表皮微形态进行了观察和比较,探讨其分类学意义,并据此编制了供试10个种类的分类检索表。结果表明:13种瘤足蕨科及其近缘类群植物气孔均仅分布在叶下表皮,气孔微下陷,气孔器椭圆形至圆形;叶上下表皮角质层纹饰复杂,类型多样,少数类群叶下表皮具毛状物;多数类群气孔外拱盖凹陷;气孔外拱盖内缘浅波状或齿状;保卫细胞两极大多数有明显的"T"型加厚。不同种间叶表皮扫描电镜特征表现出一定差异,对种的划分有一定的分类学鉴定意义。     

24种鳞毛蕨科植物叶表皮形态特征的研究

该研究利用光学显微镜对鳞毛蕨科24种植物的叶表皮形态特征进行观察。结果表明:(1)24种鳞毛蕨科植物的上表皮细胞形状为长条形或不规则形,垂周壁为深波状或浅波状,下表皮细胞均为无规则形,垂周壁均为深波状;上表皮细胞长宽比在1.5~5.7之间,下表皮细胞长宽比在2.2~3.9之间。(2)在24种鳞毛蕨科植物中共观察到8种气孔器类型,分别为不等细胞型、无规则四细胞型、极细胞型、腋下细胞型、横列型、无规则型、聚腋下细胞型和聚合极细胞型,每种植物具有2~8种气孔器类型,气孔均为下生型,多为椭圆形;气孔的长宽比在1.2~1.8之间,气孔密度在17.4~86.0个/mm~2之间,气孔指数为8.60%~37.4%。(3)通过对24种鳞毛蕨科植物的观察可将其上表皮细胞形状、垂周壁形状、上表皮细胞长宽比、主要气孔器类型及衍生类型等作为叶表皮形态特征的分类依据。(4)根据叶表皮形态特征可将24种鳞毛蕨科植物分为2类:即耳蕨类和鳞毛蕨类。该研究在一定程度上支持秦仁昌分类系统对鳞毛蕨科的划分,为鳞毛蕨科植物的系统分类及演化研究提供基础资料。     

Leaf anatomy ofDombeya andNesogordonia (Sterculiaceae), emphasizing epidermal and internal idioblasts

We studied leaf anatomy, using clearings, resin sections, and scanning electron microscopy, from mostly herbarium specimens of 123 species ofDombeya and 11 species ofNesogordonia (Sterculiaceae). Species were placed in seven idioblast categories, ranging from those without any to single and bicelled epidermal forms to multicelled nodules and single mesophyll idioblasts. Idioblast contents are possibly mucilaginous, but were not identified. In these two genera the range of foliar idioblast morphology surpasses that known previously for the entire family. Leaves are dorsiventral with mostly abaxial anomocytic stomata and typical palisade and spongy layers; paraveinal mesophyll is lacking. Miniature glandular (clavates, capitates) and nonglandular (mostly stellate) trichomes occur. Prismatic crystals predominate; epidermal prismatics and mesophyll druses are rare.     

Modification of chemical properties of cell wall polysaccharides in the inner tissues by white light in relation to the decrease in tissue tension in Pisum sativum epicotyls

When white light irradiation inhibits shoot growth in derooted pea ( Pisum sativum L. cv. Alaska) cuttings, it decreases tissue tension, a driving force for shoot growth, by making the cell wall of the inner tissues mechanically rigid. To elucidate the mechanism by which light affects the mechanical properties of the cell wall in the inner tissues, its effect on the chemical properties of the cell walls was studied by analyzing qualitatively and quantitatively cell wall polysaccharides in the epdidermis and inner tissue of pea epicotyls grown in both dark and light. The amount of polysaccharides per subhook in the cell walls of both tissues increased during a 4-h dark incubation. Light suppressed the increase in hemicellulosic (HC)-II and cellulosic polysaccharides in the inner tissues, while it did not affect the increase in other wall fractions in either the epidermal or subepidermal tissues. No light effect was observed on the neutral sugar compositions of pectin, HC-I or HC-II fractions in either of the tissues. Light increased the mass-average molecular mass of xyloglucan and rhamnoarabinogalactan in HC-II fractions in the inner tissues, while such an effect was not observed in the epidermis. These facts suggest that the light-induced decrease in the tissue tension in pea epicotyls is caused by an increase in the molecular size of xyloglucan, rhamnoarabinogalactan in the HC-II fraction and/or the suppression of the synthesis of HC-II and cellulosic polysaccharides in the inner tissues.     

Sulphur, thiols, and disulphides in the fish epidermis, with remarks on keratinization

Energy dispersive x-ray (EDX) analysis and qualitative and quantitative histochemistry were applied to study the distribution and contents of sulphur, thiols and disulphides in the epidermis of the river lamprey Lampetra fluviatilis , the lesser spotted dogfish Scyliorhinus canicula and the brown trout Salmo trutta fario . Thiols generally reacted weakly throughout the entire epidermis, whereas disulphide reactions were more distinct and differentiated. In the river lamprey, the concentrations of -S-S- groups clearly increased in the developing mucous cells from the stratum basale to the stratum superficiale; skein cells and granular cells reacted negatively to weakly. In the lesser spotted dogfish, amounts of disulphides appeared at moderate concentrations, and only goblet cells displayed a strong reaction. In the brown trout, filament cells showed low concentrations or weak reactions of disulphides, goblet cells and the most outer superficial cells stained strongly. Sulphur distribution and contents generally supported the histochemical observations in normal epidermis cells (absolute sulphur contents: 41–59 mM), only the brown trout showed high amounts of sulphur in the stratum basale (81 mM). The findings corroborate the view that there is an inverse correlation between keratinization and mucous secretion in normal fish epidermis. The sometimes distinct contents of disulphides in the outer mucous layer indicate that this system could endure higher mechanical stress than predictable from its large amounts of neutral glycoproteins.