bromochloroacetic-acid and fluorexon

Loss-of-function mutations in filaggrin are major predisposing factors for atopic dermatitis. Although various reports suggest a critical role for filaggrin in stratum corneum (SC) barrier formation, the lack of filaggrin-null (Flg(-/-)) mice has hampered detailed in vivo analysis of filaggrin's functions.. We sought to generate Flg(-/-) mice and to assess the effect of filaggrin loss on SC barrier function and percutaneous immune responses.. We generated Flg(-/-) mice using gene targeting and assessed the morphology, hydration, mechanical strength, and antigen permeability of their SC. Percutaneous immune responses were evaluated through irritant- and hapten-induced contact hypersensitivity studies and by measuring humoral responses to epicutaneous sensitization with protein antigen.. Newborn Flg(-/-) mice exhibited dry scaly skin. Despite marked decreases in natural moisturizing factor levels, which are filaggrin degradation products, SC hydration and transepidermal water loss were normal. Microscopic analyses suggested premature shedding of SC layers, and indeed, increased desquamation under mechanical stress was demonstrated. Loss of keratin patterns, which are critical for corneocyte stabilization, is likely attributable to fragility in the Flg(-/-) SC. Antigens penetrated the Flg(-/-) SC more efficiently, leading to enhanced responses in hapten-induced contact hypersensitivity and higher serum levels of anti-ovalbumin IgG(1) and IgE.. Complete filaggrin deficiency led to altered barrier integrity and enhanced sensitization, which are important factors in early-phase atopic dermatitis. Flg(-/-) mice should provide a valuable tool to further explore additional factors the dysfunction of which leads to uncontrolled inflammation in patients with atopic diseases.

Topics: Animals; Blood-Aqueous Barrier; Epithelium, Corneal; Filaggrin Proteins; Fluoresceins; Genotype; Intermediate Filament Proteins; Keratins; Mice; Mice, Knockout; Permeability; Skin

Determination of multidrug resistance (MDR) activity of tumor cells could provide important information for the personalized therapy of cancer patients. The functional calcein assay (MultiDrug Quant Assay, Solvo Biotechnology, Budaörs, Hungary) has been proven to be clinically valuable in hematological malignancies by determining the transporter activity of MDR protein 1 (MDR1, ATP-binding cassette protein [ABC] B1, P-glycoprotein-170) and MDR-related protein 1 (MRP1, ABCC1). In this study, we evaluated if the same functional test was adaptable for the analysis of MDR activity in solid tumors. For this purpose, tissue specimens of human colorectal cancer samples were subjected to limited enzymatic digestion by collagenase to provide a single-cell suspension; dead cells were excluded by 7-aminoactinomycin D staining, and epithelial cancer cells were detected by Cy5-conjugated anti-BerEP4 monoclonal antibody. The transporter functions of MDR1 and MRP1 in viable epithelial cells were assessed by flow cytometry detecting the intracellular accumulation of calcein dye after exposing cells to various MDR inhibitors. Collagenase disintegration preserved the MDR activity and the antigenicity of tumor cells. Thus using the extended calcein assay provided sufficient viable and functionally active tumor cells from surgical biopsies to determine the functional MDR activity. In conclusion, the newly described modified calcein assay may be applicable for evaluating the MDR phenotype in solid tissue specimens from colorectal forceps biopsy to surgical samples.

Topics: Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Surface; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biomarkers; Biopsy; Body Fluids; Carcinoma; Cell Separation; Cell Survival; Fluoresceins; Fluorescent Antibody Technique; Humans; Keratins; Leukemia P388; Neoplasms

A liposomal system for percutaneous absorption and transdermal drug delivery was developed and evaluated using a model apparatus connecting two chambers by a membrane. When N,N'-long chain diacyl L-cystine was incorporated into the liposomal bilayer as well as lecithin and cholesterol, liposomally entrapped calcein as the model drug transferred well across the keratin membrane functioning as the skin model.

Topics: Cellophane; Cholesterol; Cystine; Diffusion; Drug Carriers; Fluoresceins; Indicators and Reagents; Keratins; Kinetics; Lipid Bilayers; Microscopy, Electron; Models, Biological; Phosphatidylcholines