The neuronal transcription factor NeuroD and its impact on the regulation of dendricity and pigmentation of melanocytes in postinflammatory hyperpigmentation of human skin
ABSTRACT: In order to test the gene expression profile in postinflammatory hyperpigmentation, micoarray studies were performed. Totally 14 subjects ( 7 at each phase) were treated by suction blister, and samples from treated area and corresponding control area ( from the same subjects) were taken at 1 week(t1), 2 weeks(t2), 4 weeks(t3), 6 weeks(t4), and 16 weeks(t5) after treatment. Paired samples from subject 6382 at 6th week were missing for microarray hybridization. Therefore, there are 138 array chips available for statistical analysis.
Project description:To test whether there is a photoprotective benefit after different types of suberythemal repetitive UV, a 1.5 MED challenge dose was applied 1 week after the initial 2 weeks of repetitive irradiation. To determine what different mechanisms and/or factors might be involved in physiological pigmentary responses of the skin to different types of UV, we used whole human genome microarrays and immunohistochemical analyses to characterize human skin in situ to examine how melanocyte-specific proteins and paracrine melanogenic factors are regulated by repetitive exposure to suberythemal doses of different types of UV (UVA and/or UVB). Seven volunteers with skin type II-III were irradiated with UVA, UVB or UVA+UVB radiation for 2 weeks (5 times per week, 10 times total) after preliminary determination of their MEDs. A UVA+UVB challenge dose of 1.5X the MED was applied 1 week later. Biopsies were taken before the challenge dose, immediately after the challenge dose, 4 days after the challenge, and 15 weeks after the challenge.
Project description:In this study, we used immunohistochemical analysis and microarray analysis to investigate the mechanism underlying the development of age spots, focusing on the expression of specific markers associated with the functions and proliferation of melanocytes and keratinocytes 10 subjects of phototypes I or II (all female, age = 53-70) with various sizes and colors of age spots on their forearms were recruited for the study. Four mm whole skin punch biopsies were taken from age spots on the forearm of each subject and from a perilesional control area in the vicinity of each age spot.
Project description:In this study, we used microarray analysis to investigate molecular differences underlying the determination of pigmentation in various skin types, and we used immunohistochemistry to validate the expression patterns of several interesting targets that were identified. Whole skin gene expression profile were investigated in three ethnic groups:1) Caucasian 2)Asian 3)African. In each group, 4 mm whole skin punch biopsies were taken from forearm of 10 subjects, and gene expression level were tested by agilent whole human genome microarray. Samples with mRNA degraded: African_skin_03, Asian_skin_02, Asian_skin_06, and Caucasian_skin_7.
Project description:To detect time of day dependent gene expression in human epidermis suction blister samples from 20 healthy subjects were obtained at three different time points throughout the day. RNA from 20 subjects were used to perform whole genome microarray analysis. Microarrays from 19 subjects showed sufficient quality to perform analysis for differential gene expression. We detected significant differential expression levels for several canonical clock genes such as Per1, Per2, Per3, Bmal1 and Rev-Erb_alpha throughout the day. In total we identified 294 genes that showed significant circadian gene expression including several transcription factors and rate limiting enzymes. To our knowledge this is the first study to investigate genome wide circadian gene expression in human epidermis. Suction blisters of human epidermis were obtained by applying a vacuum for approximately 2.5 h. Suction blisters for each subject (19 subjects in total) were harvested at 9.30 am, 2.30 pm and 7.30 pm.
Project description:In this study we experimentally induced thyrotoxicosis in healthy volunteers to explore the effects of thyroxine excess on the plasma proteome. 16 healthy male subjects were sampled at baseline, 4 and 8 weeks under 250 µg/d thyroxine p.o., as well as 4 and 8 weeks after stopping the application.
Project description:RNA expression profiling was carried out for systemic sclerosis (scleroderma) on skin biopsies obtained from both diffuse scleroderma (dcSSc) and limited scleroderma (lcSSc) subjects patients and compared to normal subjects. The patient group comprised of 4 dcSSc and 2 lcSSc subjects, all of whom were Caucasian and belonged to the age range of 42-73 (Mean age = 61.6). Patient sample set consisted of 3 females and 3 males. Race matched female volunteers from the same geographical area were recruited as controls (n = 4) and belonged to the age range of 41-62 (Mean age = 52).
Project description:We profiled gene expression in peripheral blood cells from 28 obese patients by microarray analysis and visceral fat accumulation caused the gene expression proliles especially in circadian rhythm, inflammation, oxidative stress, and immune response. Obesity was defined as body mass index (BMI) greater than 25 kg/m2 according to the Japanese criteria and visceral fat area (VFA) was estimated by abdominal bioelectrical impedance analysis. Subjects with type 1 diabetes mellitus, autoimmune diseases, malignant diseases, and infectious diseases were excluded. Patients treated with statin and/or thiazolidinediones were also excluded.
Project description:The goal of this study is to characterize the expression profile of Epidermolysis bullosa simplex-mottled pigmentation (EBS-MP) patient compared with normal subjects using genomic expression analyses. Microarray analyses were performed with RNA isolated from skin biopsies. Robust multiarray analysis (RMA) normalization and Smyth’s moderated t test were used to select differentially expressed genes. Expression profiling comparisons show that 52 genes are differentially expressed in EBS-MP patients compared to control subjects. Difference of expression for three genes (TYR, CCL22 , and ACVR1C ) was validated using real-time reverse transcription–polymerase chain reaction. Twelve genes were related to lipid biosynthesis process, two to keratinisation and skin pigmentation, Nineteen to cell growth and apoptosis, five to immune response and fourteen to predicted or less known function cluster. To our knowledge, the distinctive pattern of gene expression that characterizes EBS-MP versus healthy skin tissue has never been reported. RNA used for the microarrays analysis was isolated from superficial 2mm punch biopsies composed of mainly epidermis with minimal dermis amounts of normal-appearing skin of one EBS-MP patient and seven healthy volunteers.
Project description:Keratinocytes are the most abundant cell type in the epidermis. They prevent desiccation and provide immunological and barrier defense against potential pathogens such as Staphylococcus aureus and Candida albicans. The study of this first line of immune defense may be hindered by invasive isolation methods and/or improper culture conditions to support stem cell maintenance and other potential mechanisms contributing to long-term subcultivation in vitro. Primary keratinocytes have been successfully isolated from blister roofs induced by negative pressure, which separates the epidermis from the dermis in vivo in human subjects. This method allows collection of pure epidermal cells without dermal contamination in a minimally invasive manner. However, the isolated keratinocytes differentiate and senesce when cultured in vitro beyond five passages. Here, we present evidence that the Rho kinase (ROCK) inhibitor Y-27632 can be used to effectively increase the proliferative capabilities of keratinocytes isolated using the suction blister method, similar to what has been previously reported for primary keratinocytes isolated using alternative methods. We show that the increase in passage number is directly correlated to delayed differentiation, and that cells passaged long term with the inhibitor retain their ability to stratify in organotypic raft cultures and respond to cytokine treatment; additionally, the late passage cells have a heterogeneous mix of differentiated and non-differentiated cells which may be predicted by a ratio of select differentiation markers. The described method presents a minimally invasive procedure for keratinocyte isolation and prolonged culture that allows analysis of keratinocyte function in both healthy volunteers and patients with dermatologic diseases. Overall design: Keratinocytes isolated using the suction blister method were grown in the absence (passages 4, 5, and 6) and presence (passages 5 and 6) of ROCK inhibitor Y-27632 or cytokines (IL17 + IFN-g)
Project description:Overall, the widely disparate transcriptomes identified prior to RT among the three clusters support the notion that at least some of the inter-individual heterogeneity in propensity for RT-induced myofiber hypertrophy is likely pre-determined. Baseline comparisons of skeletal muscle transcript profiles from 44 subjects previously clustered (K-means cluster analysis) as non-, modest-, and extreme-responders (Non, n=15; Mod, n=14; Xtr, n=15) based on the magnitudes of change in vastus lateralis myofiber cross-sectional area (CSA) following 16-wk, 3-d/wk, high-intensity RT.