Nonlesional Skin Of Patients Research Articles

Serum neurotensin (NT) is increased in psoriasis and NT induces vascular endothelial growth factor release from human mast cells

Psoriasis involves skin inflammation that often worsens with stress, but the mechanism of this effect remains obscure. We have shown that corticotropin-releasing hormone (CRH) is increased in the serum of patients with psoriasis. A peptide, neurotensin (NT), can trigger skin histamine release and augment the ability of CRH to increase skin vascular permeability. To investigate the serum level of NT, and the expression of genes for NT and NT receptor-1 (NTR-1) in lesional and nonlesional skin of patients with psoriasis, compared with normal controls. Also, to study the effect of NT on human mast cell release of vascular endothelial growth factor (VEGF), which is increased in psoriatic skin. Serum was obtained from patients with psoriasis (n = 56) and controls (n = 33); NT levels were measured with the Milliplex microbead assay. Biopsies were obtained from the lesional and nonlesional skin of patients with chronic plaque psoriasis (n = 40), who had not received any treatment for at least 15 days and were free of any systemic inflammatory diseases. Control skin samples were obtained from healthy subjects (n = 30). Expression of genes for NT and NTR-1 in the skin was evaluated by quantitative reverse transcriptase-polymerase chain reaction. LAD2 human mast cells were stimulated by NT (1 μmol L(-1)) for 24 h and VEGF was measured by enzyme-linked immunosorbent assay. Serum NT was increased in patients with psoriasis, while expression of genes for NT and NTR-1 in lesional skin was decreased compared with controls. NT induced VEGF release from mast cells and was augmented by interleukin-33. NT may play a role in psoriasis pathogenesis and its worsening by stress, at least in part through activation of skin mast cells.

Psychoneuroimmunology of Psychological Stress and Atopic Dermatitis: Pathophysiologic and Therapeutic Updates

Atopic dermatitis is a chronic inflammatory skin disease characterized by impaired epidermal barrier function, inflammatory infiltration, extensive pruritus and a clinical course defined by symptomatic flares and remissions. The mechanisms of disease exacerbation are still poorly understood. Clinical occurrence of atopic dermatitis is often associated with psychological stress. In response to stress, upregulation of neuropeptide mediators in the brain, endocrine organs, and peripheral nervous system directly affect immune and resident cells in the skin. Lesional and non-lesional skin of patients with atopic dermatitis demonstrates increased mast cells and mast cell-nerve fiber contacts. In the setting of stress, sensory nerves release neuromediators that regulate inflammatory and immune responses, as well as barrier function. Progress towards elucidating these neuroimmune connections will refine our understanding of how emotional stress influences atopic dermatitis. Moreover, psychopharmacologic agents that modulate neuronal receptors or the amplification circuits of inflammation are attractive options for the treatment of not only atopic dermatitis, but also other stress-mediated inflammatory skin diseases.

Investigation ofPropionibacterium acnesin progressive macular hypomelanosis using real‐time PCR and culture

Progressive macular hypomelanosis (PMH) is a dermatosis of unknown etiology. It has been concluded that it involves the presence of Propionibacterium acnes, a saprophyte of the pilosebaceous follicles. In our study, we investigated the presence of P. acnes in lesional and non-lesional skin of patients with PMH through quantitative real-time polymerase chain reaction (PCR) and bacterial culture from a skin fragment. An observational, exploratory study, with laboratory comparison of lesional (study group) and non-lesional skin (comparison group), in patients with PMH, was carried out with 36 patients, seen in the dermatology outpatient setting at the Oswaldo Cruz University Hospital (OCUH), Recife, Pernambuco, Brazil, between March and May 2008. All patients were submitted to a Wood's lamp examination, mycological research, and biopsies of lesional and non-lesional skin from the back. Skin fragments were submitted to a histopathology test, bacterial culture, and a quantitative real-time PCR test. The program Statistical Package for Social Sciences, version 12.0, was employed for relationship analysis with the Wilcoxon and McNemar tests. There was a significant predominance of P. acnes on lesional skin, in comparison to non-lesional skin (P<0.001), as demonstrated by culture and quantitative real-time PCR. Although P. acnes is a saprophyte, the hypothesis may be raised that this microorganism participates in the development of PMH.

Expression of GSTM4 and GSTT1 in patients with Tinea versicolor, Tinea inguinalis and Tinea pedis infections: a preliminary study

Several skin diseases are believed to be associated with oxidative stress. Defence against reactive oxygen species in the skin involves a variety of antioxidant enzymes, including glutathione-S-transferases (GSTs) catalysing the reaction between reduced glutathione, and a variety of exogenously and endogenously derived electrophilic compounds. The mammalian soluble GSTs are divided into five main classes: alpha (A), mu (M), pi (P), theta (T) and zeta (Z). To investigate the expression of GSTM4 and GSTT1 in lesional and nonlesional skin of patients with dermatophytoses and Tinea versicolor infection. Methods. Expression of GSTM4 and GSTT1 was assessed by immunohistochemistry for dermatophytoses in 15 patients with T. versicolor, 15 patients with Tinea pedis and 8 patients with Tinea inguinalis, and compared with healthy controls (n = 9). After written consent was signed by each participant, punch biopsies were excised from the centre of the lesional skin sites in patients and from the normal skin sites in controls. The relationships between expression of GSTM4 and GSTT1 isoenzymes and fungal infections were also examined. When the normal and infected tissue of these cases were compared according to their staining intensity, GSTM4 expression was found to be stronger in control epithelium than in the epithelium of patients with T. pedis, T. inguinalis or T. versicolor (P < 0.05). By contrast, expression of GSTT1 was stronger in the epithelium of patients infected with any of the three dermatophytes than in control epithelium (P < 0.05). There is a significant relationship between presence of T. versicolor, T. inguinalis and T. pedis and expression of GSTM4 and GSTT1.

Characteristics of Staphylococcus aureus colonization in patients with atopic dermatitis in Sri Lanka

Colonization of the skin of patients with atopic dermatitis (AD) by Staphylococcus aureus (SA) is associated with more severe disease. To determine the association of SA colonization patterns and densities in lesional and nonlesional skin in patients with varying severities of AD, and to determine the antibiotic sensitivity patterns of SA isolates from Sri Lanka. Skin and nasal swabs collected from 100 patients with AD and 120 controls were used to investigate the presence of SA. Severity of AD was graded using the Nottingham Eczema Severity Score. Colony counts were obtained for skin samples, and antibiotic sensitivity testing was performed in cases positive for SA. Skin colonization was seen in 57 patients (57%) but in only 10 controls (8%). Lesional skin of most patients (52/57; 91%) had SA densities of > 300 colony-forming units/cm(2) . Colonization rates with SA significantly increased with increasing age (Spearman correlation coefficient R = 0.9, P < 0.05) and increasing duration of lesions in patients with AD (Spearman R = 0.87, P < 0.05). Isolates from eight patients (13.5%) were found to be methicillin-resistant S. aureus (MRSA). Only 6 isolates (10%) were susceptible to penicillin and 22 (37%) to erythromycin, while 28 (47%) isolates had erythromycin-induced resistance to clindamycin. SA colonization rates were significantly associated with increasing age and severity of AD, and particularly with duration of lesions. Patients with severe disease were also more likely to be colonized with SA strains resistant to conventional antibiotics.

A comprehensive analysis of the COL29A1 gene does not support a role in eczema

Based on a recent positional cloning approach, it was claimed that the collagen 29A1 gene (COL29A1), which encodes an epidermal collagen, represents a major risk gene for eczema underlying a previously reported linkage to chromosome 3q21. However, thus far, not a single replication attempt has been published, and no definitive functional data have been provided. We aimed to determine whether COL29A1 polymorphisms contribute to eczema susceptibility and whether COL29A1 expression is altered in eczema. We investigated the reported association of COL29A1 variants with eczema, subtypes of eczema, and eczema-related traits in 5 independent and large study populations comprehensively phenotyped for allergic diseases: a set of 1687 German patients with eczema and 2387 population control subjects, a collection of 274 German families with eczema-diseases children, a cross-sectional population of German children (n= 3099), the Swedish population-based birth cohort Children Allergy and Milieu in Stockholm, an Epidemiologic Study (BAMSE) (n= 2033), and the European cross-sectional Prevention of Allergy-Risk Factors for Sensitization Related to Farming and Anthroposophic Lifestyle (PARSIFAL) study (n= 3113). An additional set of 19 COL29A1 coding single nucleotide polymorphisms was analyzed in BAMSE and PARSIFAL. COL29A1 expression was investigated by using in situ hybridization. We found no evidence for a relationship between COL29A1 polymorphisms and eczema. The equivalence test rejected the hypothesis of association even excluding small effects. In situ hybridization carried out on biopsy specimens from lesional and nonlesional skin of patients with eczema and from healthy control subjects did not show any differences in the cellular distribution pattern of COL29A1 expression at the mRNA level. Our results suggest that COL29A1 is unlikely to contain genetic variants that have a major effect on eczema or atopy susceptibility.

Comparative proteomic profiling of patients with atopic dermatitis based on history of eczema herpeticum infection and Staphylococcus aureus colonization

Atopic dermatitis (AD) is the most common inflammatory skin disorder in the general population worldwide, and the majority of patients are colonized with Staphylococcus aureus. Eczema herpeticum is a disseminated herpes simplex virus infection that occurs in a small subset of patients. The goal was to conduct proteomic profiling of patients with AD based on S. aureus colonization status and history of eczema herpeticum. We hoped to identify new biomarkers for improved diagnosis and prediction of eczema herpeticum and S. aureus susceptibility and to generate new hypotheses regarding disease pathogenesis. Skin taping was performed on nonlesional skin of nonatopic control subjects and on lesional and nonlesional skin of patients with AD. Subjects were classified according to the history of eczema herpeticum and S. aureus colonization. Proteins were analyzed by using mass spectrometry; diagnostic groups were compared for statistically significant differences in protein expression. Proteins related to the skin barrier (filaggrin-2, corneodesmosin, desmoglein-1, desmocollin-1, and transglutaminase-3) and generation of natural moisturizing factor (arginase-1, caspase-14, and gamma-glutamyl cyclotransferase) were expressed at significantly lower levels in lesional versus nonlesional sites of patients with AD with and without history of eczema herpeticum; epidermal fatty acid-binding protein was expressed at significantly higher levels in patients with methicillin-resistant S. aureus. This noninvasive, semiquantitative profiling method has revealed novel proteins likely involved in the pathogenesis of AD. The lower expression of skin barrier proteins and enzymes involved in the generation of the natural moisturizing factor could further exacerbate barrier defects and perpetuate water loss from the skin. The greater expression of epidermal fatty acid-binding protein, especially in patients colonized with methicillin-resistant S. aureus, might perpetuate the inflammatory response through eicosanoid signaling.

A quantitative analysis of Propionibacterium acnes in lesional and non-lesional skin of patients with progressive macular hypomelanosis by real-time polymerase chain reaction

Little is known about the etiology of progressive macular hypomelanosis, although it has been suggested that Propionibacterium acnes plays an important role. While microbiological culture is commonly employed to identify Propionibacterium acnes, new identification methods have been under investigation, amongst them polymerase chain reaction. To determine the cut-off point for the number of genome copies of Propionibacterium acnes in the lesional skin of patients with progressive macular hypomelanosis as a positive marker, employing quantitative real-time polymerase chain reaction and anaerobic culture, considered gold standard. An observational study with a comparison group, included 35 patients with dermatosis, attended at the Oswaldo Cruz University Hospital, Pernambuco, Brazil, between March and May 2008. Lesional skin was compared to non-lesional skin through positive testing with real-time polymerase chain reaction and culture. The Statistical Package for Social Sciences, version 12.0, was employed for the association analysis with the McNemar test, and the cut-off point with the ROC curve for maximum values. Propionibacterium acnes was most frequently encountered in lesional areas (p<0,025). The cut-off point of Propionibacterium acnes in lesional skin was 1,333 genome copies, with a sensitivity of 87,9% and a specificity of 100,0%. Since Propionibacterium acnes is a saprophyte, identifying the cut-off point may assist in determining its positivity in lesional skin in patients suffering with this dermatosis.

High numbers of DC-SIGN+ dendritic cells in lesional skin of cutaneous T-cell lymphoma

The role of dendritic cells (DCs) in disease progression of primary cutaneous T-cell lymphoma (CTCL) is not well understood. With their unique ability to induce primary immune responses as well as immunotolerance, DCs play a critical role in mediation of anti-tumor immune responses. Tumor-infiltrating DCs have been determined to represent important prognostic factors in a variety of human tumors. To date, only circulating DC populations have been investigated in CTCL. Therefore we analyzed the expression and tissue distribution of different DC subset markers in lesional and nonlesional skin of patients with CTCL. Twelve patients with mycosis fungoides or Sézary syndrome were included in the study. Tissue samples were obtained from lesional and nonlesional skin as a control. Immunohistochemistry was performed with different DC subset and regulatory T-cell markers and assessed using a digital image analysis system. Tissue distribution of DCs in relation to the tumor was analyzed by double immunofluorescence. We found a significant infiltration of CTCL lesions by immature CD209/DC-SIGN+ DCs with close contact to tumor cells. Matured and activated DCs were only rarely detected in lesions of CTCL. The sample size was small. The preponderance of immature CD209/DC-DIGN+ DCs in contact with regulatory T cells in lesions of CTCL points to an important role of this subset in the host's immune reaction to the malignant T cells. Since these immature DCs are known to induce immunotolerance, they might play a role in the mediation of immune escape of the proliferating clone.

Analysis of eotaxin 1/CCL11, eotaxin 2/CCL24 and eotaxin 3/CCL26 expression in lesional and non-lesional skin of patients with atopic dermatitis

Eotaxins are the chemokines which are highly selective chemotactic agents for eosinophils. The aim of our study was the evaluation of the gene expression level for eotaxin 1/CCL11, eotaxin 2/CCL24, and eotaxin 3/CCL26, both in skin changes and in uninvolved skin of atopic dermatitis (AD) patients. The study comprised 19 patients with AD and 10 healthy controls. The gene expression level for eotaxins in the skin biopsies was evaluated by the real-time quantitative PCR. The change of the gene expression level, calculated as log10 skin lesions/non-lesional skin, was 0.635 for CCL11, 0.172 for CCL24 and 0.291 for CCL26. The change of the gene expression level, calculated as log10 non-lesional skin of AD patients/healthy control, was 0.394 for CCL11, -0.216 for CCL24, and 0.229 for CCL26, while skin lesions of AD patients/healthy control, was: 0.788, -0.046, and 0.483, respectively. The mean gene expression level for CCL11, CCL24, CCL26 was higher in skin changes of AD patients than in uninvolved skin. The higher level of CCL26 in skin changes, indicates its role in their aetiology in AD. The gene expression level for CCL24 in AD patients was lower, both in involved and uninvolved skin vs. the healthy control.

Comparison of skin barrier function and sensory nerve electric current perception threshold between IgE-high extrinsic and IgE-normal intrinsic types of atopic dermatitis

Background Two types of atopic dermatitis (AD) have been proposed, with different pathophysiological mechanisms underlying this seemingly heterogeneous disorder. The extrinsic type shows high IgE levels presumably as a consequence of skin barrier damage and feasible allergen permeation, whereas the intrinsic type exhibits normal IgE levels and is not mediated by allergen-specific IgE. Objectives To investigate the relationship between pruritus perception threshold and skin barrier function of patients with AD in a comparison between the extrinsic and intrinsic types. Methods Enrolled in this study were 32 patients with extrinsic AD, 17 with intrinsic AD and 24 healthy individuals. The barrier function of the stratum corneum was assessed by skin surface hydration and transepidermal water loss (TEWL), and pruritus perception was evaluated by the electric current perception threshold (CPT) of sensory nerves upon neuroselective transcutaneous electric stimulation. Results Skin surface hydration was significantly lower and TEWL was significantly higher in extrinsic AD than intrinsic AD or normal controls. Although there was no statistically significant difference in CPT among extrinsic AD, intrinsic AD and normal controls, CPT was significantly correlated with skin surface hydration and inversely with TEWL in intrinsic AD and normal controls, but not extrinsic AD. Finally, CPT was correlated with the visual analogue scale of itch in the nonlesional skin of patients with extrinsic but not intrinsic AD. Conclusions Patients with extrinsic AD have an impaired barrier, which increases the pre-existing pruritus but rather decreases sensitivity to external stimuli. In contrast, patients with intrinsic AD retain a normal barrier function and sensory reactivity to external pruritic stimuli.

Detection of viral DNA in nonlesional skin of patients with molluscum contagiosum and on environmental fomites

Detection of viral DNA in nonlesional skin of patients with molluscum contagiosum and on environmental fomites

Enhanced expression of the antimicrobial peptide LL-37 in lesional skin of adults with atopic eczema

Atopic eczema (AE) is a common multifactorial chronic skin disease associated with a defective skin barrier and increased susceptibility to skin infections. The human cathelicidin LL-37 plays a role in the host defence of skin. Studies have demonstrated deficient expression of LL-37 in skin of AE patients. The aim of this study was to investigate the expression of LL-37 in lesional skin compared with nonlesional skin in patients with different severity of AE, patients with other eczema and healthy subjects. Twenty patients with AE, four patients with other eczema and 10 healthy subjects were included. Severity of AE was graded using SCORing of atopic dermatitis (SCORAD). Skin biopsies were taken from lesional and nonlesional skin from all patients and from skin of healthy controls. The levels of LL-37 mRNA were analysed by quantitative reverse transcriptase-polymerase chain reaction. Evaluation of dermal and epidermal protein expression of LL-37 and the degree of inflammation was performed by immunohistochemical stainings. Patients with AE and patients with other eczema had significantly (P < 0.05) higher levels of LL-37 in lesional skin than in nonlesional skin. The expression of LL-37 was not statistically associated to severity of AE valued by SCORAD. Nonlesional skin from patients did not differ from skin of healthy subjects in terms of LL-37 expression. In the presence of epidermal injury or vesicles the LL-37 peptide was always detected. Patients with AE exhibit enhanced expression of LL-37 in lesional skin compared with nonlesional, suggesting a role of LL-37 in AE that might be associated with the process of re-epithelialization.

The Increased Expression of Matrix Metalloproteinase-9 Messenger RNA in the Non-lesional Skin of Patients with Large Plaque Psoriasis Vulgaris.

A difference of the interleukin-18 (IL-18) mRNA expression among several proinflammatory genes was previously observed between large plaque (LP) psoriasis patients (more than 5 cm lesions are typical) and small plaque (SP) psoriasis patients (1~2 cm lesions are typical). Therefore, it is necessary to test whether there is any difference in the expression of the genes that activate IL-18 or the expression of genes that are induced by IL-18. To test the differential mRNA expressions of caspase-1, STAT-6, MMP-1, MMP-2, MMP-9 and TIMP-1 according to the clinical types of psoriasis vulgaris lesions in Korean patients, we have analyzed the skin samples of psoriasis vulgaris patients. The total cellular RNA of skin samples from groups of patient with LP and SP psoriasis was analyzed by performing real-time PCR (the Taqman method) to compare the differences in the mRNA expressions. The caspase-1 and STAT-6 mRNA expression levels from the SP lesional skin of the patients were increased compared with the caspase-1 and STAT-6 mRNA expression levels from SP non-lesional skin or normal skin, but these expression levels from the SP non-lesional skin were not significantly different from those of the LP non-lesional skin. Among MMP-1, MMP-2, MMP-9 and TIMP-1, the expressions of MMP-1, MMP-2 and MMP-9 mRNA were increased in the SP lesional skin compared with those of the SP non-lesional skin. The MMP-1 mRNA expressions in both the LP and SP lesional skin were increased compared with those in the normal skin (p=0.028 and p=0.007 respectively). The MMP-9 mRNA expression in the LP non-lesional skin was elevated compared with the MMP-9 mRNA expression in the SP non-lesional skin (p=0.047). The TIMP-1 mRNA expression levels from the non-lesional skin and the lesional skin of the psoriasis patients and the normal skin samples were not significantly different. The increased expression of MMP-9 mRNA in the LP non-lesional skin compared to that of the SP non-lesional skin in the psoriatic skin suggests that the increased MMP-9 mRNA expression is related to the large size type of lesion.

Expression of insulin-like growth factor-I in lesional and non-lesional skin of patients with morphoea

Morphoea (scleroderma) is a chronic disorder characterized by circumscribed sclerotic plaques with the hallmark of increased fibroblast activation and fibrosis. Through its effect on connective tissue cells and immune cells, insulin-like growth factor (IGF)-I has been found to play a role in some autoimmune connective tissue diseases and has been implicated in the pathogenesis of several fibrotic disorders. To evaluate the role of IGF-I in the pathogenesis of morphoea. The study was carried out on 15 patients with morphoea and nine healthy controls. Two 5-mm punch skin biopsies were taken from every patient (one from lesional and one from non-lesional skin) and a single biopsy was taken from the normal skin of each control. A 10-mL blood sample was also taken from each patient and control. Quantitative detection of tissue and serum levels of IGF-I was done using an enzyme-linked immunosorbent assay technique. IGF-I in lesional skin was significantly higher than in non-lesional and control skin (P = 0.001 and P = 0.021, respectively). Moreover, a significantly higher level of IGF-I was detected in patient serum when compared with control serum (P < 0.001). A direct significant correlation existed between lesional and non-lesional skin level (r = 0.618, P = 0.014), and between lesional skin level and Rodnan score (r = 0.538, P = 0.039). Despite the small sample size, this study suggests that IGF-I plays an important role in the pathogenesis of fibrosis, characteristic of morphoea. Studies on a larger number of patients with morphoea as well as on patients with systemic sclerosis are recommended. Furthermore, therapeutic trials using IGF-I antagonist (octreotide) are highly recommended in patients with morphoea.

Skin Colonization by Staphylococcus aureus in Patients With Eczema and Atopic Dermatitis and Relevant Combined Topical Therapy: A Double-Blind Multicentre Randomized Controlled Trial

Gong JQ, Lin L, Lin T, et al. Br J Dermatol. 2006;155:680–687 PURPOSE OF THE STUDY. To investigate the colonizing features of Staphylococcus aureus in the lesional and nonlesional skin of patients with eczema and atopic dermatitis (AD) in China and to compare the therapeutic effect of mupirocin plus hydrocortisone butyrate with vehicle ointment plus hydrocortisone butyrate. STUDY POPULATION. There were 327 patients with AD and eczema (177 male and 150 female); 75 were aged &amp;lt;10 years, 48 between 10 and 18 years, and 204 &amp;gt;18 years. METHODS. A multicenter, double-blind randomized trial was conducted. Eczema area and severity index scores were evaluated before the start of the trial and on the 7th, 14th, and 28th day of treatment. Swabs for bacterial isolation were taken from lesional skin before the start of the trial and on the 7th, 14th, and 28th day of treatment and from nonlesional skin only before the start of the trial. A combination topical therapy with mupirocin plus hydrocortisone butyrate ointment was used in the experimental group, with vehicle ointment plus hydrocortisone butyrate ointment as a control. RESULTS. Of 327 patients enrolled onto the study, bacteria were isolated from 74.8% of lesional and 34% of nonlesional skin samples from patients with AD, of which S aureus accounted for 79.8% and 80.5%, respectively. The colonization density of S aureus was markedly higher in lesional than in nonlesional skin and was positively correlated with lesion severity. Both groups had equivalent clearing of AD and improvement of skin lesions. The patients with severe AD improved faster with combination therapy compared with monotherapy with hydrocortisone butyrate ointment. However, the patients with severe AD were equivalent at days 14 and 28. CONCLUSIONS. This study confirmed that lesional skin of patients with AD was more frequently colonized with S aureus than was nonlesional skin. The more severe the eczema, the higher the colonization rate of S aureus. S aureus infection is related to the pathogenesis of eczema and AD. An antibiotic-corticosteroid combination and corticosteroid alone both provided good therapeutic effect in eczema and AD, and both reduced colonization by S aureus. Early combined topical therapy is beneficial to patients with moderate-to-severe eczema and AD, and it is unnecessary to use antibiotics at later stages of disease or in mild eczema or AD. REVIEWER COMMENTS. S aureus colonization has been recognized as a long-standing issue in AD. This study confirmed a high rate of colonization and correlation with AD severity. The important new finding was that treatment with topical steroids only decreased S aureus colonization without treatment of antibiotics. This is important as the rate of resistant S aureus rises. Maintaining good control of AD will keep the use of unnecessary antibiotics down. Therefore, antibiotic treatment is probably just necessary for the patients with severe AD flares.

Impaired PI3K/Akt Activation-Mediated NF-κB Inactivation Under Elevated TNF-α Is More Vulnerable to Apoptosis in Vitiliginous Keratinocytes

Levels of the cytokines IL-6, IL-1alpha, and tumor necrosis factor-alpha (TNF-alpha) are significantly higher in lesional than in non-lesional skin of patients with vitiligo. However, how cytokines affect pigmentation is not fully understood. To examine the mechanism, Western blot analysis with TNF-alpha, Fas ligand (FasL), and downstream signaling molecules such as I-kappaB, NF-kappaB, TNF-R1-associated factor 2, Akt, and PTEN (phosphatase and tension homologue) were performed for the suction-blistered depigmented and normally pigmented epidermis from 10 patients. Levels of TNF-alpha and FasL were significantly higher in the depigmented epidermis. Interestingly, phosphorylation levels of I-kappaB, NF-kappaB, and Akt were lower in the depigmented epidermis. Moreover, PTEN, which could inhibit the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway, was significantly higher in depigmented epidermis, implying that vitiliginous keratinocytes may be more susceptible to TNF-alpha-mediated apoptosis through impaired Akt and NF-kappaB activation. To test this hypothesis, cultured normal human keratinocytes were treated with TNF-alpha in the presence of a PI3K inhibitor to suppress Akt activation. Keratinocytes showing impaired Akt activation demonstrated increased apoptosis with less activation of NF-kappaB. Thus, reduced activation of NF-kappaB via impaired PI3K/Akt activation under increased TNF-alpha levels could result in increased apoptosis of vitiliginous keratinocytes.

Lymphangiogenesis and angiogenesis in non‐phymatous rosacea

Our study evaluated the expression of vascular endothelial growth factor (VEGF), CD31 and D2-40 in involved and uninvolved skin of 18 patients with rosacea. Immunostaining of facial skin specimens with VEGF, CD31 and D2-40 was compared between the lesional and the non-lesional skin of patients with erythemotelangiectatic and papulopustular rosacea. Significantly increased dermal expression of VEGF in lesional vs. non-lesional skin (88.9% and 55.6%) was observed. Dermal expression of CD31 and D2-40 was also increased in lesional vs. non-lesional skin. There was no statistically significant difference in cutaneous expression of VEGF, CD31 and D2-40 between patients with papulopustular and erythemotelangiectatic rosacea, and no correlation was found between disease duration and immunoreactivity of VEGF, CD31or D2-40. Our study showed marked immunostaining of lesional skin with VEGF, CD31 and D2-40 compared with non-lesional skin. Increased immunoreactivity of D2-40 in lesional skin is interesting, given that none of the patients had facial edema. There are no published data regarding the role of lymphangiogenesis in patients with non-phymatous rosacea; thus, our study represents a new understanding of its pathogenesis. Lack of correlation between D2-40 expression and disease duration suggests that lymphatics are involved early in the pathogenesis of rosacea and do not constitute a late event.

Bradykinin is a potent pruritogen in atopic dermatitis: A switch from pain to itch

Histamine, substance P, serotonin and bradykinin were applied by iontophoresis to lesional and visually non-lesional skin of 14 patients with atopic dermatitis, and normal skin of 15 healthy volunteers. Itch could be evoked by light stroking of skin with a cotton swab (alloknesis) in all lesional skin sites, but not in non-lesional or normal skin. Substances were applied in the same skin area before and 3 h after administration of placebo or antihistamine (olopatadine hydrochloride: H1-receptor-blocker). Intensities of itch and pain sensation and areas of flare and wheal were measured. All the substances induced significantly more intense itch in lesional skin than in non-lesional skin of patients. Even bradykinin, which evoked only weak itch and pain of similar intensities in non-lesional skin of patients and in healthy volunteers, induced intense itch in lesional skin, while the simultaneously increased pain did not suppress the itch sensation, indicating central sensitization. Histamine- and substance P-induced itch was almost completely suppressed by antihistamines, whereas bradykinin- and serotonin-induced itch was not. This suggests that substance P is a histamine-dependent pruritogen also in lesional skin under sensitized conditions but that bradykinin and serotonin are histamine-independent pruritogens in lesional skin. It is concluded that serotonin and bradykinin, classic endogenous algogens, can turn into potent histamine-independent pruritogens in lesional skin of atopic dermatitis.

Skin colonization byStaphylococcus aureusin patients with eczema and atopic dermatitis and relevant combined topical therapy: a double-blind multicentre randomized controlled trial

Staphylococcus aureus has a peculiar ability to colonize the skin of patients with eczema and atopic dermatitis (AD), and is consistently found in eczematous skin lesions in these patients. A correlation between the severity of the eczema and colonization with S. aureus has been demonstrated, and it has been determined that bacterial colonization is an important factor aggravating skin lesions. Patients colonized with S. aureus have been treated with antibiotics in several open and double-blind placebo-controlled studies, with conflicting results. To investigate the colonizing features of S. aureus in the lesional and nonlesional skin of patients with eczema and AD in China and to compare the therapeutic effect of mupirocin plus hydrocortisone butyrate with vehicle ointment plus hydrocortisone butyrate. A multicentre, double-blind randomized trial was conducted. Eczema Area and Severity Index (EASI) scores were evaluated before the start of the trial and on the 7th, 14th and 28th day of treatment. Swabs for bacterial isolation were taken from lesional skin before the start of the trial and on the 7th, 14th and 28th day of treatment, and from nonlesional skin only before the start of the trial. A combination topical therapy with mupirocin plus hydrocortisone butyrate ointment was used in the experimental group, with vehicle ointment plus hydrocortisone butyrate ointment as a control. Of 327 patients enrolled in the study, 208 had eczema and 119 had AD. Bacteria were isolated from 70.2% of lesional and 32.7% of nonlesional skin samples from patients with eczema, of which S. aureus accounted for 47.3% and 27.9%, respectively. Bacteria were isolated from 74.8% of lesional and 34.5% of nonlesional skin samples from patients with AD, of which S. aureus accounted for 79.8% and 80.5%, respectively. The colonization density of S. aureus was markedly higher in lesional than in nonlesional skin, both in patients with eczema and with AD (P < 0.01, P < 0.05), and was positively correlated with lesion severity. Considering the EASI scores before and after treatment and the final effective rate, good therapeutic effects were obtained in both the combination experimental groups and the control groups (P < 0.01), and there were no differences in the global therapeutic effect between the two groups in patients with eczema and with AD (P > 0.05). However, in patients with eczema with a clinical score of > 8 or in patients with AD with a clinical score of > 7, the therapeutic effect in the experimental groups was superior to that in the control groups (P < 0.05) on the 7th day of treatment. There were no differences between the two groups on the 14th and 28th days of treatment (P > 0.05). Following the improvement of symptoms and signs of eczema and AD, the positive rates of bacteria and S. aureus were reduced on the 7th day of treatment. This study confirmed that lesional skin of patients with eczema and AD was more frequently colonized with S. aureus than was nonlesional skin. The more severe the eczema, the higher the colonization rate of S. aureus, and S. aureus was also more often present in lesional and nonlesional skin in patients with AD than in those with eczema. Staphylococcus aureus infection is related to the pathogenesis of eczema and AD. An antibiotic-corticosteroid combination and corticosteroid alone both gave good therapeutic effect in eczema and in AD, and both reduced colonization by S. aureus. Early combined topical therapy is beneficial to patients with moderate to severe eczema and AD, and it is unnecessary to use antibiotics at later stages of disease or in mild eczema or AD.