Unaffected Skin Research Articles

018 TSST-1+ Staphylococcus aureus in bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune blistering disease that is treated with high dose immunosuppression due to lack of specific targets. Staphylococcus aureus is a commensal bacterium implicated inflammatory and autoimmune disorders because of its secretion of toxins with superantigen effects. We prospectively evaluated S. aureus colonization and its production of toxic shock syndrome toxin-1 (TSST-1) in 28 new onset BP patients. Inclusion criteria were active blistering and linear basement membrane IgG/C3 or a serum ELISA >14 for BP180 IgG. Bacterial swabs were obtained from the lesion interior, nares and unaffected skin of BP patients and nares and anatomically matched skin of 28 age- and sex-matched controls. Staphylococcal growth was assessed on blood agar, and TSST-1 production by cultured S. aureus isolates and in blister fluid was evaluated by immunoblot. S. aureus colonization of BP lesions was 3-6-fold higher than the nares or unaffected skin from the same patients (p≤0.300) and 6-fold higher than control nares or skin (p≤0.0015). Evaluation of superantigen gene profiles using PCR indicated that 96% of BP patients are colonized with a clonal strain of TSST-1+ S. aureus. Colonization and circulating levels of TSST-1 neutralizing antibodies, measured by ELISA, did not correlate. Interestingly, S. aureus colonization was not observed in patients who had received prior antibiotics. In colonized patients with severe disease, addition of anti-staphylococcal antibiotics resulted in clinical improvement and eliminated lesional colonization. This study shows that BP lesions harbor a clonal strain of TSST-1 producing S. aureus that is not evident in the general elderly population. Thus, immunosuppressive therapies should be balanced with the knowledge that S. aureus is likely present in BP lesions and the knowledge that antibiotics may play an important therapeutic role through bacterial clearance.

Hypertension due to loss of water

In the current issue of Acta Physiologica, two papers are published investigating the development of arterial hypertension under conditions of pathological loss of free water in two disease models. Kovaric et al 1 studied rats with chronic renal failure, shedding excessive free water via their remnant kidney due to a hypostenuric concentrating defect. Wild et al. 2 investigated a transgenic mouse model for psoriasis, losing free water via the psoriatic skin lesions. In both cases the animals displayed hypertension, cutaneous vasoconstriction, and metabolic adaptations towards enhanced production of urea and organic osmolytes typical for a phylogenetically ancient water conservation response termed aestivation 3 . The international consortia of investigators spearheaded by Titze, Karbach, and Kitada therefore conclude that in both cases the hypertension was due to cutaneous vasoconstriction in an attempt to limit epidermal water loss. Because in either case the hypertension was associated with loss of water, and thus with hypo- rather than hypervolemia, the authors argue, that this concept of pathogenesis of hypertension is antipodal to the classic view that relies on hypervolemia due to sodium retention.

Localized psoriasis post hand burn

The Koebner Phenomenon (KP) is the appearance of new skin lesions on previously unaffected skin secondary to trauma. This phenomenon is given the fact that the new lesions that appear are clinically and histologically identical to the patient’s underlying cutaneous disease [1].

Nuclear IL-33 Plays an Important Role in the Suppression of FLG, LOR, Keratin 1, and Keratin 10 by IL-4 and IL-13 in Human Keratinocytes

IL-33 is a chromatin-associated multifunctional cytokine implicated in the pathogenesis of atopic dermatitis (AD), an inflammatory skin disorder characterized by skin barrier dysfunction. The previous reports show that IL-33 is highly detected in the nucleus of epidermal keratinocytes in AD lesions compared with that in unaffected or normal skin. However, it is unclear whether intracellular IL-33 directly contributes to the pathogenesis of AD. T helper type 2 cytokines IL-4 and IL-13 that are elevated in AD lesions suppress keratinocyte differentiation to impair skin barrier function. We investigated whether intracellular IL-33 is involved in IL-4 and IL-13 function. In monolayer culture and living skin equivalent analyses, IL-4 and IL-13 increased the expression of full-length IL-33 in the nucleus of keratinocytes by activating the MAPK/extracellular signal‒regulated kinase kinase/extracellular signal‒regulated kinase signaling pathway, which is necessary for the inhibition of differentiation markers FLG, LOR, keratin 1, and keratin 10. The nuclear IL-33 functions as a transcription cofactor of signal transducer and activator of transcription 3, increasing the binding of phosphorylated signal transducer and activator of transcription 3 to FLG promoter, thereby inhibiting its transcription, and it inhibits the expression of transcription factor RUNX1 by signal transducer and activator of transcription 3 and signal transducer and activator of transcription 6, thereby downregulating LOR, keratin 1, and keratin 10. Thus, the elevated nuclear IL-33 in the epidermis of AD lesions may be involved in the pathogenesis of AD by inhibiting keratinocyte differentiation and skin barrier function.

Automatic Skin Tumor Detection Using Online Tiger Claw Region Based Segmentation – A Novel Comparative Technique

Skin tumor acts as a premier factor for high death rate throughout the world. It is difficult for the radiologists to segment the skin tumour cells. Various research work focus on accurate segmentation but not on the time of processing. The intention of this research work is to provide an efficient enhancement method and tumor detection from other unaltered regions. This work relies mainly on computed tomography (CT) tumor images of the skin, benign or malignant, that has been implemented efficiently. In this research paper, a novel methodology called Online Tiger Claw Region Based Segmentation (OTCRBS) is proposed which is used mainly to detect the boundary of unaffected Skin Cell, similar to tiger which uses its claws to tear off the skin of its prey during the search for its food. By using metric for the region, various properties can be formulated for the detection of anomalous skin cells. 98.68 and 97.71% accuracy is produced for procurement of benign and malignant nodule in MATLAB 2018a, respectively. Computation time was only 7.65 s. Comparative analysis is made with different segmentation methods. Experimental results establish that the proposed flow outperforms all the existing segmentation methods for the proper detection of tumor cells.

Validation of two predictive models designed to aid clinicians in identifying Stevens-Johnson syndrome/toxic epidermal necrolysis: A single institution retrospective review

Validation of two predictive models designed to aid clinicians in identifying Stevens-Johnson syndrome/toxic epidermal necrolysis: A single institution retrospective review

Cold Atmospheric Plasma (CAP) for the Treatment of Actinic Keratosis and Skin Field Cancerization: Clinical and High-Frequency Ultrasound Evaluation.

IntroductionCold atmospheric plasma (CAP) has been clinically demonstrated to be an effective treatment for actinic keratosis (AK) in a number of case series. In this study, we evaluated the efficacy of CAP in the treatment of multiple AKs and assessed morphological changes induced on the skin field of cancerization both clinically and by high-frequency ultrasound (HFUS).MethodsPatients with multiple grade I–II AKs of the scalp and/or face who were resistant or intolerant to conventional field-directed treatments were enrolled. CAP treatments were performed using a microwave-driven argon plasma jet. At baseline and 3 months after the last CAP session, performance indexes were determined using three-dimensional digital pictures and HFUS investigations were performed on a representative Olsen grade II AK and a small spot of clinically unaffected skin within the test area. ResultsTwelve patients were enrolled in the study. All clinical variables showed a statistically significant reduction after CAP. HFUS evaluation revealed that the total, epidermal and dermal thicknesses of the target AKs had not changed with treatment. CAP therapy significantly increased dermal density in both the target AK and the surrounding photodamaged skin and signficantly decreased the thickness of the subepidermal low-echogenic band in the perilesional skin, which is an ultrasound sign of photodamage. ConclusionsCold atmospheric plasma was found to be an effective treatment for patients with multiple AKs. CAP was not followed by skin atrophy. HFUS examiniation showed the CAP improved features of chronic photodamage of the dermis of the skin underlying and surrounding the AK spots.

Expression of OPN3 in fibroblasts, melanocytes, and keratinocytes of skin with facial melasma in comparison with unaffected adjacent skin

Expression of OPN3 in fibroblasts, melanocytes, and keratinocytes of skin with facial melasma in comparison with unaffected adjacent skin

Immunohistochemical Expression of Galectin-3 in Pemphigus Vulgaris.

Pemphigus vulgaris (PV) is an autoimmune bullous disorder related to immunoglobulin-G autoantibodies against desmoglein-3. Galectin-3 is one of the main elements of the immunoglobulin-E group which is essential in the cell-cell or cell-matrix adhesion. Although the presence of immunoglobulin-E autoantibodies in PV has been observed, no studies have been performed to describe the role of galectin-3 in PV. We evaluated galectin-3 expression in PV as a first step in assessing its impact in the pathogenesis of this autoimmune blistering process. In a retrospective study, 56 specimens from 45 patients diagnosed with PV were stained with antibodies to galectin-3. The percentages of nuclear and cytoplasmic galectin-3 expression as well as staining intensity were evaluated around blisters and adjacent unaffected skin. We observed a significant decrease in galectin-3 cytoplasmic and nuclear expression as well as stain intensity around blisters compared with adjacent unaffected skin. Although autoantibodies against desmogleins trigger the blister formation in PV patients, loss of galectin-3 may play a role in the extension of blister formation by initiating cell-cell disassembly at the level of the intercellular keratinocyte desmosome. We demonstrated a lower expression of galectin-3 around the blisters in PV. The pathogenesis of the blister formation may be related to lower expression of galectin-3. Additional studies are necessary to clarify the result of this outcome and determine the accurate pathogenesis of blister formation in PV.

RNA tape sampling in cutaneous lupus erythematosus discriminates affected from unaffected and healthy volunteer skin

ObjectivePunch biopsy, a standard diagnostic procedure for patients with cutaneous lupus erythematosus (CLE) carries an infection risk, is invasive, uncomfortable and potentially scarring, and impedes patient recruitment in clinical trials....

Single-cell immunopathology of systemic contact allergy associated with corticosteroids

Rationale Allergic contact dermatitis (ACD) is a classic delayed hypersensitivity reaction commonly associated with corticosteroids that can rarely manifest as a delayed rash following systemic administration. Methods A 50-year old woman developed local edema and delayed spreading rash following interarticular injection with dexamethasone and methylprednisolone acetate (MA). 6-months following this intradermal skin testing (IDST) was positive to MA and methylprednisolone sodium succinate, and IDST and patch test negative to other corticosteroids. She tolerated dexamethasone challenge. Cells were isolated from 4-mm punch biopsies from 48-hour positive IDST and unaffected skin with liberase digestion and sorted on CD3. scTCR and scRNA-seq were performed using plate-based assays (Smart-seq-2). Data was filtered using Seurat and analysed with Visual Genomics Analysis Studio (VGAS) software. Results Histopathology showed a superficial perivascular dermatitis with epidermal spongiosis in keeping with ACD. T-cells were 18x more prevalent in affected skin and those from both affected and unaffected sites were predominantly CD8+ T-cells that expressed CD45RO and polyclonal TCR alpha beta. Differentially expressed genes (DEG) in T-cells from affected skin reflected homing (CCR7,S1PR1), T-cell activation and proliferation (TNFRS9, GRAP2, ZYG11A, ZHX1), T-cell effector differentiation (IL-21R), and stress survival (EEF1A1, IFI6) without representation of markers of T-cell residency (ITGAE) or regulation. Conclusions We demonstrate insights into the immunopathogenesis of drug-induced ACD by showing that IDST+ methylprednisolone associated ACD is corticosteroid-specific and associated with a polyclonal population of primarily CD45RO+ memory CD8+ T-cells showing upregulation of markers of homing, T-cell activation, proliferation and differentiation but lacking markers of T-cell residency and regulation.

Dermal Alterations in Clinically Unaffected Skin of Pseudoxanthoma elasticum Patients

Background: Pseudoxanthoma elasticum (PXE), due to rare sequence variants in the ABCC6 gene, is characterized by calcification of elastic fibers in several tissues/organs; however, the pathomechanisms have not been completely clarified. Although it is a systemic disorder on a genetic basis, it is not known why not all elastic fibers are calcified in the same patient and even in the same tissue. At present, data on soft connective tissue mineralization derive from studies performed on vascular tissues and/or on clinically affected skin, but there is no information on patients’ clinically unaffected skin. Methods: Skin biopsies from clinically unaffected and affected areas of the same PXE patient (n = 6) and from healthy subjects were investigated by electron microscopy. Immunohistochemistry was performed to evaluate p-SMAD 1/5/8 and p-SMAD 2/3 expression and localization. Results: In clinically unaffected skin, fragmented elastic fibers were prevalent, whereas calcified fibers were only rarely observed at the ultrastructural level. p-SMAD1/5/8 and p-SMAD2/3 were activated in both affected and unaffected skin. Conclusion: These findings further support the concept that fragmentation/degradation is necessary but not sufficient to cause calcification of elastic fibers and that additional local factors (e.g., matrix composition, mechanical forces and mesenchymal cells) contribute to create the pro-osteogenic environment.

Evaluation of the neoplastic infiltration of the skin overlying canine subcutaneous soft tissue sarcomas: An explorative study.

Studies regarding the neoplastic infiltration of the skin overlying canine subcutaneous soft tissue sarcoma (sSTS) are lacking. In case of the absence of tumor infiltration, there would be the possibility of leaving this unaffected skin in place, thus simplifying surgery. The aim of the study was to investigate whether the skin overlying sSTSs is infiltrated by neoplastic cells. Dogs with sSTSs treated surgically were prospectively enrolled. After excision, the skin was dissected from the tumor along the natural surgical plane of cleavage and histologically evaluated. Twenty-nine dogs with an sSTS were included (22 grade I, 6 grade II, and 1 grade III). The sSTS-overlying skin was not tumor-infiltrated in 14/29 cases (48.3%). A higher frequency of infiltration was observed in higher grade sSTSs (grades II and III, 100%; P=.006); nevertheless, 8/22 grade I sSTSs (36%) also showed cutaneous infiltration. This infiltration involved the dermis of the skin directly in contact with the tumor (multifocal in 11 and diffuse in four cases). Although the cutaneous tumor infiltration is less frequent in grade I sSTSs and a wide excision may still be the safest treatment for any sSTS for a greater possibility of local control, this study opens the possibility to a less aggressive cutaneous excision, but still with a local curative intent, as only the skin directly in contact with the sSTS has been proven to be tumor-infiltrated. Additional studies are warranted to confirm that excision of only this skin may guarantee a complete local control, especially in lower-grade sSTSs.

Biologic Therapy of Moderate and Severe Forms of Atopic Dermatitis in Children

Atopic dermatitis (AD) is the disease with chronic inflammation, epidermal barrier dysfunction and microbial dysbiosis. AD is widespread, including pediatric population. The article discusses the disease’s pathogenesis: skin barrier deficiency, immunological causes of chronic inflammation, characteristics of normal skin microbiome and its disorders on both affected and unaffected skin of children with AD. Main principles of systemic treatment for moderate and severe forms of disease are considered. Features of targeted therapy with dupilumab (IL 4/IL 13 inhibitor) in children with moderate and severe forms of AD are discussed. The overview of the research results on the dupilumab efficacy and safety is presented.

Skin cytokine profile in patients with mycosis fungoides

Aim of the study. To determine the concentration of cytokines in the skin of patients with mycosis fungoides and correlations between their concentration and the value of the modified scale for assessing the severity of skin lesions mSWAT.
 Methods. The concentration of cytokines was simultaneously determined using xMAP technology: IL-1, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL -31, IL-33, IFN-, sCD40L, TNF- in skin biopsies of 21 patients with early (IAIIA) stages of mycosis fungoides and 4 healthy individuals. Analysis and visualization of the obtained data were carried out using R Statistical Software for MacOS (version 1.3.1056), a free open source software development environment for the R programming language.
 Results. An increase in the level of IL-4 and TNF- in the lesions in patients with mycosis fungoides compared with healthy individuals was shown (p = 0.025 and p = 0.012, respectively). Correlation analysis revealed the formation of cytokine networks in the skin of patients with mycosis fungoides that were not associated with the value of the modified scale for assessing the severity of skin lesions mSWAT. A tendency towards a negative relationship between the mSWAT and IFN scales was found (p = 0.056).
 Conclusion. None of the cytokines in lesions and visually unaffected skin from patients with mycosis fungoides were associated with mSWAT values.

Noninflammatory comedones have greater diversity in microbiome and are more prone to biofilm formation than inflammatory lesions of acne vulgaris.

The ability of Cutibacterium acnes strains to form biofilms has been correlated with their virulence. This study examined biofilm and skin microbiota in acne patients in order to understand their role in the development of acne lesions. Thin sections of punch biopsy specimens of (i) uninflamed comedones, (ii) inflammatory lesions, and (iii) uninvolved adjacent skin of acne patients were examined. Epiflourescence and confocal laser scanning microscopy were used for biofilm detection, and pyrosequencing with taxonomic classification of 16s rRNA gene amplicons was used for microbiota analysis. Of the 39 skin specimens from patients with mild-moderate acne (n=13) that were studied, nine (23%) contained biofilm. Among these specimens, biofilm was most frequently detected in comedones (55.6%) and less frequently in inflammatory papules (22.2%) and uninvolved skin (22.2%). Comedones demonstrated the highest mean alpha diversity of all the lesion subtypes. The relative abundance of Staphylococcus was significantly higher in comedones (11.400%±12.242%) compared to uninvolved skin (0.073%±0.185%, P=0.024). The microenvironment of the comedone differs from that of inflammatory lesions and unaffected skin. The increased frequency of biofilm in comedones may account for the lack of host inflammatory response to these lesions.

The erythema Q-score, an imaging biomarker for redness in skin inflammation.

Physician rating of cutaneous erythema is central to clinical dermatological assessment as well as quantification of outcome measures in clinical trials in a number of dermatologic conditions. However, issues with inter‐rater reliability and variability in the setting of higher Fitzpatrick skin types make visual erythema assessment unreliable. We developed and validated a computer‐assisted image‐processing algorithm (EQscore) to reliably quantify erythema (across a range of skin types) in the dermatology clinical setting.Our image processing algorithm evaluated erythema based upon green light suppression differentials between affected and unaffected skin. A group of four dermatologists used a 4‐point Likert scale as a human evaluation of similar erythematous patch tests. The algorithm and dermatologist scores were compared across 164 positive patch test reactions. The intra‐class correlation coefficient of groups and the correlation coefficient between groups were calculated. The EQscore was validated on and independent image set of psoriasis, minimal erythema dose testing and steroid‐induced blanching images. The reliability of the erythema quantification method produced an intra‐class correlation coefficient of 0.84 for the algorithm and 0.67 for dermatologists. The correlation coefficient between groups was 0.85. The EQscore demonstrated high agreement with clinical scoring and superior reliability compared with clinical scoring, avoiding the pitfalls of erythema underrating in the setting of pigmentation. The EQscore is easily accessible (http://lab.rockefeller.edu/krueger/EQscore), user‐friendly, and may allow dermatologists to more readily and accurately rate the severity of dermatological conditions and the response to therapeutic treatments.

18323 Diagnostic potential of five different biomechanical parameters to detect sclerotic cutaneous graft-versus-host disease

Background: The current standard for assessing skin sclerosis in chronic graft-versus-host disease (cGVHD) is based on clinician assessment. The Myoton device offers quantitative measurements of 5 soft tissue properties: stiffness, relaxation time, oscillation frequency, decrement, and creep. Previous studies revealed increased skin stiffness and an ability to differentiate sclerotic from unaffected post-transplant skin. This study further evaluates the diagnostic potential of the 5 properties.

14928 Epithelialized tracts are active mediators of inflammation in hidradenitis suppurativa

Introduction: Hidradenitis suppurativa (HS) is a chronic, inflammatory disease characterized by painful nodules, abscesses and draining sinuses, most commonly occurring in the inguinal, axillary, and submammary areas. Severe disease is associated with discharging sinus tracts and fibrotic scarring. The development of dermal tunnels and interconnecting tracts is poorly understood, and has traditionally been considered an end-stage feature of disease. Methods/Results: After receiving approval from the Rockefeller University’s Institutional Review Board, unaffected, perilesional and lesional HS skin was collected by punch biopsies.

Dynamic Leukocyte Populations Are Associated With Early- and Late-stage Lesions in Hidradenitis Suppurativa.

Hidradenitis suppurativa (HS) is a chronic inflammatory skin condition typically targeting the axillary and anogenital regions of the body. The massive inflammatory cell infiltrate produced in this cryptogenic condition has led investigators in the attempt to link particular inflammatory cell fractions and cytokines to disease development, and ultimately to disease treatment. This study qualitatively and quantitatively analyzes the white blood cell fractions of macrophages, B-lymphocytes, T-lymphocytes, plasma cells, and granulocytes in 104 HS lesions on formalin-fixed paraffin-embedded tissues using immunohistochemistry (IHC). Four dermis-associated epithelial categories were investigated from persons with HS: 15 unaffected HS skin (US), 19 distended but unruptured follicle epithelium (UF), 62 migrating stratified squamous epithelium (MSSE) from ruptured follicles, and 35 degraded migrating epithelial sheets (DMES). In addition, 27 control skin (CS) from persons without HS were evaluated. Analysis of cell counts indicated that non-migratory dermal epithelium (CS, US, and UF) stimulated very little inflammatory response. However, contrary to previous studies which indicated macrophages to be the chief inflammatory cell in HS, this study showed that plasma cells were the primary cell type present in early-stage HS lesions (MSSE), whereas granulocytes were the major cell population seen in late-stage HS lesions (DMES).