Human Skin Lesions Research Articles

Comparison of CD30L and OX40L Reveals CD30L as a Promising Therapeutic Target in Atopic Dermatitis.

Blocking IL-13 is highly efficacious in patients with Th2-biased atopic dermatitis (AD), and recent clinical data have highlighted that targeting the T cell costimulatory molecules OX40 and OX40L (TNFSF4) holds promise for future treatment of AD. We asked whether targeting another T cell costimulatory molecule, CD30L (TNFSF8), might also be a possible treatment option in AD. Single-cell RNA-seq data from human AD skin lesions was analyzed to identify pathogenic IL-13- or IL-22-producing T cells and assess expression of CD30 and its ligand in comparison to OX40 and its ligand. Additionally, a murine model of AD with repetitive exposure to house dust mite allergen was used to compare neutralizing antibodies against CD30L with those against IL-13 or OX40L. Analysis of several scRNA-seq datasets from skin lesions of AD patients showed that transcripts for CD30 or CD30L were found expressed with OX40 or OX40L in the primary T cell populations that also expressed mRNA for IL13 and/or IL22. Suggesting that this could be therapeutically relevant, mice treated prophylactically with a blocking CD30L antibody were protected from developing maximal allergen-induced AD features, including epidermal and dermal thickening, immune cell infiltration, and expression of AD-related genes, similar to mice treated with a blocking IL-13 antibody. Moreover, therapeutic neutralization of CD30L in mice with experimental AD also reduced all of the pathological skin lesion features to a comparable extent as blocking OX40L. These data suggest that targeting the CD30-CD30L axis might hold promise as a future therapeutic intervention in human AD, similar to targeting the OX40-OX40L axis.

Intravital Imaging of Disease Mechanisms in a Mouse Model of CCM Skin Lesions.

Cerebral cavernous malformation (CCM) is a disease characterized by vascular malformations that primarily develop in the brain. These malformations are prone to leak, and their rupture or thrombotic closure can cause life-threatening hemorrhages and strokes. Mouse models have been instrumental to study the disease, but most cause premature lethality, precluding the investigation of disease mechanisms through intravital microscopy. Current mouse models also do not recapitulate human CCM skin lesions. Endothelial-specific deletion of Ccm3 via systemic tamoxifen application at postnatal day 4 or 5 prolongs survival and induces vascular malformations in the mouse brain and ear skin. CCM skin lesions can also be induced by topic tamoxifen administration directly to the ear. The thin, flat morphology of the ear skin is ideal for intravital microscopy. Dextran dyes and platelet markers allow to study blood flow and blood clot formation, in living animals in real time. We report that human CCM skin lesions can be recapitulated in a mouse model and that skin lesions share hallmarks of CCM brain lesions. Intravital imaging reveals that CCM skin lesions are slow-flow malformations prone to thrombus formation. Intravital imaging of CCM skin lesions expands the toolkit of CCM research and allows longitudinal studies of lesion growth.

53005 Perceived stress can trigger psoriatic lesions in human skin in vivo, which can be suppressed by aprepitant

53005 Perceived stress can trigger psoriatic lesions in human skin in vivo, which can be suppressed by aprepitant

Experimental inoculation of pigs with monkeypox virus results in productive infection and transmission to sentinels

ABSTRACT Monkeypox virus (MPXV) is a re-emerging zoonotic poxvirus responsible for producing skin lesions in humans. Endemic in sub-Saharan Africa, the 2022 outbreak with a clade IIb strain has resulted in ongoing sustained transmission of the virus worldwide. MPXV has a relatively wide host range, with infections reported in rodent and non-human primate species. However, the susceptibility of many domestic livestock species remains unknown. Here, we report on a susceptibility/transmission study in domestic pigs that were experimentally inoculated with a 2022 MPXV clade IIb isolate or served as sentinel contact control animals. Several principal-infected and sentinel contact control pigs developed minor lesions near the lips and nose starting at 12 through 18 days post-challenge (DPC). No virus was isolated and no viral DNA was detected from the lesions; however, MPXV antigen was detected by IHC in tissue from a pustule of a principal infected pig. Viral DNA and infectious virus were detected in nasal and oral swabs up to 14 DPC, with peak titers observed at 7 DPC. Viral DNA was also detected in nasal tissues or skin collected from two principal-infected animals at 7 DPC post-mortem. Furthermore, all principal-infected and sentinel control animals enrolled in the study seroconverted. In conclusion, we provide the first evidence that domestic pigs are susceptible to experimental MPXV infection and can transmit the virus to contact animals.

An Extensive Investigation into the Use of Machine Learning Tools and Deep Neural Networks for the Recognition of Skin Cancer: Challenges, Future Directions, and a Comprehensive Review

Skin cancer poses a serious risk to one’s health and can only be effectively treated with early detection. Early identification is critical since skin cancer has a higher fatality rate, and it expands gradually to different areas of the body. The rapid growth of automated diagnosis frameworks has led to the combination of diverse machine learning, deep learning, and computer vision algorithms for detecting clinical samples and atypical skin lesion specimens. Automated methods for recognizing skin cancer that use deep learning techniques are discussed in this article: convolutional neural networks, and, in general, artificial neural networks. The recognition of symmetries is a key point in dealing with the skin cancer image datasets; hence, in developing the appropriate architecture of neural networks, as it can improve the performance and release capacities of the network. The current study emphasizes the need for an automated method to identify skin lesions to reduce the amount of time and effort required for the diagnostic process, as well as the novel aspect of using algorithms based on deep learning for skin lesion detection. The analysis concludes with underlying research directions for the future, which will assist in better addressing the difficulties encountered in human skin cancer recognition. By highlighting the drawbacks and advantages of prior techniques, the authors hope to establish a standard for future analysis in the domain of human skin lesion diagnostics.

Polarization-Based Digital Histology of Human Skin Biopsies Assisted by Deep Learning

Mueller polarimetry has proven to be a powerful optical technique to complement medical doctors in their conventional histology analysis. In this work, various degenerative and malignant human skin lesions were evaluated ex vivo using imaging Mueller polarimetry. The Mueller matrix images of thin sections of biopsies were recorded and the differential decomposition of Mueller matrices was applied pixel-wise to extract the polarization fingerprint of the specimens under study. To improve the classification accuracy, a deep learning model was created. The results indicate the sensitivity of polarimetry to different skin lesions and healthy skin zones and their differentiation, while using standard histological analysis as a ground truth. In particular, the deep learning model was found sufficiently accurate to detect and differentiate between all eight classes in the data set. Special attention was paid to the overfitting problem and the reduction of the loss function of the model. Our approach is an effort in establishing digital histology for clinical applications by complementing medical doctors in their diagnostic decisions.

Microarray Gene Expression Analysis of Lesional Skin in Canine Pemphigus Foliaceus.

Canine pemphigus foliaceus (PF) is considered the most common autoimmune skin disease in dogs; the mechanism of PF disease development is currently poorly understood. Therefore, this study aimed to characterize the molecular mechanisms and altered biological pathways in the skin lesions of canine PF patients. Using an RNA microarray on formalin-fixed, paraffin-embedded samples, we analyzed the transcriptome of canine PF lesional skin (n = 7) compared to healthy skin (n = 5). Of the 800 genes analyzed, 420 differentially expressed genes (DEGs) (p < 0.05) were found. Of those, 338 genes were significantly upregulated, including pro-inflammatory and Th17-related genes. Cell type profiling found enhancement of several cell types, such as neutrophils, T-cells, and macrophages, in PF skin compared to healthy skin. Enrichment analyses of the upregulated DEGs resulted in 78 statistically significant process networks (FDR < 0.05), including the Janus kinase signal transducer and activator of transcription (JAK-STAT) and mitogen-activated protein kinase (MAPK) signaling. In conclusion, canine PF lesional immune signature resembles previously published changes in human pemphigus skin lesions. Further studies with canine PF lesional skin using next-generation sequencing (e.g., RNA sequencing, spatial transcriptomics, etc.) and the development of canine keratinocyte/skin explant PF models are needed to elucidate the pathogenesis of this debilitating disease.

Methotrexate and electrostimulation cooperate to alleviate the relapse of psoriasiform skin inflammation by suppressing memory T cells

Methotrexate (MTX) is an immunosuppressant used to treat autoimmune diseases, including psoriasis. However, like other immunosuppressants, MTX alone does not prevent their recurrence. Electrostimulation (ES) has been utilized to treat some inflammatory disorders without any major side-effect. But it remains unknown if ES alone, or together with MTX, ameliorates autoimmune disease relapse: a sticky medical problem. In particular, the mechanisms underlying ES action remain unclear. The objective of this study was to determine an impact of ES and/or MTX on psoriasis relapse and their potential cooperation. We found that regional ES, but not MTX, ameliorated psoriasiform skin inflammation recurrence. Interestingly, treatment with both MTX and ES further prevented psoriasis recurrence compared to ES alone. Moreover, ES downregulated potassium channel Kv1.3 on T-cells and reduced CD4+/CD8+ effector memory (TEM) and CD8+ skin-resident memory T (TRM) cells, while ES plus MTX further decreased CD8+ TEM/TRM cells compared to ES alone. However, ES failed to further attenuate psoriasis recurrence or suppress T cell memory in Kv1.3-deficient mice, whereas lack of Kv1.3 itself ameliorated psoriasis relapse by shrinking T cell memory pool. Importantly, ES moderately inhibited T-cell proliferation in vitro. ES also reduced human CD8+ TRM cells and attenuated human skin lesions in humanized mice grafted with lesional skin from patients with recurrent psoriasis, with an enhanced efficacy in mice treated with both ES and MTX. Thus, ES and MTX cooperated to prevent psoriasis relapse by reducing T-cell memory via targeting potassium channel Kv1.3. Our studies may be implicated for treating human psoriasis.

170 Perceived stress can trigger psoriatic lesions in human skin in vivo, which can be suppressed by aprepitant

170 Perceived stress can trigger psoriatic lesions in human skin in vivo, which can be suppressed by aprepitant

A New Herbal Medicine Formulation with Potential Anti-scabies Properties to Treat Demodex and Sarcoptes Parasites.

Scabies is considered an external parasite notorious for its high prevalence causing severe and contagious skin lesions in humans and animals worldwide. This study has introduced a medicine to treat dogs infested with scabies (variants of Demodex, Sarcoptes, Psoroptes, Otodectes, etc.). The present study offers a no-side-effect herbal formulation to treat dogs infested with scabies. Unlike oral and injectable medicines, which take the form of an ointment and are topically applied on-site, this medicinal formulation can be easily used without concerns over its side effects or consumption dosages. This medicinal formulation requires no skin rinsing due to its herbal and high skin absorption properties, as recovery may take less than a month with a maximum of two times of application. To carry out the experiment, 25 sick dogs with various breeds and ages suspected of scabies were gathered. Following accurate morphological examinations of all the samples, a deep skin chip of the lesion site was provided, which was examined by a microscope. Then, 13 dogs (Mix, Terrier, Pug, Husky, Spitz) were infested with Demodex scabies and 12 dogs (Pittbull, Mix, Shih Tzu, Terrier, Boxer, Setter) with Sarcoptic scabies. The prepared product was topically administered at a constant 2% dosage to the bodies of all the samples. To prepare the ointment, 1 g of Borax (Na2B4O7·10H2O) was first dissolved in 35 g deionized water and heated to 70°C. Then, 45 g of liquid paraffin (CnH2n+2) was mixed with 1 g of Carvacrol (C10H14O) and 1 g of geranium (C10H18O) and stirred well to become a phase. Later, 17 g of the melted beeswax (C15H31COOC30H61) was added to the liquid paraffin compound. In the end, the aqueous phase was added to the oil phase, and the mixture process immediately began in one direction with a glass stirrer and continued until the product cooled down. Essential oils (EO) was obtained by steam distillation of fresh Thyme and Rose-Acented Geranium in a stainless steel distillation apparatus (alembic) for 3 h. The main components of the essential oils used in the formulation were performed using a Hewlett-Packard GC system interfaced with a mass spectrometer equipped with an HP5-MS capillary column (30 m, 0.32 mm, 0.25 µm film thicknesses). For GC-MS detection, electron ionization with ionization energy of 70 eV was used. To examine the presence of scabies, weekly skin sampling was performed, and the treatment continued until 30 days, when no skin chip of the scabies was noted. The findings revealed that the formulation developed no side effects and removed the daily use, as it could be administered once or twice a week. Also, complete recovery of scabies in all the breeds was found to be less than a month at most. This medicinal formulationcan be easily used without concerns over its side effects or consumption dosages. This study introduced a herbal formulation with effective herbal ingredients without any side effects to treat the sarcoptes and demodex parasites; unlike other chemical compounds, this medicinal formulation has no side effects, while some other formulations could develop side effects.

Monkeypox Detection Using Hyper-Parameter Tuned Based Transferable CNN Model

The reemergence of Monkeypox, a communicable illness resulting from the Monkeypox virus, has raised apprehensions about a potential swift global pandemic similar to the COVID-19 epidemic as COVID-19 infections diminish globally. The prompt emphasizes the criticality of prompt action within communities to mitigate the development of the phenomenon. The timely identification and accurate categorization of Monkeypox cutaneous manifestations are crucial for the successful implementation of containment strategies. This paper presents a novel methodology for detecting Monkeypox by utilizing a transferrable Convolutional Neural Network (CNN) model that has been optimized utilizing hyper-parameter tuning techniques. The proposed methodology initiates by improving the quality of the original Monkeypox images, with a specific emphasis on boosting edge details to increase visual clarity. Texture qualities are obtained through an energy layer, enhancing distinctive traits. Our methodology's cornerstone is utilizing the Hyper-parameter-based transferable Convolutional Neural Network (HPT-TCNN), specifically designed to enhance classification accuracy.&#x0D; In contrast to traditional methods, we enhance the architectural design by replacing the pooling layer with a configuration comprising three convolutional layers and one energy layer. The hyper-parameter tuning procedure is optimized by employing the Optimisation Algorithm known as MGS-ROA. In order to enhance the process of model training and validation, we have assembled the "Monkeypox Skin Lesion Dataset (MSLD)," which consists of a collection of images depicting human skin lesions produced by Monkeypox. The dataset in question is vital in evaluating and improving our methodology. In a comparison analysis conducted on other deep learning models, the suggested model has superior performance compared to other models, obtaining a notable accuracy, sensitivity, and specificity, all reaching a value of 93.60%. The outstanding performance shown in this study highlights the methodology's effectiveness in adequately classifying skin lesions associated with Monkeypox. This approach shows potential for physicians and healthcare workers since it facilitates early detection, a crucial factor in preventing the spread of Monkeypox.

B-231 Development and Evaluation of a Lyophilized Mpox Virus qPCR Assay

Abstract Background Increased cases of monkeypox virus (MPXV) have been reported to WHO from 110 countries. As of January 16, 2023, a total of 84 733 laboratory confirmed and 1354 probable cases, including 80 deaths, have been reported to WHO. The US is assessed as high-risk region. Real-time PCR, with its sensitivity and specificity, is the preferred lab test technique. Since gene mutation has been observed in MPXV, developing assays with more than one target was recommended by the FDA. Methods A lyophilized qPCR multiplex assay was developed for MPXV detection. This assay contains primers and (1) MGB probe targeting highly conserved MPXV sequence (GeneBank AF380138 F3L), (2) BHQ probe targeting non-variola orthopoxvirus (OPX) sequence, and (3) a probe targeting a selected human house-keeping gene as an internal control. Furthermore, external positive and negative controls were included. A MPXV positive clinical sample from human skin lesions was used for the assay validation. The viral copy number of the sample was determined as 1×107 copies/mL using the standard curve method. Two hundred µL of samples were processed in the KingFisherTM Flex system. The PCR reaction consisted of 5 µL of DNA sample and 20 µL reaction mix and was run on 7500 Fast Dx Real-Time PCR System (Life Technologies). The limit of detection (LoD) was determined as the concentration at which at least 95% of samples were tested positive. A total of 230 contrived samples with concentrations lower than LoD were used to determine cut-off value. Inclusivity and exclusivity studies were conducted by in silico analysis of 80 sequences of MPXV (clade I), 3026 of MPXV (clade II), 10 of camelpox virus, 93 of cowpox virus, 6 of ectromelia virus, and 116 of vaccinia virus (NCBI and GISAID accessed on 10/03/22). Clinical evaluation of the assay was conducted using 80 contrived clinical samples in unique negative skin lesion matrix, including 40 positives (five samples at 5× LOD, five at 2.5×, ten at 1.5×, and twenty at 1×), along with 40 negatives. The positive (PPA) and negative percent agreement (NPA) of the assay were calculated. The study was approved by BRANY IRB. Results The qPCR assay had a cut-off Ct value of 40, and a LoD of 500 copies/mL for both MPXV and OPX with good inclusivity and exclusivity. Clinical evaluation for MPXV yielded a PPA and NPA of 94.9% (95% CI: 83.1%–98.6%) and 100% (89.9%–100%), respectively. For OPX, the PPA and NPA were 92.3% (79.7%–97.4%) and 100% (89.9%–100%), respectively. Conclusions Both analytical and clinical performances of the developed MPXV qPCR assay were good. Lyophilized reagents contain all required components, providing longer shelf-life and easier transportation. MPXV is the only member of the OPX genus known to be circulating among humans in the US currently. This assay as designed could potentially cross-detect different non-variola OPX, such as vaccinia virus and cowpox virus. Healthcare providers should always contact the local public health authorities for more guidance. Furthermore, this study was limited to lesion swab specimens and performance evaluation with additional specimen types is warranted.

B-268 Validation of a New Real-Time PCR Test for the Detection of Mpox Virus on the BD MAXTM System

Abstract Background Enabling labs to rapidly respond to unexpected healthcare challenges is the kind of situations for which the BD MAX™ open system reagent suite is designed. Therefore, after the monkeypox outbreak in non-endemic countries on May 2022, development of a molecular assay for monkeypox detection on the BD MAXTM System was initiated. Methods VIASURE Monkeypox virus Real Time PCR Reagents for BD MAX™ System was designed for the qualitative detection of DNA from monkeypox virus in human skin lesion swabs in VTM/UTM obtained from individuals suspected of monkeypox infection. The Limit of Detection (LoD) was determined by testing a 3-fold dilution series across 5 concentrations. The lowest concentration with 100% detection was tested with an additional 20 replicates. The preliminary LoD was confirmed with a ≥95% detection with 20 replicates. Then, a fully contrived clinical evaluation was performed. Each contrived clinical specimen was prepared using a unique natural clinical specimen matrix (lesion swabs). Twenty (20) of the clinical specimens were prepared at 1-2X LoD, ten (10) were prepared at 3-5X LoD, and thirty (30) were tested as negative samples. Results The lowest detectable concentration of virus at which &amp;gt;95% of all replicates test positive (LoD) was 288 copies/mL. The results for the contrived clinical evaluation are shown in Table 1. 19/20 (≥95%) low positive specimens (1-2xLoD) and 20/20 (100%) moderate positive specimens (3-5x LoD) were detected demonstrating the functionality of the test. Conclusion VIASURE Monkeypox virus Real Time PCR Reagents for BD MAX™ System provides results in a short time and requires less than one minute of hands-on time per sample due to integration of fully automated extraction and thermocycling steps. An additional clinical testing of positive natural clinical specimens must be performed to complete evidence that this product is for the purpose and the population for which it is intended.

Protein inhibitor of activated STAT3 (PIAS3) attenuates psoriasis and associated inflammation.

Psoriasis is a common chronic inflammatory multisystem disease accompanied by hyperproliferation and inflammation of epidermal keratinocytes. Signal transducer and activator of transcription 3 (STAT3) is constitutively activated and plays an important role in epidermal keratinocytes of human psoriatic skin lesions. In this study, we investigated the effects of an endogenous STAT3 inhibitor, a protein inhibitor of activated STAT3 (PIAS3), on the proliferation and inflammation of psoriatic cells. The expression of PIAS3 in psoriatic tissues and healthy skin was analyzed using the Gene Expression Omnibus database and clinical samples. The human immortalized epidermal (HaCaT) cells were used to establish an in vitro psoriasis-like cell model. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-thethrazolium (MTS) assay was used to detect cell proliferation. Flow cytometry was used to determine apoptosis levels. Real-time PCR, western blotting, and enzyme-linked immunosorbentassay (ELISA)were used to detect the expression levels of related factors. Furthermore, a mouse model of imiquimod (IMQ)-induced psoriatic dermatitis was established to verify the in vitro experimental results. The results showed that the mRNA and protein expression levels of PIAS3 were lower in psoriatic lesions than in normal tissues. PIAS3 inhibited the proliferation and promoted apoptosis of M5-induced HaCaT cells. Simultaneously, the mRNA and protein expression levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), and keratin 17 (K17) were significantly decreased and that of p53 was increased, thereby inhibiting the inflammatory response and promoting apoptosis. PIAS3 inhibited the transcription activity of STAT3 and noncanonical nuclear factor-kappaB (NF-κB). Furthermore, PIAS3 attenuated IMQ-induced psoriasis-like inflammation in mice. Our findings suggest that PIAS3 plays an important role in psoriasis by regulating the STAT3/NF-κB signaling pathway and p53. The lack of PIAS3 may represent a novel mechanism underlying the pathogenesis of psoriasis.

A New Herbal Medicine Formulation with Potential Anti-scabies Properties to Treat Demodex and Sarcoptes Parasites

Scabies is considered an external parasite notorious for its high prevalence causing severe and contagious skin lesions in humans and animals worldwide. This study has introduced a medicine to treat dogs infested with scabies (variants of Demodex, Sarcoptes, Psoroptes, Otodectes, etc.). The present study offers a no-side-effect herbal formulation to treat dogs infested with scabies. Unlike oral and injectable medicines, which take the form of an ointment and are topically applied on-site, this medicinal formulation can be easily used without concerns over its side effects or consumption dosages. This medicinal formulation requires no skin rinsing due to its herbal and high skin absorption properties, as recovery may take less than a month with a maximum of two times of application. To carry out the experiment, 25 sick dogs with various breeds and ages suspected of scabies were gathered. Following accurate morphological examinations of all the samples, a deep skin chip of the lesion site was provided, which was examined by a microscope. Then, 13 dogs (Mix, Terrier, Pug, Husky, Spitz) were infested with Demodex scabies and 12 dogs (Pittbull, Mix, Shih Tzu, Terrier, Boxer, Setter) with Sarcoptic scabies. The prepared product was topically administered at a constant 2% dosage to the bodies of all the samples. To prepare the ointment, 1 g of Borax (Na₂B₄O₇·10H₂O) was first dissolved in 35 g deionized water and heated to 70°C. Then, 45 g of liquid paraffin (CnH2n+2) was mixed with 1 g of Carvacrol (C10H14O) and 1 g of geranium (C10H18O) and stirred well to become a phase. Later, 17 g of the melted beeswax (C15H31COOC30H61) was added to the liquid paraffin compound. In the end, the aqueous phase was added to the oil phase, and the mixture process immediately began in one direction with a glass stirrer and continued until the product cooled down. Essential oils (EO) was obtained by steam distillation of fresh Thyme and Rose-Acented Geranium in a stainless steel distillation apparatus (alembic) for 3 h. The main components of the essential oils used in the formulation were performed using a Hewlett-Packard GC system interfaced with a mass spectrometer equipped with an HP5-MS capillary column (30 m, 0.32 mm, 0.25 µm film thicknesses). For GC–MS detection, electron ionization with ionization energy of 70 eV was used. To examine the presence of scabies, weekly skin sampling was performed, and the treatment continued until 30 days, when no skin chip of the scabies was noted. The findings revealed that the formulation developed no side effects and removed the daily use, as it could be administered once or twice a week. Also, complete recovery of scabies in all the breeds was found to be less than a month at most. This medicinal formulationcan be easily used without concerns over its side effects or consumption dosages. This study introduced a herbal formulation with effective herbal ingredients without any side effects to treat the sarcoptes and demodex parasites; unlike other chemical compounds, this medicinal formulation has no side effects, while some other formulations could develop side effects.

Autologous Th2-polarized lymphocytes induce atopic dermatitis lesions in non-atopic human skin xenotransplants.

The key signals that suffice to induce atopic dermatitis (AD) in human skin remain incompletely understood. Also, current mouse models reflect human AD only unsatisfactorily. Therefore, we have asked whether a humanized AD mouse model can be developed that reflects human AD more faithfully and permit to identify key signals that suffice to induce AD lesions in previously healthy human skin in vivo. Healthy human skin from non-atopic donors was transplanted onto SCID/beige mice. After xenotransplant reinnervation by mouse sensory nerve fibers had occurred, mixed autologous human Th2 CD4+ and Tc2 CD8+ T cells that had been pretreated in vitro with IL-2, IL-4, and LPS were injected intradermally into the xenotransplants without skin barrier disruption. Injected non-atopic xenotransplants rapidly developed a morphological, functional, and immunological phenocopy of human AD lesions regarding skin barrier defects, immunopathology including intraepidermal eosinophils, mast cell activation, increase of thymic stromal lymphopoietin, eotaxin-1 and type 2 cytokine circuits, and even showed characteristic neuroimmunological abnormalities such as ß2-adrenergic receptor downregulation. The experimentally induced AD lesions in human skin responded to standard AD therapy (topical dexamethasone or tacrolimus; systemic anti-IL-4Rα antibody [dupilumab]), and relapsed when neurogenic skin inflammation was induced by exposing mice to perceived stress. This new animal model uniquely mimics the complexity of human AD and its clinical response to standard therapy and psychoemotional stressors in vivo, and shows that Th2-polarized lymphocytes associated with excessive IL-4Rα-mediated signaling suffice to induce human AD skin lesions, while atopy and epidermal barrier disruption are dispensable.

Anti-inflammatory and immunomodulatory effects of polysaccharide extracted from Wuguchong (maggot) on 2,4-dinitrochlorobenzene-induced atopic dermatitis in mice.

Atopic dermatitis (AD) is an inflammatory, heterogeneous, chronic skin disorder characterized by recurrent eczematous lesions and intense pruritus, and the pathophysiology mechanism of AD is known for immune dysregulation and inflammatory responses. Wuguchong (maggot) has been widely used in the wound field and found with pharmacological properties of the anti-inflammatory and immunomodulatory function. Recently, some polysaccharides were proven to have beneficial effects on AD skin lesions in mice and humans. However, the effect of the polysaccharide extracted from Wuguchong (PEW) on AD remains to be investigated. In the present study, we examined the anti-inflammatory and immunomodulatory effects of PEW on AD and explored the potential mechanisms. Balb/c mice were orally administrated with PEW to evaluate the therapeutic effect of PEW on 2,4-dinitrochlorobenzene (DNCB)-induced AD. Oral PEW administration significantly ameliorated the lesions and symptoms in AD mice, such as the ear thickness and ear swelling degree, epidermal and dermal thickness, and the infiltration of mast cells. In addition, PEW treatment decreased the levels of serum IgE and histamine, the frequencies of Th1 and Th17 cells, as well as the mRNA expression levels of Th1 and Th17 cytokines and nuclear transcript factors (IFN-γ, T-bet, IL-17A, and ROR-rt). Furthermore, the activation of the NF-κB pathway and the phosphorylation of MAPKs (p38, ERK, and JNK) were significantly suppressed by PEW treatment. Taken together, our study suggests that PEW exerts anti-inflammatory and immunomodulatory effects through inhibition of Th1 and Th17 responses and downregulation of NF-κB and MAPK pathways, PEW would be developed as a promising immune therapy for AD.

Application of neural network and computer vision technologies for image analysis of skin lesion

Opportunity research of using neural networks and computer vision to analyze images of skin lesion and identify features of various pathologies, including oncological neoplasms. A methodology has been developed that makes it possible to evaluate the significance of combinations of color components and spaces in feature extraction using local binary patterns (LBP) and histogram of oriented gradients (HOG) computer vision technologies to extract features of skin changes binary classification of human skin lesions. Optimization of extracted feature makes it possible to more effectively solve the problem of data separability in classification. Research reveals an accessible way to classify skin lesions on a small dataset (less than 1000 images). Research is supposed to be applied to data sequences obtained using a new unique method of multispectral processing of skin lesions. In the course of the work, data from the ISIC-19 and ISIC-20 datasets were used. Samples were formed with a limit of 1000 images for training and validating the models. Additionally, a test sample of 250 images was formed. All images were reduced to 128 × 128 pixels and converted to YCrCb, BGR, Grayscale, HSV color spaces. Features were extracted for each color channel using the HOG and LBP methods. Mathematical models, including neural networks have been used for data classification. The effectiveness of features combinations by color channels and feature extraction methods was evaluated. The preprocessed images were divided into training and validation subsets in a 70/30 ratio. The accuracy, recall, precision and f1-score metrics were used to evaluate the models. The models were evaluated using stratified cross-validation and a test dataset. Optimization of model parameters was carried out based on the loss function represented by the average of cross-validation and evaluation on the validation set. In the process of research, more than 15 000 different optimizations of model parameters were executed. The most stable results on the validation dataset were achieved using ensemble of models, which were trained on a combination of features using local binary patterns (LBP) and histogram of oriented gradients (HOG) technologies. Models which used only local binary patterns technology had the best metrics values, but these models are not recommended to be used in practice without ensemble with stronger models. The results gained can be applied for usage with an ensemble of state-of-the-art convolutional and recurrent neural networks. The proposed approach is universal and applicable both for the analysis of individual images of skin neoplasms and for the analysis of their sequences obtained by the method of multispectral image processing. The technique can be applied to datasets with a limited amount of data. The results obtained will be of interest to specialists in the fields of computer vision and medical images analysis.

Imiquimod-induced ex vivo model of psoriatic human skin via interleukin-17A signalling of T cells and Langerhans cells.

Several in vitro models have been developed to study the mechanisms involved in psoriasis and to screen new antipsoriatic drugs. However, most of them use single-cell or reconstructed human skin models that did not have complex anatomy of human skin. Thus, this study aimed to create a new model of psoriasis-like dermatitis using human skin under in vitro conditions. To perform this, human skin explants were topically treated with imiquimod (IMQ) or vehicle for 2, 3 or 6 consecutive days. Some explants were treated with an anti-psoriatic drug or antibody anti-interleukin-17A (IL-17A). Topical application of IMQ increased total epidermal area, epidermal proliferation and keratinocyte differentiation at 3, 4 or 7 days. The protein levels of CD3 were augmented in the IMQ-treated human skin explants at 7 days reflecting the activation of T cells. Topical IMQ promoted a higher protein and mRNA levels of IL-17A in human skin ex vivo. Immunofluorescence analysis showed CD207-positive Langerhans cells (LCs) and CD3-positive T cells expressing IL-17A in IMQ-treated human skin explants at 7 days. In addition, administration of antibody anti-IL-17A or an anti-psoriatic drug inhibited IMQ-induced increase in the epidermal thickness in ex vivo human skin at 7 days. In conclusion, topical IMQ application promotes epidermal changes in ex vivo human skin that resemble to human psoriatic skin lesions. Moreover, IMQ-induced production of IL-17 by LCs and T cells is critical to development of psoriasis-like inflammation in our model. This new model is suitable for in vitro screening of antipsoriatic drugs.

Upregulated integrin α11 in the stroma of cutaneous squamous cell carcinoma promotes skin carcinogenesis

Integrin α11β1 is a collagen-binding integrin that is needed to induce and maintain the myofibroblast phenotype in fibrotic tissues and during wound healing. The expression of the α11 is upregulated in cancer-associated fibroblasts (CAFs) in various human neoplasms. We investigated α11 expression in human cutaneous squamous cell carcinoma (cSCC) and in benign and premalignant human skin lesions and monitored its effects on cSCC development by subjecting α11-knockout (Itga11−/− ) mice to the DMBA/TPA skin carcinogenesis protocol. α11-deficient mice showed significantly decreased tumor cell proliferation, leading to delayed tumor development and reduced tumor burden. Integrin α11 expression was significantly upregulated in the desmoplastic tumor stroma of human and mouse cSCCs, and the highest α11 expression was detected in high-grade tumors. Our results point to a reduced ability of α11-deficient stromal cells to differentiate into matrix-producing and tumor-promoting CAFs and suggest that this is one causative mechanism underlying the observed decreased tumor growth. An unexpected finding in our study was that, despite reduced CAF activation, the α11-deficient skin tumors were characterized by the presence of thick and regularly aligned collagen bundles. This finding was attributed to a higher expression of TGFβ1 and collagen crosslinking lysyl oxidases in the Itga11-/- tumor stroma. In summary, our data suggest that α11β1 operates in a complex interactive tumor environment to regulate ECM synthesis and collagen organization and thus foster cSCC growth. Further studies with advanced experimental models are still needed to define the exact roles and molecular mechanisms of stromal α11β1 in skin tumorigenesis.