A qPCR to quantify Wolbachia from few Onchocerca volvulus microfilariae as a surrogate for adult worm histology in clinical trials of antiwolbachial drugs

The filarial nematode Onchocerca volvulus causes onchocerciasis (river blindness), a neglected tropical disease affecting 21 million people, mostly in Sub-Saharan Africa. Targeting the endosymbiont Wolbachia with antibiotics leads to permanent sterilization and killing of adult worms. The gold standard to assess Wolbachia depletion is the histological examination of adult worms in nodules beginning at 6 months post-treatment. However, nodules can only be used once, limiting the time points to monitor Wolbachia depletion. A diagnostic to longitudinally monitor Wolbachia depletion from microfilariae (MF) at more frequent intervals < 6 months post-treatment would accelerate clinical trials of antiwolbachials. We developed a TaqMan qPCR amplifying the single-copy gene wOvftsZ to quantify Wolbachia from as few as one MF that had migrated from skin biopsies and compared quantification using circular and linearized plasmids or synthetic dsDNA (gBlock®). qPCR for MF from the rodent nematode Litomosoides sigmodontis was used to support the reproducibility and validate the principle. The qPCR using as few as 2 MF from O. volvulus and L. sigmodontis reproducibly quantified Wolbachia. Use of a linearized plasmid standard or synthesized dsDNA resulted in numbers of Wolbachia/MF congruent with biologically plausible estimates in O. volvulus and L. sigmodontis MF. The qPCR assay yielded a median of 48.8 (range 1.5–280.5) Wolbachia/O. volvulus MF. The qPCR is a sensitive tool for quantifying Wolbachia in a few MF from skin biopsies and allows for establishing the qPCR as a surrogate parameter for monitoring Wolbachia depletion in adult worms of new antiwolbachial candidates.

TDP-43 Cytoplasmic Translocation in the Skin Fibroblasts of ALS Patients

Diagnosis of ALS is based on clinical symptoms when motoneuron degeneration is significant. Therefore, new approaches for early diagnosis are needed. We aimed to assess if alterations in appearance and cellular localization of cutaneous TDP-43 may represent a biomarker for ALS. Skin biopsies from 64 subjects were analyzed: 44 ALS patients, 10 healthy controls (HC) and 10 neurological controls (NC) (Parkinson’s disease and multiple sclerosis). TDP-43 immunoreactivity in epidermis and dermis was analyzed, as well as the percentage of cells with TDP-43 cytoplasmic localization. We detected a higher amount of TDP-43 in epidermis (p < 0.001) and in both layers of dermis (p < 0.001), as well as a higher percentage of TDP-43 cytoplasmic positive cells (p < 0.001) in the ALS group compared to HC and NC groups. Dermal cells containing TDP-43 were fibroblasts as identified by co-labeling against vimentin. ROC analyses (AUC 0.867, p < 0.001; CI 95% 0.800–0.935) showed that detection of 24.1% cells with cytoplasmic TDP-43 positivity in the dermis had 85% sensitivity and 80% specificity for detecting ALS. We have identified significantly increased TDP-43 levels in epidermis and in the cytoplasm of dermal cells of ALS patients. Our findings provide support for the use of TDP-43 in skin biopsies as a potential biomarker.

A new case of scalp angiosarcoma revealed by eyelid edema

Cutaneous angiosarcoma is a rare and highly aggressive neoplasm with poor prognosis. Owing to its variable presentation, it may be a challenging clinical and histological diagnosis. Herein, we describe a particular case of cutaneous angiosarcoma of the head and neck in a 71-year-old male with skin type V who presented with solid, progressive eyelid edema. A histological examination of skin biopsies first concluded cutaneous metastasis of an undifferentiated carcinomatous tumor. Immunostaining was essential to reach the correct diagnosis. The treatment of cutaneous angiosarcoma remains unsatisfactory. We sincerely hope that the prognosis of cutaneous angiosarcoma will be improved with the use of targeted therapies based on current genetic studies as it has been for melanoma.

Expression Patterns of Clock Gene mRNAs and Clock Proteins in Human Psoriatic Skin Samples

Psoriasis is a systemic inflammatory skin disorder that can be associated with sleep disturbance and negatively influence the daily rhythm. The link between the pathomechanism of psoriasis and the circadian rhythm has been suggested by several previous studies. However, there are insufficient data on altered clock mechanisms in psoriasis to prove these theories. Therefore, we investigated the expression of the core clock genes in human psoriatic lesional and non-lesional skin and in human adult low calcium temperature (HaCaT) keratinocytes after stimulation with pro-inflammatory cytokines. Furthermore, we examined the clock proteins in skin biopsies from psoriatic patients by immunohistochemistry. We found that the clock gene transcripts were elevated in psoriatic lesions, especially in non-lesional psoriatic areas, except for rev-erbα, which was consistently downregulated in the psoriatic samples. In addition, the REV-ERBα protein showed a different epidermal distribution in non-lesional skin than in healthy skin. In cytokine-treated HaCaT cells, changes in the amplitude of the bmal1, cry1, rev-erbα and per1 mRNA oscillation were observed, especially after TNFα stimulation. In conclusion, in our study a perturbation of clock gene transcripts was observed in uninvolved and lesional psoriatic areas compared to healthy skin. These alterations may serve as therapeutic targets and facilitate the development of chronotherapeutic strategies in the future.

U lympho tế bào T dạng viêm mỡ dưới da ở trẻ em

TÓM TẮT U lympho tế bào T dạng viêm mỡ dưới da (Subcutaneous panniculitis - like T - cell lymphoma - SPTCL) là một u lympho hiếm gặp với tổn thương chủ yếu ở bề mặt da và tổ chức mỡ dưới da. SPTCL có thể xảy ra ở cả người lớn và trẻ nhỏ, độ tuổi trung bình được chẩn đoán là 30 tuổi, nữ nhiều hơn nam. Trong SPTCL, bệnh nhân có nhiều nốt tổn thương ở mô mỡ dưới da và có thể gây loét bề mặt da. Tổn thương này được biết đến như viêm mỡ dưới da. Cơ chế bệnh sinh chưa rõ ràng nhưng có sự tham gia của yếu tố di truyền. Chẩn đoán SPTCL vẫn còn là thách thức, chủ yếu dựa vào sinh thiết da với sự bộc lộ của các dấu ấn hóa mô miễn dịch (HMMD). Điều trị SPTCL chưa thống nhất, phụ thuộc vào triệu chứng nhưng thường sử dụng steroid hoặc các thuốc ức chế miễn dịch (ƯCMD). Một số bệnh nhân với SPTCL tiến triển cần điều trị hóa chất. Về tiên lượng, SPTCL thường tiến triển chậm, tuy nhiên khoảng 20% bệnh nhân có hội chứng thực bào máu (Hemophagocytic lymphohistiocytosis - HLH), những bệnh nhân này kém đáp ứng với điều trị và có tiên lượng xấu hơn. Chúng tôi báo cáo một bé trai 13 tháng tuổi vào viện vì sốt và nhiều khối tổn thương dưới da. Bệnh nhân được chẩn đoán viêm mô tế bào và điều trị kháng sinh 2 tuần nhưng bệnh không thuyên giảm. Sau đó, bệnh nhân được sinh thiết tổn thương dưới da, dựa trên kết quả nhuộm HMMD, chúng tôi kết luận phù hợp với SPTCL. ABSTRACT SUBCUTANEOUS PANNICULITIS - LIKE T - CELL LYMPHOMA IN A CHILD Subcutaneous panniculitis - like T - cell lymphoma (SPTCL) is a rare type of lymphoma that mainly affects the skin and subcutaneous fat tissue. SPTCL can occur in both adults and children. The average age at diagnosis is 30 years old, with more women than men. In SPTCL, the patient has multiple nodules in the fat layer under the skin, and these can cause swelling. This is known as panniculitis. The pathogenesis is unknown, but genetic factors may be involved. Diagnosis of SPTCL is challenging, mainly based on skin biopsies with staining of immunohistochemical markers. Treatment of SPTCL is inconsistent, depending on symptoms but usually using steroids or immunosuppressive drugs (ACEIs). Some patients with advanced SPTCL require chemotherapy. In terms of patients who have hemophagocytic lymphohistiocytosis (HLH), which is less responsive to treatment and has a worse prognosis. We report a 13 - month - old boy admitted to the hospital with fever and multiple subcutaneous lesions. The patient was diagnosed with cellulitis and treated with antibiotics for 2 weeks, but the disease did not improve. Then, the patient had a skin lesion biopsy. Based on the results of immunohistochemical staining, we concluded that it was suitable for SPTCL. Keywords: Subcutaneous panniculitis - like T - cell lymphoma, Immunohistochemical, Hemophagocytic lymphohistiocytosis.

A Systematic Review of Clinical Studies on Mucocutaneous Manifestations of COVID-19: Virus-Related and Drug-Related

Coronavirus could affect almost any part of the body including the skin. In this systematic review, the primary skin lesions resulting from the direct activity of the virus or the medications used for treatment and the changes in the behavior of the virus regarding the occurrence of these symptoms over time were assessed. PubMed/MEDLINE, Embase, PsycINFO, TRIP Cochrane, Cochrane Skin were searched for all published articles from February 19 to July 1, 2020, which met the inclusion criteria. Thirty-six related articles were extracted. Twenty-eight studies reported virus-related mucocutaneous eruptions and 8 articles, the drug-reactions. Data of 583 patients were included. Skin lesions of COVID-19 could be caused by both the virus itself or the influence of drugs used for the treatment. Morbilliform rashes, urticaria, and acral-vasculopathic cutaneous lesions were at the forefront of primary COVID-dependent skin lesions with no significant change during time, Also, Hydroxychloroquine, lopinavir/ritonavir, paracetamol, and antibiotics were reported as the main causes of drug-induced rashes. Since dermatologic manifestations may occur prior or simultaneously/after other COVID clinical symptoms, so they may helpful in patients’ early diagnosis or prediction of internal organ involvements via histopathologic evaluations of skin biopsies especially about vasculopathic and vasculitic, respectively.

SFRP4 Expression Is Linked to Immune-Driven Fibrotic Conditions, Correlates with Skin and Lung Fibrosis in SSc and a Potential EMT Biomarker

Secreted Frizzled Receptor Protein 4 (SFRP4) has been shown to be increased in Scleroderma (SSc). To determine its role in immune-driven fibrosis, we analysed SSc and sclerotic Chronic Graft Versus Host Disease (sclGVHD) biosamples; skin biopsies (n = 24) from chronic GVHD patients (8 with and 5 without sclGVHD), 8 from SSc and 3 healthy controls (HC) were analysed by immunofluorescence (IF) and SSc patient sera (n = 77) assessed by ELISA. Epithelial cell lines used for in vitro Epithelial-Mesenchymal-Transition (EMT) assays and analysed by Western Blot, RT-PCR and immunofluorescence. SclGVHD skin biopsies resembled pathologic features of SSc. IF of fibrotic skin biopsies indicated the major source of SFRP4 expression were dermal fibroblasts, melanocytes and vimentin positive/caveolin-1 negative cells in the basal layer of the epidermis. In vitro studies showed increased vimentin and SFRP4 expression accompanied with decreased caveolin-1 expression during TGFβ-induced EMT. Additionally, SFRP4 serum concentration correlated with severity of lung and skin fibrosis in SSc. In conclusion, SFRP4 expression is increased during skin fibrosis in two different immune-driven conditions, and during an in vitro EMT model. Its serum levels correlate with skin and lung fibrosis in SSc and may function as biomarker of EMT. Further studies are warranted to elucidate the role of SFRP4 in EMT within the pathogenesis of tissue fibrosis.

An Analysis of IL-10, IL-17A, IL-17RA, IL-23A and IL-23R Expression and Their Correlation with Clinical Course in Patients with Psoriasis

Being one of the most common dermatological inflammatory disorders, psoriasis is a frequent subject of research. It is considered to be a T cell-dependent immune disease whose pathogenesis is influenced by cytokines, such as IL-10, IL-17A, IL-17RA, IL-23A and IL-23R. The present study examines whether the expression of selected genes is correlated with the clinical course of psoriasis, assessed by the PASI, BSA and DLQI scales. Skin biopsies and blood from 60 patients with psoriasis and 24 healthy controls were obtained for RNA isolation. These were subjected to RT-PCR for IL-10, IL-17A, IL-17RA, IL-23A and IL-23R genes. The results were presented as an RQ value. IL-17A and IL-23R expression levels were higher in psoriatic skin compared to controls, while IL-10 expression was lower. A positive correlation was also found between RQ for IL-23A and PASI index. Psoriatic skin is characterised by elevated expression of IL-17A and IL-23R and decreased expression of IL-10. This indicates that the selected cytokines may be one of the factors involved in the pathogenesis and pathomechanism of psoriasis, but more studies need to be made before we can elucidate the exact reason for the unbalance in cytokine expression levels.

Thalidomide Exerts Anti-Inflammatory Effects in Cutaneous Lupus by Inhibiting the IRF4/NF-ҡB and AMPK1/mTOR Pathways

Thalidomide is effective in patients with refractory cutaneous lupus erythematosus (CLE). However, the mechanism of action is not completely understood, and its use is limited by its potential, severe side-effects. Immune cell subset analysis in thalidomide’s CLE responder patients showed a reduction of circulating and tissue cytotoxic T-cells with an increase of iNKT cells and a shift towards a Th2 response. We conducted an RNA-sequencing study using CLE skin biopsies performing a Therapeutic Performance Mapping System (TMPS) analysis in order to generate a predictive model of its mechanism of action and to identify new potential therapeutic targets. Integrating RNA-seq data, public databases, and literature, TMPS analysis generated mathematical models which predicted that thalidomide acts via two CRBN-CRL4A- (CRL4CRBN) dependent pathways: IRF4/NF-ҡB and AMPK1/mTOR. Skin biopsies showed a significant reduction of IRF4 and mTOR in post-treatment samples by immunofluorescence. In vitro experiments confirmed the effect of thalidomide downregulating IRF4 in PBMCs and mTOR in keratinocytes, which converged in an NF-ҡB reduction that led to a resolution of the inflammatory lesion. These results emphasize the anti-inflammatory role of thalidomide in CLE treatment, providing novel molecular targets for the development of new therapies that could avoid thalidomide’s side effects while maintaining its efficacy.