Skin Biopsy Specimens Research Articles

Disease activity in chronic inflammatory demyelinating polyneuropathy: association between circulating B-cell subsets, cytokine levels, and clinical outcomes.

Chronic inflammatory demyelinating polyneuropathy (CIDP), a common and treatable autoimmune neuropathy, is frequently misdiagnosed. The aim of this study is to evaluate the relationship between immunological markers and clinical outcome measures in a mixed cohort of patients with typical CIDP and CIDP variants at different disease stages. Twenty-three typical, 16 multifocal and five distal CIDP patients were included. Twenty-five sex and age-matched healthy controls and 12 patients with Charcot-Marie-Tooth type 1A (CMT1A) disease served as controls. Peripheral B-cell populations were analyzed by flow cytometry. IL6, IL10, TNFA mRNA and mir-21, mir-146a, and mir-155-5p expression levels were evaluated by real-time polymerase chain reaction in peripheral blood mononuclear cells (PBMC) and/or skin biopsy specimens. Results were then assessed for a possible association with clinical disability scores and intraepidermal nerve fiber densities (IENFD) in the distal leg. We detected a significant reduction in naive B cells (P ≤ 0.001), plasma cells (P ≤ 0.001) and regulatory B cells (P < 0.05), and an elevation in switched memory B cells (P ≤ 0.001) in CIDP compared to healthy controls. CMT1A and CIDP patients had comparable B-cell subset distribution. CIDP cases had significantly higher TNFA and IL10 gene expression levels in PBMC compared to healthy controls (P < 0.05 and P ≤ 0.01, respectively). IENFDs in the distal leg showed a moderate negative correlation with switched memory B-cell ratios (r = -0.51, P < 0.05) and a moderate positive correlation with plasma cell ratios (r = 0.46, P < 0.05). INCAT sum scores showed a moderate positive correlation with IL6 gene expression levels in PBMC (r = 0.54, P < 0.05). Altered B-cell homeostasis and IL10 and TNFA gene expression levels imply chronic antigen exposure and overactivity in the humoral immune system, and seem to be a common pathological pathway in both typical CIDP and CIDP variants.

Dystrophic calciphylaxis in panniculitis: features of the clinical picture and diagnosis

The clinical presentation of lobular panniculitis (PN) associated with calciphylaxis (CP, calcification) can vary widely and may be associated with joint and internal organs involvement, making the diagnosis of the disease difficult.Objective: to evaluate the frequency and significance of CP in patients with PN using long-term prospective follow-up.Material and methods. From 2018 to 2023, at the V.A. Nasonova Research Institute of Rheumatology 217 patients with referral diagnosis "erythema nodosum" or "panniculitis" were examined. In 19.3% of cases (9 men and 33 women aged 37 to 72 years) CP was confirmed with an average disease duration of 56.3±11.2 months. Clinical examination of patients was performed according to the standards recommended by the Russian Association of Rheumatologists. International criteria were used to confirm the diagnosis of systemic lupus erythematosus (SLE), idiopathic inflammatory myopathies (IIM), systemic sclerosis (SS), and lipodermatosclerosis (LDS). In 12 patients with indurations, pathological examination of biopsy specimens of skin and subcutaneous fatty tissue from the area of induration was performed, which allowed confirming the diagnosis of idiopathic lobular PN (ILPN) in 3 cases. Four grades of calcification were distinguished according to the size and depth of the calcifications. In addition, considering the type of radiological changes and clinical manifestations, four subtypes of CP were identified: mousse-like, stone-like, mesh-like and lamellar-like.Results and discussion. In the study group, the ratio of women to men was 3.6:1, and the mean age was 43.8±7.6 years. On clinical examination we determined, in 60% of cases CP predominantly stone-like subtype (71.4%) of first grade (47.6%), which was significantly more frequently located on the upper and/or lower extremities and/or trunk (57.1%; p=0.05). Using clinical, laboratory and instrumental data, we confirmed the development of CP in ILPN (n=3), SLE (n=3), LDS (n=21), IIM (n=5), SS (n=1), and idiopathic CP (n=9) with a mean disease duration of 8.7±2.4 years.An increase in ESR and CRP levels occurred in different diseases, while urinary syndrome was associated with SLE (66.6%) and an increase in creatinine phosphokinase with IIM. Decreased calcium and 25-hydroxyvitamin D levels and increased phosphorus and parathyroid hormone levels were found in many patients studied.Conclusion. In the absence of clear diagnostic criteria for CP in patients with PN, early diagnosis is critical for the development of an effective multidisciplinary treatment plan.

Neutrophil extracellular traps formation in the lesional skin of various types of pyoderma gangrenosum.

Pyoderma gangrenosum (PG) is a chronic neutrophilic disorder characterized by recurrent painful ulcers. Aseptic inflammation by neutrophils plays an essential role, and neutrophil extracellular traps (NETs) formation can contribute to the pathogenesis of PG. Seventy-five patients were diagnosed as having PG in our department, among which 58 ulcerative, 4 bullous, 3 pustular and 10 vegetative type. We examined the 20 skin biopsy specimens (11 ulcerative, 3 bullous, 2 pustular and 4 vegetative type), and local NETs formation in various types of PG was compared among each type. The biopsied specimens were double labelled for myeloperoxidase, citrullinated histone H3. Immunofluorescent images indicated that the histopathologic location and depth of NETs formation in PG varied by the clinical subtypes. In ulcerative PG, NETs formation was observed in the upper to deep dermis. In bullous PG, NETs formation was mainly observed in the epidermis. Pustular type showed NETs formation in the epidermis near the pustules, and in vegetative type, showed NETs formation mainly in the upper dermis. These results indicate that NETting neutrophils play an important role in the pathogenesis of various forms of PG, although the location and depth of NETs formation in the skin lesion of PG differ depending on each type. Further studies are necessary to examine what factors identify different clinical features of PG.

Histopathological differences between vitiligo and lichen sclerosus et atrophicus using quantitative immunohistochemical analysis.

Lichen sclerosus et atrophicus (LS) is rare skin condition characterized by the presence of whitish patches primarily affecting the genital and perianal areas, though it can occur other parts of the body. LS may result in skin depigmentation without textural changes and should be differentiated from vitiligo. However, the histopathological features of hypopigmentation during vitiligo and LS have rarely been compared and have not been precisely described using quantitative immunohistochemical analysis. This study, therefore, aimed to investigate and compare the pigmentary characteristics of LS and vitiligo lesions using histochemical and immunohistochemical staining. We included 31 and 46 patients diagnosed with LS and vitiligo, respectively, at Ajou University Hospital between March 2009 and March 2020 in this study. Their medical charts and skin biopsy specimens were retrospectively reviewed. Additionally, Fontana-Masson staining for melanin and immunohistochemical staining for Melan-A, NKI/beteb, tyrosinase, and microphthalmia-associated transcription factor was performed. The melanin content, as well as the number of melanocytes was, in general, significantly higher in the epidermis of patients in the LS group compared with that in the vitiligo group. However, 22.6% of LS tissues showed less melanin pigmentation, 25.8% of LS specimens exhibited a lower number of melanocytes, and 29.0% of LS specimens demonstrated less melanocyte activity when compared with the average of vitiligo specimens. As lower melanin pigmentation and the near absence number of melanocytes were also observed in several LS specimens, both the clinical and histological findings must be comprehensively reviewed to differentiate vitiligo from LS.

Utility of C3d and C4d immunohistochemical staining in formalin-fixed skin or mucosal biopsy specimens in diagnosis of bullous pemphigoid and mucous membrane pemphigoid

Bullous pemphigoid (BP) and mucous membrane pemphigoid (MMP) sometimes have overlapping clinical, histopathological, and direct immunofluorescence (DIF) features in the early stages. Complement deposition is an intrinsic component of the patho-mechanism of BP in contrast to MMP. Hence immunohistochemistry (IHC) for C3d and C4d may be helpful in differentiating the two disorders. Seventy-four patients of BP and 18 patients of MMP along with 10 negative controls were enrolled in this study. C3d and C4d IHC was performed in formalin-fixed skin biopsy specimens. C3d IHC staining in BP/MMP had a sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 59.2%/41.2%, 100%/100%, 100%/100%, 25.6%/50.0%, respectively. C4d IHC staining in BP/MMP had a sensitivity, specificity, PPV and NPV of 26.8%/17.6%, 100%/100%, 100%/100% and 16.1%/41.7%, respectively. Receiver operator analysis showed utility of C3d in diagnosing both BP [Area under curve (AUC) = 0.8, p = 0.0001] and MMP (AUC = 0.71; p = 0.001). C4d was useful in diagnosis of BP (AUC = 0.5; p = 0.0001), but not MMP (AUC = 0.6; p = 0.064). Hence, C3d is a better diagnostic modality for BP as compared to C4d, whereas C3d and C4d have lower diagnostic importance in MMP. C3d IHC can be employed in diagnosing BP when a second biopsy for direct immunofluorescence (DIF) is not possible or where a facility for IF microscopy does not exist.

Gene fusions in superficial mesenchymal neoplasms: Emerging entities and useful diagnostic adjuncts.

Cutaneous mesenchymal neoplasms are diagnostically challenging because of their overlapping morphology, and, often, the limited tissue in skin biopsy specimens. Molecular and cytogenetic techniques have identified characteristic gene fusions in many of these tumor types, findings that have expanded our understanding of disease pathogenesis and motivated development of useful ancillary diagnostic tools. Here, we provide an update of new findings in tumor types that can occur in the skin and superficial subcutis, including dermatofibrosarcoma protuberans, benign fibrous histiocytoma, epithelioid fibrous histiocytoma, angiomatoid fibrous histiocytoma, glomus tumor, myopericytoma/myofibroma, non-neural granular cell tumor, CIC-rearranged sarcoma, hybrid schwannoma/perineurioma, and clear cell sarcoma. We also discuss recently described and emerging tumor types that can occur in superficial locations and that harbor gene fusions, including nested glomoid neoplasm with GLI1 alterations, clear cell tumor with melanocytic differentiation and ACTIN::MITF translocation, melanocytic tumor with CRTC1::TRIM11 fusion, EWSR1::SMAD3-rearranged fibroblastic tumor, PLAG1-rearranged fibroblastic tumor, and superficial ALK-rearranged myxoid spindle cell neoplasm. When possible, we discuss how fusion events mediate the pathogenesis of these tumor types, and we also discuss the related diagnostic and therapeutic implications of these events.

003 Utility of plucked hair direct immunofluorescence as a possible replacement for conventional skin biopsy direct immunofluorescence in immunological activity monitoring and prediction of relapse in pemphigus

Abstract Pemphigus vulgaris (PV) is a potentially lethal chronically relapsing autoimmune bullous disorder with intraepithelial lesions affecting the skin and mucous membranes. The prediction of clinical relapse has been studied with conventional skin biopsy and plucked hair direct immunofluorescence (DIF; Daneshpazhooh M, Naraghi ZS, Ramezani A et al. Direct immunofluorescence of plucked hair for evaluation of immunologic remission in PV. J Am Acad Dermatol 2011; 65:173–7) which can be of use in timing treatment withdrawal and helping with better management. The aims of our study were to determine the association of plucked hair root DIF positivity status at baseline with clinical relapse within the study duration, and to determine the correlation between skin biopsy and plucked hair DIF. Seventy-eight consecutive patients diagnosed with PV in clinical remission on therapy, on minimal therapy or off therapy, were included after obtaining informed consent. Three-monthly follow-up was carried out for 12 months. Immunological monitoring with plucked hair DIF was done at baseline and repeated every 3 months for 1 year or a relapse was experienced, whichever occurred earlier. Conventional skin biopsy DIF was performed at baseline, at 3 months and at the time of relapse. In patients receiving adjuvant therapy during their present remission episode, discontinuation of therapy was done at inclusion, at 3 months and at 6 months, and occurrence of relapse in relation to the time point of discontinuation was assessed. Of 78 patients, 22 had disease relapse and 56 remained in complete remission. Clinical relapse during the study duration was significantly more frequent in patients who had higher mean Autoimmune Bullous Skin Disorder Intensity Score during the preceding active disease episode (P = 0.033), pruritus during the present clinical remission episode (P = 0.007) and early discontinuation (within 3 months of study inclusion) of concomitant adjuvant therapy (P = 0.007). Immunological parameters associated with clinical relapse were baseline IgG (P = 0.005) and C3 positivity (P = 0.047) in plucked hair, greater intensity of IgG and C3 positivity at baseline in plucked hair (IgG, P = 0.002; C3, P = 0.033) and skin biopsy specimens (IgG, P = 0.004; C3, P = 0.0.002), and positive hair DIF at the time of adjuvant therapy withdrawal (P = 0.017). Plucked hair DIF is a reliable technique to assess immunological remission in patients with PV who are in clinical remission and helps to guide treatment according to immunological remission status.

Cytophagic Histiocytic Panniculitis with Undetermined Autoimmune Disease: A Rare Case Report

Cytophagic histiocytic panniculitis is a rare disease, associated with either nonmalignant conditions or subcutaneous panniculitis-like T-cell lymphoma (SPTL), and often also associated with hemophagocytic lymphohistiocytosis (HLH). We report the case of a 63-year-old woman who developed a multiple tender, indurated, erythmatous lesions over the abdomen, back, trunk and limbs. Histopathologic findings from skin biopsy specimens revealed significant lobular panniculitis with benign histiocytes showing hemophagocytosis. ANA by IF was positive. Though it is very rare, to confirm diagnosis of CHP by histopathology is very important as CHP and SPLT may span a clinicopathologic spectrum in which there is a natural disease progression from CHP to fatal SPTL.

Systemic complications of intravitreal bevacizumab: a case report and literature review

Background: Systemic complications of intravitreal bevacizumab (IVB) injections have been previously reported. We aimed to summarize the systemic complications reported in online primary studies. Moreover, we describe a patient experiencing simultaneous renal and cutaneous drug-induced adverse effects, with exacerbation of chronic renal insufficiency and granulomatous skin lesions, after receiving several IVB injections to manage bilateral ischemic branch retinal vein occlusion (BRVO). Case Presentation: A 69-year-old Hispanic diabetic man with chronic renal insufficiency due to polyclonal gammopathy received several IVB injections to treat bilateral ischemic BRVO. One week after the sixth injection, the patient developed acute-on-chronic renal failure and multiple rounded maculopapular, erythematous, and ulcerated skin lesions. Renal and skin biopsy specimens revealed granulomatous drug-induced responses in both organs, and granulomatous diseases of infectious and oncological sources were ruled out. We performed an electronic search of the PubMed/MEDLINE database with no language or time restrictions using the keywords “intravitreal bevacizumab” or “intravitreal Avastin” combined with “systemic side effects,” “systemic complications,” or “systemic adverse,” or “systemic adverse event.” The search yielded 147 articles published over almost two decades. After screening and assessment, we selected and summarized 40 primary studies that mentioned IVB-related systemic complications. Conclusions: IVB-induced systemic complications, such as arteriothrombotic events, venous thrombotic events, and hypertension, are rare but potentially serious. Care should be taken when administering multiple doses of intravitreal IVB to patients with pre-existing kidney dysfunction. Bevacizumab-related toxicity must be considered in cases of sudden deterioration of renal function and / or unexpected granulomatous skin lesions in oncologic or chronically polymedicated patients.

Acellular Dermal Matrix Cover Improves Skin Growth during Tissue Expansion by Affecting Distribution of Mechanical Forces.

Biological cover over tissue expander prostheses has been introduced to provide soft-tissue support for tissue expanders during breast reconstruction. However, its impact on mechanically induced skin growth remains unknown. This study investigates the hypothesis that covering the tissue expander with acellular dermal matrix (ADM) affects mechanotransduction without compromising the efficacy of tissue expansion. Tissue expansion, with and without use of ADM, was performed on a porcine model. The tissue expanders were inflated twice with 45 mL of saline, and the full-thickness skin biopsy specimens were harvested from expanded and control unexpanded skin 1 week and 8 weeks after the final inflation. Histologic evaluation, immunohistochemistry staining, and gene expression analysis were performed. Skin growth and total deformation were evaluated using isogeometric analysis. The authors' results demonstrate that use of ADM as a biological cover during tissue expansion does not impede mechanotransduction that leads to skin growth and blood vessel formation. Isogeometric analysis revealed similar total deformation and growth of expanded skin with and without a biological cover, confirming that its use does not inhibit mechanically induced skin growth. In addition, the authors found that use of an ADM cover results in more uniform distribution of mechanical forces applied by the tissue expander. These results suggest that ADM improves mechanically induced skin growth during tissue expansion by facilitating a more uniform distribution of mechanical forces applied by the tissue expander. Therefore, the use of a biological cover has potential to improve outcomes in tissue expansion-based reconstruction.

An Overview of 10 Years of Activity of a Molecular Laboratory for Buruli Ulcer Diagnosis at a Field Hospital in Benin.

Buruli ulcer (BU) is a neglected tropical disease caused by Mycobacterium ulcerans. Early diagnosis is crucial to prevent morbidity. In November 2012, a field laboratory fully equipped for the rapid on-site quantitative PCR (qPCR) diagnosis of M. ulcerans was established at the Buruli ulcer treatment center (CDTLUB) center in Pobè Benin, a region where BU is endemic. We describe its first 10 years of activity and its gradual evolution into an expert laboratory for BU diagnosis. From 2012 to 2022, the laboratory analyzed 3,018 samples from patients attending consultations for suspected BU at the CDTLUB in Pobè. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. Since 2019, the laboratory has also received and analyzed 570 samples from other centers. The laboratory confirmed the diagnosis of BU by qPCR for 39.7% samples: M. ulcerans DNA was detected in 34.7% of swabs, 47.2% of all fine needle aspiration samples (FNA) and 44.6% of all skin biopsy specimens. Positive Ziehl-Neelsen staining results were obtained for 19.0% samples. Bacterial load, estimated by qPCR, was significantly greater for the Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples, and detection rates were highest for FNA samples. Overall, 26.3% of the samples received from other centers were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado, Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success. Optimal patient care depends on the close proximity of a molecular biology structure to BU treatment centers. Finally, FNA should be promoted among caregivers. IMPORTANCE Here, we describe the first 10 years of activity at a field laboratory established at the Buruli ulcer treatment center (CDTLUB) in Pobè, Benin, a country in which Mycobacterium ulcerans is endemic. Between 2012 and 2022, the laboratory analyzed 3,018 samples from patients consulting the CDTLUB of Pobè with a suspected clinical BU. Ziehl-Neelsen staining and qPCR targeting the IS2404 sequence were performed. In total, 39.7% of samples tested positive by qPCR and 19.0% tested positive by Ziehl-Neelsen staining. Detection rates were highest for FNA samples, and the bacterial loads estimated by qPCR were significantly higher for Ziehl-Neelsen-positive samples than for Ziehl-Neelsen-negative samples. Since 2019, the laboratory has also analyzed 570 samples received from outside the CDTLUB of Pobè, 26.3% of which were positive for BU. Most of these samples were sent by the CDTLUBs of Lalo, Allada, and Zagnanado in Benin. The establishment of the laboratory in the CDTLUB of Pobè has been a huge success, with major benefits for both the medical staff and patients. Our findings illustrate that the usefulness and feasibility of having a diagnostic center in rural Africa, where the disease is endemic, is a key part of optimal patient care, and that FNA should be promoted to increase detection rates.

Morphological features of skin changes in the undetermined clinical picture of psoriasis and atopic dermatitis in children

Absrtact Introduction The literature describes atypical clinical forms of psoriasis and atopic dermatitis in children that combine features of both diseases. In such cases, dermatologists resort to biopsy of plaque skin elements in the complex diagnosis. However, the histological picture in these biopsy specimens is uncertain and does not allow to make an unequivocal judgment about the correspondence of changes to one or another disease. The purpose of work was to compare pathomorphological changes in biopsy specimens of plaque skin elements in children with undetermined clinical picture of the disease with changes in typical forms of psoriasis and atopic dermatitis. Materials and methods A morphometric study of 10 plaque skin biopsy specimens from children (mean age 9.2±1.11 years) with an indeterminate clinical picture of the disease was performed. The obtained data were compared with two control groups of adult patients with typical clinical pictures of psoriasis (n=20; mean age 34.35±3.58 years) and atopic dermatitis (n=10; mean age 41.33±7.35 years). Results A comparative morphometric analysis of skin biopsy specimens revealed the fewest differences between the analyzed parameters of the squamous epithelium of the patients in the study group and the group with psoriasis. In all three groups, the main mass of the inflammatory infiltrate in the dermis was composed of lymphocytes and macrophages whose numerical density differed in psoriasis and atopic dermatitis at different levels of the dermis, while in the study group it had intermediate values. The number of neutrophils and eosinophils in the study group had minimal values. Discussion Morphological changes in the epidermis of the study group were closest to those in psoriasis, while the cellular composition of the inflammatory infiltrate of the dermis did not correspond to any of the control groups and had either intermediate values (in terms of lymphocytes and macrophages) or minimal values (in terms of eosinophils and neutrophils).

The neglected twin: Nummular eczema is a variant of atopic dermatitis with codominant TH2/TH17 immune response

Nummular eczema (NE) is a common chronic inflammatory skin disease characterized by multiple, pruritic, discoid-shaped lesions. Since the underlying immune mechanisms are not fully understood, it is unclear whether NE should be regarded as variant of atopic dermatitis (AD) or a distinct disease. We compared the clinical, histopathologic, and molecular signatures of NE with that of type 2 and type 3 skin diseases. We performed bulk RNA sequencing as well as histologic and clinical studies in lesional and nonlesional skin biopsy specimens from NE (n= 50), AD (n= 47), and psoriasis (n= 90) patients. NE displayed typical hallmarks of AD, such as an impaired epidermal barrier, microbial colonization, spongiosis, and eosinophil infiltration, but also aspects of psoriasis, including increased epidermal thickness, number of Ki-67+ cells, and neutrophilic infiltration. At the gene expression level, neutrophil-attracting cytokines (IL19, CXCL8, CXCL5) were upregulated, whereas TH2-related cytokines (IL13, CCL17, CCL18, CCL26, CCL27) were similarly expressed in NE compared to AD. Principal component analysis of transcriptome data from lesional skin showed that AD and NE cluster together distinct of psoriasis. In line with this, an established molecular classifier identified NE as AD rather than psoriasis. Finally, we demonstrated clinical and molecular efficacy of dupilumab treatment in NE. NE shows overlapping type 2 and type 3 immune signatures, while type 2 immunity predominates and should be the primary target of specific therapeutic interventions. This supports the view of NE as a variant of AD.

Fibreglass dermatitis: diagnostic challenges and occupational interventions.

Fibreglass dermatitis is a common occupationally acquired irritant contact dermatitis, where small spicules of fibreglass lodging in the stratum corneum result in mechanical irritation. We present two patients, an air-conditioning ducting worker and an injection moulding machine operator, who both presented with generalized pruritus. In the first case, polarized microscopy of a skin biopsy specimen demonstrated rare small spicules, with a diameter of 1 µm, lodged in the stratum corneum. In the second case, skin tape stripping demonstrated fibreglass particles, not found on skin biopsy. Proper work practices, personal hygiene and use of impervious barrier materials were recommended. The first patient did not return for follow-up, and the second patient's dermatitis resolved after handling of fibreglass-containing material was eliminated from his job scope. In conclusion, we present two cases of fibreglass dermatitis to illustrate the challenges in diagnosis and highlight strategies for prevention.

The role of skin biopsy in the detection of exposure to endocrine disrupting chemicals in Mediterranean cetaceans

Use of skin biopsy is proposed as a sensitive non-lethal technique for the hazard assessment of Mediterranean cetaceans exposed to endocrine disrupting chemicals (EDCs). EDCs are a structurally diverse group of compounds that may adversely affect the health of humans and wildlife or their progeny, by interaction with the endocrine system. In the Mediterranean environment top predators accumulate high concentrations of polyhalogenated aromatic hydrocarbons (PHAHs) and toxic metals, incurring high toxicological hazard. In this paper, the hypothesis that Mediterranean cetaceans are potentially at risk due to PHAH-EDCs is investigated using skin biopsy samples. Benzo-a-pyrene monoxigenase (BPMO) activity in skin biopsies was used as a potential indicator of exposure to different organochlorines (OCs) known to have endocrine disrupting properties. The main objective of this paper was to use this non-destructive ecotoxicological tool to define the potential hazard to Mediterranean odontocete and mysicete species, comparing the present data with values detected in other cetaceans from heavily polluted areas, affected by pseudohermaphroditism and other reproductive dysfunction. Subcutaneous tissue consisting of skin and blubber was obtained from striped dolphins (Stenella coeruleoalba), bottlenose dolphins (Tursiops truncatus), common dolphins (Delphinus delphis) and fin whales (Balaenoptera physalus) in the Mediterranean basin. Sampling was performed in the western Ligurian Sea, between Corsica and the French-Italian coast, and in the Ionian Sea. High concentrations of DDT metabolites and PCB congeners (known as Endocrine Disruptors) were detected in the different species. Significant differences in BPMO induction and OC levels were found between odontocetes and mysticetes. Differences in organochlorine bioaccumulation and consequently potential risk due to endocrine disruptors were primarily related to different positions in the marine food web. A statistical correlation was found between BPMO activity and organochlorine (op’DDT, a potent estrogen and antiandrogen and pp’DDE, a potent antiandrogen) levels in skin biopsy specimens of the endangered Mediterranean population of common dolphin. Several conclusions on the potential risk to Mediterranean cetaceans can be drawn from comparison of the levels of OC-EDs detected in Mediterranean odontocetes with those in white whales (Delphinapterus leucas) of the St Lawrence estuary and bowhead whales (Balaena mysticetus) affected by pseudohermaphroditism and other reproductive dysfunction. Finally, these results suggest that BPMO induction may be an early sign of exposure to EDCs such as OCs and a warning of the possibility of transgenerational effects through exposure of future generations via the placenta and milk.

Immunohistochemical Analysis of Adhesion Molecules E-Selectin, Intercellular Adhesion Molecule-1, and Vascular Cell Adhesion Molecule-1 in Inflammatory Lesions of Atopic Dermatitis.

E-selectin, ICAM-1 (intercellular adhesion molecule-1), and VCAM-1 (vascular cell adhesion molecule-1) play a role in atopic dermatitis (AD). This study aimed to evaluate their expression in skin biopsy specimens of patients diagnosed with AD using an optimized computer program. A descriptive analysis and comparison of digitally measured surface area and cell number were performed. The number of E-selectin-positive cells did not vary between the groups. In patients with AD, decreases of 1.2-fold for ICAM-1- and 1.3-fold for VCAM-1- positive cells were observed. The E-selectin-positive epidermal surface area increased (p < 0.001), while ICAM1 and VCAM1 decreased 2.5-fold and 2-fold, respectively, compared to controls. In the AD-affected skin, the E-selectin-positive endothelial area was 3.5-fold larger (p < 0.001), and the ICAM1-positive area was almost 4-fold larger (p < 0.001). E-selectin and ICAM-1 were expressed in the control dermis moderately and weakly, respectively. A strong E-selectin signal was detected in the AD-affected skin macrophages and a strong ICAM-1 signal in the dermal vessel endothelium. In the endothelial cells of AD-affected skin, no VCAM-1 signal could be found. E-selectin, ICAM-1, and VCAM-1 expression show significant disease-specific changes between AD-affected and control skin. The combination of digital analysis and a pathologist's evaluation may present a valuable follow-up of AD activity parameters.

Mycobacterium leprae and host immune transcriptomic signatures for reactional states in leprosy.

Mycobacterium leprae transcriptomic and human host immune gene expression signatures that demonstrate a plausible association with type I (T1R) and type II reactions (T2R) aid in early diagnosis, prevention of nerve damage and consequent demyelinating neuropathy in leprosy. The aim of the study is to identify M. leprae and host-associated gene-expression signatures that are associated with reactional states in leprosy. The differentially expressed genes from the whole transcriptome of M. leprae were determined using genome-wide hybridization arrays with RNA extracted from skin biopsies of 20 T1R, 20 T2R and 20 non reactional controls (NR). Additionally, human immune gene-expressions were profiled using RT2-PCR profiler arrays and real-time qPCRs. The RNA quality was optimal in 16 NR, 18 T1R and 19 T2R samples. Whole transcriptome expression array of these samples revealed significant upregulation of the genes that encode integral and intrinsic membrane proteins, hydrolases and oxidoreductases. In T1R lesional skin biopsy specimens, the top 10 significantly upregulated genes are ML2064, ML1271, ML1960, ML1220, ML2498, ML1996, ML2388, ML0429, ML2030 and ML0224 in comparison to NR. In T2R, genes ML2498, ML1526, ML0394, ML1960, ML2388, ML0429, ML0281, ML1847, ML1618 and ML1271 were significantly upregulated. We noted ML2664 was significantly upregulated in T1R and repressed in T2R. Conversely, we have not noted any genes upregulated in T2R and repressed in T1R. In both T1R and T2R, ML2388 was significantly upregulated. This gene encodes a probable membrane protein and epitope prediction using Bepipred-2.0 revealed a distinct B-cell epitope. Overexpression of ML2388 was noted consistently across the reaction samples. From the host immune gene expression profiles, genes for CXCL9, CXCL10, CXCL2, CD40LG, IL17A and CXCL11 were upregulated in T1R when compared to the NR. In T2R, CXCL10, CXCL11, CXCL9, CXCL2 and CD40LG were upregulated when compared to the NR group. A gene set signature involving bacterial genes ML2388, ML2664, and host immune genes CXCL10 and IL-17A can be transcriptomic markers for reactional states in leprosy.

Clinical, radiological, and molecular analyses of neuronal intranuclear inclusion disease with polyglycine inclusions

Neuronal intranuclear inclusion disease (NIID) is a clinically complex neurological disorder that appears sporadically or autosomally. Expansions of intronic GGC trinucleotide repeats in the NOTCH2 N-terminal-like C (NOTCH2NLC) gene cause NIID. In this study, to clarify the clinical characteristics useful for the differential diagnosis of NIID, clinical data of neurological examination, neuroimaging, and nerve conduction studies of six NIID patients diagnosed by pathological or genetic investigations were analyzed. Clinically useful characteristics for diagnosing NIID include general hyporeflexia, episodic disturbance of consciousness, sensory disturbance, miosis, and dementia. Furthermore, neuroimaging findings, such as leukoencephalopathy in T2-weighted magnetic resonance imaging and a linear high intensity of subcortical U-fibers in diffusion-weighted imaging (DWI), as well as decreased motor nerve conduction velocity, are especially important biomarkers for NIID. However, it is necessary to remember that these features may not always be present, as shown in one of the cases who did not have a DWI abnormality in this study. This study also investigated whether expanded GGC repeats were translated into polyglycine. Immunohistochemical analysis using a custom antibody raised against putative C-terminal polypeptides followed by polyglycine of uN2CpolyG revealed that polyglycines were localized in the intranuclear inclusions in skin biopsy specimens from all six patients, suggesting its involvement in the pathogenesis of NIID.

Kikuchi‐Fujimoto disease presenting with papular lesions on the elbows and knees

Kikuchi-Fujimoto disease is a self-limited febrile lymphadenitis of unknown etiology, possibly associated with cutaneous lesions. We report a case of Kikuchi-Fujimoto disease presenting with papular lesions limited to the elbows and knees. The skin biopsy specimen showed spongiosis and pseudo-Pautrier abscesses in addition to the typical histopathological findings of Kikuchi-Fujimoto disease. Although necrosis was not observed in the lymph node biopsy specimen, histopathological and immunohistochemical findings were consistent with Kikuchi-Fujimoto disease in the proliferating phase. Kikuchi-Fujimoto disease (KFD) is a self-limited febrile lymphadenitis of unknown etiology, predominantly affecting the cervical lymph nodes of young adults.1 Nodal lesions show paracortical necrosis with non-neutrophilic karyorrhexis, infiltration of CD8+ T cells and myeloperoxidase (MPO)+ histiocytes, and CD123+ plasmacytoid dendritic-cell clusters, although necrosis may be absent depending on the disease phases.1, 2 Cutaneous lesions have been reported in up to 40% of cases.3 The skin manifestations are variable, but often present on the upper body.4-6 We report a case of KFD with papular lesions limited to the elbows and knees. A 20-year-old man presented with a 2-week history of fever. Physical examination revealed non-tender lymphadenopathy of bilateral posterior cervical, axillary, and inguinal lymph nodes and non-pruritic erythematous papules on the elbows and knees (Figure 1A,B). Laboratory tests showed elevated levels of C-reactive protein (3.62 mg/dL) and serum lactate dehydrogenase (318 U/L; reference range, 124–222 U/L) and decreased white blood cell count (1700/μL). Anti-nuclear antibodies were negative. Computed tomography did not detect central lymph node swelling. The skin biopsy specimen showed interface dermatitis, papillary dermal edema, and dermal infiltrate (Figure 1C). In the epidermis, spongiosis and pseudo-Pautrier abscesses were present (Figure 1D). The dermal lymphohistiocytic infiltrate was associated with karyorrhexis in the absence of neutrophils (Figure 1E). Most lymphocytes were positive for CD3 and contained both CD4+ and CD8+ cells, although most of the intraepidermal T cells were CD8+ cells expressing TIA1, perforin, and granzyme B. The dermal infiltrate contained numerous CD68+ (clones KP-1 and PG-M1) histiocytes (Figure 1F) that were comprised of CD163+, MPO+, and CD123+ cells. The lymph node biopsy specimen showed paracortical lymphohistiocytic infiltrate with karyorrhexis in the absence of neutrophils (Figure 1G). CD8+ T cells predominated over CD4+ cells (Figure 1H,I). CD68+, CD163+ histiocytes were abundant, and most of them expressed MPO (Figure 1J,K). Clusters of CD123+ plasmacytoid dendritic cells were present (Figure 1L). Although necrosis was not observed in the lymph node biopsy specimen, the diagnosis of KFD in the proliferative phase was made based on the histopathological and immunohistochemical findings.1, 2, 4-6 Only supportive therapies with an oral non-steroidal anti-inflammatory drug and topical corticosteroid were given. Fever, lymphadenopathy, and cutaneous lesions resolved spontaneously in 5 weeks after the disease onset. Cutaneous lesions of KFD are characterized by superficial and deep dermal lymphohistiocytic infiltrate, non-neutrophilic karyorrhexis, interface dermatitis, and papillary dermal edema.4-6 Although spongiosis was observed in some cases,5, 6 presence of pseudo-Pautrier abscesses has not been reported. The reason why the cutaneous lesions in our case were limited to the elbows and knees is unknown, but their localization and the spongiotic epidermal changes suggest involvement of exogenous stimuli. Differential diagnosis of KFD mainly includes SLE and lymphomas.1 In our case, serum anti-nuclear antibodies were negative, and clinical signs suggestive of SLE other than fever and lymphadenopathy were absent. Histopathological and immunohistochemical findings of skin and lymph node biopsy specimens excluded both SLE and lymphomas. In patients with cutaneous eruptions, fever, and lymphadenopathy, KFD should be considered. Immunohistochemical studies are useful for accurate diagnosis, especially in atypical cases. We thank K. Gunshin for her technical assistance. The authors declare no conflict of interest. Approval of the research protocol: N/A. Informed consent: Written informed consent was obtained from the patient. Registration No. of the study/trial: N/A. Animal studies: N/A. The data that support the findings of this study are available from the corresponding author, K.K., upon reasonable request.

Coexistence of psoriasiform stratum corneum and underlying lichenoid features in skin biopsies of cutaneous reactions to anti-PD1.

Coexistence of psoriasiform stratum corneum and underlying lichenoid features in skin biopsies of cutaneous reactions to anti-PD1.