Discrimination of normal and cancerous human skin tissues based on laser-induced spectral shift fluorescence microscopy.

A homemade spectral shift fluorescence microscope (SSFM) is coupled with a spectrometer to record the spectral images of specimens based on the emission wavelength. Here a reliable diagnosis of neoplasia is achieved according to the spectral fluorescence properties of ex-vivo skin tissues after rhodamine6G (Rd6G) staining. It is shown that certain spectral shifts occur for nonmelanoma/melanoma lesions against normal/benign nevus, leading to spectral micrographs. In fact, there is a strong correlation between the emission wavelength and the sort of skin lesions, mainly due to the Rd6G interaction with the mitochondria of cancerous cells. The normal tissues generally enjoy a significant red shift regarding the laser line (37 nm). Conversely, plenty of fluorophores are conjugated to unhealthy cells giving rise to a relative blue shift i.e., typically SCC (6 nm), BCC (14 nm), and melanoma (19 nm) against healthy tissues. In other words, the redshift takes place with respect to the excitation wavelength i.e., melanoma (18 nm), BCC (23 nm), and SCC (31 nm) with respect to the laser line. Consequently, three data sets are available in the form of micrographs, addressing pixel-by-pixel signal intensity, emission wavelength, and fluorophore concentration of specimens for prompt diagnosis.

Ichthyosis, psoriasiform dermatitis, and recurrent fungal infections in patients with biallelic mutations in PERP.

BACKGROUND: Germline autosomal dominant and autosomal recessive mutations in PERP, encoding p53 effector related to PMP-22 (PERP), a component of epidermal desmosomes, have been associated with a spectrum of keratodermas. Monoallelic nonsense mutations cause Olmsted syndrome with severe periorificial keratoderma and palmoplantar keratoderma (PPK). Biallelic recessive frameshift and missense mutations are associated with milder forms of the disease, including generalised erythrokeratoderma and PPK. OBJECTIVES: To add new insights into the genotype-phenotype correlations as a consequence of PERP mutations and to provide a comprehensive review of the literature. METHODS: Among 26 previously unresolved families within a cohort of 180 extended Iranian families with syndromic or non-syndromic ichthyosis, two families with shared clinical features were examined by whole-exome sequencing and genome-wide homozygosity mapping. Mycological and dermatopathological studies were performed to further characterise their atypical phenotypic presentations. RESULTS: In two unrelated multiplex consanguineous families affected by ichthyosis, two novel biallelic PERP variants, NM_022121.5, c.89T > C, p.Leu30Pro and c.466G > C, p.Gly156Arg, located inside of genomic homozygosity regions of the probands were detected. Interestingly, some patients had areas of scaly psoriasiform plaques on the background of generalised ichthyosis that appeared during active cutaneous fungal infections. Mycological examinations of these lesions revealed infections caused by Candida albicans, Epidermophyton floccosum, or Trichophyton rubrum. Histopathology of the psoriasiform lesions shared some features with psoriasis, which when combined with clinical presentation, led to incorrect diagnosis of guttate psoriasis or pustular psoriasis. CONCLUSIONS: PERP variants in ichthyosis patients can confer susceptibility to recalcitrant cutaneous fungal infections. Additionally, patients with episodic psoriasiform dermatitis in the setting of keratoderma should be considered for PERP genotyping and cutaneous fungal examinations.

Evaluation of Th17 associated antigen in Old World Cutaneous Leishmaniasis: A comparative study in acute versus chronic human cutaneous Leishmaniasis using immunohistochemistry.

There are little information about Th17 cells and cutaneous Leishmaniasis (CL), due to an important effect of Th17 cells on immune response, it is worth to explore the role of Th17 on CL. The purpose of this study was to assess Th17 population in patients with acute vs. chronic CL lesions in comparison with skin samples collected from healthy volunteers in an endemic region of Old World CL. A total of 49 patients with clinical manifestations of chronic (n=16) and acute (n=33) CL lesions were recruited. The clinical diagnosis of CL was confirmed by direct smear or PCR. Biopsy specimens from prelesional skin of non-infectious lesions of 30 healthy individuals were used as control. Tissue sections of 3µm thickness were prepared and used for immunohistochemistry (IHC) analysis with primary antibody specific for Th17 associated antigen (CD161). For IHC, Envision+ (DakoCytomation) system was used and developed by using diaminobenzidine (DakoCytomation). The mean age of 33 patients with acute CL and the mean age of 16 patients with chronic CL were accordingly 45.24±16.43 and 33.56±15.87. In acute and chronic CL the mean (±standard deviation) and median (±interquartile range) were accordingly 2.92±2.21, 2.56±2.9 and 2.1±1.99, 1.54±2.81. In healthy controls the mean (±standard deviation) and median (±interquartile range) were 0.72±0.41 and 0.61±0.58 respectively. With pairwise comparison of acute, chronic and control groups, there were significant difference between acute and control (P value < 0.001), chronic and control (P value = 0.043). The results showed that there was an increasing cellular response of Th17 in both acute and chronic CL patients. Th17 was significantly higher in patients with acute and chronic CL lesions in comparison with healthy control group. However, there was no significant difference between acute and chronic infection concerning to Th17 cells.

Evaluation of Th17 associated antigen in Old World Cutaneous Leishmaniasis: A comparative study in acute versus chronic human cutaneous Leishmaniasis using immunohistochemistry

@#There are little information about Th17 cells and cutaneous Leishmaniasis (CL), due to an important effect of Th17 cells on immune response, it is worth to explore the role of Th17 on CL. The purpose of this study was to assess Th17 population in patients with acute vs. chronic CL lesions in comparison with skin samples collected from healthy volunteers in an endemic region of Old World CL. A total of 49 patients with clinical manifestations of chronic (n=16) and acute (n=33) CL lesions were recruited. The clinical diagnosis of CL was confirmed by direct smear or PCR. Biopsy specimens from prelesional skin of non-infectious lesions of 30 healthy individuals were used as control. Tissue sections of 3μm thickness were prepared and used for immunohistochemistry (IHC) analysis with primary antibody specific for Th17 associated antigen (CD161). For IHC, Envision+ (DakoCytomation) system was used and developed by using diaminobenzidine (DakoCytomation). The mean age of 33 patients with acute CL and the mean age of 16 patients with chronic CL were accordingly 45.24±16.43 and 33.56±15.87. In acute and chronic CL the mean (±standard deviation) and median (±interquartile range) were accordingly 2.92±2.21, 2.56±2.9 and 2.1±1.99, 1.54±2.81. In healthy controls the mean (±standard deviation) and median (±interquartile range) were 0.72±0.41 and 0.61±0.58 respectively. With pairwise comparison of acute, chronic and control groups, there were significant difference between acute and control (P value < 0.001), chronic and control (P value = 0.043). The results showed that there was an increasing cellular response of Th17 in both acute and chronic CL patients. Th17 was significantly higher in patients with acute and chronic CL lesions in comparison with healthy control group. However, there was no significant difference between acute and chronic infection concerning to Th17 cells.

Diagnostic accuracy of BP180 NC16a and BP230-C3 ELISA in serum and saliva of patients with bullous pemphigoid.

BACKGROUND: Bullous pemphigoid (BP) is a subepidermal blistering disease, characterized by autoantibodies directed against BP180 and BP230. Collecting saliva is an easy and painless way of obtaining biological samples, and can be used for diagnosis of autoimmune diseases. AIM: To compare the diagnostic accuracy of serum and salivary BP180-NC16a and BP230-C3 in the initial diagnosis of BP. METHODS: We assessed 50 patients newly diagnosed with BP and 50 healthy controls. The diagnosis of BP was confirmed based on clinical, histopathological and immunofluorescence findings. Serum and saliva samples were collected from both groups, and BP180 and BP230 titres were assessed using commercially available ELISA kits. RESULTS: Using serum, the sensitivity of the serum BP180 and BP230 ELISA assays was 88% and 48%, respectively, and the specificity of both was 96%. Using saliva with the cutoff value proposed by the manufacturer, sensitivity was 56.2% and 14.6%, and specificity was 98% and 100%, respectively. Using the best calculated cutoff for saliva, sensitivity increased to 87.5% and 77.1%, and specificity to 96% and 62%, respectively. There was a significant correlation between serum and saliva BP180 levels and the severity of skin disease. Both serum and saliva BP230 levels were significantly higher in patients with mucosal involvement. CONCLUSION: Serum BP180 NC16a ELISA is a sensitive and specific test for the initial diagnosis of BP, whereas serum BP230-C3 ELISA is highly specific, but less sensitive. Saliva may be a noninvasive and convenient alternative for use in the BP180 NC16a ELISA to diagnose BP.

Diagnostic accuracy of punch biopsy in subtyping basal cell carcinoma.

BACKGROUND: Basal cell carcinoma (BCC) is the most common skin cancer in humans. The histological subtype reported by punch biopsy may influence the type of treatment. Few studies have investigated the accuracy of punch biopsy in diagnosing the true BCC subtype. OBJECTIVE: To determine the accuracy, sensitivity and specificity of punch biopsy in BCC subtype diagnosis. METHODS: In this retrospective study, 333 biopsy specimens and excisions were reviewed. Histological subtypes present in the initial biopsy were compared with tumour subtypes of the total excision. RESULTS: The concordance between the BCC subtype present in the biopsy specimen and in the subsequent excision specimen was 72.3%. The most common BCC patterns were nodular (158, 47.5%) and mixed subtype (90, 27%). Most mixed tumours contained one or more aggressive subtype (63/90, 70%). In 47/120 (39.1%) aggressive tumours (14.1% of the total), punch biopsy failed to correctly identify the aggressive component. The most commonly missed aggressive subtype was mixed aggressive including nodular/micronodular and nodular/infiltrative (30/47, 63.8%). In 45/213 (21.1%) non-aggressive BCCs (13.5% of total cases), punch biopsy incorrectly reported an aggressive subtype. The most commonly misidentified non-aggressive subtype was nodular (39/45, 86.6). The sensitivity and specificity of punch biopsy in diagnosing aggressive vs. non-aggressive BCC subtypes 60.8% (95% CI, 51.9-69.1) and 78.9% (95% CI, 72.8-83.8), respectively. The positive and negative predictive values were 61.9% and 78.1%, respectively. CONCLUSION: Punch biopsy has serious pitfalls in differentiating aggressive and non-aggressive BCC subtypes. Dermatologists should consider the possibility of aggressive components within non-aggressive BCCs reported using punch biopsy.