Newly Developed Psoriasis in a Patient with Telangiectasia Macularis Eruptiva Perstans and Systemic Mastocytosis Treated with Interferon.

The authors present a rare case of a patient with telangiectasia macular eruptiva perstans, with confirmed D816V mutation which later progressed to systemic mastocytosis confirmed by trepanobiopsy. First-line treatment - phototherapy - had to be stopped, and systemic treatment with interferon alpha-2a was initiated. The treatment was successful with regression of skin lesions as well as mast cell infiltrates in the bone marrow. However, the treatment was complicated by the onset of psoriasis lesions.

Detection of BRAFV600E Mutation in Melanoma Patients by Digital PCR of Circulating DNA.

AIMS: About 50% of melanomas have the BRAFV600E mutation. This mutation is an attractive therapeutic target. The aims of our study were to detect BRAFV600E mutations within circulating cell-free DNA in plasma ("liquid biopsy") by a droplet digital PCR (ddPCR) method, and to investigate how well the Breslow-Clark score can be predicted by ddPCR. MATERIALS AND METHODS: We analyzed 113 patients with malignant melanoma. ddPCR was performed using the QX200 system (BIO-RAD®, Hercules). All samples were tested in duplicate. Besides the results of the liquid biopsy, we have collected data on gender and age of the patients, as well as the mitotic activity of the tumor; the tumor subtype and localization, and the Breslow-Clark score. The limit of detection (LoD) was determined by the method of Tzonev. The LoD was found to be five events per well. RESULTS: The BRAFV600E mutation was detected in 37 of 113 samples. A moderate predictive accuracy of the Breslow-Clark score can be attained with the mitotic activity and the type of melanoma as the most important predictors. CONCLUSION: Our results show that ddPCR is a highly sensitive method and could be used for a routine laboratory detection of the BRAFV600E mutation as well as for follow-up monitoring to determine the treatment response in patients with malignant melanomas.

Endothelial Dysfunction in Children with Juvenile Psoriatic Arthritis.

BACKGROUND: To evaluate the presence of endothelial dysfunction in Slovak children with juvenile psoriatic arthritis in the absence of classic cardiovascular risk factors in order to assess its relationship to the disease activity and disability. METHODS: 25 juvenile psoriatic arthritis patients (JPSA) and 25 healthy controls aged 6-19 years were enrolled into this study. In all subjects vascular measurements over a period of three years (January 2013 - January 2016) were performed, in accordance with the guidelines for ultrasonographic evaluation of FMD% (flow-mediated endothelial dependent vasodilatation) of the brachial artery. The measured items were compared to the variables reflecting the disease activity and disability. RESULTS: Significantly lower FMD% values in patients with JPSA when compared to healthy controls {mean(SD), median, range: 5.49% (3.77), 3.55, 0.3-13.0 vs. 9.28% (1.72), 9.3, 6.4-13.1} (p < 0.001) have been documented. Strong correlations between FMD% values and disease duration (p < 0.01), non-specific inflammatory markers levels (p < 0.001) or functional disability (p < 0.01) have been observed. Significantly lower FMD% values in patients with an early disease onset (JPSA onset < 5 years of age) when compared to the rest of JPSA group {mean (SD), median, range: 4.39% (2.47), 4.45, 0.9-13.2 vs. 6.38% (1.42), 6.3, 3.2-12.1} (p < 0.01) have also been detected. CONCLUSION: Study is the only one addressing endothelial dysfunction development in Slovak children with psoriatic arthritides. We state that endothelial dysfunction is present in these patients even during childhood and in the absence of classic cardiovascular risk factors. Its development seems to be related to an early disease onset as well as to the increased disease activity and disability. Potential genetic predictors have also been identified.

Droplet digital PCR for detection of BRAF V600E mutation in formalin-fixed, paraffin-embedded melanoma tissues: a comparison with Cobas® 4800, Sanger sequencing, and allele-specific PCR.

Cutaneous melanoma has the worst prognosis of all skin cancers. Although emerging targeted therapies, such as B-Raf kinase inhibitor vemurafenib, improve prognosis they require an accurate and sensitive means of detecting the pathogenic BRAF V600E mutation. We compared the sensitivity of four BRAF V600E detection methods in formalin-fixed, paraffin-embedded melanoma biopsies from 87 consecutive melanoma patients with Breslow stage I-V disease (staging based on the depth of tumor of invasion). The methods assessed were the widely used Cobas® 4800 system based on real-time PCR amplification, Sanger sequencing, allele-specific PCR (AS-PCR), and droplet digital PCR (ddPCR). The BRAF V600E mutation was found in 8 (9.2%), 23 (26.4%), 23 (26.4%) and 31 (35.6%) biopsies, respectively. The limit of detection (LoD) was determined by three different methods: Poisson confidence limits, calibration regression and Tzonev's method. Pair-wise agreement between the methods was as follows: Cobas vs. Sanger, P = 0.33; Cobas® 4800 vs. AS-PCR, P = 0.33; Cobas® 4800 vs. ddPCR, P = 0.65; Sanger vs. AS-PCR, P = 1; Sanger vs. ddPCR, P = 0.08; AS-PCR vs. ddPCR, P = 0.06. Multinomial logistic regression was used for predictive modeling of the Breslow-Clark score; ddPCR emerged as the best predictor, the other predictors were mitotic activity, type of malignant melanoma and patient's age. Our results demonstrate that ddPCR is the most sensitive method of detecting the BRAF V600E mutation.