Repair of Defects of the Nasal Tip After Mohs Surgery.

INTRODUCTION: Mohs Micrographic Surgery (MMS) is a treatment option for high-risk facial nonmelanoma skin cancer with high cure rates. Especially on the nasal tip, the tissue sparing properties of MMS are appealing. The nasal tip is a common location of nonmelanoma skin cancer and can be a challenging anatomical structure for reconstructive surgery due to its prominent location in the face, the shortage of spare tissue, as well as the stiffness and composition of different skin types, cartilage and bone. OBJECTIVES: The aim of the present paper is to review and demonstrate how reconstruction of the nasal tip can be done successfully to improve the care for patients undergoing MMS in this area. METHODS: Using selected literature on the area and the surgeons experience, each method of repair are described including their individual advantages and challenges. Pictures and consent were selected from one patient who underwent each repair method and three photos are presented in this paper: one after tumor resection, one immediately after repair, and one minimum 6 months post-surgery. RESULTS: We present eight surgical methods as well as pictures from previous surgeries. CONCLUSIONS: The results demonstrate obtainable results using very different surgical methods and the importance of an individualized approach to repairing cutaneous defects of the nasal tip.

Topical Brimonidine Delays Ultraviolet Radiation-Induced Squamous Cell Carcinoma in Hairless Mice.

We investigated whether topical brimonidine delayed or enhanced the development of squamous cell carcinoma (SCC) when ultraviolet radiation (UVR) was applied to a well-established murine model. Hairless female mice (n = 125) were randomized into five groups and treated as follows: 1% brimonidine cream before UVR (Group 1), 0.33% brimonidine gel before UVR (Group 2), 1% brimonidine cream after UVR (Group 3), UVR only (control; Group 4) and 1% brimonidine cream only (control; Group 5). For each animal, the first four tumors were recorded and followed until three tumors reached 4 mm or one tumor reached 12 mm in diameter. All animal experiments continued for up to 365 days or until death. Application of 1% brimonidine cream before UVR delayed tumor development relative to control mice treated with UVR alone (P = 0.000006). However, when 0.33% brimonidine gel was applied before UVR (P = 0.313) or 1% brimonidine cream was applied after UVR (P = 0.252), there was no significant delay in tumor development relative to control mice treated with UVR alone. The development of the second and third tumors followed a similar pattern. Topical 1% brimonidine cream applied before UVR exposure delayed SCC development in hairless mice. In contrast, when brimonidine was applied after UVR there was no significant delay in tumor development. These results suggest that the 1% brimonidine cream probably absorbed the UVR, and therefore, a delay in tumor formation was only seen when brimonidine was applied before irradiation. However, there can be multiple reasons for this delay in photocarcinogenesis.

Few X-ray and PUVA treatments accelerate photocarcinogenesis in hairless mice.

PUVA is a treatment that combines oral methoxypsoralen (8-MOP) with ultraviolet radiation A (UVA). It is used for severe psoriasis and the early stages of T-cell lymphoma. X-rays are an effective treatment for skin cancers. Both treatments are in higher doses used to treat skin malignancies and simultaneously increase the risk of keratinocyte cancer. The main objective of this study was to test whether a few PUVA or X-ray treatments could delay the development of ultraviolet radiation (UVR)-induced skin tumors in a well-established hairless mouse model. Three groups of immunocompetent mice (total, N = 75) were included in the study. All groups were UVR-exposed during the study period. In addition, one group was treated with PUVA and another group was treated with X-rays at days 45, 52, 90 and 97. A control group was treated with UVR only. We recorded when the first, second and third skin tumors were induced in each mouse. Skin tumors developed significantly earlier in both the PUVA and X-ray groups (median, 188 days) than in the control mice (median, 215 days; p < 0.001). Therefore, a few X-ray and PUVA treatments both significantly accelerated the development of skin tumors in hairless mice, compared to UVR controls. Neither treatment showed a delay of UVR-induced skin tumors and caution should be exercised before applying these treatments to sun-damaged skin.

A Skin Cancer Prophylaxis Study in Hairless Mice Using Methylene Blue, Riboflavin, and Methyl Aminolevulinate as Photosensitizing Agents in Photodynamic Therapy.

The high incidence of sunlight-induced human skin cancers reveals a need for more effective photosensitizing agents. In this study, we compared the efficacy of prophylactic photodynamic therapy (PDT) when methylene blue (MB), riboflavin (RF), or methyl aminolevulinate (MAL) were used as photosensitizers. All mice in four groups of female C3.Cg/TifBomTac hairless immunocompetent mice (N = 100) were irradiated with three standard erythema doses of solar-simulated ultraviolet radiation (UVR) thrice weekly. Three groups received 2 × 2 prophylactic PDT treatments (days 45 + 52 and 90 + 97). The PDT treatments consisted of topical administration of 16% MAL, 20% MB, or 20% RF, and subsequent illumination that matched the photosensitizers' absorption spectra. Control mice received no PDT. We recorded when the first, second, and third skin tumors developed. The pattern of tumor development after MB-PDT or RF-PDT was similar to that observed in irradiated control mice (p > 0.05). However, the median times until the first, second, and third skin tumors developed in mice given MAL-PDT were significantly delayed, compared with control mice (256, 265, and 272 vs. 215, 222, and 230 days, respectively; p < 0.001). Only MAL-PDT was an effective prophylactic treatment against UVR-induced skin tumors in hairless mice.

Shot-noise limited, supercontinuum-based optical coherence tomography.

We present the first demonstration of shot-noise limited supercontinuum-based spectral domain optical coherence tomography (SD-OCT) with an axial resolution of 5.9 µm at a center wavelength of 1370 nm. Current supercontinuum-based SD-OCT systems cannot be operated in the shot-noise limited detection regime because of severe pulse-to-pulse relative intensity noise of the supercontinuum source. To overcome this disadvantage, we have developed a low-noise supercontinuum source based on an all-normal dispersion (ANDi) fiber, pumped by a femtosecond laser. The noise performance of our 90 MHz ANDi fiber-based supercontinuum source is compared to that of two commercial sources operating at 80 and 320 MHz repetition rate. We show that the low-noise of the ANDi fiber-based supercontinuum source improves the OCT images significantly in terms of both higher contrast, better sensitivity, and improved penetration. From SD-OCT imaging of skin, retina, and multilayer stacks we conclude that supercontinuum-based SD-OCT can enter the domain of shot-noise limited detection.

[Diagnostics and treatment of lentigo maligna melanoma in Denmark].

The incidence of maligna melanoma has doubled in Denmark over the latest 30 years. Lentigo maligna melanomas (LMM) account for 10-26% of the melanomas in head and neck. LMM most often strike people over the age of 70 years and frequently occur in sun-exposed skin areas. LMM can be difficult to diagnose, since its clinical features are similar to benign skin lesions, solar lentigo and seborrhoeic keratosis. This review presents various methods for diagnosing and treating LMM. The specific method is chosen in dialogue with the patients, and overall health is an important factor when choosing an appropriate treatment.

[Melanonychia].

Melanonychia is seen, when melanin is incorporated in the nail, which gives a band of dark discolouration of the nail. Nail discolouration is a common cause for patients to seek medical attention, and it is often benign. Melanonychia can be seen due to melanocytic proliferation (ungual naevi), benign hyperplasia (lentigo) and hypermelanosis (infections, traumas). However, subungual melanoma also commonly presents with melanonychia and is often overseen, leading to a worse prognosis. It is therefore important systematically to examine all nail discolourations to find the cause and rule out malignancy.

[Mohs surgery for basal cell carcinoma].

Micrographic surgery is currently the only technique which ensures complete removal of basal cell carcinomas. The major limitation is the high set-up cost, which is particularly connected with specialized training of surgeons, technicians and the set-up of a histology facility for frozen tissue sectioning and staining. In the long run, however, the cost of Mohs surgery per patient does not exceed that of conventional surgery. The technique is very safe and has multiple advantages over any other treatment modality. It achieves the highest cure rates, it is minimally invasive, it is tissue-sparing and it enables the optimal closure of the surgical defect. Mohs surgery is cost-effective, especially when dealing with poorly demarcated, high-risk, facial tumours, where it should be considered as the first choice of treatment.

[Basal cell carcinoma surgery].

Basal cell carcinoma (BCC) is the most common type of cancer in humans. The lifetime risk of developing BCC is 30%. The scope of the treatment is total eradication of the tumour followed by preservation of functionality and optimal cosmetic results. The risk of recurrence after treatment is highest in the tumours with aggressive growth patterns, in tumours located in the facial H-zone and in recurrent tumours. The optimal treatment of high-risk BCCs is surgical excision or radiotherapy. Low-risk tumours may be treated with cryotherapy, curettage, photodynamic therapy or topical agents.

Pigmentary Markers in Danes--Associations with Quantitative Skin Colour, Nevi Count, Familial Atypical Multiple-Mole, and Melanoma Syndrome.

To investigate whether pigmentation genes involved in the melanogenic pathway (melanogenesis) contributed to melanoma predisposition, we compared pigmentary genetics with quantitative skin pigmentation measurements, the number of atypical nevi, the total nevus count, and the familial atypical multiple mole and melanoma (FAMMM) syndrome. We typed 32 pigmentary SNP markers and sequenced MC1R in 246 healthy individuals and 116 individuals attending periodic control for malignant melanoma development, 50 of which were diagnosed with FAMMM. It was observed that individuals with any two grouped MC1R variants (missense, NM_002386:c. 456C > A (p.TYR152*), or NM_002386:c.83_84insA (p.Asn29Glnfs*14) had significantly (p<0.001) lighter skin pigmentation of the upper-inner arm than those with none or one MC1R variant. We did not observe any significant association of the MC1R variants with constitutive pigmentation measured on the buttock area. We hypothesize that the effect of MC1R variants on arm pigmentation is primarily reflecting the inability to tan when subjected to UVR. A gender specific effect on skin pigmentation was also observed, and it was found that the skin pigmentation of females on average were darker than that of males (p<0.01). We conclude that MC1R variants are associated with quantitative skin colour in a lightly pigmented Danish population. We did not observe any association between any pigmentary marker and the FAMMM syndrome. We suggest that the genetics of FAMMM is not related to the genetics of the pigmentary pathway.

Favourable results of Mohs micrographic surgery for basal cell carcinoma.

INTRODUCTION: Basal cell carcinoma (BCC) is the most common malignant neoplasm with an annual incidence approaching 200/100,000 person-years. Mohs micrographic surgery (MMS) is widely used in North America and in Europe for treatment of BCC. This technique ensures radical tumour removal, sparing of the surrounding healthy skin, and it also offers higher cure rates than standard tumour excision with a predefined margin of healthy skin. The superiority of MMS relies on the fact that the entire (100%) margin of the excised tissue is examined microscopically for residual tumour in contrast to the traditional histopathological examination, in which 2% of the margin is examined. METHODS: In Denmark, MMS was first introduced by us in 2012. In the present study, we retrospectively included all patients who underwent MMS from May 2012 to June 2015. RESULTS: A total of 231 patients with 263 BCC were included. The mean age was 66.1 years. The most common localisations were the forehead (31.3%), the nose (31.0%) and the cheek (14.7%). Primary BCC comprised 54.0%; the remaining cases were relapses, most frequently after curettage (36.9%), radiotherapy (18.9%) and photodynamic therapy (11.7%). MMS leads to 40% smaller skin defects than standard excisions with 4 or 6 mm margins. Closure of skin defects was achieved by side-to-side closure in 49% and by local flaps in 40%. There were no relapses during the observation time. The safety, cosmetic and functional outcome were excellent. CONCLUSIONS: We recommend that MMS be included in the Danish BCC treatment guidelines, especially for high-risk BCC in the face, in line with standard practice in Europe and the United States. FUNDING: none. TRIAL REGISTRATION: not relevant.

Incidence of In Situ and Invasive Melanoma in Denmark From 1985 Through 2012: A National Database Study of 24,059 Melanoma Cases.

IMPORTANCE: In Denmark, the incidence of malignant melanoma (MM) has doubled during the past 25 years, with an incidence of 29.5 and 31.7 per 100,000 person-years in 2012 for men and women, respectively. Understanding the nature of this increase in incidence is important to optimize prevention, early diagnosis, and treatment of in situ and invasive melanoma in Denmark. OBJECTIVE: To describe changes over time in the incidence and clinical and pathologic characteristics of in situ and invasive melanoma in Denmark from 1985 through 2012. DESIGN, SETTING, AND PARTICIPANTS: We used the official national Danish Melanoma Group database to describe all eligible, prospectively registered cases of in situ and invasive melanoma in Denmark from January 1, 1985, through December 31, 2012. Data analyses were performed from April 1, 2012, through January 31, 2013. MAIN OUTCOMES AND MEASURES: Estimated annual percentage changes (EAPCs) for men and women in European age-standardized incidence, age at diagnosis, and tumor region for in situ melanoma and MM. For MM only, melanoma type, Breslow thickness, ulceration, and mortality. RESULTS: We included 3299 cases of in situ melanoma and 20,760 cases of MM. The incidence (95% CI) of MM increased by 4.5% (3.6%-5.3%) for men and 4.3% (3.5%-5.2%) for women, which was especially pronounced in patients older than 60 years (EAPCs, 5.8% [4.7%-6.8%] and 4.8% [3.8%-5.9%], respectively), in thin (Breslow thickness, <0.75 mm) melanoma (EAPCs, 6.6% [5.0%-8.2%] and 6.1% [6.0%-7.1%], respectively), and in superficially spreading MM (EAPCs, 5.2% [4.3%-6.2%] and 4.7% [3.9%-5.7%], respectively). We found no significant EAPC in the incidence of melanomas with Breslow thickness greater than 2.00 mm in women, and relative ulceration rates (95% CI) declined in both sexes (EAPCs, -3.3% [-4.0% to -2.6%] in men and -3.4% [-4.0% to -2.8%] in women). More proximal tumor location occurred over time (P < .001). Incidence of in situ melanoma (95% CI) greatly increased (EAPCs, 14.0% [12.2%-15.8%] in men and 11.6% [10.2%-13.2%] in women) with changes over time in age and region (defined by codes in the International Statistical Classification of Diseases, Tenth Revision) similar to those for MM. Mortality related to MM increased in men (EAPC, 0.6% [0.1% to 1.2%]), whereas mortality in women (EAPC, -0.4% [-1.0% to 0.3%]) remained stable. CONCLUSIONS AND RELEVANCE: This study confirms a worldwide increase in melanoma incidence. Results may indicate the importance of secondary melanoma prevention in Denmark. Future efforts could intensify primary prevention aimed at young adults, adolescents, and children and maintain and target secondary prevention at the population older than 60 years.

miR-125b induces cellular senescence in malignant melanoma.

BACKGROUND: Micro RNAs (miRs) have emerged as key regulators during oncogenesis. They have been found to regulate cell proliferation, differentiation, and apoptosis. Mir-125b has been identified as an oncomir in various forms of tumours, but we have previously proposed that miR-125b is a suppressor of lymph node metastasis in cutaneous malignant melanoma. Our goal was therefore to further examine this theory. METHODS: We used in-situ-hybridization to visualise miR-125b expression in primary tumours and in lymph node metastasis. Then using a miRVector plasmid containing a miR-125b-1 insert we transfected melanoma cell line Mel-Juso and then investigated the effect of the presence of a stable overexpression of miR-125b on growth by western blotting, flow cytometry and ß-galactosidase staining. The tumourogenicity of the transfected cells was tested using a murine model and the tumours were further examined with in-situ-hybridization. RESULTS: In primary human tumours and in lymph node metastases increased expression of miR-125b was found in single, large tumour cells with abundant cytoplasm. A stable overexpression of miR-125b in human melanoma cell line Mel-Juso resulted in a G0/G1 cell cycle block and emergence of large cells expressing senescence markers: senescence-associated beta-galactosidase, p21, p27 and p53. Mel-Juso cells overexpressing miR-125b were tumourigenic in mice, but the tumours exhibited higher level of cell senescence and decreased expression of proliferation markers, cyclin D1 and Ki67 than the control tumours. CONCLUSIONS: Our results confirm the theory that miR-125b functions as a tumour supressor in cutaneous malignant melanoma by regulating cellular senescence, which is one of the central mechanisms protecting against the development and progression of malignant melanoma.

Down-regulation of microRNAs controlling tumourigenic factors in follicular thyroid carcinoma.

The molecular determinants of thyroid follicular nodules are incompletely understood and assessment of malignancy is a diagnostic challenge. Since microRNA (miRNA) analyses could provide new leads to malignant progression, we characterised the global miRNA expression in follicular adenoma (FA) and follicular carcinoma (FC). Comparison of carcinoma and adenoma with normal thyroid revealed 150 and 107 differentially expressed miRNAs respectively. Most miRNAs were down-regulated and especially miR-199b-5p and miR-144 which were essentially lost in the carcinomas. Integration of the changed miRNAs with differentially expressed mRNAs demonstrated an enrichment of seed sites among up-regulated transcripts encoding proteins implicated in thyroid tumourigenesis. This was substantiated by the demonstration that pre-miR-199b reduced proliferation when added to cultured follicular thyroid carcinoma cells. The down-regulated miRNAs in FC exhibited a substantial similarity with down-regulated miRNAs in anaplastic carcinoma (AC) and by gene set enrichment analysis, we observed a significant identity between target mRNAs in FC and transcripts up-regulated in AC. To examine the diagnostic potential of miRNA expression pattern in distinguishing malignant from benign nodules we employed a supervised learning algorithm and leave-one-out-cross-validation. By this procedure, FA and FC were identified with a negative predicted value of 83% (data generated by microarray platform) and of 92% (data generated by qRT-PCR platform). We conclude that follicular neoplasia is associated with major changes in miRNA expression that may promote malignant transformation by increasing the expression of transcripts encoding tumourigenic factors. Moreover, miRNA profiling may facilitate the diagnosis of carcinoma vs adenoma.

Are all melanomas dangerous?

The increased incidence of cutaneous malignant melanoma, together with only minor changes in mortality, has brought into question the existence of a melanoma epidemic. The discrepancy between incidence and mortality suggests that most newly diagnosed melanomas have indolent behaviour. This review summarizes the most recent epidemiological findings regarding the incidence of cutaneous malignant melanoma, mortality, Breslow thickness and clinical stage. Studies published between 2005 and 2010 with more than 2,000 test subjects were included in this review. These studies all report an increase in incidence of melanoma during the last few decades, with by far the highest increase in tumours at a very early stage (T1 or IA). Little or no change was seen in mortality. However, increases in both mortality and incidence of thick melanomas were found in the oldest subgroups, especially in men. These findings indicate the existence of a certain degree of overdiagnosis of melanoma. They also indicate the existence of two different types of epidemic, for younger and older subgroups.

MicroRNA miR-125b induces senescence in human melanoma cells.

MicroRNAs (miRNAs) are small noncoding RNA molecules involved in gene regulation. Aberrant expression of miRNA has been associated with the development or progression of several diseases, including cancer. In a previous study, we found that the expression of miRNA-125b (miR-125b) was two-fold lower in malignant melanoma producing lymph node micrometastases than in nonmetastasizing tumors. To get further insight into the functional role of miR-125b, we assessed whether its overexpression or silencing affects apoptosis, proliferation, or senescence in melanoma cell lines. We showed that overexpression of miR-125b induced typical senescent cell morphology, including increased cytoplasmatic/nucleus ratio and intensive cytoplasmatic ß-galactosidase expression. In contrast, inhibition of miR-125b resulted in 30-35% decreased levels of spontaneous apoptosis. We propose that downregulation of miR-125b in an early cutaneous malignant melanoma can contribute to the increased metastatic capability of this tumor.

The microRNA molecular signature of atypic and common acquired melanocytic nevi: differential expression of miR-125b and let-7c.

MicroRNAs (miRNAs) are small non-coding RNAs, which regulate gene expression through base pairing with mRNA and which are crucially involved in carcinogenesis (the so-called oncomiRs). We compared the miRNA signature between acquired melanocytic nevi showing clinical atypia (atypic nevi, AN) and common acquired nevi (common nevi, CN). We obtained miRNA profiles from 41 biopsies (22 AN and 19 CN) and showed that AN could be differentiated from CN on the basis of the expression of 36 miRNAs (false discovery rate <0.05). OncomiRs were present in this group, and we further confirmed the differential expression of miR-125b and let-7c by qRT-PCR. Our data suggest that miRNAs are functionally involved in the pathogenesis of nevi and possibly malignant melanoma.

A simple procedure for routine RNA extraction and miRNA array analyses from a single thyroid in vivo fine needle aspirate.

CONTEXT: microRNA (miRNA) expression profiling and classification of tissue obtained from fine-needle aspirates (FNA) could be a major improvement of the preoperative diagnosis of thyroid nodules. OBJECTIVE: Before this can be clinically implemented, a robust and non-toxic method for obtaining sufficient quantity and quality of RNA from single in vivo FNA has to be established. RNAlater is a non-toxic RNA-stabilizing agent. However, due to the high density of RNAlater, pelleting of the tissue samples is difficult, and results in low recovery of RNA that is insufficient for subsequent miRNA array expression analysis. We therefore developed a simple centrifugation method for capturing tissue stored in RNAlater on a 0.45-µm filter. DESIGN: FNA from 24 patients with a solitary cold thyroid nodule was stored in Trizol, liquid nitrogen, or RNAlater. The tissue stored in RNAlater was either pelleted by centrifugation or captured on the 0.45-µm filters. RNA was extracted using the Trizol method. To validate results, FNA from additional 30 patients were analyzed based on the modified RNAlater protocol. MAIN OUTCOME: Capturing FNA tissue samples on the filters increased the RNA yield 10 fold and maintained RNA purity, permitting miRNA array expression profiling and allowing comparable levels of known miRNA-clusters regardless of preservation technique. Results were confirmed in an additional 30 patients. CONCLUSION: The modified RNAlater protocol is well suited for isolating RNA from single thyroid in vivo FNA in a clinical setting. Furthermore this permits shipping of FNA samples at room temperature from peripheral centers to a centralized array core facility.

Downregulation of miR-125b in metastatic cutaneous malignant melanoma.

This study aimed to identify microRNA species involved in the earliest metastatic event in cutaneous malignant melanoma (MM). Samples from 28 patients with MM [stage T2 (tumor), M0 (distant metastasis)] were grouped by the presence of micrometastasis in the sentinel lymph nodes (N0/N1). Melanoma cells were harvested from primary, cutaneous MM tumors by laser-capture microdissection, and microRNA expression profiles were obtained by the microarray technique. Results were validated by quantitative reverse transcription PCR. We found that miR-125b was downregulated in the primary cutaneous melanomas that produced early metastases (T2, N1, M0) compared with the sentinel lymph node-negative (T2, N0, M0) melanomas. MiR-125b has earlier been found to be downregulated in other tumor types and in atypic naevi compared with the common acquired naevi. In conclusion, miR-125b may be involved in an early progression of cutaneous MM.