Rhabdomyosarcoma in a congenital pigmented nevus

A 7-month-old boy had a giant pigmented lesion involving the trunk and thighs that exhibited many hyperpigmented hairy and verrucous nevi. One of the nevi ulcerated and on histological examination consisted of pleomorphic rhabdomyosarcoma cells that stained for muscle-specific actin (HHF-35), desmin, and myoglobin. Around the tumor, in the dermis, benign pigmented nevus cells were observed. The occurrence of malignant tumors, other than malignant melanoma, in pigmented nevi is rarely described.

Resultados del tratamiento de nevos melanocíticos con láser

Introducción A pesar de que los nevos melanocíticos son un motivo de consulta frecuente en nuestra población no existen estudios a nivel de Colombia acerca de su tratamiento, a nivel mundial existe muy poca literatura al respecto por lo que hay un vacío conceptual en este campo. Objetivos Evaluar los cambios en cuanto a la presencia de pigmento y cicatrización, en los nevos melanocíticos adquiridos tratados con láser, basados en la experiencia de un solo centro en Bogotá. Materiales y métodos Es un estudio observacional de antes y después, en una cohorte histórica, de 90 casos de nevos melanocíticos adquiridos, tratados con láser en Uniláser Medica, en los que se evaluó la presencia de pigmento, cicatrización, y otras variables, con un control realizado a no menos de 3 meses de la intervención. Resultados Se encontró un rango de edad entre los 18 -51 años, promedio 27,59 años; fototipo de III-V; en el 32% de los casos, solo fue requerida una sesión de láser de Co2 y Erbio, para el aclaramiento completo de la misma. La duración del eritema en el 54,4% los casos fue de 1 a 3 meses. En un 64,4% quedó pigmento residual al control, pero de éstos casos el 48,2% fue entre un 5 a un 10% del inicial. El 58,9% hizo cicatriz, de éstos el 63% fue estética. La satisfacción por parte de los pacientes es alta a pesar de la persistencia pigmentaria y/o la presencia de cicatriz. Discusión El tratamiento de nevos melanocíticos adquiridos con láser es una opción terapéutica que genera cambios estadísticamente significativos en cuanto a pigmento, cicatriz estética y alta satisfacción por parte de los pacientes. Se requieren estudios, analíticos, para determinar eficacia del tratamiento.

Role of ROC1 protein in the control of cyclin D1 protein expression in skin melanomas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Indicators for the total number of melanocytic naevi : an adjunct for screening campaigns. Observational study on 292 patients.

BACKGROUND: The presence of multiple melanocytic naevi is a strong risk factor for melanoma. Use of the whole body naevus count to identify at-risk patients is impractical. OBJECTIVES: To (i) identify a valid anatomical predictor of total naevus count; (ii) determine the number of naevi that most accurately predict total naevus count above 25, 50 and 100; and (iii) evaluate determinants of multiple melanocytic naevi and atypical naevi. METHODS: Clinical data from 292 consecutive Spanish patients consulting for skin lesions requiring debriding were collected throughout 2009 and 2010. Correlations between site-specific and whole body naevus counts were analysed. Cut-offs to predict total naevus counts were determined using the area under the receiver operating characteristic curve. RESULTS: The studied population was young (median age 31 years, interquartile range 28-43). The naevus count on the right arm correlated best with the total nevus count (R(2) 0·80 for men, 0·86 for women). Presence of at least five naevi on the right arm was the strongest determinant of a total naevus count above 50 [odds ratio (OR) 34·4, 95% confidence interval (CI) 13·9-85·0] and of having at least one atypical naevus (OR 5·7, 95% CI 2·4-13·5). Cut-off values of 6, 8 and 11 naevi on the right arm best predicted total naevus count above 25, 50 and 100, respectively. CONCLUSIONS: Our results support the arm as a practical and reliable site to estimate the total naevus count when screening or phenotyping large populations. Threshold values for the number of naevi on the arm are proposed to help identify patients for melanoma screening.

Epidermal Growth Factor Gene (EGF) Polymorphism and Risk of Melanocytic Neoplasiay

A common single nucleotide polymorphism (SNP) in the 5' untranslated region (5'UTR) of the epidermal growth factor (EGF) gene modulates the level of transcription of this gene and hence is associated with serum levels of EGF. This variant may be associated with melanoma risk, but conflicting findings have been reported. An Australian melanoma case-control sample was typed for the EGF+61A>G transversion (rs4444903). The sample comprised 753 melanoma cases from 738 families stratified by family history of melanoma and 2387 controls from 645 unselected twin families. Ancestry of the cases and controls was recorded, and the twins had undergone skin examination to assess total body nevus count, degree of freckling and pigmentation phenotype. SNP genotyping was carried out via primer extension followed by matrix-assisted laser desorption time of flight (MALDI-TOF) mass spectroscopy. The EGIF+61 SNP was not found to be significantly associated with melanoma status or with development of nevi or freckles. Among melanoma cases, however, G homozygotes had thicker tumors (p=0.05), in keeping with two previous studies. The EGF polymorphism does not appear to predispose to melanoma or nevus development, but its significant association with tumor thickness implies that it may be a useful marker of prognosis.

Melanoma mouse model implicates metabotropic glutamate signaling in melanocytic neoplasia

To gain insight into melanoma pathogenesis, we characterized an insertional mouse mutant, TG3, that is predisposed to develop multiple melanomas. Physical mapping identified multiple tandem insertions of the transgene into intron 3 of Grm1 (encoding metabotropic glutamate receptor 1) with concomitant deletion of 70 kb of intronic sequence. To assess whether this insertional mutagenesis event results in alteration of transcriptional regulation, we analyzed Grm1 and two flanking genes for aberrant expression in melanomas from TG3 mice. We observed aberrant expression of only Grm1. Although we did not detect its expression in normal mouse melanocytes, Grm1 was ectopically expressed in the melanomas from TG3 mice. To confirm the involvement of Grm1 in melanocytic neoplasia, we created an additional transgenic line with Grm1 expression driven by the dopachrome tautomerase promoter. Similar to the original TG3, the Tg(Grm1)EPv line was susceptible to melanoma. In contrast to human melanoma, these transgenic mice had a generalized hyperproliferation of melanocytes with limited transformation to fully malignant metastasis. We detected expression of GRM1 in a number of human melanoma biopsies and cell lines but not in benign nevi and melanocytes. This study provides compelling evidence for the importance of metabotropic glutamate signaling in melanocytic neoplasia.

High frequency of BRAF mutations in nevi

To evaluate the timing of mutations in BRAF (v-raf murine sarcoma viral oncogene homolog B1) during melanocytic neoplasia, we carried out mutation analysis on microdissected melanoma and nevi samples. We observed mutations resulting in the V599E amino-acid substitution in 41 of 60 (68%) melanoma metastases, 4 of 5 (80%) primary melanomas and, unexpectedly, in 63 of 77 (82%) nevi. These data suggest that mutational activation of the RAS/RAF/MAPK pathway in nevi is a critical step in the initiation of melanocytic neoplasia but alone is insufficient for melanoma tumorigenesis.

A meta-analysis of pigmentary characteristics, sun sensitivity, freckling and melanocytic nevi and risk of basal cell carcinoma of the skin

Objective: To calculate pooled risk estimates of the association between pigmentary characteristics and basal cell carcinoma (BCC) of the skin. Methods: We searched three electronic databases and reviewed the reference lists of the retrieved articles until July 2012 to identify eligible epidemiologic studies. Eligible studies were those published in between 1965 and July 2012 that permitted quantitative assessment of the association between histologically-confirmed BCC and any of the following characteristics: hair colour, eye colour, skin colour, skin phototype, tanning and burning ability, and presence of freckling or melanocytic nevi. We included 29 studies from 2236 initially identified. We calculated summary odds ratios (ORs) using weighted averages of the log OR, using random effects models. Results: We found strongest associations with red hair (OR 2.02; 95% CI: 1.68, 2.44), fair skin colour (OR 2.11; 95% CI: 1.56, 2.86), and having skin that burns and never tans (OR 2.03; 95% CI: 1.73, 2.38). All other factors had weaker but positive associations with BCC, with the exception of freckling of the face in adulthood which showed no association. Conclusions: Although most studies report risk estimates that are in the same direction, there is significant heterogeneity in the size of the estimates. The associations were quite modest and remarkably similar, with ORs between about 1.5 and 2.5 for the highest risk level for each factor. Given the public health impact of BCC, this meta-analysis will make a valuable contribution to our understanding of BCC.

Digital Photography in the Diagnosis and Follow-up of Ocular Diseases

Purpose: The aim of the present study was to develop and test new digital imaging equipment and methods for diagnosis and follow-up of ocular diseases. Methods: The whole material comprised 398 subjects (469 examined eyes), including 241 patients with melanocytic choroidal tumours, 56 patients with melanocytic iris tumours, 42 patients with diabetes, a 52-year old patient with chronic phase of VKH disease, a 30-year old patient with an old blunt eye injury, and 57 normal healthy subjects. Digital 50° (Topcon TRC 50 IA) and 45° (Canon CR6-45NM) fundus cameras, a new handheld digital colour videocamera for eye examinations (MediTell), a new subtraction method using the Topcon Image Net Program (Topcon corporation, Tokyo, Japan), a new method for digital IRT imaging of the iris we developed, and Zeiss photoslitlamp with a digital camera body were used for digital imaging. Results: Digital 50° red-free imaging had a sensitivity of 97.7% and two-field 45° and 50° colour imaging a sensitivity of 88.9-94%. The specificity of the digital 45°-50° imaging modalities was 98.9-100% versus the reference standard and ungradeable images that were 1.2-1.6%. By using the handheld digital colour video camera only, the optic disc and central fundus located inside 20° from the fovea could be recorded with a sensitivity of 6.9% for detection of at least mild NPDR when compared with the reference standard. Comparative use of digital colour, red-free, and red light imaging showed 85.7% sensitivity, 99% specificity, and 98.2 % exact agreement versus the reference standard in differentiation of small choroidal melanoma from pseudomelanoma. The new subtraction method showed growth in four of 94 melanocytic tumours (4.3%) during a mean ±SD follow-up of 23 ± 11 months. The new digital IRT imaging of the iris showed the sphincter muscle and radial contraction folds of Schwalbe in the pupillary zone and radial structural folds of Schwalbe and circular contraction furrows in the ciliary zone of the iris. The 52-year-old patient with a chronic phase of VKH disease showed extensive atrophy and occasional pigment clumps in the iris stroma, detachment of the ciliary body with severe ocular hypotony, and shallow retinal detachment of the posterior pole in both eyes. Infrared transillumination imaging and fluorescein angiographic findings of the iris showed that IR translucence (p=0.53), complete masking of fluorescence (p=0.69), presence of disorganized vessels (p=0.32), and fluorescein leakage (p=1.0) at the site of the lesion did not differentiate an iris nevus from a melanoma. Conclusions: Digital 50° red-free and two-field 50° or 45° colour imaging were suitable for DR screening, whereas the handheld digital video camera did not fulfill the needs of DR screening. Comparative use of digital colour, red-free and red light imaging was a suitable method in the differentiation of small choroidal melanoma from different pseudomelanomas. The subtraction method may reveal early growth of the melanocytic choroidal tumours. Digital IRT imaging may be used to study changes of the stroma and posterior surface of the iris in various diseases of the uvea. It contributed to the revealment of iris atrophy and serous detachment of the ciliary body with ocular hypotony together with the shallow retinal detachment of the posterior pole as new findings of the chronic phase of VKH disease. Infrared translucence and angiographic findings are useful in differential diagnosis of melanocytic iris tumours, but they cannot be used to determine if the lesion is benign or malignant.

Monozygotic twins discordant for phacomatosis pigmentovascularis: evidence for the concept of twin spotting.

The term phacomatosis pigmentovascularis (PPV) refers to the occurrence of vascular nevi with melanocytic or epidermal nevi. We report on monozygotic twins (MZTs) discordant for phacomatosis cesioflammea (PPV type II) providing evidence for the mechanism of twin spotting in the development of PPV. The affected twin had a nevus flammeus on the right arm and the right maxilla, and a pigmented area on the trunk in keeping with a persistent, aberrant Mongolian spot. The affected twin had bilateral ocular melanocytosis with abnormal scleral pigmentation, iris mamillations, increased pigmentation of the trabecular meshwork, and increased fundal pigmentation and secondary glaucoma. DNA testing confirmed monozygosity. This case of MZTs discordant for PPV supports the hypothesis that PPV results from mosaicism due to a post-zygotic mutational event and the concept of twin spotting.

Mfge8 Expression In Conjunctival Melanocytic Proliferations

Purpose: Milk fat globule epidermal growth factor-8 (MFGE8) is a secreted phosphatidylserine-binding protein that has been involved in phagocytosis, as well as in VEGF dependent neovascularization. In a study evaluating protein expression in membrane rafts of cutaneous melanoma at different stages of progression, MFGE8 expression was only identified in membrane rafts of metastatic cutaneous melanoma cell lines. Furthermore, MFGE8, identified at higher level in the vertical growth phase of cutaneous melanoma, promoted tumor growth in vivo, enhanced invasion in vitro and metastatic spread in a mouse model. The purpose of this study was to assess the expression of MFGE8 in conjunctival melanocytic proliferations.Methods: MFGE8 expression was assessed by immunohistochemistry in 66 melanocytic conjunctival proliferations including 21 conjunctival naevi, 20 Primary Acquired Melanosis (PAM) including (4 PAM without atypia and 16 PAM with atypia) and 25 conjunctival melanomas. Expression was independently assessed by 2 pathologists. Relevant clinico-pathological data were retrieved. Statistical anaylis was performed using JUMP 8 software.Results: The concordance between the 2 pathologists had an 87,5% agreement on the first independent assessment of MFGE8 expression. Complete agreement was further reached after joint revision of discordant cases. In the naevi, MFGE8 expression was found in only 4 cases (3 subepithelial cases and 1 composed combined naevus). In the PAM group, MFGE8 was identified in 1 PAM without atypia and 10 PAM with atypia. In the melanoma group, MFGE8 expression was observed in 68% of cases. The expression of MFGE8 in the conjunctival melanocytic proliferation was significantly higher in the melanoma (p=0,0009) and in the PAM (p=0,0169) than in naevi. Within the PAM subgroup, we found no significant correlation between MFGE8 expression and the presence of atypia in the respective specimen examined so far.Conclusions: We demonstrate a significant higher expression of MFGE8 in conjunctival melanoma compared to benign melanocytic lesions, suggesting that this protein may play a role in tumor progression of conjunctival melanocytic proliferations. Further experimental studies should be performed to better characterize MFGE8 involvement in conjunctival melanoma tumorigenesis.

Comparación: injerto libre o cierre primario más lente de contacto, en resección de tumores de conjuntiva limbar

INTRODUCCION: Existe controversia en cuanto a la técnica quirúrgica para el manejo de tumores del limbo conjuntival. El uso de cierre primario con uso de lente de contacto puede ofrecer una mejor cicatrización y tener ventajas adicionales sobre la técnica tradicional con el uso de plastia. OBJETIVOS: Comparar los resultados en cuanto a grado de dolor, picadas, prurito, porcentaje de epitelización y cicatrización, comodidad del paciente, grado de quemosis y tiempo de retorno a actividades diarias en ambas técnicas quirúrgicas. MATERIALES Y METODOS: Experimento clínico controlado aleatorizado en dos grupos: Al primer grupo se le realizó cirugía de resección de la lesión más plastia. Al segundo grupo se le practicó la resección de la lesión cierre primario y lente de contacto. El seguimiento se realizó al primer y cuarto día, y cada semana durante el primer mes de postoperatorio. Se utilizó el SPSS 20.0 ® para análisis estadístico de datos y se utilizó estadística no paramétrica. RESULTADOS: Se conto con 10 pacientes por grupo. El dolor y porcentaje de cicatrización al primer día postoperatorio fueron mayores en el grupo usando lente de contacto (p=0.048). Al cuarto día postquirúrgico se encontró un mayor porcentaje de cicatrización en el grupo usando lente de contacto. (p=0.075). CONCLUSIONES: El cierre por afrontamiento con uso de lente de contacto mostró dolor y picadas mayores al primer y cuarto día postoperatorio. Pero la epitelización y cicatrización fueron tempranas con un retorno corto a actividades cotidianas.

Growth of melanocytic cells is associated with down-regulation of protein kinase C α,δ and ε isoforms: Possible role of diacylglycerol

Protein kinase C (PKC) down-regulation has been shown to correlate with the growth of murine melanocytic cells in culture (Brooks, G., Wilson, R. E., Dooley, T. P., Goss, M. W., and Hart, I. R. (1991) Cancer Res. 51, 3281-3288). We now show that PKC alpha, delta, epsilon, and zeta isoforms are present at the protein level in quiescent, non-transformed Mel-ab melanocytes, maintained in the absence of phorbol ester. Proliferation of Mel-ab cells, achieved by incubation in the continual presence of phorbol 12,13-dibutyrate, was associated with a down-regulation of the PKC alpha, delta, and epsilon isozymes. Examination of two transformed syngeneic lines (the B16 murine melanoma and the long terminal repeat Ras.2 line), that grew in the absence of exogenous phorbol esters, showed that PKC alpha protein levels were either partially down-regulated or unaffected, the PKC delta and epsilon isoforms were down-regulated completely, and the levels of PKC zeta protein remained unaltered relative to quiescent Mel-ab cells. Basal levels of total diacylglycerol were elevated 5-fold in B16 melanoma cells compared with levels found in quiescent or proliferating Mel-ab melanocytes and appear to arise largely from the breakdown of phosphatidylinositol phospholipids accompanied by a significant rise in phospholipase C activity. Hourly treatments of quiescent Mel-ab melanocytes with the synthetic diacylglycerol analogue, 1,2-dioctanoyl-sn-glycerol, for 24 h, resulted in an induction of DNA synthesis which was associated with a significant down-regulation of PKC levels mediated largely via post-translational rather than transcriptional mechanisms. These results show for the first time that specific isoforms of PKC are down-regulated at the protein level during proliferation of murine melanocytic cells and suggest that the constitutive down-regulation of PKC in transformed melanoma cells may arise as a consequence of elevated endogenous phosphatidylinositol-derived diacylglycerol levels.

Effects of biologically active tumour-promoting and non-promoting phorbol esters on in vivo growth of melanocytic cells

Sapintoxin A (SAP A), a naturally occurring biologically active but non-promoting phorbol ester, acts as an effective in vitro mitogen for freshly derived human melanocytes. Seven days after addition of 50 nM SAP A there was a four to fivefold increase in melanocyte number over that observed in untreated control cultures comparable to that achieved with a 50 nM concentration of 12-0-tetradecanoylphorbol 13-acetate (TPA). The fluorescent stage 2 promoter sapintoxin D (SAP D) also supported the growth of these cells, with a 50 nM dose producing an increase in cell number comparable to that observed with 200 nM TPA. Similar results were obtained with an established, but non-tumorigenic, line of murine melanocytes. The same compounds exerted a potent anti-proliferative effect against transformed melanocyte lines of murine and human origin associated with morphological alterations and an increase in melanin production consistent with induced cytodifferentiation.