Confocal Raman Microscopy (CRM) is used to study the cell internalization of poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) fabricated by emulsion techniques with either poly(ethylene imine) (PEI) or bovine serum albumin (BSA) as surface stabilizers. HepG2 cells were exposed to PEI and BSA stabilized PLGA NPs. Spontaneous Confocal Raman Spectra taken in one and the same spot of exposed cells showed bands arising from the cellular environment as well as bands characteristic for PLGA, proving that the PLGA NPs have been internalized. It was found that PLGA NPs preferentially colocalize with lipid bodies. The results from Raman spectroscopy are compared with flow cytometry and confocal scanning laser microscopy (CLSM) data. The advantages of CRM as a label-free technique over flow cytometry and CLSM are discussed. Additionally, cell viability studies by means of quick cell counting solution and MTT tests in several cell lines show a generally low toxicity for both PEI and BSA stabilized PLGA NPs, with BSA stabilized PLGA NPs having an even lower toxicity than PEI stabilized.

The frequency of nevus-associated melanoma (NAM) is widely variable among different reports. Recently, it has been estimated to be around 29% of all diagnosed melanomas. 1,2 The aim of this observational retrospective study was to analyze dermoscopic, reflectance confocal microscopy (RCM), and histologic features of a case series of 22 NAM. 2 | MATERIAL AND ME THODS We retrospectively reviewed a database recording the melanomas diagnosed in the Pathology Unit of Alcorcon Foundation Hospital between September 2011 and 2018, obtaining a total of 240 melanomas. In 45 of them (18.75%), an associated nevus was found, and we selected for the study 22 cases with good quality dermoscopic

Background The management of melanocytic lesions with peripheral globules (MLPGs) is usually age-dependent and can be challenging in high-risk melanoma patients.Objectives To evaluate clinical, dermoscopic and reflectance confocal microscopy (RCM) features of MLPG in patients under digital dermoscopic surveillance. To know whether dermoscopic or RCM findings correlate with histologic diagnosis and the accuracy of the dermoscopy-RCM compared with histopathology.Methods During 24 months, we prospectively enrolled MLPG in patients under digital dermoscopy follow-up. All were evaluated by dermoscopy and RCM and excised for histologic examination. Results We enrolled 154 patients, mean age 42.45 years (18.78-73.19). Three melanomas and 19 dysplastic naevi (DNs) were diagnosed. There were no significant differences in the age of the patients (P = 0.662). MLPGs with diameter of 6 mm or more and asymmetry in two axes were associated with melanoma (P = 0.01, P = 0.003). Patients with more than one MLPG were less likely to have melanoma. Blue-grey and red colours were more frequent in melanoma (P = 0.013 and P = 0.000). Different sizes and shapes of PG were associated with DN and melanoma (P = 0.000 and P = 0.001). In a new lesion, PG in <25% of the circumference was related to malignancy (P = 0.010). RCM signs of malignancy were related to melanoma: pagetoid cells (P = 0.000), non-edged papillae (P = 0.001), atypical junctional thickenings (P = 0.000) and atypical cells at the dermal-epidermal junction (P = 0.000). Dense irregular nests were associated to melanoma (P = 0.019). Dermoscopy and confocal evaluation were able to diagnose 100% of melanomas and 84.21% of DNs. The kappa coefficient between dermoscopy-RCM vs. histology was 0.76. ConclusionsWe recommend to excise a MLPG when it presents asymmetry in two axes, 6 or more mm, new lesion with PG in less than the 25% of the circumference, irregular size and shape PGs and irregular dense nests on RCM, regardless of the patient's age.

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Digital dermoscopy (DD) follow up is a useful strategy for monitoring patients at high risk of melanoma. Reflectance confocal microscopy (RCM) is a valuable second-level examination after dermoscopy for the evaluation of difficult to diagnose lesions. The aim of this study was to assess the value of RCM in routine DD monitoring of patients at high risk of melanoma. In this prospective study, performed over 22 months, changing melanocytic lesions were detected by DD and excised. RCM imaging was performed before surgical excision of all the lesions, and histopathology used as the gold standard diagnostic test. Eighty-seven lesions, including 11 thin melanomas, were studied. Dermoscopic evaluation at follow up revealed a significant association between melanoma and asymmetry in two axes (P = 0.035). Enlargement and other changes in structure or color did not significantly differ between nevi and melanomas. Widespread pagetoid cells, non-edged papillae, irregular and dishomogeneous junctional clusters, and sheet-like structures were significantly associated with malignancy (P < 0.001). RCM allowed accurate diagnosis of melanoma in 10 of 11 cases (90.9%). The remaining case was classified as a dysplastic nevus. Forty-six lesions (52.8%) in which RCM revealed no atypia were deemed unnecessarily removed. This study was limited by sample size. In conclusion, combined DD and RCM facilitates the recognition of thin malignant melanomas and reduces unnecessary excisions.

The diversity and utility of laser procedures have increased over the recent years and nowadays, applications for medical and cosmetic reasons have increased considerably. Problematic intraoral and cutaneous hirsutisms have been described as a consequence of complex reconstruction usually after oncology surgery. We present three patients in whom hair removal laser was performed on grafts and flaps in different compromised anatomical areas: oral cavity, penis, and auricular pavilion. All three patients were men; in two of them the hairy graft was a consequence after oncologic surgery reconstruction whereas the third patient presented hair in his auricular pavilion after cochlear implant due to a congenital ear malformation. In all the patients, neodymium:yttrium, aluminum, garnet laser (Nd:YAG) (1,064 nm) laser was performed with excellent aesthetic and functional outcomes with only three sessions. Hair removal laser is a well‐accepted and effective method of achieving permanent decrease in hair density. Several lasers have been used successfully, including the long‐pulse Alexandrite (755 nm), the long‐pulse diode (810 nm), and the Nd:YAG laser (1,064 nm). There is currently no standard protocol for laser use on hairy grafts or flaps and there is limited published data regarding skin graft revision to enhance aesthetics and function.