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Francesco Spagnolo,1 Paola Ghiorzo,2,3 Laura Orgiano,4 Lorenza Pastorino,2,3 Virginia Picasso,4 Elena Tornari,4 Vincenzo Ottaviano,4 Paola Queirolo4 1Department of Plastic and Reconstructive Surgery, IRCCS Azienda Ospedaliera Universitaria San Martino, IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy; 2Department of Internal Medicine and Medical Specialties (DiMI), University of Genoa, Genova, Italy; 3Genetics of Rare Cancers, IRCCS Azienda Ospedaliera Universitaria San Martino, IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy; 4Department of Medical Oncology, IRCCS Azienda Ospedaliera Universitaria San Martino, IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy Abstract: BRAF inhibitors vemurafenib and dabrafenib achieved improved overall survival over chemotherapy and have been approved for the treatment of BRAF-mutated metastatic melanoma. More recently, the combination of BRAF inhibitor dabrafenib with MEK inhibitor trametinib has shown improved progression-free survival, compared to dabrafenib monotherapy, in a Phase II study and has received approval by the US Food and Drug Administration. However, even when treated with the combination, most patients develop mechanisms of acquired resistance, and some of them do not achieve tumor regression at all, because of intrinsic resistance to therapy. Along with the development of BRAF inhibitors, immunotherapy made an important step forward: ipilimumab, an anti-CTLA-4 monoclonal antibody, was approved for the treatment of metastatic melanoma; anti-PD-1 agents achieved promising results in Phase I/II trials, and data from Phase III studies will be ready soon. The availability of such drugs, which are effective regardless of BRAF status, has made the therapeutic approach more complex, as first-line treatment with BRAF inhibitors may not be the best choice for all BRAF-mutated patients. The aim of this paper is to review the systemic therapeutic options available today for patients affected by BRAF V600-mutated metastatic melanoma, as well as to summarize the mechanisms of resistance to BRAF inhibitors and discuss the possible strategies to overcome them. Moreover, since the molecular analysis of tumor specimens is now a pivotal and decisional factor in the treatment strategy of metastatic melanoma patients, the advances in the molecular detection techniques for the BRAF V600 mutation will be reported. Keywords: melanoma, BRAF, vemurafenib, dabrafenib, resistance, BRAF inhibitor

Background: The Italian Society of Human Genetics'(SIGU) recommendations on genetic counseling and testing for hereditary melanoma state that clinical genetic testing can be offered to Italian melanoma families with at least two affected members. Objective: In the framework of a cooperative study, we sought to establish the frequency of cyclin-dependent kinase inhibitor 2,4 mutations in melanoma families that underwent clinical genetic Counseling and testing in accordance with die SIGU recommendations at 9 centers in different Italian regions. Methods: inhibitor 2A testing was conducted by direct sequencing and multiplex ligation-dependent probe amplification analysis in melanoma families with at least two affected members. Results: A total of 33% (68/204) of the families harbored cyclin-dependent kinase inhibitor 2A Mutations. In the 145 families with two affected members the mutation frequency was 25%. Three novel mutations, L94P, A86T, and c.407dupG, were identified among the cases and not in 200 controls. Limitations: We were unable to perform separate analyses for individual centers, as in some cases the number of families was too small. Conclusions: The availability of clinical genetic testing for melanoma to families with just two affected members in the same branch is justified in Italy in terms of the likelihood of identifying a mutation. (J Am Acad Dermatol 20001:775-82.)

© 2016, Springer Science+Business Media New York.The delivery and controlled release of drugs in the human body is one of the main research fields of nanotechnology applied to medicine. An ideal drug carrier should be able to deliver drug molecules to the site of action and to interact specifically with target cells. In this perspective, different organic and inorganic nanosystems have been proposed and tested. One of the most interesting techniques for the synthesis of drug carriers is represented by the layer-by-layer self-assembly (LbL) of nanoengineered shells onto sacrificial templates. However, this technique has been mostly applied for the fabrication of hollow microcapsules, while targeting single cells would require the fabrication of nanocapsules. LbL-based nanocapsules have been proposed in the literature using inorganic nanoparticles or polymeric ones as sacrificial templates, requiring the use of organic solvents for their removal. In the view of a perspective application of such nanocapsules, the use of biocompatible and biodegradable reagents plays a pivotal role. In this respect, recently, the possibility of synthesizing in a highly controlled way calcium carbonate nanoparticles has opened new perspectives for such kind of carrier systems.

Aim: To determine the epithelial expression of ß-catenin and survivin in sporadic (primary, and recurrent) and nevoid basal cell carcinoma syndrome (NBCCS) keratocystic odontogenic tumour (KCOT) in order to assess activation of the ß-catenin pathway and evidence of apoptotic inhibition, processes that may contribute to the known differences in their biological behaviour. Materials and Methods: Sections from 40 cases of KCOT (19 sporadic/primary; 9 sporadic/recurrent and 12 NBCCS-associated) were immunohistochemically stained for ß-catenin and survivin. The extent and intensity of immunoreactivity within the lining epithelium was assessed, using semi-quantitative scales, independently by two pathologists who were blinded to the clinical-pathological data. Data were analysed using Kruskal-Wallis test and, for pair-wise comparisons, Mann-Whitney test with Bonferroni correction. Results: All cystic epithelial linings stained for ß- catenin and survivin but there were differences in the pattern and intensity of staining among KCOT types. Sporadic primary KCOT showed weaker staining for ß- catenin (P=0.0003) and survivin (P

© 2015 AIP Publishing LLC. Nanoengineered polymeric capsules are currently widely used in the drug design sector. In this paper, we present data, allowing to consider this objects as important elements of bio-computers, allowing, in particular, targeted delivery and smart release of the "main program" (special molecular preparations) to zones, where the adequate functioning of the system can be affected by undesirable events.

The MC1R gene is a key regulator of skin pigmentation. We aimed to evaluate the association between MC1R variants and the risk of sporadic cutaneous melanoma (CM) within the M-SKIP project, an international pooled-analysis on MC1R, skin cancer and phenotypic characteristics. Data included 5,160 cases and 12,119 controls from 17 studies. We calculated a summary odds ratio (SOR) for the association of each of the nine most studied MC1R variants and of variants combined with CM by using random-effects models. Stratified analysis by phenotypic characteristics were also performed. Melanoma risk increased with presence of any of the main MC1R variants: the SOR for each variant ranged from 1.47 (95%CI: 1.17-1.84) for V60L to 2.74 (1.53-4.89) for D84E. Carriers of any MC1R variant had a 66% higher risk of developing melanoma compared with wildtype subjects (SOR; 95%CI: 1.66; 1.41-1.96) and the risk attributable to MC1R variants was 28%. When taking into account phenotypic characteristics, we found that MC1R-associated melanoma risk increased only for darker-pigmented Caucasians: SOR (95%CI) was 3.14 (2.06-4.80) for subjects with no freckles, no red hair and skin Type III/IV. Our study documents the important role of all the main MC1R variants in sporadic CM and suggests that they have a direct effect on melanoma risk, independently on the phenotypic characteristics of carriers. This is of particular importance for assessing preventive strategies, which may be directed to darker-pigmented Caucasians with MC1R variants as well as to lightly pigmented, fairskinned subjects. © 2014 UICC.

Introduzione: Le differenze geografiche nell'incidenza del melanoma e nella penetranza delle mutazioni di CDKN2A, principale gene di suscettibilità al melanoma, sono tali che non esistono criteri per indirizzare un paziente al test genetico applicabili in modo uniforme a livello internazionale. Leachman et al. hanno proposto che due eventi oncologici (melanoma e/o adenocarcinoma pancreatico, associato anch'esso a mutazioni di CDKN2A) in un singolo paziente o in un famiglia siano considerati criterio sufficiente per il test genetico nelle aree a bassa incidenza di melanoma(1). L'Italia è inclusa fra queste aree sulla base dei dati liguri in cui mutazioni da effetto fondatore sono state riscontrate anche nel 40% dei casi familiari di melanoma. Lo studio basato sulle raccomandazioni SIGU per l'ammissione al test per il melanoma familiare ha riscontrato mutazioni di CDKN2A nel 25% dei casi familiari con due soli affetti (2), che aumentavano in presenza di casi di melanoma multiplo (MPM). Altri geni di predisposizione al melanoma sono stati identificati di recente ma la penetranza delle nuove mutazioni non è ancora stata chiarita. Scopo principale del nostro studio è stato di valutare su base nazionale la prevalenza di mutazione in CDKN2A, CDK4 e MITF in casi di MPM per stabilire se anche in assenza di familiarità lo sviluppo di MPM possa essere considerato criterio sufficiente di avvio al test genetico. Materiali e metodi: 587 casi di melanoma multiplo e un numero uguale di casi di melanoma singolo e controlli sono stati consecutivamente reclutati nei centri partecipanti e analizzati per CDKN2A, CDK4 e MITF. Risultati Mutazioni di CDKN2A sono state riscontrate nel 19% dei casi di melanoma multiplo e nel 4.4% dei casi di melanoma singolo, compresi melanomi in situ. Nei casi multipli familiari il rate di mutazione varia dal 36.6% al 58.8% mentre nei casi sporadici dall'8.2% al 17.6% nei casi con due o con 3 o più melanomi, rispettivamente. La mutazione MITF E318K è stata riscontrata nel 3% dei casi di melanoma multiplo. Conclusioni I pazienti italiani con due melanomi, anche in situ, e senza familiarità dovrebbero essere indirizzati alla valutazione genetica. Bibliografia 1.Leachman, S. A. et al. Selection criteria for genetic assessment of patients with familial melanoma. Journal of the American Academy of Dermatology 61, (2009) 2.Bruno, W. et al. Clinical genetic testing for familial melanoma in Italy: A cooperative study. J. Am. Acad. Dermatol. 61, 775-782 (2009)

© 2015 IEEE. The paper reports the temporal dependence of the pore formation in polymeric capsules. The results are important for the drug design and programming of biochemical computers.

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Chitosan has been used for drug delivery applications for many years, and pectin has been recently used for several biomedical applications too. Both chitosan and pectin have shown to possess mucoadhesive properties. We report the assembly and characterization of biocompatible and biodegradable multilayers based on the layer-by-layer (LbL) deposition of chitosan and pectin. The dependence of the multilayer properties on the degree of esterification of pectin, as well as on the assembly pH, is characterized, and the mucoadhesiveness of the multilayers is estimated at oral and gastric pHs. The LbL assembly of chitosan and pectin for the fabrication of hollow capsules is carried out for the first time, and its potential use for drug delivery applications opens new prospects for the production of novel drug delivery materials. Chitosan/pectin multilayers and nanostructured capsules are developed by the layer-by-layer electrostatic self-assembly technique. The influence of the pH assembly and of the pectin degree of charge on the multilayer properties is characterized, as well as the interaction with the mucoadhesive protein mucin. The biocompatible, biodegradable, and mucoadhesive capsules are stable and can be regarded as novel drug delivery materials.

The main aim of TASNANO project is to focus on the development of a web-based system that allows cooperative work between partners. Even if some commercial tools become available, the specificities of nanotechnology applications suggest implementing an ad-hoc tool. This approach permits to update information every moment, so that not only the latest versions of the documents are always available, but also the raw experimental data are shared in a protected environment. Organizing big amount of data begins with modelling of conceptual schemes, which individualizes entities involved into the project and links between them. The rules’ application of logic modeling leads to the production of a logic scheme. The model allows the data base design, which can be enlarged and enriched preserving coherence and avoiding redundancy. Data and information can be directly treated on the web in accordance with latest modifications. [Article copies are available for purchase from InfoSci-on-Demand.com]

Electrospray mass spectrometry coupled to liquid chromatography was utilized to measure two PKC neuronal substrates, B-50/GAP-43 and neurogranin, in single rat brain areas. Aliquots of perchloric acid extracts were directly injected and mass spectra recorded. At elution times of 14.2 and 27.0 min two molecular species of MW 7450 and 23 602 Da were observed. These values are in excellent agreement for the expected MW for rat neurogranin and B-50/GAP-43. The presence of molecular species shifted by 80 mass units in both cases indicates that these proteins are present in phosphorylated forms in cortical and hippocampal extracts.