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BackgroundWe previously reported rare regressive genetic trajectories of KRAS pathogenic mutations as a specific hallmark of the genuine oligometastatic status in colorectal cancer (CRC).MethodsSurvival and prognostic impact of disease extent in 140 metastatic CRC patients were evaluated through the Kaplan–Meyer curves and the Log-Rank test. KRAS mutations were assessed through the Illumina NovaSeq 6000 platform and TruSight™ Oncology 500 kit. HLA typing was carried out by PCR with sequence-specific oligonucleotides. Lymphocyte densities in tumors were expressed as cells per square millimeter. NKs isolated and CD8+ from NK-depleted PBMCs were characterized through flow cytometry. CD107a externalization was evaluated as NKs/CD8 cytotoxicity toward human colon cancer cells HT29, SW620, HCT116, and LS174T carrying different KRAS mutations.ResultsThe oligometastatic status was a strong and independent variable for survival (HR: 0.08 vs. polymetastatic disease; 95% CI: 0.02–0.26; p KRAS was observed in 3 patients: patient (PAT)2, PAT5, and PAT8. PAT2 and PAT5 presented the highest levels of GrzB+ lymphocytes in the tumor cores of the metastases (120 ± 11.2 and 132 ± 12.2 cells/mm2, respectively). Six out of 9 patients (67%), including PAT2 and PAT5, expressed HLA-C7. Twopatients (PAT2 and PAT5) presented high CD3+/CD8+-dependent cytotoxicity against HLA-C7+ SW620 cells (p.G12V-mutated cells), which was consistent with their observed mutational regression (p.G12V/p.G13D in primary→p.G13D in metastatic tumor).ConclusionsWe provide evidence that CD3+/CD8+ lymphocytes from oligometastatic CRC patients display differential cytotoxicity against human colon cancer cells carrying KRAS mutations. This could provide an interesting basis for monitoring oligometastatic disease and developing future adoptive immunotherapies.

L���obiettivo del lavoro �� proporre una precisa categorizzazione dei molteplici strumenti utilizzabili, nell���ambito di una strategia di heritage marketing, per gestire in termini manageriali la narrazione della storia dell���impresa e dei suoi prodotti e/o marchi. A tal fine, �� stata realizzata una multiple case study research su 20 imprese ultracentenarie italiane che si sono distinte per le attivit�� di valorizzazione del proprio patrimonio storico. Secondo un approccio qualitativo di tipo narrativo, ciascun caso �� stato costruito integrando fonti orali (interviste), testuali (materiale aziendale) e visive (visite e osservazione diretta). In base alla diversa natura della narrazione, viene proposta una declinazione dell���heritage marketing mix in quattro categorie di strumenti, il cui utilizzo integrato e contestuale pu�� offrire un contributo fondamentale alla valorizzazione dell���identit�� racchiusa nel patrimonio storico aziendale, con particolare riferimento alle imprese del settore agroalimentare. Il lavoro costituisce un ���follow up��� di un precedente contributo pubblicato su questa Rivista, e mira a correggere la prospettiva d���analisi che era stata allora utilizzata, alla luce degli sviluppi pi�� recenti di un percorso di ricerca che non si �� mai interrotto e che risulta tuttora incompiuto. A conoscenza degli autori, si tratta della pi�� dettagliata declinazione dell���heritage marketing mix presente in letteratura. The aim of the paper is to propose a precise categorization of the multiple tools that can be used, as part of a heritage marketing strategy, to manage the narration of the history of the company and its products and/or its brands according to a strategic perspective. To this aim, a multiple case study research has been carried out on 20 Italian companies that have distinguished themselves for the enhancement of their historical heritage. According to a qualitative narrative approach, each case was built by integrating oral (interviews), textual (company material) and visual (visits and direct observation) sources. Based on the different nature of the narrative used, a definition of the heritage marketing mix is proposed, consisting in four categories of tools, whose integrated and contextual use can offer a fundamental contribution to the enhancement of the identity embedded in the company���s historical heritage, with particular reference to companies operating in the food and beverage industries. This paper is a ���follow up��� of a previous contribution published in this Journal, and aims to correct the analysis perspective that was then used, in light of the most recent developments of a research path that has never been interrupted and which is still unfinished. To the best of our knowledge, this is the most detailed definition of the heritage marketing mix that it is possible to find in the literature., IL CAPITALE CULTURALE. Studies on the Value of Cultural Heritage, Supplementi (10/2020) Cibo e vino: rappresentazioni, identit�� culturali e co-creazione di sviluppo sostenibile / Food and Wine: representations, cultural identities and co-creation for sustainable development

Additional file 2: Figure S2. Mass spectra of the triply charged hACE2 tryptic peptide 342-357 in the glycated (panel A) and non-glycated (panel B) form. hACE2 is for human Angiotensin Converting enzyme 2 type.

La rivista Clio intitola il numero 46 del 2016 alle cittadinanze. L’indagine tematica è ripartita nelle sezioni Dossier, Regards complémentaires, Documents, Actualité de la recherche, Portrait, Varia. A partire dal mondo greco e romano, gli studi affrontano aspetti della questione ‘cittadinanza’ al femminile, declinati nella diversità delle prospettive e delle coordinate di tempo e spazio. Filo conduttore del volume, come nell’introduzione di Pascale Barthélémy e di Violaine Sebillotte Cuchet...

Table S1: Primer sequences for SYBR Green RT-qPCR. (PPTX 68 kb)

Table S3. Detailed characteristics of 28 VHL-WT-RCC patients. (PPTX 71 kb)

Table S2. Detailed characteristics of 23 VHL-MUT-RCC patients. (PPTX 71 kb)

Somatostatin analogues and mTOR inhibitors have been used as medical therapy in lung carcinoids with variable results. No data are available on dopamine agonists as treatment for lung carcinoids. The main aim of the current study was to evaluate the effect of the combined treatment of somatostatin analogue octreotide and the dopamine agonist cabergoline with mTOR inhibitors in an in vitro model of typical lung carcinoids: the NCI-H727 cell line. In NCI-H727 cell line, reverse transcriptase-quantitative polymerase chain reaction and immunofluorescence were assessed to characterize the expression of the somatostatin receptor 2 and 5, dopamine receptor 2 and mTOR pathway components. Fifteen typical lung carcinoids tissue samples have been used for somatostatin receptor 2, dopamine receptor 2, and the main mTOR pathway component p70S6K expression and localization by immunohistochemistry. Cell viability, fluorescence-activated cell sorting analysis and western blot have been assessed to test the pharmacological effects of octreotide, cabergoline and mTOR inhibitors, and to evaluate the activation of specific cell signaling pathways in NCI-H727 cell line. NCI-H727 cell line expressed somatostatin receptor 2, somatostatin receptor 5 and dopamine receptor 2 and all mTOR pathway components at messenger and protein levels. Somatostatin receptor 2, dopamine receptor 2, and p70S6K (non phosphorylated and phosphorylated) proteins were expressed in most typical lung carcinoids tissue samples. Octreotide and cabergoline did not reduce cell viability as single agents but, when combined with mTOR inhibitors, they potentiate mTOR inhibitors effect after long-term exposure, reducing Akt and ERK phosphorylation, mTOR escape mechanisms, and increasing the expression DNA-damage-inducible transcript 4, an mTOR suppressor. In conclusion, the single use of octreotide and cabergoline is not sufficient to block cell viability but the combined approach of these agents with mTOR inhibitors might reduce the mTOR inhibitors-induced escape mechanisms and/or activate the endogenous mTOR suppressor, potentiating the effect of the mTOR inhibitors in an in vitro model of typical lung carcinoids.

The paper aims to provide additional knowledge about the relationship between store environment and consumer behavior by investigating the influence of selected environmental variables (internal design and tenant-mix) on consumers' emotional states and shopping outcomes. The proposed framework is based on model proposed by Mehrabian and Russell (1974) and is tested through a survey in two shopping centres in the city of Benevento. Results show that the selected environmentalvariables play a significant role in influencing consumers' shopping outcomes and that such relationship is moderated by the different nature of emotional states. On the basis of results, authors discuss the contribution of the study to marketing literature and the implications for the management of retail and service firms.

Il presente lavoro intende offrire un contributo al dibattito sempre più attuale circa il ruolo della cultura quale risorsa distintiva per lo sviluppo economico e sociale del Paese. In particolare, partendo dall’impostazione teorica di Throsby sul capitale culturale, è approfondito il legame tra cultura e creatività nel Paese e l’immagine dello stesso nel panorama internazionale. A seguire, si evidenziano gli elementi di criticità che, nella gestione dell’ampio e unico patrimonio culturale italiano, hanno determinato una progressiva perdita di competitività del nostro Paese sia in termini di attrazione dei flussi turistici internazionali che di immagine-Paese. Infine, viene proposto un nuovo approccio alla gestione del patrimonio culturale che, ribaltando le logiche “classiche”, suggerisce l’adozione di logiche di gestione strategica e di marketing territoriale come strumenti per la valorizzazione del patrimonio artistico-culturale e per il territorio che lo ospita. The paper aims to contribute to the increasingly relevant debate about the role of culture as a distinctive resource for the economic and social development of a country. Specifically, adopting the theoretical framework of the cultural capital by Throsby, it has been investigated the relationship between culture and creativity in our country and its imagine in the international scenario. To follow, we highlight the critical elements that, considering the management of the wide and unique Italian cultural heritage, have determined the gradual decline in competitiveness of the country, both in terms of attractiveness of the international touristic flows and country-image. In conclusion, we propose a new approach to the management of the cultural heritage that, turning the classical approach over, suggests the adoption of methods of strategic management and territorial marketing as tools for the valorisation of the cultural heritage of a place., IL CAPITALE CULTURALE. Studies on the Value of Cultural Heritage, N° 11 (2015)

1. Introduzione. - 2. Strategie al plurale: l’importanza dell’implementazione. - 2.1. Progettazione delle strutture e comportamento organizzativo: una frontiera non eludibile. - 2.1.1. Le persone lungo la frontiera design/behaviour: il contributo degli studi di critical management. - 3. Comportamento organizzativo: personalità, posizione e ruolo. - 3.1. Personalità. - 3.2. Posizione. - 3.3. Ruolo. - 3.4. Interpretare posizione e ruolo come atto linguistico. - 4. Le competenze. - 4.1. Sul concetto di competenza tra teoria e pratica. - 4.1.1. Competenza per competere: il ruolo della capacità simbolica. - 4.1.2. Concezioni di competenza e vantaggio competitivo. - 5. Gestione Strategica del cambiamento organizzativo. - 5.1. Strategia e struttura. - 5.1.1. Allineamento strategia e struttura. - 5.2. Il difficile allineamento strategia/struttura: quando le persone fanno la differenza. - 6. Resistenze al cambiamento. - 6.1. Le potenzialità insite nelle resistenze: il ruolo della management education. - 6.1.1. Umanesimo del management e retorica del linguaggio manageriale. - 6.2. Cambiamento, spinte e resistenze: il ruolo della tecnologia. - 6.2.1. La portata strategica della tecnologia.

Con el propósito de indagar los discursos que circulan en la cultura del fitness he construido los siguientes ejes de trabajo: 1) discursos que atraviesan lo corporal y las prácticas de los clientes, 2) discursos que permiten comercializar productos, prácticas y lineamientos y 3) estrategias discursivas que se despliegan en función de ampliar el mercado del fitness y generar la adherencia de los usuarios a los productos que esta cultura promueve., Facultad de Humanidades y Ciencias de la Educación

This paper is aimed to discuss the actual commitment of Italian universities towards their third mission of economic and social development, providing an innovative perspective of analysis. We apply Critical Discourse Analysis to assess how Italian universities engagement with the stakeholder community is interpreted, implemented, communicated and disseminated within and outside the academic organizations. One of the most significant points in the evolution of organizational studies is the growing interest in language, whose construction is a key, alternative to traditional, to understand the organizational and inter-organizational phenomena. Critical Discourse Analysis is based on the assumption that language is not neutral with respect to organizational and inter-organizational dynamics, but it contributes to their production. In management studies, discourse analysis is a new heuristic strategy for those who are interested in the processes of social construction of organizational phenomena that have as their central element the text. Critical Discourse Analysis tries to unite, and determines the relationship between, three levels of analysis: (a) the actual text; (b) the discursive practices; and (c) the larger social context that bears upon the text and the discursive practices. We made our survey focusing on the texts published by each one of the 84 Italian universities on their websites. The first level of analysis was a lexical-statistical study of the texts with the main aim of observing the adopted terminology (vocabulary). At a second level, and on the basis of the previously performed analysis, we employed multidimensional statistic techniques, in order to study the discursive practices. Discourse study starts from concordances analysis, that deals with the identification of some local contexts containing word-types of specific interest. The final result is the reconstruction of the major sense behavior models. At a macro level, an exploratory study of the social context has been realized in order to link the texts and the discursive practices to the background conditions. Such analysis partially contributed to fill a gap in the literature, that is the study of the entrepreneurial university model from the discursive profile in the specific Italian context.

Additional file 1: Figure S1. In vivo Scheme of treatment. A. Scheme of treatment for syngeneic murine models: Peptide R was intraperitoneally administered 5 day/week for 2, Anti-PD-1 was intraperitoneally administered at Day 1–4–7-10-13. B. Scheme of treatment for human PES43 melanoma xenograft: Peptide R54 was intraperitoneally administered 5 day/week for 3 weeks, Anti-PD-1 nivolumab was intraperitoneally administered at Day 1–4–7-10-13-16-19. Figure S2. CXCR4 antagonist Pep R54 in combination with nivolumab reduced plasma lactate. Reduction in plasma lactate was revealed in Pep R54 + nivolumab treated animals as compared to untreated mice at 44 days post-treatment. Lactate dehydrogenase (LDH) blood level from retro-orbital plexus (100 μl) blood sampling (plasma concentration mg/dL, mean ± SD). Figure S3. A. Peptide R54 in combination with nivolumalb reduced PES43 lung nodules. Representative PES43 metastasis in athymic mouse lung. Metastatic nodules were evaluated 8 weeks after PES43 subcutaneous injection (3/5 untreated mice, 1/4 nivolumab, 1/6 Pep R54, and 0/4 Pep R54 + nivolumab treated mice). Upper: H&E of lungs from PES43 xenograft mice: Lower: IHC for MELAN-MART1. B. Peptide R54 in combination with nivolumab reduced CXCR4-, P-ERK downstream signaling and Ki67 in PES43 xenograft. B. Representative IHC pictures (magnification 400x) for P-ERK downstream signaling pathway and Ki67 with membrane/cytoplasmic and nuclear localization respectively.

Additional file 1: Figure S1. In vivo Scheme of treatment. A. Scheme of treatment for syngeneic murine models: Peptide R was intraperitoneally administered 5 day/week for 2, Anti-PD-1 was intraperitoneally administered at Day 1–4–7-10-13. B. Scheme of treatment for human PES43 melanoma xenograft: Peptide R54 was intraperitoneally administered 5 day/week for 3 weeks, Anti-PD-1 nivolumab was intraperitoneally administered at Day 1–4–7-10-13-16-19. Figure S2. CXCR4 antagonist Pep R54 in combination with nivolumab reduced plasma lactate. Reduction in plasma lactate was revealed in Pep R54 + nivolumab treated animals as compared to untreated mice at 44 days post-treatment. Lactate dehydrogenase (LDH) blood level from retro-orbital plexus (100 μl) blood sampling (plasma concentration mg/dL, mean ± SD). Figure S3. A. Peptide R54 in combination with nivolumalb reduced PES43 lung nodules. Representative PES43 metastasis in athymic mouse lung. Metastatic nodules were evaluated 8 weeks after PES43 subcutaneous injection (3/5 untreated mice, 1/4 nivolumab, 1/6 Pep R54, and 0/4 Pep R54 + nivolumab treated mice). Upper: H&E of lungs from PES43 xenograft mice: Lower: IHC for MELAN-MART1. B. Peptide R54 in combination with nivolumab reduced CXCR4-, P-ERK downstream signaling and Ki67 in PES43 xenograft. B. Representative IHC pictures (magnification 400x) for P-ERK downstream signaling pathway and Ki67 with membrane/cytoplasmic and nuclear localization respectively.

Background: Blood culturing remains the mainstream tool to inform an appropriate treatment in hospital-acquired bloodstream infections and to diagnose any bacteremia. Methods: A retrospective investigation on the prevalence of Gram-negative bacteria (GNB) and their resistance in hospitalized patients by age, sex, and units from blood cultures (BCs) was conducted from January 2018 to April 2020 at Sant’Elia hospital, Caltanissetta, southern Italy. We divided the patient age range into four equal intervals. Results: Multivariate demographic and microbiological variables did not show an association between bacteria distributions and gender and age. The distribution by units showed a higher prevalence of Klebsiella pneumoniae and Acinetobacter baumannii in the intensive care unit (ICU) and Escherichia coli in the non-intensive care units (non-ICUs). The analysis of antibiotic resistance showed that E. coli was susceptible to a large class of antibiotics such as carbapenem and trimethoprim-sulfamethoxazole. K. pneumoniae showed a significant susceptibility to colistin, tigecycline, and trimethoprim-sulfamethoxazole. From the survival analysis, patients with E. coli had a higher survival rate. Conclusions: The authors stress the importance of the implementation of large community-level programs to prevent E. coli bacteremia. K. pneumoniae and E. coli susceptibility patterns to antibiotics, including in the prescription patterns of general practitioners, suggest that the local surveillance and implementation of educational programs remain essential measures to slow down the spread of resistance and, consequently, increase the antibiotic lifespan.

Synthetic nucleic acid interactors represent an exciting research field due to their biotechnological and potential therapeutic applications. The translation of these molecules into drugs is a long and difficult process that justifies the continuous research of new chemotypes endowed with favorable binding, pharmacokinetic and pharmacodynamic properties. In this scenario, we describe the synthesis of two sets of homo-thymine nucleopeptides, in which nucleobases are inserted in a peptide structure, to investigate the role of the underivatized amino acid residue and the distance of the nucleobase from the peptide backbone on the nucleic acid recognition process. It is worth noting that the CD spectroscopy investigation showed that two of the reported nucleopeptides, consisting of alternation of thymine functionalized L-Orn and L-Dab and L-Arg as underivatized amino acids, were able to efficiently bind DNA and RNA targets and cross both cell and nuclear membranes.

The chemokine receptor CXCR4 is overexpressed and functional in colorectal cancer. To investigate the role of CXCR4 antagonism in potentiating colon cancer standard therapy, the new peptide CXCR4 antagonist Peptide R (Pep R) was employed. Human colon cancer HCT116 xenograft-bearing mice were treated with chemotherapeutic agents (CT) 5-Fluorouracil (5FU) and oxaliplatin (OX) or 5FU and radio chemotherapy (RT-CT) in the presence of Pep R. After two weeks, CT plus Pep R reduced by 4-fold the relative tumor volume (RTV) as compared to 2- and 1.6-fold reductions induced, respectively, by CT and Pep R. In vitro Pep R addition to CT/RT-CT impaired HCT116 cell growth and further reduced HCT116 and HT29 clonal capability. Thus, the hypothesis that Pep R could target the epithelial mesenchyme transition (EMT) process was evaluated. While CT decreased ECAD and increased ZEB-1 and CD90 expression, the addition of Pep R restored the pretreatment expression. In HCT116 and HT29 cells, CT/RT-CT induced a population of CD133+CXCR4+ cells, supposedly a stem-resistant cancer cell population, while Pep R reduced it. Taken together, the results showed that targeting CXCR4 ameliorates the effect of treatment in colon cancer through inhibition of cell growth and reversal of EMT treatment-induced markers, supporting further clinical studies.

Chemoattraction through the CXCR4-CXCL12 axis has been shown to be an important mechanism to direct circulating tumor cells toward distant sites. The objective of this work was to prepare a fake metastatic niche made up of a gel loaded with CXCL12. The gel is designed to create a steep concentration gradient of the chemokine in the proximity of the site of administration/injection, aimed to divert and capture circulating CXCR4+ tumor cells. To this aim, different thermoresponsive gels based on methylcellulose (MC) or poloxamers, loaded with CXCL12, with or without hyaluronic acid (HA) were designed and their mechanical properties correlated with the ability to attract and capture in vitro CXCR4+ cells. Results of in vitro cell studies showed that all prepared gels induced CEM tumor cell migration whereas only gels based on MC embedded with CXCL12 are able to capture them.

In this study a novel auditory version of the Single Category Implicit Association Test (SC-IAT-A) was developed to investigate (a) the valence of adults’ associations to infant cries and laughs, (b) moderation of implicit associations by gender and empathy, and (c) the robustness of implicit associations controlling for auditory sensitivity. Eighty adults (50% females) were administered two SC-IAT-As, the Empathy Quotient, and the Weinstein Noise Sensitivity Scale. Adults showed positive implicit associations to infant laugh and negative ones to infant cry; only the implicit associations with the infant laugh were negatively related to empathy scores, and no gender differences were observed. Finally, implicit associations to infant cry were affected by noise sensitivity. The SC-IAT-A is useful to evaluate the valence of implicit reactions to infant auditory cues and could provide fresh insights into understanding processes that regulate the quality of adult–infant relationships.

// Sara Santagata 1 , Maria Napolitano 1 , Crescenzo D'Alterio 1 , Sonia Desicato 2 , Salvatore Di Maro 3 , Luciana Marinelli 3 , Alessandra Fragale 4 , Maria Buoncervello 4 , Francesco Persico 5 , Lucia Gabriele 4 , Ettore Novellino 3 , Nicola Longo 5 , Sandro Pignata 2 , Sisto Perdona 2 and Stefania Scala 1 1 Functional Genomics, Istituto Nazionale per lo Studio e la Cura dei Tumori, Fondazione “G. Pascale”-IRCCS, 80131 Naples, Italy 2 Uro-Gynecological Department, Istituto Nazionale per lo Studio e la Cura dei Tumori, Fondazione “G. Pascale”-IRCCS, 80131 Naples, Italy 3 Department of Pharmacy, University of Naples Federico II, 80131 Naples, Italy 4 Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanita, 00161 Rome, Italy 5 Urology Division, University Federico II, 80131 Naples, Italy Correspondence to: Stefania Scala, email: s.scala@istitutotumori.na.it , scalaste@gmail.com Keywords: CXCR4, immune suppression, renal cell carcinoma, T regulatory cells, tumor microenvironment Received: May 07, 2017 Accepted: June 25, 2017 Published: August 19, 2017 ABSTRACT With the intent to identify biomarkers in renal cell carcinoma (RCC) the functional status of T-regulatory cells (Tregs) was investigated in primary RCC. Tregs were isolated from tumoral-(TT), peritumoral tissue-(PT) and peripheral blood-(PB) of 42 primary RCC patients and function evaluated through effector T cells (Teff) proliferation, cytokines release and demethylation of Treg Specific Region (TSDR). The highest value of Tregs was detected in TT with the uppermost amount of effector-Tregs-(CD4 + CD25 hi FOXP3 hi CD45RA - ). PB-RCC Tregs efficiently suppress Teff proliferation compared to healthy donor (HD)-Tregs and, at the intrapatient evaluation, TT-derived Tregs were the most suppressive. Higher demethylation TSDR was detected in TT- and PB-RCC Tregs vs HD-Tregs ( P

We previously reported the discovery of a CXCL12-mimetic cyclic peptide (2) as a selective CXCR4 antagonist showing promising in vitro and in vivo anticancer activity. However, further development of this peptide was hampered by its degradation in biological fluids as well as by its low micromolar affinity for the receptor. Herein, extensive chemical modifications led to the development of a new analogue (10) with enhanced potency, specificity, and plasma stability. A combined approach of Ala-amino acid scan, NMR, and molecular modeling unraveled the reasons behind the improved binding properties of 10 vs 2. Biological investigations on leukemia (CEM) and colon (HT29 and HCT116) cancer cell lines showed that 10 is able to impair CXCL12-mediated cell migration, ERK-phosphorylation, and CXCR4 internalization. These outcomes might pave the way for the future preclinical development of 10 in CXCR4 overexpressing leukemia and colon cancer.

Development of distant metastasis relies on interactions between cancer and stromal cells. CXCL12, also known as stromal-derived factor 1α (SDF-1α), is a major chemokine constitutively secreted in bone marrow, lymph nodes, liver and lung, playing a critical role in the migration and seeding of neoplastic cells. CXCL12 activates the CXCR4 receptor that is overexpressed in several human cancer cells. Recent evidence reveals that tumors induce pre-metastatic niches in target organ producing tumor-derived factors. Pre-metastatic niches represent a tumor growth-favoring microenvironment in absence of cancer cells. A commercially available dermal filler, hyaluronic acid (HA) -based gel, loaded with CXCL12 (CLG) reproduced a “fake” pre-metastatic niche. In vitro, B16-hCXCR4-GFP, human cxcr4 expressing murine melanoma cells efficiently migrated toward CLG. In vivo, CLGs and empty gels (EGs) were subcutaneously injected into C57BL/6 mice and 5 days later B16-hCXCR4-GFP cells were intravenously inoculated. CLGs were able to recruit a significantly higher number of B16-hCXCR4-GFP cells as compared to EGs, with reduced lung metastasis in mice carrying CLG. CLG were infiltrated by higher number of CD45-positive leukocytes, mainly neutrophils CD11b+Ly6G+ cells, myeloid CD11b+Ly6G- and macrophages F4/80. CLG recovered cells recapitulated the features of B16-hCXCR4-GFP (epithelial, melanin rich, MELAN A/ S100/ c-Kit/CXCR4 pos; α-SMA neg). Thus a HA-based dermal filler loaded with CXCL12 can attract and trap CXCR4+tumor cells. The CLG trapped cells can be recovered and biologically characterized. As a corollary, a reduction in CXCR4 dependent lung metastasis was detected.

Cationic nucleopeptides belong to a family of synthetic oligomers composed by amino acids and nucleobases. Their capability to recognize nucleic acid targets and to cross cellular membranes provided the basis for considering them as novel non-covalent delivery agents for nucleic acid pharmaceuticals. Herein, starting from a 12-mer nucleopeptide model, the number of cationic residues was modulated in order to obtain new nucleopeptides endowed with high solubility in acqueous medium, acceptable bio-stability, low cytotoxicity and good capability to bind nucleic acid. Two candidates were selected to further investigate their potential as nucleic acid carriers, showing higher efficiency to deliver PNA in comparison with RNA. Noteworthy, this study encourages the development of nucleopeptides as new carriers to extend the known strategies for those nucleic acid analogues, especially PNA, that still remain difficult to drive into the cells.

C-X-C chemokine receptor 4 (CXCR4) is over-expressed in multiple human cancers and correlates with tumor aggressiveness, poor prognosis and increased risk for distant metastases. Imaging agents for CXCR4 are thus highly desirable. We developed a novel CXCR4-targeted near-infrared (NIR) fluorescent probe (Peptide R-NIR750) conjugating the new developed CXCR4 peptidic antagonist Peptide R with the NIR fluorescent dye VivoTag-S750. Specific CXCR4 binding was obtained in cells overexpressing human CXCR4 (B16-hCXCR4 and human melanoma cells PES43), but not in CXCR4 low expressing cells (FB-1). Ex vivo evaluation demonstrated that PepR-NIR750 specifically detects B16-hCXCR4-derived subcutaneous tumors and lung metastases. Fluorescence Molecular Tomography (FMT) in vivo imaging was performed on mice carrying subcutaneous CHO and CHO-CXCR4 tumors. PepR-NIR750 accumulates only in CXCR4-positive expressing subcutaneous tumors. Additionally, an intense NIR fluorescence signal was detected in PES43-derived lung metastases of nude mice injected with PepR-NIR750 versus mice injected with VivoTag-S750. With a therapeutic intent, mice bearing PES43-derived lung metastases were treated with Peptide R. A the dramatic reduction in PES43-derived lung metastases was detected through a decrease of the PepR-NIR750 signal. PepR-NIR750 is a specific probe for non-invasive detection of human high CXCR4-expressing tumors and metastatic lesion and thus a valuable tool for cancer molecular imaging.

asda Copyright © 2015 John Wiley & Sons, Ltd.

A neoadjuvant clinical trial was previously conducted in patients with resectable colorectal cancer liver metastases (CRLM). At a median follow up of 28 months, 20/33 patients were dead of disease, 8 were alive with disease and 5 were alive with no evidence of disease. To shed further insight into biological features accounting for different outcomes, the expression of CXCR4-CXCL12-CXCR7, TLR2-TLR4, and the programmed death receptor-1 (PD-1)/programmed death-1 ligand (PD-L1) was evaluated in excised liver metastases. Expression profiles were assessed through qPCR in metastatic and unaffected liver tissue of 33 CRLM neoadjuvant-treated patients. CXCR4 and CXCR7, TLR2/TLR4, and PD-1/PD-L1 mRNA were significantly overexpressed in metastatic compared to unaffected liver tissues. CXCR4 protein was negative/low in 10/31, and high in 21/31, CXCR7 was negative/low in 16/31 and high in 15/31, CXCL12 was negative/low in 14/31 and high in 17/31 CRLM. PD-1 was negative in 19/30 and positive in 11/30, PD-L1 was negative/low in 24/30 and high in 6/30 CRLM. Stromal PD-L1 expression, affected the progression-free survival (PFS) in the CRLM population. Patients overexpressing CXCR4 experienced a worse PFS and cancer specific survival (CSS) (p = 0.001 and p = 0.0008); in these patients, KRAS mutation identified a subgroup with a significantly worse CSS (p < 0.01). Thus, CXCR4 and PD-L1 expression discriminate patients with the worse PFS within the CRLM evaluated patients. Within the CXCR4 high expressing patients carrying Mut-KRAS in CRLM identifies the worst prognostic group. Thus, CXCR4 targeting plus anti-PD-1 therapy should be explored to improve the prognosis of Mut-KRAS-high CXCR4-CRLMs.

Il volume, incentrato sulle pratiche narrative nei contesti della cura, si pone l’obiettivo di mettere a confronto pensieri, studi, interessi di ricerca ed interventi formativi di un gruppo di ricercatori e professionisti che da anni operano nell’ambito socio-educativo e sanitario secondo un orientamento umanistico alla cura, restituita nella complessità e profondità esistenziale e dentro la più ampia cornice delle Medical Humanities. Il lavoro intende, quindi, sollevare interrogativi e riflessioni attraverso cui interpellare criticamente il pensare e l’agire della cura nei vari ambiti (in particolare, quello medico-sanitario) in una prospettiva pedagogica di ampio respiro, che possa essere in grado di rimettere al centro le nozioni di narrazione e di narratività. Attraverso l’utilizzo di differenti dispositivi formativi – l’arte, il cinema, il teatro, la letteratura e i new media – il testo evidenzia sia l’importanza di una formazione degli operatori dal punto di vista relazionale sia la potenza trasformativa che il raccontarsi e il raccontare possono avere su quanti – come i pazienti di una struttura ospedaliera – attraversano particolari momenti di difficoltà e di disagio.

Il saggio si sofferma sulla cultura visuale quale strumento di conoscenza del sé professionale per cui lo studente è incoraggiato a sviluppare competenze nelle ricerche culturali all’interno del complesso dibattitto che investe lo spazio sociale dei media, dell’arte, della costruzione dell’immaginario, e più in generale ad essere in grado di dispiegare queste competenze nel proprio ambito professionale. Le metodologie didattiche che si servono di supporti audio e video appaiono essere quelle che maggiormente stimolano le riflessioni degli studenti, favorendo il recupero e l’elaborazione di aspetti emotivi della propria esperienza formativa e il potenziamento di competenze riflessive.

The chemokine CXCL12 activates CXCR4, initiating multiple pathways that control immune cell trafficking, angiogenesis and embryogenesis; CXCR4 is also overexpressed in multiple tumors affecting metastatic dissemination. While there has been great enthusiasm for exploiting the CXCR4-CXCL12 axis as a target in cancer therapy, to date the promise has yet to be fulfilled. A new class of CXCR4-antagonist cyclic pep- tides was recently developed and the compound named Peptide R was identified as the most active. With the intent to improve the efficacy and biodistribution of Peptide R, stealth liposomes decorated with Peptide R were developed (PL-Peptide R). In vitro PL-Peptide R efficiently inhibited CXCR4-dependent migration and in vivo it significantly reduced lung metastases and increased overall survival in B16-CXCR4 injected C57BL/6 mice. To evaluate if PL-Peptide R could also be a drug delivery system for CXCR4 expressing tumors, the PL-Peptide R was loaded with doxorubicin (DOX) (PL-Peptide R-DOX). PL-Peptide R-DOX efficiently delivered DOX to CXCR4 expressing cell lines with a consequent decrease in the DOX IC50 efficient dose. In vivo, B16-CXCR4 injected C57BL/6 mice treated with PL-Peptide R-DOX developed fewer lung metastases compared to PL-DOX treated mice. This work provides the proof-of-concept to prevent metastasis by using combined nanomedicine.

Deregulation of mTOR and IGF pathways is frequent in hepatocellular carcinoma (HCC), thus mTOR and IGF1R represent suitable therapeutic targets in HCC. The aim of this study was to evaluate the effects of mTOR inhibitors (mTORi) and OSI-906, blocker of IGF1R/IR, on HCC cell proliferation, viability, migration and invasion, and alpha-fetoprotein (α-FP) secretion. In HepG2 and HuH-7 we evaluated, the expression of mTOR and IGF pathway components; the effects of Sirolimus, Everolimus, Temsirolimus and OSI-906 on cell proliferation; the effects of Sirolimus, OSI-906, and their combination, on cell secretion, proliferation, viability, cell cycle, apoptosis, invasion and migration. Moreover, intracellular mechanisms underlying these cell functions were evaluated in both cell lines. Our results show that HepG2 and HuH-7 present with the same mRNA expression profile with high levels of IGF2. OSI-906 inhibited cell proliferation at high concentration, while mTORi suppressed cell proliferation in a dose-time dependent manner in both cell lines. The co-treatment showed an additive inhibitory effect on cell proliferation and viability. This effect was not related to induction of apoptosis, but to G0/G1 phase block. Moreover, the co-treatment prevented the Sirolimus-induced AKT activation as escape mechanism. Both agents demonstrated to be differently effective in inhibiting α-FP secretion. Sirolimus, OSI-906, and their combination, blocked cell migration and invasion in HuH-7. These findings indicate that, co-targeting of IGF1R/IR and mTOR pathways could be a novel therapeutic approach in the management of HCC, in order to maximize antitumoral effect and to prevent the early development of resistance mechanisms.