Synthesis of the fungal metabolite YWA1 and related constructs as tools to study MelLec-mediated immune response to Aspergillus infections

Funding Sources Funding was provided by the Wellcome Trust (102705, 097377), the Medical Research Council Centre for Medical Mycology and the University of Aberdeen (MR/N006364/1). ACKNOWLEDGMENT We thank Ian Fraser Flow Cytometry and Microscopy and Histology facilities, University of Aberdeen.Peer reviewedPostprin

Design, synthesis, radiosynthesis and biological evaluation of Fenretinide analogues as anticancer and metabolic syndromepreventive agents

We thank the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement N° 675417 (PET3D project) for financial support of the project and the studentship of I.P. We also thank the British Heart Foundation for the project grant PG/16/90/32518.Peer reviewedPostprin

An assay system to evaluate riboflavin/UV-A corneal phototherapy efficacy in a porcine corneal organ culture model

The purpose of this study is to investigate the response of a porcine corneal organ cultures to the riboflavin/UV-A phototherapy in the injury healing of induced lesions. A porcine corneal organ culture model has been established. Corneal alterations in the stroma were valuated setting an assay system, based on an automated image analysis method able to quantify the damaged (brightness values), within of the 24 regions of interest (ROIs) in which the corneal section has been divided and integrating the data analysis with a multi-aspect approach. Three group of corneas have been analyzed: (healthy, injured and injured-and-treated group). Our study revealed a significant effect of the riboflavin/UV-A phototherapy in the injury healing of porcine corneas after induced lesions. The injured corneas had significant differences of brightness values in comparison to treated (p< 0.00) and healthy (p<0.001) corneas whereas the treated and healthy corneas showed not significant difference (p = 0.995). Riboflavin/UV-A phototherapy shows a significant effect in the restoring the brightness values of damaged corneas to the values of healthy corneas, suggesting treatment restores the injury healing of corneas after lesions. Our assay system may be compared to clinical diagnostic method such as the OCT imaging for in vivo damaged ocular structures investigations

An Assay System to Evaluate Riboflavin/UV-A Corneal Phototherapy Efficacy in a Porcine Corneal Organ Culture Model

The purpose of this study was to investigate the response of porcine corneal organ cultures to riboflavin/UV-A phototherapy in the injury healing of induced lesions. A porcine corneal organ culture model was established. Corneal alterations in the stroma were evaluated using an assay system, based on an automated image analysis method able to (i) localize the holes and gaps within the stroma and (ii) measure the brightness values in these patches. The analysis has been performed by dividing the corneal section in 24 regions of interest (ROIs) and integrating the data analysis with a "multi-aspect approach." Three group of corneas were analyzed: healthy, injured, and injured-and-treated. Our study revealed a significant effect of the riboflavin/UV-A phototherapy in the injury healing of porcine corneas after induced lesions. The injured corneas had significant differences of brightness values in comparison to treated (p < 0.00) and healthy (p < 0.001) corneas, whereas the treated and healthy corneas showed no significant difference (p = 0.995). Riboflavin/UV-A phototherapy shows a significant effect in restoring the brightness values of damaged corneas to the values of healthy corneas, suggesting treatment restores the injury healing of corneas after lesions. Our assay system may be compared to clinical diagnostic methods, such as optical coherence tomography (OCT) imaging, for in vivo damaged ocular structure investigations

Tumour-associated macrophages correlate with microvascular bed extension in colorectal cancer patients

Tumour-associated macrophages (TAMs) represent pivotal components of tumour microenvironment promoting angiogenesis, tumour progression and invasion. In colorectal cancer (CRC), there are no conclusive data about the role of TAMs in angiogenesis-mediated tumour progression. In this study, we aimed to evaluate a correlation between TAMs, TAM immunostained area (TAMIA) microvascular density (MVD), endothelial area (EA) and cancer cells positive to VEGF-A (CCP-VEGF-A) in primary tumour tissue of locally advanced CRC patients undergone to radical surgery. A series of 76 patients with CRC were selected and evaluated by immunohistochemistry and image analysis. An anti-CD68 antibody was employed to assess TAMs and TAMIA expression, an anti-CD34 antibody was utilized to detect MVD and EA expression, whereas an anti-VEGF-A antibody was used to detect CCP-VEGF-A; then, tumour sections were evaluated by image analysis methods. The mean&nbsp;±&nbsp;S.D. of TAMs, MVD and CCP-VEGF-A was 65.58&nbsp;±&nbsp;21.14, 28.53&nbsp;±&nbsp;7.75 and 63%&nbsp;±&nbsp;37%, respectively; the mean&nbsp;±&nbsp;S.D. of TAMIA and EA was 438.37&nbsp;±&nbsp;124.14μ2 and 186.73&nbsp;±&nbsp;67.22μ2, respectively. A significant correlation was found between TAMs, TAMIA, MVD and EA each other (r&nbsp;ranging from 0.69 to 0.84; P ranging from 0.000 to 0.004). The high level of expression of TAMs and TAMIA in tumour tissue and the significant correlation with both MVD and EA illustrate that TAMs could represent a marker that plays an important role in promoting angiogenesis-mediated CRC. In this context, novel agents killing TAMs might be evaluated in clinical trials as a new anti-angiogenic approach

Efficacy of conventional versus innovative therapies for treating skin wounds in veterinary medicine

open16siINTRODUCTION: The skin is the largest organ of mammals. The loss of skin integrity may induce important dysfunctions or even death. For superficial wounds, the endogenous healing mechanisms in combination with traditional wound care are sufficient to achieve functional repair. In contrast, in larger wounds, like third and fourth degree burns, chronic wound or deep ulcers it is difficult to obtain the restitutio ad integrum and fibrosis and/or scar tissue develops1,2. The aim of this study was to verify the efficacy of conventional and innovative topic treatments on skin regeneration, induced experimentally in sheep. To achieve this goal different types of investigations (clinical, molecular, histological, immunohistochemical) were performed. METHODS: Six skin lesions (4x4cm) were surgically created on the back of six healthy adult sheep; every single wound was destined, in a randomized way, to one of the following treatments: Acemannan gel, Manuka Honey, hyaluronic acid, Plasma3 (ionized gas), allogeneic mesenchymal stem cells isolated from peripheral blood (PB-MSCs). The sixth wound was the placebo. Biopsies were collected with a surgical punch (0,6x0,6 cm) at time T0, T15 and T40 days. Lesions were clinically evaluated considering the presence and color of wound fluid, the state of hydration, the wound surface/surroundings and other parameters. Histological examinations considered crust formation, re-epithelization and epidermal thickness, dermis edema, extension of granulation tissue, acute and chronic inflammation. Immunohistochemistry for evaluation of inflammation, vascularization and cell proliferation was performed using CD3, CD20, MHCII, von Willebrand factor (vWF) and KI67 antibodies. Furthermore, Real time-PCR investigated genes as V ascular endothelial growth factors (VEGF), Transforming growth factor beta 1(TGFβ1), Vimentin (VIM), Collagen 1α1 (Col1α1) and hair Keratin (hKER). RESULTS: Clinically, the lesions treated with plasma healed more rapidly respect to other treatments and a reduced bacterial load was observed. At T7 wounds treated with stem cells and plasma were less macerated than lesions treated with other therapies. At T15 the wounds treated with hyaluronic acid showed a normal state of hydration while lesions treated with Manuka Honey exhibited a normal hydration from the third week only (Acemannan gel at fourth week). From the second week onwards all wounds did not show presence of fluid and exhibited a dry and clean secondary layer. All lesions, excluded wounds treated with acemannan gel, presented a red (hyaluronic acid and plasma) and dark red (Manuka Honey, PB-MSCs) granulation tissue starting from the first week. Molecular analysis showed a correspondence between clinical and molecular/histologic results. For instance, VEGF mRNA expression confirms angiogenetic events observed at histological level while TGF-β, CD3 and CD20 mRNA/protein expression indicated the presence/absence of inflammation in the used treatments. DISCUSSION & CONCLUSIONS: Innovative therapies led to surprising results regarding regeneration of mammalian skin. Indeed, on the basis of clinical analysis, wounds treated with plasma and MSC healed more rapidly. Further examinations are ongoing in order to elucidate possible mechanisms explaining these differences. REFERENCES: 1S.Y. Broeckx, S. Maes, T. Martinello, et al (2014) Equine epidermis: a source of epithelial-like stem/progenitor cells with in vitro and in vivo regenerative capacities Stem Cells Dev, pp 1134-48. 2J.H. Spaas, C. Gomiero, S.Y. Broeckx, et al (2016) Wound healing markers after autologous and allogeneic epithelial-like stem cell treatment Cytotherapy 2016 (in press). 3E. Martines, M. Zuin, R. Cavazzana, et al. (2009) A novel plasma source for sterilization of living tissues, New J. Phys. 11, 115014.openPatruno, MARCO VINCENZO; Gomiero, Chiara; Martinello, Tiziana; Perazzi, Anna; Gemignani, F; DE BENEDICTIS, GIULIA MARIA; Ferro, Silvia; Zuin, M; Martines, E; Cordaro, Luigi; Brun, Paola; Maccatrozzo, Lisa; Broeckx, Sy; Spaas, Jh; Chiers, K; Iacopetti, IlariaPatruno, MARCO VINCENZO; Gomiero, Chiara; Martinello, Tiziana; Perazzi, Anna; Gemignani, F; DE BENEDICTIS, GIULIA MARIA; Ferro, Silvia; Zuin, M; Martines, E; Cordaro, Luigi; Brun, Paola; Maccatrozzo, Lisa; Broeckx, Sy; Spaas, Jh; Chiers, K; Iacopetti, Ilari

Mutable Collagenous Tissue Isolated from Echinoderms Leads to the Production of a Dermal Template That Is Biocompatible and Effective for Wound Healing in Rats

The mutable collagenous tissue (MCT) of echinoderms possesses biological peculiarities that facilitate native collagen extraction and employment for biomedical applications such as regenerative purposes for the treatment of skin wounds. Strategies for skin regeneration have been developed and dermal substitutes have been used to cover the lesion to facilitate cell proliferation, although very little is known about the application of novel matrix obtained from marine collagen. From food waste we isolated eco-friendly collagen, naturally enriched with glycosaminoglycans, to produce an innovative marine-derived biomaterial assembled as a novel bi-layered skin substitute (Marine Collagen Dermal Template or MCDT). The present work carried out a preliminary experimental in vivo comparative analysis between the MCDT and Integra, one of the most widely used dermal templates for wound management, in a rat model of full-thickness skin wounds. Clinical, histological, and molecular evaluations showed that the MCDT might be a valuable tool in promoting and supporting skin wound healing: it is biocompatible, as no adverse reactions were observed, along with stimulating angiogenesis and the deposition of mature collagen. Therefore, the two dermal templates used in this study displayed similar biocompatibility and outcome with focus on full-thickness skin wounds, although a peculiar cellular behavior involving the angiogenesis process was observed for the MCDT

Progettazione e sintesi di derivati a nucleo 1,2,4-triazolico inibitori di proteine chinasi recettoriali coinvolte nell'eziopatogenesi del tumore tiroideo

Questo lavoro di tesi è stato dedicato alla progettazione ed alla sintesi di derivati eteroci- clici a nucleo 1,2,4-triazolico, pensati come inibitori di proteine chinasi recettoriali coinvolte nell’eziopatogenesi del tumore tiroideo, in particolare RET e VEGFR2. Il tumore tiroideo rappresenta circa il 95% di tutte le neoplasie endocrine. E’ una patologia eterogenea che può essere classificata in carcinoma tiroideo differenziato (DTC) (95% dei casi), carcinoma tiroideo indifferenziato (anaplastico) (ATC), e carcinoma tiroideo midollare (MTC). In commercio esistono già inibitori chinasici usati per il trattamento di questa neoplasia, ma questi sono caratterizzati da effetti collaterali che portano spesso alla sospensione del- la terapia. Da qui la necessità di sviluppare farmaci nuovi e più sicuri. A questo scopo, ci si rivolge sempre più spesso allo sviluppo di inibitori multi-target. Infatti, sebbene lo sviluppo del tumore sia generalmente promosso da una alterata attività di una specifica proteina chinasi, l’inibizione selettiva dell’oncogene identificato può risultare inefficace. La natura eterogenea del tumore e, soprattutto, la capacità di cross-talk tra le molte proteine chinasi cellulari rendono necessario inibire contemporaneamente più classi diverse. Solo il blocco dell’intero processo di trasduzione del segnale, dall’esterno all’interno della cellula, assicu- ra una inibizione efficace della proliferazione cellulare del tumore ed anche della sua va- scolarizzazione. Inoltre, l’uso di un inibitore multi-target consente di ridurre, o addirittura prevenire, i fenomeni di resistenza che generalmente accompagnano l’uso di farmaci se- lettivi per una specifica chinasi e che sono determinati dalla capacità della cellula tumorale di attivare vie biochimiche alternative regolate da proteine chinasi diverse. Grazie ad uno studio di virtual screening, effettuato dal nostro gruppo di ricerca in collabo- razione con l’Università di Napoli, è stato individuato il derivato 1,2,4-triazolico DP01920 come nuovo ed efficace inibitore di proteine tirosina chinasi recettoriali coinvolte nell’ezio- patogenesi del tumore tiroideo e tra queste, in particolare, RET e VEGFR2, maggiormente coinvolti nello sviluppo del tumore tiroideo. ! DP01920 rappresenta pertanto un ottimo punto di partenza per lo sviluppo di una nuova classe di inibitori chinasi multitarget a nucleo 1,2,4-triazolico, da studiare come candidati farmaci per il trattamento del tumore tiroideo

Influence of temperature, time and different media on mesenchymal stromal cells shipped for clinical application

Cell-based therapies, such as the use of mesenchymal stromal cells (MSCs), are becoming popular in veterinary medicine. When MSCs are not cryopreserved, they are shipped in suspension, but no previous studies have analyzed MSC viability during delivery. Here, the impact of several experimental shipping conditions on the number of equine blood-derived (ePB-MSC) and canine adipose-derived (cA-MSC) MSCs were evaluated. Among the different parameters tested, only time and temperature influenced MSC number during the experimental shipping conditions. Cells were monitored over different time intervals for gene expression of typical MSC markers and to evaluate acquired resistance to apoptosis and beta-galactosidase activity. Overall, these results indicate that ePB-MSC and cA-MSC should be delivered in phosphate buffered saline at room temperature and within 9-12h