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Context: Tumor acidity represents a major cause of chemoresistance. Proton pump inhibitors (PPIs) can neutralize tumor acidity, sensitizing cancer cells to chemotherapy.Objective: To compare the anti-tumor efficacy of different PPIs in vitro and in vivo.Materials and methods: In vitro experiments PPIs anti-tumor efficacy in terms of cell proliferation and cell death/apoptosis/necrosis evaluation were performed. In vivo PPIs efficacy experiments were carried out using melanoma xenograft model in SCID mice.Results: Lansoprazole showed higher anti-tumor effect when compared to the other PPIs. The lansoprazole effect lasted even upon drug removal from the cell culture medium and it was independent from the lipophilicity of the PPIs formulation.Discussion: These PPIs have shown different anti-tumoral efficacy, and the most effective at low dose was lansoprazole.Conclusion: The possibility to contrast tumor acidity by off-label using PPIs opens a new field of oncology investigation.

Magnetic resonance imaging (MRI) and spectroscopy (MRS) offer powerful approaches for detecting physiological and metabolic alterations in malignancies and help investigate underlying molecular mechanisms. Research on epithelial ovarian carcinoma (EOC), the gynaecological malignancy with the highest death rate characterised by frequent relapse and onset of drug resistance, could benefit from application of these molecular imaging approaches. In this study, MRI/MRS were used to characterise solid tumour models obtained by subcutaneous (s.c.) or intraperitoneal (i.p.) implantation of human SKOV3.ip cells in severe combined immunodeficiency (SCID) mice. In vivo MRI/ MRS, ex vivo magic-angle-spinning (MAS), and in vitro 1 H-NMR measurements were carried out at 4.7T, 9.4T, and 9.4/16.5T, respectively. MRI evaluation was performed by T1-, T2-, and diffusion-weighted (DW) multislice spin-echo imaging. The in vivo 1 H spectra of all tumour models showed a prominent resonance of total choline-containing metabolites (tCho). Quantitative in vivo MRS of both i.p. and s.c. SKOV3.ip xenografts showed that the mean tCho content was in the 2.9-4.5mM range, with a mean PCho/tCho ratio of 0.990.01 [23 examinations, 14–34days post injection (dpi)], in good agreement with ex vivo and in vitro analyses. Myo-inositol ranged between 11.7 and 17.0mM, with a trend towards higher values in i.p. xenografts at 14–16 dpi. The average apparent diffusion coefficient (ADC) values of SKOV3.ip xenografts [1.640.11 (n=9, i.p.) and 1.580.03 x10 -3 mm 2 /s (n=7, s.c.)] were in agreement with values reported for tumours from patients with EOC, while the mean vascular signal fraction (VSF) was lower (≤ 4%), probably due to the more rapid growth of preclinical models. Both s.c. and i.p. xenografts are valuable preclinical models for monitoring biochemical and physiopathological changes associated with in vivo EOC tumour growth and response to therapy, which may serve as the basis for further clinical development of noninvasive MR approaches. Copyright © 2011 John Wiley & Sons, Ltd. Supporting information may be found in the online version of this paper.

Non-destructive mapping of the leaf nitrogen index (NBI1) at bunch closure was carried out at the 'Fattoria di Magliano' winery (South Tuscany, Italy) on two consecutive years, 2013 and 2014, on Vermentino cv. using the Multiplex fluorescence optical sensor. Measurements were performed manually on a 1 ha vineyard using a 15m x 15m grid, detecting leaves with similar age of three adjacent vines on two contiguous rows. The spatial heterogeneity of leaf nitrogen (N) content, based on the calibration of NBI1 against destructive analyses, was reported as a colorimetric map with 1 x 1 m grid resolution, after kriging. The 2013 map showed lower N levels compared to the 2014 map with a range of leaf N between 1.64% and 2.14%. The effect of a winter N-fertilization was nicely evidenced by the 2014 N map, showing the recovery in leaf N up to 2.64%. Just before the 2014 harvest, the heterogeneity of grape maturity on the same vineyard was evaluated by mapping the fluorescence-based Chl index, using a 15m x 15m grid. For each point, six bunches from three adjacent vines on two contiguous rows were detected. The grape soluble solids content (SSC), estimated by means of a calibration of the Chl index, ranged between 13.8 and 21.2°Bx. Two areas with lower and higher SSC were estimated by segmentation based on the median (16.9°Bx), and confirmed by the destructive SSC determination on the related grape samplings, which resulted to be 16.3°Bx and 19.4°Bx, respectively. Our work was aimed to show the utility of a non-destructive optical sensor in precision viticulture to identify distribution and heterogeneity in leaf nitrogen and in grape berries technological maturity. The sensor could be use to split the vineyard in subzones managed differently, to correct N deficiencies only where needed reducing fertilizer leakage and to perform selective harvest either as single pick or as separate picks.

This paper presents the results of the work conducted in cooperation with ANAS, which is in charge of the Italian road and highway network of national interest. The cooperation also is framed into the Italian Technical Committee on Strategic Theme 4 — Infrastructure for the specific topic of “Balancing of environmental and engineering aspects in management of road networks.” The paper presents an improved model, the EST (Energy Screening of Tunnels) model, to account for both the consumption of electricity and the performance of tunnel lighting systems. The models were developed so that each tunnel was represented on an adequate (x, y) plane that was defined by two indices. The main improvements of the model include the introduction of boundary values of the indices derived from International Standards and from resolutions of the Italian Authority of Electrical Energy and Gas.

Intrinsic resistance to cytotoxic drugs has been a main issue in cancer therapy for decades. Microenvironmental acidity is a simple while highly efficient mechanism of chemoresistance, exploited through impairment of drug delivery. The latter is achieved by extracellular protonation and/or sequestration into acidic vesicles. This study investigates the importance of extracellular acidosis and nanovesicle (exosome) release in the resistance of human tumour cell to cisplatin (CisPt); in parallel to proton pump inhibitors (PPI) ability of interfering with these tumour cell features. The results showed that CisPt uptake by human tumour cells was markedly impaired by low pH conditions. Moreover, exosomes purified from supernatants of these cell cultures contained various amounts of CisPt, which correlated to the pH conditions of the culture medium. HPLC-Q-ICP-MS analysis revealed that exosome purified from tumour cell culture supernatants contained CisPt in its native form. PPI pre-treatment increased cellular uptake of CisPt, as compared to untreated cells, in an acidic-depend manner. Furthermore, it induced a clear inhibition of exosome release by tumour cells. Human tumours obtained from xenografts pretreated with PPI contained more CisPt as compared to tumours from xenografts treated with CisPt alone. Further analysis showed that in vivo PPI treatment induced a clear reduction in the plasmatic levels of tumour-derived exosomes which also contained lower level of CisPt. Altogether, these findings point to the identification of a double mechanism that human malignant melanoma use in resisting to a dreadful cellular poison such as cisplatin. This framework of resistance includes both low pH-dependent extracellular sequestration and an exosome-mediated elimination. Both mechanisms are markedly impaired by proton pump inhibition, leading to an increased CisPt-dependent cytotoxicity.

IL PROBLEMA DELLA CERTEZZA SI SVILUPPA SIA IN AMBITO GIURIDICO CHE IN AMBITO POLITICO, SEGUANDO E RINNOVANDO EQUILIBRI SEMPRE INSTABILI E LEGATI ALLA LEGITTIMITA'. DOPO AVER TRACCIATO L'EVOLVERSI DEL PARADIGMA DELLA CERTEZZA FINO ALLA MODERNITA', SI CERCA DI COMPRENDERNE LA CRISI ATTUALE E LE POSSIBILITA' DI NUOVI EQUILIBRI PER IL FUTURO.

Persistent infection with high risk genotypes of human papillomavirus (HPV) is the cause of cervical cancer, one of most common cancer among woman worldwide, and represents an important risk factor associated with other anogenital and oropharyngeal cancers in men and women. Here, we designed a therapeutic vaccine based on integrase defective lentiviral vector (IDLV) to deliver a mutated nononcogenic form of HPV16 E7 protein, considered as a tumor specific antigen for immunotherapy of HPV-associated cervical cancer, fused to calreticulin (CRT), a protein able to enhance major histocompatibility complex class I antigen presentation (IDLV-CRT/E7). Vaccination with IDLV-CRT/E7 induced a potent and persistent E7-specific T cell response up to 1 year after a single immunization. Importantly, a single immunization with IDLV-CRT/E7 was able to prevent growth of E7-expressing TC-1 tumor cells and to eradicate established tumors in mice. The strong therapeutic effect induced by the IDLV-based vaccine in this preclinical model suggests that this strategy may be further exploited as a safe and attractive anticancer immunotherapeutic vaccine in humans.

Magnetic resonance imaging (MRI) and spectroscopy (MRS) offer powerful approaches for detecting physiological and metabolic alterations in malignancies and help investigate underlying molecular mechanisms. Research on epithelial ovarian carcinoma (EOC), the gynaecological malignancy with the highest death rate characterised by frequent relapse and onset of drug resistance, could benefit from application of these molecular imaging approaches. In this study, MRI/MRS were used to characterise solid tumour models obtained by subcutaneous (s.c.) or intraperitoneal (i.p.) implantation of human SKOV3.ip cells in severe combined immunodeficiency (SCID) mice. In vivo MRI/MRS, ex vivo magic-angle-spinning (MAS), and in vitro (1)H-NMR measurements were carried out at 4.7 T, 9.4 T, and 9.4/16.5 T, respectively. MRI evaluation was performed by T1-, T2-, and diffusion-weighted (DW) multislice spin-echo imaging. The in vivo (1)H spectra of all tumour models showed a prominent resonance of total choline-containing metabolites (tCho). Quantitative in vivo MRS of both i.p. and s.c. SKOV3.ip xenografts showed that the mean tCho content was in the 2.9-4.5 mM range, with a mean PCho/tCho ratio of 0.99 ± 0.01 [23 examinations, 14-34 days post injection (dpi)], in good agreement with ex vivo and in vitro analyses. Myo-inositol ranged between 11.7 and 17.0 mM, with a trend towards higher values in i.p. xenografts at 14-16 dpi. The average apparent diffusion coefficient (ADC) values of SKOV3.ip xenografts [1.64 ± 0.11 (n = 9, i.p.) and 1.58 ± 0.03 x10(-3) mm(2)/s (n = 7, s.c.)] were in agreement with values reported for tumours from patients with EOC, while the mean vascular signal fraction (VSF) was lower (≤ 4%), probably due to the more rapid growth of preclinical models. Both s.c. and i.p. xenografts are valuable preclinical models for monitoring biochemical and physiopathological changes associated with in vivo EOC tumour growth and response to therapy, which may serve as the basis for further clinical development of noninvasive MR approaches.

Background The HPV16 E7 protein is both a tumour-specific and a tumour-rejection antigen, the ideal target for developing therapeutic vaccines for the treatment of HPV16-associated cancer and its precursor lesions. E7, which plays a key role in virus-associated carcinogenesis, contains 98 amino acids and has two finger-type structures which bind a Zn++ ion. The ability of an Escherichia coli-produced E7-preparation, assembled into particles, to induce protective immunity against a HPV16-related tumour in the TC-1-C57BL/6 mouse tumour model, was evaluated. Methods E7 was expressed in E. coli, purified via a one-step denaturing protocol and prepared as a soluble suspension state after dialysis in native buffer. The presence in the E7 preparation of particulate forms was analysed by non-reducing SDS-PAGE and negative staining electron microscopy (EM). The Zn++ ion content was analysed by mass-spectrometry. Ten μg of protein per mouse was administered to groups of animals, once, twice or three times without adjuvant. The E7-specific humoral response was monitored in mice sera using an E7-based ELISA while the cell-mediated immune response was analysed in mice splenocytes with lymphoproliferation and IFN-γ ELISPOT assays. The E7 immunized mice were challenged with TC-1 tumour cells and the tumour growth monitored for two months. Results In western blot analysis E7 appears in multimers and high molecular mass oligomers. The EM micrographs show the protein dispersed as aggregates of different shape and size. The protein appears clustered in micro-, nano-aggregates, and structured particles. Mice immunised with this protein preparation show a significant E7-specific humoral and cell-mediated immune response of mixed Th1/Th2 type. The mice are fully protected from the tumour growth after vaccination with three E7-doses of 10 μg without any added adjuvant. Conclusions This report shows that a particulate form of HPV16 E7 is able to induce, without adjuvant, an E7-specific tumour protection in C57BL/6 mice. The protective immunity is sustained by both humoral and cell-mediated immune responses. The E. coli-derived HPV16 E7 assembled in vitro into micro- and nanoparticles represents not only a good substrate for antigen-presenting cell uptake and processing, but also a cost-effective means for the production of a new generation of HPV subunit vaccines.