1,721,000 research outputs found
Chairperson EUTROC- Mayo Conference-Berlin - Preliminary characterization of pharmacodynamic biomarkers for LR-peptide growth inhibition of ovarian cancer (OC) cell models.
No full textPreliminary characterization of pharmacodynamic biomarkers for LR-peptide growth inhibition of ovarian cancer (OC) cell models.
M.R.Amorosoa,b, G.Marvertib, F.Genovese, DS. Matassab, J.HEllemanc, E.Bernsc, F.Espositob*, MP Costib*.
aDept. Biomedical Sciences and cDept. Life Science, Via Campi 287-183, University of Modena and Reggio Emilia, 41125, Modena, Italy; Dept. of Medical Oncology, Erasmus University Medical Center - Cancer Center, PO Box 2040, 3000 CA, Rotterdam, Netherlands. bDepartment of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Via Pansini 5, Naples 80131, Italy.
LR-peptide is an inhibitor of human Thymidylate synthase (TS) through the stabilization of its inactive form as shown in drug-protein interaction experiments (1). Our previous findings demonstrated an inhibition of ovarian cancer cell growth at 2-4 μM in four cis-platin sensitive and resistant ovarian cancer cell-lines, i.e. A2780 and A2780/CP, 2008 and C13. The level of inhibition caused by LR-peptide is similar to that of paclitaxel, a first line drug in OC therapy. Mass spectrometry differential proteomic studies were performed on A2780 OC cell models in which untreated cells were examined in comparison with LR-peptide treated cells (2). Bioinformatic analysis of the results led to the identification of at least 10 proteins that were modulated upon LR-peptide treatment. Among them TRAP1 an antiapoptotic mitochondrial HSP75 protein was observed. This result was the starting point for a deeper understanding of the role of the combined TRAP1/DHFR/hTS modulation in ovarian cancer cells. We first studied the role of TRAP1 alone in OC cell models. We treated a panel of ovarian cancer cells (including IGROV1 and COV504) with paclitaxel, whose cytotoxic activity involves the activation of ER (endoplasmic reticulum) stress pathways. TRAP1 shows a protective role from ER stress, as reported previously (3). We observed that upon 1-hour exposure to a sub-lethal concentration of paclitaxel, cells expressing higher TRAP1 levels (IGROV1) showed a weak activation of ER stress sensors. such as phospho PERK, phospho eIf2alpha and Grp78/BiP. COV504 cell line express lower level of TRAP1, are more sensitive to paclitaxel treatment, showing an hyperactivation of ER stress markers. Higher concentrations of paclitaxel for longer times led to apoptotic processes as confirmed by stronger activation of the ER-stress induced caspase 12, in COV cells than in IGROV cell line. These data confirm the protective role of TRAP1 against paclitaxel induced ER stress, offering a possible mechanism of drug resistance in ovarian cancer.
1.Cardinale D. et al., PNAS 2011.
2. Proceedings EPS32 Athen 2013 p_466
3. Amoroso MR, et al.Cell Death Differ. 2012
Acknowledgement
This work has been supported by AIRC-DROC 10474 project to MPC
Metodo per la funzionalizzazione sito specifica di molecole proteiche
No full textViene qui fornito un metodo in soluzione per le modifiche chimiche sito-specifiche di proteine, sfruttando l’affinità di cavità strutturate di proteine (come le cavità catalitiche degli enzimi) verso miscele di inibitori (ir)reversibili/(co)substrati o ligandi per mascherare i residui coinvolti nella formazione del complesso, permettendo la funzionalizzazione chemoselettiva di residui opportunamente selezionati al di fuori della tasca. Pertanto, grazie al mascheramento di residui di cavità strutturate, è possibile studiare ed esplorare siti allosterici a bassa affinità sia chimicamente, come porzioni funzionalizzabili, che funzionalmente. Alcune delle applicazioni in campo diagnostico, analitico e terapeutico dell’enzima ingegnerizzato risultante vengono qui discusse. Questa piattaforma di coniugazione potrebbe portare alla progettazione di un kit di coniugazione per eseguire modifiche chimiche sito-specifiche di proteine per applicazioni di ricerca, diagnostiche e terapeutiche
Storiografia civile genovese: l'opera di Filippo Casoni
No full textStudio sulla figura e l'opera dello storico genovese Filippo Cason
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
An HPLC and UHPLC-HRMS approach to study PSMA-11 instability in aqueous solution
No full textBackgroundThe stability of precursors and reagents is of utmost importance for developing a robust radiolabelling method that provides high and constant radiochemical yield and radiochemical purity.While performing the QC of the [Ga-68]Ga-PSMA-11 injectable solutions according to Ph. Eur. Monograph that has recently been published, a trend to the instability of the standard PSMA-11, the same used as a precursor for [Ga-68]Ga-PSMA-11 radiosynthesis, has been observed. This instability led to the formation of a side product in a time-dependent manner. The formation of this compound, besides making the implementation of the Ph. Eur. analytical method more difficult, negatively influenced the radiochemical yield and the radiochemical purity by increasing gallium-68 in colloidal and ionic forms.ResultsThe nature of the side product was investigated by adding chelators, such as EDTA, to PSMA-11 solutions and using the combination of UHPLC-HRMS. The results led to the definition of the side product structure, as Fe-nat-PSMA-11, from the combination of the high-affinity chelator HBED-CC, present in the molecule of PSMA-11, and environmental Fe (III).ConclusionsStrategies to reduce the risk of low radiolabeling yields and to increase the stability of the PSMA-11 in an aqueous solution were also discussed
Fragment Based Discovery of Thymidylate Synthase Dimeric Interface Inhibitors Through Mass Spectrometry. Invited lecture to the Fragment Based Lead Discovery track.
No full textFragment-based drug design has been applied to Thymidylate synthase.The strategy applied is Mass Spectrometry related. identification of small molecule library that can bind to the protein represents the starting point for further drug design of a novel class of TS inhibitors with high potential as anticancer agents
EVALUATION OF LR STABILITY BY LC CHIP Q-TOF AND QUANTITATIVE DETERMINATION OF LR PEPTIDE CELL PENETRATION BY LC-MS/MS
No full textIn the present work the degradation profile of an anticancer peptide in different biological matrixes like DMEM
(Dulbecco’s Modified Eagle Medium) and cell lysates by LC Chip Q-TOF was shown. Subsequently, an LC-MS/MS method
for the quantitative analysis of LR in cell lysates was developed and fully validated
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