Σύνθεση βιοδραστικών παραγώγων χαλκόνης με φαρμακολογικό ενδιαφέρον

Στην παρούσα εργασία μελετάται η σύνθεση παραγώγων χαλκόνης. Αρχικά, εξηγούνται τι είναι οι χαλκόνες, τα δομικά τους χαρακτηριστικά και η φυσική τους αφθονία. Στη συνέχεια γίνεται μία βιβλιογραφική επισκόπηση των μεθόδων σύνθεσής τους, περιγράφονται οι βιολογικές δράσεις τους και παρουσιάζονται γνωστά παράγωγα με θεραπευτικές ιδιότητες. Στο τελευταίο μέρος της εργασίας, περιγράφεται η σύνθεση χαλκονών και στη συνέχεια, η σύνθεση, από αυτά, νέων παραγώγων χαλκόνης. Συγκεκριμένα, τα παράγωγα που συντέθηκαν ανήκουν στις κατηγορίες των θειοσεμικαρβαζονών, θειοκαρβοϋδραζονών, θειοκαρβαμιδίων, πυραζολίων, θειονών και θειαζολίων. Τέλος, γίνεται η ταυτοποίηση των χαλκονών και των παραγώγων τους με φασματοσκοπία πυρηνικού μαγνητικού συντονισμού (NMR 1H και 13C) και φασματομετρία μάζας (MS).In the present work the synthesis of chalcone derivatives is studied. In the introductory part, chalcones’ definition, chemical structure and existence are described and their natural abundance is mentioned. Chemical synthesis, biological applications and therapeutic properties of this class of compounds are reviewed as well. Then, chalcones’ synthesis and possible reaction mechanisms are described in details along with the synthesis of numerous new derivatives, namely thiosemicarbazones, thiocarbohydrazones, thiocarbamides, pyrazoles, thiones and thiazoles. All these new derivatives are characterized by Nuclear Magnetic Resonance Spectroscopy (NMR 1H and 13C) and Mass Spectrometry (MS)

Το σώμα στη μεταβίβαση: Η συμμετοχή του στην εγκαθίδρυση της θηλυκότητας

Στην παρούσα κλινική εννοιολογική εργασία μελετάται η διαδικασία των διεργασιών προσέγγισης της θηλυκής έμφυλης ταυτότητας (female gender identity) κατά τη διάρκεια της ψυχοδυναμικής ψυχοθεραπείας. Στο θεωρητικό μέρος της εργασίας, επιλέγονται ψυχαναλυτικά κείμενα, σχετικά με τις διεργασίες διαμόρφωσης της θηλυκής έμφυλης ταυτότητας, από την πρώτη επαφή της ψυχανάλυσης με τη θηλυκή σεξουαλικότητα, έως και σήμερα. Ιδιαίτερη μνεία γίνεται στον ρόλο του πρωταρχικού αντικειμένου, δηλαδή της μητέρας, στην ψυχοσεξουαλική ανάπτυξη του κοριτσιού και τη διαμόρφωση της δικής του σχέσης με το σώμα του. Στη συνέχεια, διερευνάται η έννοια της «θηλυκής θεραπευτικής δυάδας», ως μια μορφή επαναβίωσης της ομοερωτικής σχέσης του κοριτσιού με τη μητέρα. Η ενότητα αυτή επικεντρώνεται σε κείμενα αναφορικά με την αναβίωση πρώιμων εμπειριών, συναισθημάτων και αντικειμενοτρόπων σχέσεων στη θεραπεία γυναικών από γυναίκες. Σε περιπτώσεις πρωιμότερων ασθενών δε, όπως ασθενών οριακής οργάνωσης, δύναται να εγκατασταθεί μια μορφή συμβιωτικής μεταβίβασης, ανάλογη της πρωταρχικής σχέσης, καθώς και να εκδηλωθούν στοιχεία του ψυχικού περιεχομένου της ασθενούς μέσα στις σωματικές αισθήσεις και εκδηλώσεις της θεραπεύτριας. Στο κλινικό μέρος της εργασίας, παρατίθεται υλικό από τη ψυχοδυναμική ψυχοθεραπεία νεαρής γυναίκας (Ν=1), για την οποία ζητήματα σχετικά με τη θηλυκότητα και το σώμα ανάγονται σε πρωταρχικά. Τα κλινικά δεδομένα προέρχονται αφενός από το ατομικό ιστορικό της ασθενούς και αφετέρου από γραπτές αναφορές της θεραπεύτριας, οι οποίες συντάχθηκαν κατά τη διάρκεια της θεραπείας. Το υλικό επικεντρώνεται στις σχέσεις της με τα αντικείμενα, την προοιδιπόδεια καθήλωση καθώς και σε ασυνείδητες προσυμβολικές φαντασιώσεις της, συχνά αντιληπτές μέσω των ισχυρών ενσώματων αντιμεταβιβαστικών εκδηλώσεων που προκαλούν. Η ψυχική λειτουργία της ασθενούς και οι δυσκολίες στις διεργασίες ταύτισης με το θηλυκό μητρικό αντικείμενο, θα προσεγγιστούν μέσω υποδοχής και κατανόησης της αμοιβαίας -ως επί το πλείστον σωματικής- αλληλεπίδρασης του θηλυκού θεραπευτικού ζεύγους.This conceptual-clinical study examines pathways of approaching female gender identity during psychodynamic psychotherapy processes. The theoretical framework consists of an array of psychoanalytic texts on female gender identity processes and offers a timeline of the psychoanalytic tangencies with female sexuality up to the present day. A certain emphasis is given to the role of the mother as a primary object, regarding the girl’s psychosexual development and her own body conception. Furthermore, this study explores the concept of “female therapeutic dyad” as a form of representation of a homoerotic relation between girl and mother. This part focuses on texts regarding the resurgence of primary experiences, emotions, and object-relation connotations in therapeutic relationships between female analysts and female analysands. Considering cases of particular patients, such as patients with borderline personality, a symbiotic transference corresponding to the primary relation, might be established. Additionally, fragments of the patient’s psychic content can be displayed as embodied senses and perceptions of the therapist. The clinical part of this dissertation emerges from the psychodynamic psychotherapy of a young woman (N=1) whose issues regarding body and femininity are being perceived as primary. The clinical data employed consist of the patient’s personal history along with clinical notes recorded during the therapeutic process. In particular, the clinical material focuses on the patient’s relation to objects, the pre-oedipal fixation, and her unconscious pre-symbolic phantasies, often understood through the prism of powerful embodied countertransference manifestations. The psychic function of the patient and difficulties with the female maternal object identification processes, will be approached through a reception and an intelligibility of the mutual -mostly embodied- interaction of the female therapeutic dyad

Targeting Hsp27/eIF4E interaction with phenazine compound: A promising alternative for castration-resistant prostate cancer treatment

The actual strategy to improve current therapies in advanced prostate cancer involves targeting genes activated by androgen withdrawal, either to delay or prevent the emergence of the castration-refractory phenotype. However, these genes are often implicated in several physiological processes, and long-term inhibition of survival proteins might be accompanied with cytotoxic effects. To avoid this problem, an alternative therapeutic strategy relies on the identification and use of compounds that disrupt specific protein-protein interactions involved in androgen withdrawal. Specifically, the interaction of the chaperone protein Hsp27 with the initiation factor eIF4E leads to the protection of protein synthesis initiation process and enhances cell survival during cell stress induced by castration or chemotherapy. Thus, in this work we aimed at i) identifying the interaction site of the Hsp27/eIF4E complex and ii) interfere with the relevant protein/protein association mechanism involved in castration-resistant progression of prostate cancer. By a combination of experimental and modeling techniques, we proved that eIF4E interacts with the C-terminal part of Hsp27, preferentially when Hsp27 is phosphorylated. We also observed that the loss of this interaction increased cell chemo-and hormone-sensitivity. In order to find a potential inhibitor of Hsp27/eIF4E interaction, BRET assays in combination with molecular simulations identified the phenazine derivative 14 as the compound able to efficiently interfere with this protein/protein interaction, thereby inhibiting cell viability and increasing cell death in chemo- and castration-resistant prostate cancer models in vitro and in vivo

Repair or destruction: an intimate liaison between ubiquitin ligases and molecular chaperones in proteostasis

Cellular differentiation, developmental processes, and environmental factors challenge the integrity of the proteome in every eukaryotic cell. The maintenance of protein homeostasis, or proteostasis, involves folding and degradation of damaged proteins, and is essential for cellular function, organismal growth, and viability [1, 2]. Misfolded proteins that cannot be refolded by chaperone machineries are degraded by specialized proteolytic systems. A major degradation pathway regulating cellular proteostasis is the ubiquitin/proteasome-system (UPS), which regulates turnover of damaged proteins that accumulate upon stress and during aging. Despite the large number of structurally unrelated substrates, ubiquitin conjugation is remarkably selective. Substrate selectivity is mainly provided by the group of E3 enzymes. Several observations indicate that numerous E3 ubiquitin ligases intimately collaborate with molecular chaperones to maintain the cellular proteome. In this Review, we provide an overview of specialized quality control E3 ligases playing a critical role in the degradation of damaged proteins. The process of substrate recognition and turnover, the type of chaperones they team up with, and the potential pathogeneses associated with their malfunction will be further discusse

Diagnostic, prognostic and predictive value of cell-free miRNAs in prostate cancer : A systematic review

Publisher Copyright: © 2016 Endzeliņš et al.Prostate cancer, the second most frequently diagnosed cancer in males worldwide, is estimated to be diagnosed in 1.1 million men per year. Introduction of PSA testing substantially improved early detection of prostate cancer, however it also led to overdiagnosis and subsequent overtreatment of patients with an indolent disease. Treatment outcome and management of prostate cancer could be improved by the development of non-invasive biomarker assays that aid in increasing the sensitivity and specificity of prostate cancer screening, help to distinguish aggressive from indolent disease and guide therapeutic decisions. Prostate cancer cells release miRNAs into the bloodstream, where they exist incorporated into ribonucleoprotein complexes or extracellular vesicles. Later, cell-free miRNAs have been found in various other biofluids. The initial RNA sequencing studies suggested that most of the circulating cell-free miRNAs in healthy individuals are derived from blood cells, while specific disease-associated miRNA signatures may appear in the circulation of patients affected with various diseases, including cancer. This raised a hope that cell-free miRNAs may serve as non-invasive biomarkers for prostate cancer. Indeed, a number of cell-free miRNAs that potentially may serve as diagnostic, prognostic or predictive biomarkers have been discovered in blood or other biofluids of prostate cancer patients and need to be validated in appropriately designed longitudinal studies and clinical trials. In this review, we systematically summarise studies investigating cell-free miRNAs in biofluids of prostate cancer patients and discuss the utility of the identified biomarkers in various clinical scenarios. Furthermore, we discuss the possible mechanisms of miRNA release into biofluids and outline the biological questions and technical challenges that have arisen from these studies.publishersversionPeer reviewe

Phospholipase D inhibitors reduce human prostate cancer cell proliferation and colony formation

BACKGROUND: Phospholipases D1 and D2 (PLD1/2) hydrolyse cell membrane glycerophospholipids to generate phosphatidic acid, a signalling lipid, which regulates cell growth and cancer progression through effects on mTOR and PKB/Akt. PLD expression and/or activity is raised in breast, colorectal, gastric, kidney and thyroid carcinomas but its role in prostate cancer (PCa), the major cancer of men in the western world, is unclear. METHODS: PLD1 protein expression in cultured PNT2C2, PNT1A, P4E6, LNCaP, PC3, PC3M, VCaP, 22RV1 cell lines and patient-derived PCa cells was analysed by western blotting. PLD1 protein localisation in normal, benign prostatic hyperplasia (BPH), and castrate-resistant prostate cancer (CRPC) tissue sections and in a PCa tissue microarray (TMA) was examined by immunohistochemistry. PLD activity in PCa tissue was assayed using an Amplex Red method. The effect of PLD inhibitors on PCa cell viability was measured using MTS and colony forming assays. RESULTS: PLD1 protein expression was low in the luminal prostate cell lines (LNCaP, VCaP, 22RV1) compared with basal lines (PC3 and PC3M). PLD1 protein expression was elevated in BPH biopsy tissue relative to normal and PCa samples. In normal and BPH tissue, PLD1 was predominantly detected in basal cells as well in some stromal cells, rather than in luminal cells. In PCa tissue, luminal cells expressed PLD1. In a PCa TMA, the mean peroxidase intensity per DAB-stained Gleason 6 and 7 tissue section was significantly higher than in sections graded Gleason 9. In CRPC tissue, PLD1 was expressed prominently in the stromal compartment, in luminal cells in occasional glands and in an expanding population of cells that co-expressed chromogranin A and neurone-specific enolase. Levels of PLD activity in normal and PCa tissue samples were similar. A specific PLD1 inhibitor markedly reduced the survival of both prostate cell lines and patient-derived PCa cells compared with two dual PLD1/PLD2 inhibitors. Short-term exposure of PCa cells to the same specific PLD1 inhibitor significantly reduced colony formation. CONCLUSIONS: A new specific inhibitor of PLD1, which is well tolerated in mice, reduces PCa cell survival and thus has potential as a novel therapeutic agent to reduce prostate cancer progression. Increased PLD1 expression may contribute to the hyperplasia characteristic of BPH and in the progression of castrate-resistant PCa, where an expanding population of neuroendocrine-like cells express PLD1.British Journal of Cancer advance online publication, 14 November 2017; doi:10.1038/bjc.2017.391 www.bjcancer.com

Localisation and function of the a1A-adrenoceptor in caveolae of human prostate cancer cells

Le cancer de la prostate est un réel problème de santé publique, essentiellement dû à une rechute des patients par échappement hormonal après traitement. Lors de sa progression, le cancer de la prostate, initialement androgéno-dépendant, évolue vers un état androgéno-indépendant supposant que des facteurs autres que les androgènes régulent la croissance et la survie des cellules cancéreuses. Il existe des évidences de l’implication des neurotransmetteurs et leurs récepteurs couplés aux protéines G dans le cancer de la prostate. De plus, la cavéoline-1, constituant majeur des cavéoles membranaires, a été associée à la progression maligne du cancer de la prostate. La localisation privilégiée des récepteurs couplés aux protéines G au sein des cavéoles favoriserait la transmission des signaux aboutissant à l’échappement hormonal des cellules cancéreuses androgéno-indépendantes. Je me suis alors intéressée à la localisation et au rôle du récepteur adrénergique a1A dans les cellules prostatiques androgéno-indépendantes DU145. J’ai tenté de mettre en évidence l’implication des cavéoles dans une fonction de survie et/ou de croissance du récepteur. Nos résultats démontrent que le récepteur adrénergique a1A peut être associé à la cavéoline-1, au sein de cavéoles dans les cellules androgéno-indépendantes. Nous avons aussi mis en évidence que la stimulation du récepteur participe à la protection de ces cellules vis-àvis de stimuli apoptotiques et ce via les cavéoles. Enfin, le profil d’expression du récepteur a1A et de la cavéoline-1 suggère qu’il y ait une corrélation positive avec l’état pathologique de la prostate.Prostate cancer has become a real public health issue, mainly due to patients’ relapse by hormone-refractory disease after treatment. During its progression, initially androgendependent prostate cancer evolves in an androgen-independent state supposing that factors others than androgens regulate growth and survival of cancer cells. There is growing evidence concerning the involvement of neurotransmitters and their G-protein coupled receptors in prostate cancer. Moreover, overexpression of a major caveolae constituent, caveolin-1 has been associated with aggressive prostate cancer. Privileged localisation of G-protein coupled receptors in caveolae could enhance signalling pathways leading to hormone-refractory mechanisms in androgen-independent prostate cancer cells. I was therefore interested in determining the localisation and functional role of the a1A-adrenoceptor in the DU145 androgen-independent prostate cells. I attempted to demonstrate the involvement of caveolae in a growth/survival function of this receptor. Our results showed that the a1A-adrenoceptor can be associated with caveolin-1 in caveolae of these androgen-independent cells. We also demonstrated that a1A-adrenoceptor stimulation mediated through caveolae contributes to the protection of these cells from apoptotic stimuli. Finally, the expression of a1A-adrenoceptor and caveolin-1 seems to be positively correlated with prostate pathological state

Localisation et fonction du récepteur adrénergique a1A au sein de cavéoles des cellules cancéreuses prostatiques humaines

Le cancer de la prostate est un réel problème de santé publique, essentiellement dû à une rechute des patients par échappement hormonal après traitement. Lors de sa progression, le cancer de la prostate, initialement androgéno-dépendant, évolue vers un état androgéno-indépendant supposant que des facteurs autres que les androgènes régulent la croissance et la survie des cellules cancéreuses. Il existe des évidences de l implication des neurotransmetteurs et leurs récepteurs couplés aux protéines G dans le cancer de la prostate. De plus, la cavéoline-1, constituant majeur des cavéoles membranaires, a été associée à la progression maligne du cancer de la prostate. La localisation privilégiée des récepteurs couplés aux protéines G au sein des cavéoles favoriserait la transmission des signaux aboutissant à l échappement hormonal des cellules cancéreuses androgéno-indépendantes. Je me suis alors intéressée à la localisation et au rôle du récepteur adrénergique a1A dans les cellules prostatiques androgéno-indépendantes DU145. J ai tenté de mettre en évidence l implication des cavéoles dans une fonction de survie et/ou de croissance du récepteur. Nos résultats démontrent que le récepteur adrénergique a1A peut être associé à la cavéoline-1, au sein de cavéoles dans les cellules androgéno-indépendantes. Nous avons aussi mis en évidence que la stimulation du récepteur participe à la protection de ces cellules vis-àvis de stimuli apoptotiques et ce via les cavéoles. Enfin, le profil d expression du récepteur a1A et de la cavéoline-1 suggère qu il y ait une corrélation positive avec l état pathologique de la prostate.Prostate cancer has become a real public health issue, mainly due to patients relapse by hormone-refractory disease after treatment. During its progression, initially androgendependent prostate cancer evolves in an androgen-independent state supposing that factors others than androgens regulate growth and survival of cancer cells. There is growing evidence concerning the involvement of neurotransmitters and their G-protein coupled receptors in prostate cancer. Moreover, overexpression of a major caveolae constituent, caveolin-1 has been associated with aggressive prostate cancer. Privileged localisation of G-protein coupled receptors in caveolae could enhance signalling pathways leading to hormone-refractory mechanisms in androgen-independent prostate cancer cells. I was therefore interested in determining the localisation and functional role of the a1A-adrenoceptor in the DU145 androgen-independent prostate cells. I attempted to demonstrate the involvement of caveolae in a growth/survival function of this receptor. Our results showed that the a1A-adrenoceptor can be associated with caveolin-1 in caveolae of these androgen-independent cells. We also demonstrated that a1A-adrenoceptor stimulation mediated through caveolae contributes to the protection of these cells from apoptotic stimuli. Finally, the expression of a1A-adrenoceptor and caveolin-1 seems to be positively correlated with prostate pathological state.LILLE1-Bib. Electronique (590099901) / SudocSudocFranceF