Polimorfismo dei geni GSTP1, NQO1 e MDR1 e risposta alla chemioterapia nel trattamento del Mieloma Multiplo.

Il Mieloma Multiplo (MM) è una neoplasia con un’incidenza annua è di tre nuovi casi per 100.000 abitanti, mentre l’età media di insorgenza è di 70 anni, con una prevalenza nel sesso maschile. La prognosi dei pazienti di MM è piuttosto variabile e la sopravvivenza oscilla da 20 a 60 mesi. La terapia varia a secondo dello stadio clinico della malattia, ma uno dei regimi maggiormente usati è denominato DAV o VAD-like (dal nome dei farmaci impiegati, che sono vincristina, adriblastina e desametasone), che produce una buona risposta, senza danneggiare le cellule sane del midollo osseo, che sono destinate all’autotrapianto. Tuttavia una discreta percentuale di pazienti risulta refrattario alle dosi standard, e possono manifestare effetti collaterali, anche molto gravi. La variabilità interindividuale nella risposta clinica e nella sensibilità alla tossicità del farmaco, comune a molti regimi chemioterapici, può essere associata ad alterazioni genetiche, tra cui alcune mutazioni puntiformi (SNPs) in grado di alterare i livelli delle proteine coinvolte nel meccanismo d’azione del farmaco o nella sua biotrasformazione. La relazione tra la risposta alla chemioterapia e il genotipo del paziente può fornire utili informazioni che possono essere usate per predire l’efficacia de trattamento e/o la tossicità. Lo scopo del presente lavoro di tesi è di valutare la risposta individuale alla terapia DAV, in pazienti affetti da MM, in relazione al polimorfismo genetico di due enzimi metabolici Glutatione-S-Trasferasi-P1 (GSTP1 105Val) e NAD(P)H-Chinone:Ossidoreduttasi (NQO1 ProSer187) e del trasportatore ABCB1 (C3435T). Il GSTP1 appartiene alla famiglia delle Glutatione-S-Transferasi, un gruppo di enzimi della fase II di detossificazione, attraverso la coniugazione con il glutatione, di numerosi xenobiotici e chemioterapici. La variante GSTP1 105Val è associata ad una bassa stabilità termica ed ad una alterata attività catalitica substrato-specifica rispetto all’allele selvatico. Il gene NQO1 codifica per la NAD(P)H-Chinone:Ossidoreduttasi, un flavoenzima citosolico di fase I nel metabolismo degli xenobiotici e la sua variante 187Ser è associata ad una perdita di attività catalitica. Il gene MDR1 (o ABCB1) codifica per la glicoproteina-P (gp-P), un membro della superfamiglia dei trasportatori di membrana (ABC). La gp-P è coinvolta nel trasporto attivo di un ampio numero di molecole anfipatiche attraverso le membrane lipidiche. Il principale ruolo fisiologico è di protezione della cellula, e quindi dell’organismo, contro gli xenobiotici compresi i farmaci, come ad esempio le antracicline e gli alcaloidi della vinca. Il polimorfismo C3435T di MDR1 è la sola variante che è stata associata ad un’espressione alterata della proteina in vari tessuti e, generalmente, risulta in una diminuzione dei livelli di espressione. Il gruppo di pazienti reclutati prevalentemente dall’U.O. di Ematologia dell’Università Pisana, con la collaborazione delle U.O. di Parma e Reggio Calabria, è composto da 115 individui affetti da MM e sottoposti a regime DAV. Dei partecipanti allo studio sono state raccolte le informazioni sulle variabili demografiche (età, sesso) e cliniche (diagnosi, trattamento e follow-up) e campioni di sangue, da cui è stato estratto il DNA per l’analisi del genotipo con la metodica Taqman. L’analisi del genotipo non ha messo in evidenza alcun effetto significativo per i polimorfismi di NQO1 e MDR1 in relazione alla risposta alla chemioterapia, al trapianto e alla sopravvivenza a 30 mesi. Mentre la variante GSTP1 105Val risulta statisticamente associata con un esito peggiore della chemioterapia di induzione, ma non ha effetto sulla sopravvivenza. Questo risultato indica che GSTP1 potrebbe essere un "marker predittivo" della risposta a DAV nel trattamento del Mieloma Multiplo

IL MOVIMENTO DELL’APPARIRE TRA ESSERE E NULLA: LE RADICI FENOMENOLOGICHE DELLA CONCEZIONE DELLA VITA DI EUGEN FINK

This paper investigates some essential steps in Eugen Fink’s conception of life, in order to show its phenomenological roots. First of all, it is explained how the conception of individual life interpreted as self-understanding builds the foundations of the ontological project movement, which consists precisely in the vitality - i.e. the problematization towards cosmological difference - of those thoughts of Being that constitute our pre-comprehension. Secondly, the structure of the movement of the world is described in its main characteristics; moreover, the relationship between the life of the individual as ontological understanding and the life of the world as a continuous process of appearing is deepened. Finally, an attempt is made to find the mediation between the two instances, individual and cosmic, through the reference to the experience of the phenomenon of death as an index of the totality of Nothingness

The Effect of Solids Residence Time on Phosphorus Uptake in Co-precipitation Systems Targeting Low Phosphorus Concentrations

Chemical phosphorus (P) removal is generally considered to be necessary when attempting to achieve ultra low P concentrations (i.e. < 50 μg/L) in wastewater treatment. However, the impact of aging of chemical solids (i.e. hydrous ferric oxides (HFO)) and thereby solids residence times (SRT) typical of conventional wastewater treatment processes on P removal is not well understood. This study characterized the uptake of P in co-precipitation systems under varying influent P concentrations and methods of P addition (i.e. steady state versus transient dosing); identified the impact of SRT on P removal, HFO floc structure, equilibrium adsorption of P to HFO and on the dynamics of P removal under steady state and transient conditions; and described the dynamic behaviour of P sorption onto HFO floc by the development of a model. The study employed lab scale continuous flow sequencing batch reactor (SBR) pilots that received the discharge of a flash mix tank. The pilots were fed with a synthetic natural water and operated over a range of SRTs (i.e. 2.8-26.6 days). Operation at different influent P concentrations and methods of P addition (i.e. steady state vs. transient dosing) was evaluated. Batch sorption testing with solids from the SBRs was also conducted. Under steady state operating conditions the majority of P removal occurred in the flash tank (94% with 3.4 mg influent P/L; 83% with 6.4 mg influent P/L) with an additional smaller fraction of removal in the SBRs (additional 3.3 – 4.8% removal with 3.4 mg influent P/L; 5.5 - 8.8% with 6.4 mg influent P/L). Soluble P uptake was higher at SRTs ≤ 7.4 days with 3.4 mg influent P/L and SRTs ≤ 14.3 days with 6.4 mg influent P/L. The floc morphology (i.e. open vs. compact floc structure) in the flash solids was found to be different from that of the SBR aged solids using SEM analysis. Particle size distributions were found to be the same in all systems supporting the hypothesis that changes in floc morphology were more responsible for differences in P removal than floc size. Batch sorption studies indicated that fresh HFO had a higher sorption capacity in comparison to aged (2.8, 7.4, 10.8 and 22.8 day) HFO. P desorption from HFO solids was found to be negligible supporting chemisorption as the mechanism of P adsorption. P adsorption onto HFO solids was determined to be best described by the Freundlich isotherm. An equilibrium model was found to adequately describe P adsorption onto HFO solids of different ages. Modelling showed that fresh HFO contributed more to P sorption than aged HFO in each SBR implying that the fresh flash solids were more responsible for the observed P uptake in the SBRs in comparison to the aged SBR solids. Transient studies showed that P removal in the SBRs and batch sorption tests was characterized by an initial fast period of removal followed by a period of slower removal until pseudo-equilibrium was reached. A model was developed to describe the dynamic behaviour of P sorption onto three different solid types (i.e. steady state dosed SBR solids, transient dosed SBR solids, and preformed batch solids). Overall, the calibrated model was found to provide a good description of P removal in the SBRs and batch testing. The model was able to reflect the different process conditions (i.e. mixing) in experiments conducted in the SBRs and the batch tests. This was reflected in the estimated values of the rate coefficients obtained from the batch tests and the SBRs. It was found that the same rate coefficient could be used to describe P adsorption onto HFO floc of different ages (i.e. 2.8-26.6 days)

A Combination of Aqueous Extraction and Polymeric Membranes as a Sustainable Process for the Recovery of Polyphenols from Olive Mill Solid Wastes

Polyamide commercial membranes in flat-sheet configuration and with molecular weight cut-o (MWCO) in the range of ultrafiltration (UF) to nanofiltration (NF) were tested for the recovery of phenolic compounds fromclarified olive mill solid waste (OMSW) aqueous extracts. The performance of selected membranes was evaluated in terms of productivity (permeate flux) and selectivity towards biologically active compounds (such as phenolic compounds, flavanols, and hydroxycinnamic acids derivatives) and total antioxidant activity (TAA) as a function of transmembrane pressure (TMP). NF membranes produced higher permeate fluxes and a lower fouling index in comparison with UF membranes. Retention of bioactive compounds was also significantly higher for NF membranes than for UF membranes. In particular, membranes with MWCO in the range 150–500 Da showed rejection towards flavanols and hydroxycinnamic acid derivatives of about 100%. On the other hand, the rejection towards TAA and total polyphenols was of about 90% and 72%, respectively. Therefore, NF retentate fractions appear of practical interest for the production of food additives and food supplements due to their high antioxidant activit

IL MOVIMENTO DELL'APPARIRE TRA ESSERE E NULLA: LE RADICI FENOMENOLOGICHE DELLA CONCEZIONE DELLA VITA DI EUGEN FINK

This paper investigates some essential steps in Eugen Fink's conception of life, in order to show its phenomenological roots. First of all, it is explained how the conception of individual life interpreted as self-understanding builds the foundations of the ontological project movement, which consists precisely in the vitality - i.e. the problematization towards cosmological difference - of those thoughts of Being that constitute our pre-comprehension. Secondly, the structure of the movement of the world is described in its main characteristics; moreover, the relationship between the life of the individual as ontological understanding and the life of the world as a continuous process of appearing is deepened. Finally, an attempt is made to find the mediation between the two instances, individual and cosmic, through the reference to the experience of the phenomenon of death as an index of the totality of Nothingness

Purification of artichoke polyphenols by using membrane filtration and polymeric resins

The present study aimed at evaluating the potential of an integrated process based on the use of membrane technology and adsorbent resins for the recovery, concentration and purification of phenolic compounds from artichoke wastewaters. In particular, artichoke wastewaters coming from the blanching step were pre-treated by ultrafiltration (UF) in order to remove suspended solids and macromolecular compounds. The UF permeate was submitted to a nanofiltration (NF) process producing a concentrated fraction enriched in phenolic and sugar compounds. Three different macroporous resins were tested through adsorption/desorption methods to produce purified phenolic fractions with high antioxidant activity. Samples produced in UF, NF and adsorption desorption tests were assayed for phenolic composition (chlorogenic acid and apigenin 7-O-glucoside), sugar composition (fructose, glucose and sucrose) and antioxidant activity. Among the three different tested resins, the S 7968 offered the best performance in terms of adsorption/desorption ratio for chlorogenic acid, with a total adsorption/desorption yield (TADY) of 63.39%; for the apigenin 7-O-glucoside the S 7968 and the S 2328 resins showed a TADY in the range 68.31-78.45%. (c) 2015 Elsevier B.V. All rights reserved.Authors acknowledge the Vicerrectorado de Investigacion of the "Universitat Politecnica de Valencia" for the financial support (project 1965) from the call "Proyectos de Nuevas Lineas de Investigacion Mul-tidisciplinares (PAID05-11)".Conidi, C.; Rodríguez López, AD.; Garcia-Castello, EM.; Cassano, A. (2015). Purification of artichoke polyphenols by using membrane filtration and polymeric resins. Separation and Purification Technology. 144(1):153-161. https://doi.org/10.1016/j.seppur.2015.02.025S153161144

Food industry by-products valorization and new ingredients: cases of study

The concern about food and beverages is gaining importance for the general public in terms of health and more environmentally sustainable food products. Healthy foods imply the awareness on their safety, nutritional characteristics, and the potential inclusion of nutritive complements such as antioxidants, vitamins, and proteins, which promote a benefit to the consumer's health. Also, organic foods, with less added chemicals such as pesticides, are more demanded recently. The environmentally sustainable food production has to reconsider the wastes as by-products that can be transformed to provide valuable compounds (antioxidants, fiber, fuels, etc.) that could be used as new products or raw materials in the food industry or even applied in other sectors such as pharmaceutical, polymer, and energy industries. In this chapter, selected successful case studies in which food wastes are transformed into new products by using different separation and purification technologies will be shown. Furthermore, the use of different wild vegetables from natural environments as a source of valuable compounds and new ingredients will be described.info:eu-repo/semantics/publishedVersio

Measurement of the tt production cross-section in the ⊤ + jets final state at √s = 7 TeV with the ATLAS detector at the LHC

This thesis contributes to the area of top physics with the measurement of the top pairs (t-tbar) production cross-section via strong interaction in proton-proton collisions at the LHC. The aim is to measure the t-tbar production cross-section in the semi-leptonic decay channel with in the final state a hadronically decaying tau lepton (t-tbar → τ + jets). This represents one of the most challenging experimental final state, due to the difficulty of reconstructing and identifying the hadronically decaying τ and due to the presence of more than one neutrino as the source of missing transverse energy (ETmiss ). The final state contains various additional jets, two of them originating from a b-quark. Jet reconstruction with precise jet energy scale estimation and efficient and well calibrated tagging of b-quark jets constitute other challenging experimental issues. The measurement is done using 2.05 fb-1 of LHC data produced at the center-of-mass energy of √s = 7 TeV, collected by the ATLAS detector during 2011. The cross-section is extracted with a profile likelihood fit of the transverse mass of the leptonically decaying W, combining the information of the 1-prong tau, 3-prong tau and electron channels. The cross-section in the tau and electron channel can vary independently. Systematic uncertainties are implemented as nuisance parameters in the fit and are constrained by the data improving the precision of the measurement. The result of the t-tbar → τ + jets cross-section measurement is : σ t-tbar→ τ +jets = 205 ± 11 (stat) ± 39 (syst) pb. It is compatible with the result of another measurement of the t-tbar → τ + jets cross-section at √s =7 TeV done with a with a sample of 1.7 fb-1 and based on a very different technique. Our measurement achieves a better precision with a relative error about 30% smaller. Both measurements are compatible with the theoretical calculation. We measured also the cross-section in the t-tbar → e + jets channel, which resulted in the value of σ t-tbar → e+jets = 178 ± 14 (stat+syst) pb , in good agreement with the ATLAS combined measurement in the electron and muon channels. The ratio of the t-tbar cross-section in the electron and the tau + jets channels is measured to be 0.86 +0.13 -0.11 compatible with 1 as expected in the Standard Model. As future perspective, we discussed possible improvements of the method developed in this thesis that could be achieved with a higher statistics sample like the 25 fb-1 of 8 TeV data.La presente tesis constituye una aportación a la área de la física del quark top proporcionando la medida de la sección eficaz de producción de pares top-antitop (t-tbar) vía interacción fuerte en colisiones protón-protón en el LHC. El objetivo de la tesis es él de medir la sección eficaz de producción de t-tbar en el canal de desintegración semi-leptónico con en el estado final un leptón tau que a su vez se desintegra de manera hadrónica (t-tbar → τ + jets). Este representa uno de los estados finales más desafiantes desde el punto de vista experimental, debido a la dificultad de reconstruir y identificar el leptón τ en su modo de desintegración hadrónico y debido también a la presencia de más de un neutrino como fuente de la energía transversa faltante (ETmiss ). Además, el estado final consta de otros jets, dos de los cuales producidos por quarks de tipo b. La reconstrucción de los jets con una estimación precisa de su escala de energía y la puesta a punto de un método para etiquetar los jets de tipo b (b-tagging) que sea eficiente y bien calibrado, constituyen otro desafío experimental. La medida está hecha con 2.05 fb-1 de datos del LHC producidos a la energía de centro de masa de √s = 7 TeV y coleccionados por el detector ATLAS durante el 2011. La sección eficaz está extraída con un profile likelihood fit de la masa transversa del bosón W que se desintegra leptónicamente, combinando la información de los canales de tau 1-prong y 3-prong y de electrón. La sección eficaz de los canales de tau se deja variar de manera independiente con respecto a la sección eficaz del canal de electrón. Los errores sistemáticos están implementados como nuisance parameters en el fit y están constreñidos por los datos: de tal manera se obtiene una mejora en la precisión de la medida. El resultado de la medida de la sección eficaz del proceso t-tbar → τ + jet es: σ t-tbar→ τ +jets = 205 ± 11 (stat) ± 39 (syst) pb. Este resultado es compatible con él de otra medida de sección eficaz del mismo estado final a √s =7 TeV hecho con una muestra de 1.7 fb-1 y basado en un procedimiento muy distinto. Nuestra medida alcanza una mejor precisión con un error relativo un 30% aproximadamente más bajo. Por otra parte, ambas medidas son compatibles con el valor calculado teóricamente. Hemos medido también la sección eficaz del canal t-tbar → e + jets, la cual resulta: σ t-tbar → e+jets = 178 ± 14 (stat+syst) pb , en buen acuerdo con la medida combinada de ATLAS en los canales de electrón y muón. El ratio de la sección eficaz de t-tbar en el canal de electrón y de tau resulta ser 0.86 +0.13 -0.11 , lo cual es compatible con 1, como se espera en el Modelo Estándard. Con respecto a las perspectivas para el futuro, se ha discutido una posible mejora del método desarrollado en esta tesis que se podría conseguir con una muestra con estadística más elevada, por ejemplo la muestra de 25 fb-1 de datos producidos a la energía de centro de masa de 8 TeV

Zeb2 DNA-Binding Sites in Neuroprogenitor Cells Reveal Autoregulation and Affirm Neurodevelopmental Defects, Including in Mowat-Wilson Syndrome

Functional perturbation and action mechanism studies have shown that the transcription factor Zeb2 controls cell fate decisions, differentiation, and/or maturation in multiple cell lineages in embryos and after birth. In cultured embryonic stem cells (ESCs), Zeb2’s mRNA/protein upregulation is necessary for the exit from primed pluripotency and for entering general and neural differentiation. We edited mouse ESCs to produce Flag-V5 epitope-tagged Zeb2 protein from one endogenous allele. Using chromatin immunoprecipitation coupled with sequencing (ChIP-seq), we mapped 2432 DNA-binding sites for this tagged Zeb2 in ESC-derived neuroprogenitor cells (NPCs). A new, major binding site maps promoter-proximal to Zeb2 itself. The homozygous deletion of this site demonstrates that autoregulation of Zeb2 is necessary to elicit the appropriate Zeb2-dependent effects in ESC-to-NPC differentiation. We have also cross-referenced all the mapped Zeb2 binding sites with previously obtained transcriptome data from Zeb2 perturbations in ESC-derived NPCs, GABAergic interneurons from the ventral forebrain of mouse embryos, and stem/progenitor cells from the post-natal ventricular-subventricular zone (V-SVZ) in mouse forebrain, respectively. Despite the different characteristics of each of these neurogenic systems, we found interesting target gene overlaps. In addition, our study also contributes to explaining developmental disorders, including Mowat-Wilson syndrome caused by ZEB2 deficiency, and also other monogenic syndromes.</p

Transcriptional repressor ZEB2 promotes terminal differentiation of CD8⁺ effector and memory T cell populations during infection

ZEB2 is a multi-zinc-finger transcription factor known to play a significant role in early neurogenesis and in epithelial-mesenchymal transition-dependent tumor metastasis. Although the function of ZEB2 in T lymphocytes is unknown, activity of the closely related family member ZEB1 has been implicated in lymphocyte development. Here, we find that ZEB2 expression is up-regulated by activated T cells, specifically in the KLRG1(hi) effector CD8(+) T cell subset. Loss of ZEB2 expression results in a significant loss of antigen-specific CD8(+) T cells after primary and secondary infection with a severe impairment in the generation of the KLRG1(hi) effector memory cell population. We show that ZEB2, which can bind DNA at tandem, consensus E-box sites, regulates gene expression of several E-protein targets and may directly repress Il7r and Il2 in CD8(+) T cells responding to infection. Furthermore, we find that T-bet binds to highly conserved T-box sites in the Zeb2 gene and that T-bet and ZEB2 regulate similar gene expression programs in effector T cells, suggesting that T-bet acts upstream and through regulation of ZEB2. Collectively, we place ZEB2 in a larger transcriptional network that is responsible for the balance between terminal differentiation and formation of memory CD8(+) T cells