Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Efecto de las Estatinas sobre el Peptidoma Plasmático y Urinario de Pacientes Trasplantados Renales

Estudiem l’efecte de l’atorvastatina sobre perfil lipídic, funció renal, marcadors d’inflamació, peptidoma plasmàtic i urinari de 54 pacients trasplantats renals. L’estudi proteòmic es basa en tecnologia d’esferes magnètiques combinada amb espectrometria de masses MALDI-TOF. L’atorvastatina va millorar el perfil lipídic dels pacients i va provocar canvis significatius al peptidoma plasmàtic. Es va observar la disminució en plasma de pèptids derivats de fragments del quininogen i de C4. Atès que bradiquinina i C4a, que s’alliberen de quininogen i C4, son potents mediadors de la inflamació, els nostres resultats poden aclarir els efectes antiinflamatoris atribuïts a les estatines.

Análisis del perfil de péptidos en sangre y orina en pacientes con nefropatía IgA y su asociación con las lesiones histológicas según la clasificación de Oxford

La nefropatía IgA (NIgA) es causa importante de insuficiencia renal crónica. Se propone estudiar el perfil proteómico en sangre/orina de pacientes con NIgA y su asociación con las lesiones histológicas según la clasificación de Oxford. Se incluyeron pacientes con NIgA entre 2006-2009, evaluando las lesiones histológicas según la Clasificación de Oxford. Se realizó análisis proteómico mediante microesferas magnéticas y espectrometría de masas (MALDI-TOF MS). Encontramos una asociación significativa entre péptidos en sangre/orina y las lesiones histológicas. Uromodulina, alfa-1-antitripsina y bradiquinina, mostraron una asociación significativa con la lesión tubulointersticial y glomerulosclerosis. Los péptidos m/z (1769,1898,1913,1945,2378,2491,2977,3004,3389,3406,4752,5337,9289) se asociaban con una peor función renal. Palabras claves: Clasificación de Oxford, Nefropatía IgA, perfil de péptidos.

Esfingolipids i senyal•lització cel.lular: l'(E)-2-hexadecenal com a possible lípid bioactiu

L’esfingosina-1-fosfat (S1P) és un lípid bioactiu amb funcions crucials en la biologia cel•lular. Entre aquestes, la seva activitat mitogènica i citoprotectora són les més estudiades. L’S1P és catabolitzada intracel•lularment mitjançant l’esfingosina-1-fosfat liasa (SGPL1) per generar (E)-2-hexadecenal i fosforiletanolamina. L’objectiu d’aquest projecte és explorar si l’(E)-2-hexadecenal és realment un catabòlit innocu o bé si, pel seu caràcter acceptor de Michael, és capaç de reaccionar amb pèptids o proteïnes específics. Aquesta interacció podria traduïr-se en funcions biològiques determinades, algunes de les quals són possiblement atribuïdes a l’esfingosina-1-fosfat com a tal. Per poder explorar el potencials adductes proteïcs amb l’aldehid, s’han emprat, sobre cèl•lules HeLa que sobreexpressen SGPL1, sondes anàlegs a esfingosina i esfinganina (i els seus derivats fosforil•lats) que presenten una funció azida en la posició omega de la cadena esfingoide. Aquestes, mitjançant química click sense coure, s’han fet reaccionar amb una molècula que presenta un dibenzociclooctí unit a biotina DBCObiotina). Després d’aïllar les proteïnes així biotinilades amb una reïna d’estreptavidina, aquestes es van separar per electroforesi. Les bandes proteïques observades es van extreure del gel i es van digerir amb tripsina, per posteriorment analitzar els pèptids per MALDI-TOF, el que permetria l’identificació de proteïnes a partir de “peptide mass fingerprinting”. Lamentablement, a la fi d’aquest contracte, encara no s’ha pogut identificar cap proteïna que s’uneixi a l’aldehid alliberat per la reacció de l’esfingosina-1- fosfat liasa. No obstant, durant aquest temps s’ha millorat el mètode per detectar aquests adductes proteïcs. Per això, si la recerca continua en aquesta línia, properament es podria saber amb certesa si existeixen o no aquestes interaccions covalents entre determinades proteïnes i l’(E)-2-hexadecenal.

Conjugation of a Ru(II) Arene Complex to Neomycin or to Guanidinoneomycin Leads to Compounds with Differential Cytotoxicities and Accumulation between Cancer and Normal Cells

A straightforward methodology for the synthesis of conjugates between a cytotoxic organometallic ruthenium(II) complex and amino- and guanidinoglycosides, as potential RNA-targeted anticancer compounds, is described. Under microwave irradiation, the imidazole ligand incorporated on the aminoglycoside moiety (neamine or neomycin) was found to replace one triphenylphosphine ligand from the ruthenium precursor [(η6-p-cym)RuCl(PPh3)2]+, allowing the assembly of the target conjugates. The guanidinylated analogue was easily prepared from the neomycin-ruthenium conjugate by reaction with N,N′-di-Boc-N″-triflylguanidine, a powerful guanidinylating reagent that was compatible with the integrity of the metal complex. All conjugates were purified by semipreparative high-performance liquid chromatography (HPLC) and characterized by electrospray ionization (ESI) and matrix-assisted laser desorptionionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and NMR spectroscopy. The cytotoxicity of the compounds was tested in MCF-7 (breast) and DU-145 (prostate) human cancer cells, as well as in the normal HEK293 (Human Embryonic Kidney) cell line, revealing a dependence on the nature of the glycoside moiety and the type of cell (cancer or healthy). Indeed, the neomycinruthenium conjugate (2) displayed moderate antiproliferative activity in both cancer cell lines (IC50 ≈ 80 μM), whereas the neamine conjugate (4) was inactive (IC50 ≈ 200 μM). However, the guanidinylated analogue of the neomycinruthenium conjugate (3) required much lower concentrations than the parent conjugate for equal effect (IC50 = 7.17 μM in DU-145 and IC50 = 11.33 μM in MCF-7). Although the same ranking in antiproliferative activity was found in the nontumorigenic cell line (3 2 > 4), IC50 values indicate that aminoglycoside-containing conjugates are about 2-fold more cytotoxic in normal cells (e.g., IC50 = 49.4 μM for 2) than in cancer cells, whereas an opposite tendency was found with the guanidinylated conjugate, since its cytotoxicity in the normal cell line (IC50 = 12.75 μM for 3) was similar or even lower than that found in MCF-7 and DU-145 cancer cell lines, respectively. Cell uptake studies performed by ICP-MS with conjugates 2 and 3 revealed that guanidinylation of the neomycin moiety had a positive effect on accumulation (about 3-fold higher in DU-145 and 4-fold higher in HEK293), which correlates well with the higher antiproliferative activity of 3. Interestingly, despite the slightly higher accumulation in the normal cell than in the cancer cell line (about 1.4-fold), guanidinoneomycinruthenium conjugate (3) was more cytotoxic to cancer cells (about 1.8-fold), whereas the opposite tendency applied for neomycinruthenium conjugate (2). Such differences in cytotoxic activity and cellular accumulation between cancer and normal cells open the way to the creation of more selective, less toxic anticancer metallodrugs by conjugating cytotoxic metal-based complexes such as ruthenium(II) arene derivatives to guanidinoglycosides.

Conjugation of a Ru(II) Arene Complex to Neomycin or to Guanidinoneomycin Leads to Compounds with Differential Cytotoxicities and Accumulation between Cancer and Normal Cells

A straightforward methodology for the synthesis of conjugates between a cytotoxic organometallic ruthenium(II) complex and amino- and guanidinoglycosides, as potential RNA-targeted anticancer compounds, is described. Under microwave irradiation, the imidazole ligand incorporated on the aminoglycoside moiety (neamine or neomycin) was found to replace one triphenylphosphine ligand from the ruthenium precursor [(η6-p-cym)RuCl(PPh3)2]+, allowing the assembly of the target conjugates. The guanidinylated analogue was easily prepared from the neomycin-ruthenium conjugate by reaction with N,N′-di-Boc-N″-triflylguanidine, a powerful guanidinylating reagent that was compatible with the integrity of the metal complex. All conjugates were purified by semipreparative high-performance liquid chromatography (HPLC) and characterized by electrospray ionization (ESI) and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and NMR spectroscopy. The cytotoxicity of the compounds was tested in MCF-7 (breast) and DU-145 (prostate) human cancer cells, as well as in the normal HEK293 (Human Embryonic Kidney) cell line, revealing a dependence on the nature of the glycoside moiety and the type of cell (cancer or healthy). Indeed, the neomycin-ruthenium conjugate (2) displayed moderate antiproliferative activity in both cancer cell lines (IC50 ≈ 80 μM), whereas the neamine conjugate (4) was inactive (IC50 ≈ 200 μM). However, the guanidinylated analogue of the neomycin-ruthenium conjugate (3) required much lower concentrations than the parent conjugate for equal effect (IC50 = 7.17 μM in DU-145 and IC50 = 11.33 μM in MCF-7). Although the same ranking in antiproliferative activity was found in the nontumorigenic cell line (3 2 > 4), IC50 values indicate that aminoglycoside-containing conjugates are about 2-fold more cytotoxic in normal cells (e.g., IC50 = 49.4 μM for 2) than in cancer cells, whereas an opposite tendency was found with the guanidinylated conjugate, since its cytotoxicity in the normal cell line (IC50 = 12.75 μM for 3) was similar or even lower than that found in MCF-7 and DU-145 cancer cell lines, respectively. Cell uptake studies performed by ICP-MS with conjugates 2 and 3 revealed that guanidinylation of the neomycin moiety had a positive effect on accumulation (about 3-fold higher in DU-145 and 4-fold higher in HEK293), which correlates well with the higher antiproliferative activity of 3. Interestingly, despite the slightly higher accumulation in the normal cell than in the cancer cell line (about 1.4-fold), guanidinoneomycin-ruthenium conjugate (3) was more cytotoxic to cancer cells (about 1.8-fold), whereas the opposite tendency applied for neomycin-ruthenium conjugate (2). Such differences in cytotoxic activity and cellular accumulation between cancer and normal cells open the way to the creation of more selective, less toxic anticancer metallodrugs by conjugating cytotoxic metal-based complexes such as ruthenium(II) arene derivatives to guanidinoglycosides.