Person:
Marciello, Marzia

First Name

Marzia

Last Name

Marciello

URI

https://hdl.handle.net/20.500.14352/77512

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Farmacia

Department

Química en Ciencias Farmacéuticas

Area

Química Física

Identifiers

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Search Results

Now showing 1 - 10 of 24

Fine Modulation of the Catalytic Properties of Rhizomucor miehei Lipase Driven by Different Immobilization Strategies for the Selective Hydrolysis of Fish Oil
(Molecules, 2020) Yousefi, Maryam; Marciello, Marzia; Guisan, José Manuel; Fernandez-Lorente, Gloria; Mohammadi, Mehdi; Filice, Marco
Functional properties of each enzyme strictly depend on immobilization protocol used for linking enzyme and carrier. Different strategies were applied to prepare the immobilized derivatives of Rhizomucor miehei lipase (RML) and chemically aminated RML (NH2-RML). Both RML and NH2-RML forms were covalently immobilized on glyoxyl sepharose (Gx-RML and GxNH2-RML), glyoxyl sepharose dithiothreitol (Gx-DTT-RML and Gx-DTT-NH2-RML), activated sepharose with cyanogen bromide (CNBr-RML and CNBr-NH2-RML) and heterofunctional epoxy support partially modified with iminodiacetic acid (epoxy IDA-RML and epoxy-IDA-NH2-RML). Immobilization varied from 11% up to 88% yields producing specific activities ranging from 0.5 up to 1.9 UI/mg. Great improvement in thermal stability for Gx-DTT-NH2-RML and epoxy-IDA-NH2-RML derivatives was obtained by retaining 49% and 37% of their initial activities at 70 ◦C, respectively. The regioselectivity of each derivative was also examined in hydrolysis of fish oil at three different conditions. All the derivatives were selective between cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) in favor of EPA. The highest selectivity (32.9 folds) was observed for epoxy-IDA-NH2-RML derivative in the hydrolysis reaction performed at pH 5 and 4 ◦C. Recyclability study showed good capability of the immobilized biocatalysts to be used repeatedly, retaining 50–91% of their initial activities after five cycles of the reaction.
Multifunctional Silica-Based Nanoparticles with Controlled Release of Organotin Metallodrug for Targeted Theranosis of Breast Cancer
(Cancers, 2020) Ovejero Paredes, Karina; Díaz García, Diana; García Almodóvar, Victoria; Lozano Chamizo, Laura; Marciello, Marzia; Díaz-Sánchez, Miguel; Prashar, Sanjiv; Gómez-Ruiz, Santiago; Filice, Marco
Three different multifunctional nanosystems based on the tethering onto mesoporous silica nanoparticles (MSN) of different fragments such as an organotin-based cytotoxic compound Ph3Sn{SCH2CH2CH2Si(OMe)3} (MSN-AP-Sn), a folate fragment (MSN-AP-FA-Sn), and an enzyme-responsive peptide able to release the metallodrug only inside cancer cells (MSN-AP-FA-PEP-S-Sn), have been synthesized and fully characterized by applying physico-chemical techniques. After that, an in vitro deep determination of the therapeutic potential of the achieved multifunctional nanovectors was carried out. The results showed a high cytotoxic potential of the MSN-AP-FA-PEP-S-Sn material against triple negative breast cancer cell line (MDA-MB-231). Moreover, a dose-dependent metallodrug-related inhibitory effect on the migration mechanism of MDA-MB-231 tumor cells was shown. Subsequently, the organotin-functionalized nanosystems have been further modified with the NIR imaging agent Alexa Fluor 647 to give three different theranostic silica-based nanoplatforms, namely, MSN-AP-Sn-AX (AX-1), MSN-AP-FA-Sn-AX (AX-2), and MSN-AP-FA-PEP-S-Sn-AX (AX-3). Their in vivo potential as theranostic markers was further evaluated in a xenograft mouse model of human breast adenocarcinoma. Owing to the combination of the receptor-mediated site targeting and the specific fine-tuned release mechanism of the organotin metallodrug, the nanotheranostic drug MSN-AP-FA-PEP-S-Sn-AX (AX-3) has shown targeted diagnostic ability in combination with enhanced therapeutic activity by promoting the inhibition of tumor growth with reduced hepatic and renal toxicity upon the repeated administration of the multifunctional nanodrug.
Effect of Nanoclustering and Dipolar Interactions in Heat Generation for Magnetic Hyperthermia
(Langmuir, 2016) Coral Coral, Diego Fernando; Mendoza Zélis, Pedro; Marciello, Marzia; Puerto Morales, María Del ; Craievich, Aldo; Sánchez, Francisco Homero; Fernández van Raap, Marcela Beatriz
Biomedical magnetic colloids commonly used in magnetic hyperthermia experiments often display a bidisperse structure, i.e., are composed of stable nanoclusters coexisting with well-dispersed nanoparticles. However, the influence of nanoclusters in the optimization of colloids for heat dissipation is usually excluded. In this work, bidisperse colloids are used to analyze the effect of nanoclustering and long-range magnetic dipolar interaction on the magnetic hyperthermia efficiency. Two kinds of colloids, composed of magnetite cores with mean sizes of around 10 and 18 nm, coated with oleic acid and dispersed in hexane, and coated with meso-2,3-dimercaptosuccinic acid and dispersed in water, were analyzed. Small-angle X-ray scattering was applied to thoroughly characterize nanoparticle structuring. We proved that the magnetic hyperthermia performances of nanoclusters and single nanoparticles are distinctive. Nanoclustering acts to reduce the specific heating efficiency whereas a peak against concentration appears for the well-dispersed component. Our experiments show that the heating efficiency of a magnetic colloid can increase or decrease when dipolar interactions increase and that the colloid concentration, i.e., dipolar interaction, can be used to improve magnetic hyperthermia. We have proven that the power dissipated by an ensemble of dispersed magnetic nanoparticles becomes a nonextensive property as a direct consequence of the long-range nature of dipolar interactions. This knowledge is a key point in selecting the correct dose that has to be injected to achieve the desired outcome in intracellular magnetic hyperthermia therapy.
Useful Oriented Immobilization of Antibodies on Chimeric Magnetic Particles: Direct Correlation of Biomacromolecule Orientation with Biological Activity by AFM Studies
(Langmuir, 2014) Marciello, Marzia; Filice, Marco; Olea, David; Velez, Marisela; Guisan, José M.; Mateo, César
The preparation and performance of a suitable chimeric biosensor basedon antibodies (Abs) immobilized on lipase-coated magnetic particles by means of astanding orienting strategy are presented. This novel system is based on hydrophobicmagnetic particles coated with modified lipase molecules able to orient and furtherimmobilize different Abs in a covalent way without any previous site-selective chemicalmodification of biomacromolecules. Different key parameters attending the process werestudied and optimized. The optimal preparation was performed using a controlled loading(1 nmol Ab g−1chimeric support) at pH 9 and a short reaction time to recover abiological activity of about 80%. AFM microscopy was used to study and confirm the Abs-oriented immobilization on lipase-coated magnetic particles and thefinal achievement of ahighly active and recyclable chimeric immune sensor. This direct technique wasdemonstrated to be a powerful alternative to the indirect immunoactivity assay methodsfor the study of biomacromolecule-oriented immobilizations.
Immobilisation and stabilisation of β-galactosidase from Kluyveromyces lactis using a glyoxyl support
(International DairyJournal, 2013) Bernal Zuloaga, Claudia Patricia; Marciello, Marzia; Mesa Cadavid, Mónica ; Sierra, María Ligia ; Fernández-Lorente, Gloria ; Mateo González, César ; Guisan Seijas, José Manuel
β-galactosidase from Kluyveromyces lactis was covalently immobilised on a Glyoxyl Sepharose (GS) support by multi-point attachment. The enzyme immobilisation process was very efficient; the supports immobilised almost all the protein responsible for the catalytic activity in a short period of time, retaining approximately 82% of the activity in the case of the optimal immobilised preparations. Stability of the GS derivatives varied as a function of enzyme-support incubation time. The optimal immobilised preparation was produced after 2 h of incubation with the support at alkaline pH. This derivative, obtained by multi-point covalent attachment, was 100-fold more stable at pH 7 and 50 °C than the cyanogen bromide Sepharose derivative obtained by a one-point covalent immobilisation method. Stabilisation was also observed under a wide range of experimental conditions. This method allowed the immobilisation of 9000 IU enzyme g−1 of support, resulting in highly active and stable derivatives suitable for industrial processes.
Hydrolysis of fish oil by hyperactivated rhizomucor miehei lipase immobilized by multipoint anion exchange
(Biotechnology progress, 2011) Filice, Marco ; Marciello, Marzia; Betancor, Lorena ; Carrascosa Santiago, Alfonso Vicente ; Guisan Seijas, José Manuel; Fernández‐Lorente, Gloria
Rhizomucor miehei lipase (RML) is greatly hyperactivated (around 20- to 25-fold toward small substrates) in the presence of sucrose laurate. Hyperactivation appears to be an intramolecular process because it is very similar for soluble enzymes and covalently immobilized derivatives. The hyperactivated enzyme was immobilized (in the presence of sucrose laurate) on cyanogen bromide-activated Sepharose (very mild covalent immobilization through the amino terminal residue), on glyoxyl Sepharose (intense multipoint covalent immobilization through the region with the highest amount of Lys residues), and on different anion exchangers (by multipoint anionic exchange through the region with the highest density of negative charges). Covalent immobilization does not promote the fixation of the hyperactivated enzyme, but immobilization on Sepharose Q retains the hyperactivated enzyme even in the absence of a detergent. The hydrolysis of fish oils by these hyperactivated enzyme derivatives was sevenfold faster than by covalently immobilized derivatives and three and a half times faster than by the enzyme hyperactivated on octyl-Sepharose. The open structure of the hyperactivated lipase is fairly exposed to the medium, and no steric hindrance should interfere with the hydrolysis of large substrates. These new hyperactivated derivatives seem to be more suitable for hydrolysis of oils by RML immobilized inside porous supports. In addition, the hyperactivated derivatives are fairly stable against heat and organic cosolvents. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011.
Project number: 270
Jugando con métodos interactivos para incentivar el aprendizaje de la física y fisicoquímica
(2023) Marciello, Marzia; Alvear Jiménez, Alexis Fernando; Contreras Cáceres, Rafael; Díaz García, Diana; Fernández Fernández, María de la Cabeza; Filice, Marco; García Almodóvar, Victoria; Gómez Ruiz, Santiago; Laurenti, Marco; Lozano Chamizo, Laura; Méndez Gonzalez, Diego; Ovejero Paredes, Karina; Sanchez Sanchez, Visitación
La sociedad actual se presenta cada día más tecnificada y los estudiantes están creciendo en ella desarrollando distintas capacidades cognitivas que resultan casi imprescindible comprender y atender para poder aumentar su interés en el aprendizaje de diferentes temas. Esta realidad está llevando los profesores a reconfigurar sus estrategias pedagógicas, actualizando los métodos educativos con el fin de mejorar la calidad de la educación adaptando los contenidos que se imparten con las necesidades de la sociedad. Por ello, cada vez es más frecuente recurrir a nuevas herramientas como las TIC (Tecnologías de la Información y las Comunicaciones) y a aplicaciones lúdicas que apoyen y fomenten el proceso de aprendizaje. La aplicación de las TICs en la enseñanza, ha supuesto un avance en la transmisión y recepción del conocimiento para incentivar a los alumnos al aprendizaje de aquellas asignaturas que suponen una mayor dificultad durante su proceso formativo. Asimismo, la gamificación, que consiste en incorporar elementos de juegos como las recompensas y la competición para animar a las estudiantes a realizar tareas que normalmente se consideran aburridas, está ganando cada día más atención sobre todo gracias a su innegable carácter motivacional. De hecho, el juego es un activador en la atención y surge como alternativa para complementar los esquemas de la enseñanza tradicional. Debido a su naturaleza intrínseca, el aprendizaje de la física y de la fisicoquímica (tanto en campo farmacéutico como en general) suele resultar un proceso complicado y muchas veces difícil de entender. Los métodos tradicionales de enseñanza de la ciencia en general, y de la física y fisicoquímica en particular, hacen que estas disciplinas fundamentales sean vistas por los alumnos como algo abstracto y poco estimulante. El objetivo de la actividad propuesta en este proyecto es convertir el estudio de dichas materias en algo más interesante y participativo, motivando a los estudiantes mediante juegos basados en herramientas TIC y generando finalmente un mejor conocimiento gracias a la competición lúdica, al aprendizaje colaborativo y a la autoevaluación. En este sentido, nuestra propuesta se ha centrado en actividades interactivas basadas en la gamificación para el desarrollo de un aprendizaje proactivo de asignaturas complejas permitiendo al mismo tiempo una autoevaluación tanto a los estudiantes como a los profesores. De hecho, los alumnos, gracias a la resolución de preguntas en forma de juego han podido valorar su nivel de aprendizaje durante el desarrollo del curso y el profesor ha podido comprobar si sus clases son los suficientemente claras y/o cuales argumentos debería de profundizar y/o explicar de otra manera para obtener una mayor comprensión por parte de los estudiantes.
Thermally sensitive gels based on chitosan derivatives for the treatment of oral mucositis
(European Journal of Pharmaceutics and Biopharmaceutics, 2010) Rossi, S. ; Marciello, Marzia; Bonferoni, M.C. ; Ferrari, F. ; Sandri, G. ; Dacarro, C. ; Grisoli, P. ; Caramella, C.
The aim of the present work was the development of a thermally sensitive mucoadhesive gel based on chitosan derivatives for the treatment of oral mucositis. Trimethyl chitosan (TMC) and methylpyrrolidinone chitosan (MPC) were considered. They were mixed with glycerophosphate (GP) according to different polymer/GP molar ratios and characterized for gelation properties by means of rheological analysis in comparison with chitosan. The influence of molecular weight and substitution degree (SD) of TMC on gelation temperature and time was investigated. The mucoadhesive properties of the mixtures were also assessed using porcine buccal mucosa. The best properties were shown by TMC with high MW and low SD mixed with GP according to 1:2 molar ratio. Such mixture was loaded with benzydamine hydrochloride, an anti-inflammatory drug with antimicrobial properties and subjected to in vitro drug release and wash away test. The formulation based on TMC/GP mixture was able to prolong drug release and to withstand the removal physiological mechanisms. The antimicrobial properties of both vehicle and formulation were investigated. Also in absence of drug, TMC/GP mixture was characterized by antimicrobial properties.
Synthesis of heterogeneous enzyme–metal nanoparticle biohybrids in aqueous media and their applications in C–C bond formation and tandem catalysis
(Chemical Communications, 2013) Filice, Marco; Marciello, Marzia; Puerto Morales, María Del ; Palomo, José María
The straightforward synthesis of novel enzyme–metalNP nanobiohybrids in aqueous medium was developed. These new nanobiohybrids were excellent multivalent catalysts combining both activities in various sets of synthetic reactions even at ultra-low concentrations (ppb amount).
Attomolar detection of hepatitis C virus core protein powered by molecular antenna-like effect in a graphene field-effect aptasensor
(2023) Palacio, Irene; Moreno, Miguel; Náñez, Almudena; Purwidyantri, Agnes; Domingues, Telma; Cabral, Patricia D.; Borme, Jérôme; Marciello, Marzia; Mendieta-Moreno, Jesús Ignacio; Torres-Vázquez, Beatriz; Martínez, José Ignacio; López, María Francisca; García-Hernández, Mar; Vázquez, Luis; Jelínek, Pavel; Alpuim, Pedro; Briones, Carlos; Martín-Gago, José Ángel
Biosensors based on graphene field-effect transistors have become a promising tool for detecting a broad range of analytes. However, their performance is substantially affected by the functionalization protocol. In this work, we use a controlled in-vacuum physical method for the covalent functionalization of graphene to construct ultrasensitive aptamer-based biosensors (aptasensors) able to detect hepatitis C virus core protein. These devices are highly specific and robust, achieving attomolar detection of the viral protein in human blood plasma. Such an improved sensitivity is rationalized by theoretical calculations showing that induced polarization at the graphene interface, caused by the proximity of covalently bound molecular probe, modulates the charge balance at the graphene/aptamer interface. This charge balance causes a net shift of the Dirac cone providing enhanced sensitivity for the attomolar detection of the target proteins. Such an unexpected effect paves the way for using this kind of graphene-based functionalized platforms for ultrasensitive and real-time diagnostics of different diseases.