Development of enzyme-functionalized hybrid mesoporous nanodevices for advanced chemical communication

Tesis por compendio[ES] La presente tesis doctoral se centra en el diseño, síntesis y caracterización de varios nanodispositivos híbridos orgánico-inorgánicos, utilizando como soporte nanopartículas de sílice mesoporosa equipadas con enzimas y puertas moleculares, los cuales muestran capacidades comunicativas además de la evaluación de diferentes estrategias de comunicación. El primer capítulo incluye un resumen de diferentes conceptos sobre los que se fundamentan los estudios realizados tales como nanotecnología, materiales de sílice mesoporosa, materiales con puertas moleculares que reaccionan a estímulos específicos, partículas Janus y biocomputación. Finalmente, se incluyen conceptos básicos acerca de la comunicación química, materiales y estrategias empleados hasta ahora y ejemplos representativos. A continuación, en el segundo capítulo, se presentan los objetivos generales de esta tesis doctoral que son abordados en los siguientes capítulos experimentales. El tercer capítulo muestra un sistema de biocomputación para liberación basado en nanopartículas Janus de oro-sílice mesoporosa capaces de comunicarse con el entorno procesando la información e imitando la función lógica booleana propia de un demultiplexer y que resulta en la liberación controlada de la carga. Se muestra que dicho nanodispositivo puede llevar a cabo sus funciones en medios complejos como en células cancerígenas. En el cuarto capítulo, se presenta un modelo circular de comunicación dentro de una red de tres nanopartículas diferentes basado en el intercambio jerárquicamente programado de mensajes químicos. La parte mesoporosa del nanodispositivo 1 (S1βgal) es cargada con la especie fluorescente [Ru(bpy)3]Cl2 y tapada con cadenas de oligo(etilenglicol) que contienen puentes disulfuro y que funcionan como puertas moleculares, mientras que la enzima β-galactosidasa es unida a la parte del oro. En la nanopartícula 2 (S2galox), la enzima galactosa oxidasa es inmovilizada en la cara del oro mientras que la sílice mesoporosa es cargada con 4-(bromometil)benzoato de metilo y los poros tapados con un derivado de arilboronato autoinmolante sensible a H2O2 que forma un complejo huéspedanfitrión con β-ciclodextrina. Finalmente, el nanodispositivo 3 (S3est) es funcionalizado con la enzima esterasa en la parte del oro, cargada con la especie reductora hidroclururo de tris(2-carboxietil)fosfina (TCEP) en la parte mesoporosa y tapada con una nanoválvula supramolecular que responde a pH (βciclodextrina:benzimidazol). En el quinto capítulo, se muestra un modelo interactivo de comunicación química entre una nanopartícula Janus abiótica y un organismo vivo (Saccharomyces cerevisiae). En particular, el nanodispositivo está basado en nanopartículas funcionalizadas con glucosa oxidasa en la parte del oro, cargadas con el genotóxico fleomicina y tapadas con la puerta molecular sensible a pH (βciclodextrina:benzimidazol). El microorganismo usado en el estudio es una levadura modificada que expresa GFP bajo el control del promotor del gen RNR3; la transcripción de dicho gen es inducida con la exposición a agentes que dañan el ADN. La ruta de comunicación interactiva empieza con la adición de sacarosa (estímulo de entrada) la cual es hidrolizada en glucosa por la invertasa localizada en el espacio periplásmico de las levaduras y que difunde al nanodispositivo donde es trasformada en el correspondiente ácido por la glucosa oxidasa de la parte del oro. La bajada local de pH da lugar a la apertura de la nanoválvula sensible a pH del nanovehículo y con ello a la liberación de fleomicina (mensaje de vuelta) que induce la expresión de GFP (señal de salida) en las levaduras. En el sexto capítulo, proponemos una estrategia para establecer una comunicación lineal entre dos microorganismos diferentes que no interactúan entre ellos mediada por un nanodispositivo que actúa como traductor químico. Finalmente, las conclusiones generales de la presente tesis doctoral son expuestas en el capítulo siete. El estudio de las capacidades comunicativas de los nanodispositivos mesoporosos funcionalizados con enzimas permite la construcción de estrategias de cooperación entre diferentes entidades que permiten funcionalidades que van más allá que aquellas llevadas a cabo por agentes individuales.[CA] La present tesi doctoral es centra en el disseny, síntesi i caracterització de diversos nanodispositius híbrids orgànic-inorgànics, utilitzant com a suport nanopartícules de sílice mesoporosa equipades amb enzims i portes moleculars, i que mostren capacitats comunicatives a més de l’avaluació de diferents estratègies de comunicació. El primer capítol inclou un resum de diferents conceptes sobre els quals es fonamenten els estudis realitzats com ara nanotecnologia, materials de sílice mesoporosa, materials amb portes moleculars que reaccionen a estímuls específics, partícules Janus i biocomputació. Finalment, s’inclouen conceptes bàsics sobre la comunicació química, materials i estratègies utilitzades fins ara i exemples representatius. A continuació, en el segon capítol, es presenten els objectius generals d’aquesta tesi doctoral que són abordats en els següents capítols experimentals. El tercer capítol mostra un sistema de biocomputació per alliberament basat en nanopartícules Janus d’or-sílice mesoporosa capaços de comunicar-se amb l’entorn processant la informació i imitant la funció lògica booleana pròpia d’un demultiplexer i que resulta en l’alliberament controlat de la càrrega. Es mostra que aquest nanodispositiu pot dur a terme les seves funcions en mitjans complexos com en cèl·lules canceroses. En el quart capítol, es presenta un model circular de comunicació dins d’una xarxa de tres nanopartícules diferents basat en l’intercanvi jeràrquicament programat de missatges químics. La part mesoporosa del nanodispositiu 1 (S1βgal) es carrega amb l’espècie fluorescent [Ru(bpy)3]Cl2 i es tapa amb cadenes d’oligo(etilenglicol) que contenen ponts disulfur i que funcionen com portes moleculars, mentre que l’enzim β-galactosidasa s’immobilitza a la part de l’or. A la nanopartícula 2 (S2galox), l’enzim galactosa oxidasa s’immobilitza a la cara de l’or mentre que la sílice mesoporosa es carrega amb 4-(bromometil)benzoat de metil i els porus són tapats amb un derivat d’arilboronat autoimmolant sensible a H2O2 que forma un complex hoste-amfitrió amb β-ciclodextrina. Finalment, el nanodispositu 3 (S3est) es funcionalitza amb l’enzim esterasa en la part de l’or, es carrega amb l’espècie reductora hidroclurur de tris (2-carboxietil) fosfina (TCEP) a la part mesoporosa i es tapa amb una nanoválvula supramolecular que respon a pH (β-ciclodextrina:benzimidazol). En el cinqué capítol, es mostra un model interactiu de comunicació química entre una nanopartícula Janus abiòtica i un organisme viu (Saccharomyces cerevisiae). En particular, el nanodispositiu està basat en nanopartícules funcionalitzades amb glucosa oxidasa en la part de l’or, carregades amb el genotòxic fleomicina i tapades amb la porta molecular sensible a pH (βciclodextrina:benzimidazol). El microorganisme utilitzat en l’estudi és un rent modificat que expressa GFP sota el control del promotor del gen RNR3; la transcripció d’aquest gen és induïda amb l’exposició a agents que danyen l’ADN. La ruta de comunicació interactiva comença amb l’addició de sacarosa (estímul d’entrada) la qual és hidrolitzada en glucosa per la invertasa localitzada en l’espai periplasmàtic dels rents i que difon al nanodispositiu on és transformada en el corresponent àcid per la glucosa oxidasa de la part de l’or. La baixada local de pH dona lloc a l’obertura de la nanoválvula sensible a pH del nanovehicle i amb això l’alliberament de fleomicina (missatge de tornada) que indueix l’expressió de GFP (senyal de sortida) en el rent. En el sisé capítol, proposem una estratègia per establir una comunicació lineal entre dos microorganismes diferents que no interactuen entre ells facilitada per un nanodispositiu que actua com a traductor químic. Finalment, les conclusions generals de la present tesi doctoral són exposades en el capítol set. L’estudi de les capacitats comunicatives dels nanodispositius mesoporosos funcionalitzats amb enzims permet la construcció d’estratègies de cooperació entre diferents entitats que permeten funcionalitats que van més enllà que aquelles dutes a terme per agents individuals. Esperem que els resultats obtinguts inspiren aplicacions futures en diferents àrees com ara biomedicina, nanorobots, materials que imiten la naturalesa i tecnologies de la informació.[EN] This PhD Thesis is focused on the design, synthesis and characterization of several hybrid organic-inorganic nanodevices using mesoporous silica nanoparticles equipped with enzymes and molecular gates which display communication capabilities as well as the design and evaluation of different communication strategies. The first chapter includes an overview of the different concepts which lay the foundations of the presented studies such as nanotechnology, mesoporous silica materials, stimuli-responsive gated materials, Janus particles and biocomputing. Basic concepts of chemical communication, materials and enabling technologies employed so far and representative examples in this field are also included. Next, in the second chapter, the general objectives of this PhD Thesis that are addressed in the following experimental chapters are presented. The third chapter shows a biocomputing delivery system based on Janus gold-mesoporous silica nanoparticles capable of chemically communicating with the environment and processing the information mimicking a demultiplexer Boolean logic function which results in a programmed cargo release. Finally, it is shown that such nanodevice is operative in complex media such as cancer cells. In the fourth chapter, it is presented a circular model of communication within a network of three different nanoparticles based on the hierarchically programmed exchange of chemical messages. The mesoporous face of nanodevice 1 (S1βgal) is loaded with the fluorescent dye [Ru(bpy)3]Cl2 and capped with disulfidecontaining oligo(ethylene glycol) chains acting as gatekeepers, whereas the enzyme β-galactosidase is attached to the gold face. In nanoparticle 2 (S2galox), the enzyme galactose oxidase is immobilized on the Au face, while the mesoporous silica is loaded with methyl 4-(bromomethyl)benzoate and the mesopores capped with a H2O2-sensitive self-immolative arylboronate derivative which forms a host-guest complex with β-cyclodextrin. Finally, the nanodevice 3 (S3est) is functionalized with the enzyme esterase on the Au face, loaded with the reductive species tris(2- carboxyethyl)phosphine hydrochloride (TCEP) in the mesoporous face and capped with a pH-responsive supramolecular nanovalve (β-cyclodextrin:benzimidazole). In the fifth chapter, it is showed an interactive model of chemical communication between an abiotic Janus nanoparticle and a living organism (Saccharomyces cerevisiae). In particular, the nanodevice is based on Janus goldmesoporous silica nanoparticles functionalized with glucose oxidase on the Au face, loaded with the genotoxin phleomycin and capped with a pH-responsive (βcyclodextrin:benzimidazole) gatekeeper. The microorganism used in the studies is an engineered budding yeast that expresses GFP under the control of the RNR3 promoter; RNR3 gene transcription is induced upon exposure to DNA-damaging agents. The interactive communication pathway starts with the addition of sucrose (input) which is hydrolyzed into glucose by invertase located in periplasmic space of yeasts and diffuses to the nanodevice where it is transformed into the corresponding acid by glucose oxidase on the Au face. The local drop in pH leads to uncapping of the pH-sensitive nanovalve in the nanocarrier and the release of phleomycin (feedback messenger) that induces GFP expression (output) in yeasts. In the sixth chapter, we propose a strategy to establish linear communication between two different non-interacting microorganisms mediated by a nanodevice which acts as a chemical “nanotranslator”. Finally, the general conclusions from this PhD Thesis are presented in chapter seven. The study of communication capabilities of enzyme-functionalized mesoporous nanodevices enables the construction of strategies of cooperation between different entities allowing sophisticated functionalities that go beyond those carried out by individual agents. We hope that the obtained results inspire future applications in different areas such as biomedicine, nanorobots, life-like materials and information technologies.The authors wish to thank the Spanish Government (projects RTI2018-100910-B-C41 and RTI2018-101599-B-C22 (MCUI/AEI/FEDER, UE), CTQ2017-87954-P), the Generalitat Valenciana (PROMETEO 2018/024), the Comunidad de Madrid (IND2017/BMD7642) and CIBER-BBN (NANOCOMMUNITY project) for support.De Luis Fernández, B. (2021). Development of enzyme-functionalized hybrid mesoporous nanodevices for advanced chemical communication [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/171506TESISCompendi

Heparan sulfate proteoglycans undergo differential expression alterations in left sided colorectal cancer, depending on their metastatic character

Abstract Background Heparan sulfate proteoglycans (HSPGs) are complex molecules which play a role in the invasion and growth and metastatic properties of cancerous cells. In this work we analyze changes in the patterns of expression of HSPGs in left sided colorectal cancer (LSCRC), both metastatic and non-metastatic, and the results are also compared with those previously obtained for right sided tumors (RSCRCs). Methods Eighteen LSCRCs were studied using qPCR to analyze the expression of both the proteoglycan core proteins and the enzymes involved in heparan sulfate chain biosynthesis. Certain HSPGs also carry chondroitin sulfate chains and so we also studied the genes involved in its biosynthesis. The expression of certain genes that showed significant expression differences were also analysed using immunohistochemical techniques. Results Changes in proteoglycan core proteins were dependent on their location, and the main differences between metastatic and non-metastatic tumors affected cell-surface glypicans, while other molecules were quite similar. Glypicans were also responsible for the main differences between RS- and LS- malignances. Regarding the biosynthesis of heparan sulfate chains, differential alterations in transcription depending on the presence or not of metastasis affected genes involved in the modification of uronic acid (epimerization and 2-O sulfation), and some isoforms responsible for sulfation of glucosamine (NDST1, HS6ST1). Moreover, in RSCRCs differences were preferentially found in the expression of genes involved in C6 and C3 sulfation of glucosamine, but not in NDSTs or SULFs. Finally, synthesis of chondroitin sulfate showed some alterations, which affected various steps, including polimerization and the modification of chains, but the main variations dependent on the presence of metastases were epimerization and 6C sulfation; however, when compared with RSCRCs, the essential divergences affected polymerization of the chains and the 6C sulfation of the galactosamine residue. Conclusions We evidenced alterations in the expression of HSPGs, including the expression of cell surface core proteins, many glycosiltransferases and some enzymes that modify the GAG chains in LSCRCs, but this was dependent on the metastatic nature of the tumor. Some of these alterations are shared with RSCRCs, while others, focused on specific gene groups, are dependent on tumor localization

Use of Artificial Neural Networks as a Predictive Method to Determine Moisture Resistance of Particle and Fiber Boards Under Cyclic Testing Conditions (UNE-EN 321)

Determining internal bond strength and thickness swelling after cyclic aging tests in humid conditions is essential to assess moisture resistance of particle and fiber boards. However, because operating procedures for these types of tests take at least 3 wk, their use in daily finished product control is impractical. To solve this problem, an artificial neural network was used as a predictive method to determine these values from the board properties of thickness, density, and moisture content in conjunction with thickness swelling and internal bond strength values obtained before the aging cycle. Using 113 boards, an artificial neural network was designed consisting of two separate feedforward multilayer perceptrons applying the hyperbolic tangent as the transfer function. Training was conducted through supervised learning after the input data had been normalized. In the testing group, the network attained a determination coefficient of 0.94 for internal bond strength and 0.92 for thickness swelling

A 1-to-2 demultiplexer hybrid nanocarrier for cargo delivery and activation

[EN] A biocomputing strategy implemented in hybrid nanocarriers for controlled cargo delivery is described. The nanodevice consists of enzyme-functionalized Janus Au-mesoporous silica nanoparticles, which behave as an electronic demultiplexer (DEMUX). The nanocarrier is capable of reading molecular information from the environment (lactose) and selecting one of two possible outputs (galactose production or 4-methylumbellilferone release and activation) depending on the presence of an addressing input NAD(+).The authors wish to thank the Spanish Government (projects RTI2018-100910-B-C41 (MCUI/AEI/FEDER, UE), CTQ2017-87954-P), the Generalitat Valenciana (PROMETEO 2018/024), the Comunidad de Madrid (IND2017/BMD-7642) and CIBER-BBN (NANOCOMMUNITY project) for support.De Luis-Fernández, B.; García-Fernández, A.; Llopis-Lorente, A.; Villalonga, R.; Sancenón Galarza, F.; Martínez-Máñez, R. (2020). A 1-to-2 demultiplexer hybrid nanocarrier for cargo delivery and activation. Chemical Communications. 56(69):9974-9977. https://doi.org/10.1039/d0cc03803bS997499775669Soto, F., & Chrostowski, R. (2018). Frontiers of Medical Micro/Nanorobotics: in vivo Applications and Commercialization Perspectives Toward Clinical Uses. Frontiers in Bioengineering and Biotechnology, 6. doi:10.3389/fbioe.2018.00170Zhang, X., Chen, L., Lim, K. H., Gonuguntla, S., Lim, K. W., Pranantyo, D., … Soh, S. (2019). The Pathway to Intelligence: Using Stimuli‐Responsive Materials as Building Blocks for Constructing Smart and Functional Systems. Advanced Materials, 31(11), 1804540. doi:10.1002/adma.201804540Mailloux, S., & Katz, E. (2014). Biocomputing, Biosensing and Bioactuation Based on Enzyme Biocatalyzed Reactions. Biocatalysis, 1(1). doi:10.2478/boca-2014-0002Katz, E. (2018). Boolean Logic Gates Realized with Enzyme‐catalyzed Reactions – Unusual Look at Usual Chemical Reactions. ChemPhysChem, 20(1), 9-22. doi:10.1002/cphc.201800900Erbas-Cakmak, S., Kolemen, S., Sedgwick, A. C., Gunnlaugsson, T., James, T. D., Yoon, J., & Akkaya, E. U. (2018). Molecular logic gates: the past, present and future. Chemical Society Reviews, 47(7), 2228-2248. doi:10.1039/c7cs00491eSu, H., Xu, J., Wang, Q., Wang, F., & Zhou, X. (2019). High-efficiency and integrable DNA arithmetic and logic system based on strand displacement synthesis. Nature Communications, 10(1). doi:10.1038/s41467-019-13310-2Orbach, R., Willner, B., & Willner, I. (2015). Catalytic nucleic acids (DNAzymes) as functional units for logic gates and computing circuits: from basic principles to practical applications. Chemical Communications, 51(20), 4144-4160. doi:10.1039/c4cc09874aArugula, M. A., Bocharova, V., Halámek, J., Pita, M., & Katz, E. (2010). Enzyme-Based Multiplexer and Demultiplexer. The Journal of Physical Chemistry B, 114(15), 5222-5226. doi:10.1021/jp101101bAndréasson, J., Straight, S. D., Bandyopadhyay, S., Mitchell, R. H., Moore, T. A., Moore, A. L., & Gust, D. (2007). A Molecule-Based 1:2 Digital Demultiplexer. The Journal of Physical Chemistry C, 111(38), 14274-14278. doi:10.1021/jp074429pTuran, I. S., Gunaydin, G., Ayan, S., & Akkaya, E. U. (2018). Molecular demultiplexer as a terminator automaton. Nature Communications, 9(1). doi:10.1038/s41467-018-03259-zOrbach, R., Remacle, F., Levine, R. D., & Willner, I. (2014). DNAzyme-based 2:1 and 4:1 multiplexers and 1:2 demultiplexer. Chemical Science, 5(3), 1074. doi:10.1039/c3sc52752bLuo, C., Sun, J., Sun, B., & He, Z. (2014). Prodrug-based nanoparticulate drug delivery strategies for cancer therapy. Trends in Pharmacological Sciences, 35(11), 556-566. doi:10.1016/j.tips.2014.09.008Moreira, J., Hamraz, M., Abolhassani, M., Bigan, E., Pérès, S., Paulevé, L., … Schwartz, L. (2016). The Redox Status of Cancer Cells Supports Mechanisms behind the Warburg Effect. Metabolites, 6(4), 33. doi:10.3390/metabo6040033Adekola, K., Rosen, S. T., & Shanmugam, M. (2012). Glucose transporters in cancer metabolism. Current Opinion in Oncology, 24(6), 650-654. doi:10.1097/cco.0b013e328356da72Jerez, G., Kaufman, G., Prystai, M., Schenkeveld, S., & Donkor, K. K. (2009). Determination of thermodynamic pKavalues of benzimidazole and benzimidazole derivatives by capillary electrophoresis. Journal of Separation Science, 32(7), 1087-1095. doi:10.1002/jssc.200800482Guo, Z. (2017). The modification of natural products for medical use. Acta Pharmaceutica Sinica B, 7(2), 119-136. doi:10.1016/j.apsb.2016.06.003Llopis-Lorente, A., de Luis, B., García-Fernández, A., Jimenez-Falcao, S., Orzáez, M., Sancenón, F., … Martínez-Máñez, R. (2018). Hybrid Mesoporous Nanocarriers Act by Processing Logic Tasks: Toward the Design of Nanobots Capable of Reading Information from the Environment. ACS Applied Materials & Interfaces, 10(31), 26494-26500. doi:10.1021/acsami.8b05920Llopis-Lorente, A., Díez, P., Sánchez, A., Marcos, M. D., Sancenón, F., Martínez-Ruiz, P., … Martínez-Máñez, R. (2017). Interactive models of communication at the nanoscale using nanoparticles that talk to one another. Nature Communications, 8(1). doi:10.1038/ncomms15511Tregubov, A. A., Nikitin, P. I., & Nikitin, M. P. (2018). Advanced Smart Nanomaterials with Integrated Logic-Gating and Biocomputing: Dawn of Theranostic Nanorobots. Chemical Reviews, 118(20), 10294-10348. doi:10.1021/acs.chemrev.8b0019

Análisis de los recursos naturales y prácticas campesinas en ecosistemas de altura

El objetivo de este trabajo fue analizar la relación entre percepciones y prácticas agrícolas frente a los efectos del cambio climático desde el contexto agro-socio-económico y la estructura hidrológica en el ecosistema de altura Cochapata en Urcuquí-Ecuador. La estrategia metodológica responde al paradigma cualitativo, método de estudio de caso. Se levantó información primaria mediante lectura del paisaje, entrevistas y observación participante. Se encontró que el territorio está ocupado por industrias madereras cuyas especies arbóreas se caracterizan por excesiva demanda hídrica. El subpáramo está habitado por campesinos en condición de sobrevivencia que usan la tierra para obtener bienes de sustento familiar, valiéndose de su fuerza de trabajo. Se evidenció que la distribución del agua es de carácter hidrosocial, pues excluye al campesinado del derecho a riego y abrevadero. Respecto a los efectos del clima cambiante, los pobladores perciben en su cotidianidad olas de calor y variaciones en las temporadas de lluvias, sin embargo, no se encontraron prácticas agrícolas con intención adaptativa al fenómeno climático. Se concluye que la vulnerabilidad del ecosistema no guarda relación directa con el cambio climático, esta responde a las fallas históricas en la distribución de los bienes ecosistémicos tierra y agua, al desconocimiento del sujeto campesino por parte del Estado y de la sociedad, y al excesivo extractivismo implantado durante las últimas décadas

Predictive value and dynamic risk stratification of high sensitive basal or stimulated thyroglobulin assay in a long-term thyroid carcinoma cohort

Producción CientíficaPurpose: To evaluate the predictive value of the rhTSH thyroglobulin stimulation test (rhTSH-Tg) compared to basal high- sensitive thyroglobulin (hs-Tg) under TSH suppressive therapy at 12 months after the completion of initial treatment to predict the long-term response and Dynamic Risk Stratification (DRS) at the last follow-up visit in a long-term DTC cohort. Methods: Prospective study in 114 DTC patients (77.2% women, mean age 46.4 ± 14.1 years old, median/IQR evolution 6.7[3.1–8.0] years) from 2013 to 2020 undergoing total thyroidectomy and radioiodine ablation in whom hs-Tg and rhTSH- Tg was performed 12 months after completing initial treatment. Pearson correlation, receiving operating characteristics (ROC) and DRS at initial and last follow-up visit were analyzed. Results: hs-Tg and rhTSH-Tg show a strong positive linear correlation (r = 0.864, p < 0.001). The diagnostic performance of initial hs-Tg and rhTSH-Tg levels were evaluated via ROC-AUC as a predictor of excellent response (ER) in the last follow-up visit. Hs-Tg showed a better AUC (0.969, 95%CI = 0.941–0.997) than rhTSH-Tg (0.944, 95%IC = 0.905–0.984; p < 0.001). The hs-Tg and rhTSH-Tg cutoff point of highest sensitivity (S) and specificity (E) was 0.110 and 0.815 ng/dl, respectively. Hs-Tg showed a higher diagnostic accuracy than rhTSH-Tg (S = 100% vs 96.8%, E = 84.3% vs 84.3%, NPV = 100% vs 98.6%, PPV = 70.5% vs 69.7%; p < 0.05). The DRS based on initial hs-Tg showed better ability to predict ER (93.3% vs 86.7%) and biochemical incomplete response (53.3%vs13.3%) in the last follow-up visit compared to rhTSH-Tg. Conclusions: Both initial hs-Th and rhTSH-Tg were good predictors of long-term ER. In patients with hs-Tg, the rhTSH-test did not provide relevant prognosis information. An ER after initial treatment was associated with a very high NPV at subsequent follow-up.Publicación en abierto financiada por el Consorcio de Bibliotecas Universitarias de Castilla y León (BUCLE), con cargo al Programa Operativo 2014ES16RFOP009 FEDER 2014-2020 DE CASTILLA Y LEÓN, Actuación:20007-CL - Apoyo Consorcio BUCL

Targeted bacterial conjugation mediated by synthetic cell-to-cell adhesions

Genetic interventions on microbiomes, for clinical or biotechnological purposes, remain challenging. Conjugation-based delivery of genetic cargo is still unspecific and limited by low conjugation rates. Here we report an approach to overcome these problems, based on a synthetic bacterial adhesion system. Mating assemblers consist on a synthetic adhesion formed by the expression on the surface of donor and target cells of specific nanobodies (Nb) and their cognate antigen (Ag). The Nb?Ag bridge increased 1?3 logs transfer of a variety of plasmids, especially in liquid media, confirming that cell-cell docking is a main determinant limiting mating efficiency. Synthetic cell-to-cell adhesion allows efficient conjugation to targeted recipients, enhancing delivery of desired genes to a predefined subset of prey species, or even specific pathogenic strains such as enterohemorrhagic Escherichia coli (EHEC), within a bacterial community. The synthetic conjugation enhancer presented here optimizes plasmid delivery by selecting the target hosts with high selectivity.FUNDING: Spanish Science and Innovation Ministry (MCIN) [PID2020-117923GB-I00 to F.d.l.C.]; MCIN/AEI and FEDER [BIO2017-89081-R]; MCIN/AEI and NextGeneration EU/PRTR [PLEC2021-007739 to L.A.F.]. Conflict of interest statement. None declared ACKNOWLEDGEMENTS: We thank R. Fernandez-López for providing us with the BWmKate2 strain and, along with M. P. Garcillan-Barcia, for helpful discussions. We also thank E. Zechner for providing the pAR106 plasmid, and I. Rosenshine for EHEC strains and V. Campa for technical assistance in setting up the microscopy assay

Hybrid Mesoporous Nanocarriers Act by Processing Logic Tasks: Toward the Design of Nanobots Capable of Reading Information from the Environment

[EN] Here, we present the design of smart nano-devices capable of reading molecular information from the environment and acting accordingly by processing Boolean logic tasks. As proof of concept, we prepared Au-mesoporous silica (MS) nanoparticles functionalized with the enzyme glucose dehydrogenase (GDH) on the Au surface and with supramolecular nanovalves as caps on the MS surface, which is loaded with a cargo (dye or drug). The nanodevice acts as an AND logic gate and reads information from the solution (presence of glucose and nicotinamide adenine dinucleotide (NADI), which results in cargo release. We show the possibility of coimmobilizing GDH and the enzyme urease on nanoparticles to mimic an INHIBIT logic gate, in which the AND gate is switched off by the presence of urea. We also show that such nanodevices can deliver cytotoxic drugs in cancer cells by recognizing intracellular NAD(+) and the presence of glucose.A.L.-L. is grateful to "La Caixa" Banking Foundation for his Ph.D. grant. A.G.-F. and B.de L. thank the Spanish government for FPU fellowships. The authors are grateful to the Spanish Government (MINECO Projects MAT2015-64139-C4-1, CTQ2014-58989-P, and CTQ2015-71936-REDT) and the Generalitat Valencia (Project PROMETEOII/2014/047 and PROMETEOII/2014/061) for support. The Comunidad de Madrid (S2013/MIT-3029, Programme NANOAVANSENS) is also gratefully acknowledged.Llopis-Lorente, A.; De Luis-Fernández, B.; García-Fernández, A.; Jiménez-Falcao, S.; Orzaez, M.; Sancenón Galarza, F.; Villalonga, R.... (2018). Hybrid Mesoporous Nanocarriers Act by Processing Logic Tasks: Toward the Design of Nanobots Capable of Reading Information from the Environment. ACS Applied Materials & Interfaces. 10(31):26494-26500. https://doi.org/10.1021/acsami.8b05920S2649426500103

Beneficial effects of melatonin on liver fibrosis: A systematic review of current biological evidence

[EN] Hepatic fibrosis is a reversible response to either acute or chronic cellular injury froma wide variety of etiologies, characterized by excessive deposition of extracellularmatrix resulting in liver dysfunction and cirrhosis. Melatonin (N‐acetyl‐5‐methoxytryptamine), the main product secreted by the pineal gland, is a multitaskingindolamine with important physiological functions such as anti‐inflammatory andantioxidant actions, modulation of circadian rhythms, and immune system enhance-ment. Among the numerous biological activities of melatonin, its antifibrotic effectshave received increasingly more attention. In this study, we performed a systematicreview of publications of the last 10 years evaluating the mechanisms of action ofmelatonin against liver fibrosis. The study protocol was registered at PROSPERO(CRD42022304744). Literature research was performed employing PubMed,Scopus, and Web of Science (WOS) databases, and after screening, 29 articleswere included. Results from the selected studies provided denoted the usefulactions of melatonin on the development, progression, and evolution of liver fibrosis.Melatonin antifibrotic effects in the liver involved the reduction of profibrogenicmarkers and modulation of several cellular processes and molecular pathways,mainly acting as an antioxidant and anti‐inflammatory agent. In addition, theindolamine influenced different molecular processes, such as hepatocyte apoptosis,modulation of autophagy and mitophagy, restoration of circadian rhythms, andmodulation of microRNAs, among others. Although some limitations have beenfound regarding variability in the study design, the findings here summarized displaythe potential role of melatonin in ameliorating the development of liver fibrosis andits possible progression to liver cirrhosis and hepatocarcinomaS