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The construction of a novel enzyme-controlled nanomachine with multiple release mechanisms for on-command delivery is described. This nanodevice was assembled by modifying mesoporous silica nanoparticles with 2-(benzo[d]thiazol-2-yl)phenyl 4-aminobenzoate moieties, and further capped with beta-cyclodextrin-modified glucose oxidase neoglycoenzyme. The device released the encapsulated payload in the presence of H2O2 and acidic media. The use of glucose as an input chemical signal also triggered cargo release through the enzymatic production of gluconic acid and hydrogen peroxide, and the subsequent disruption of the gating mechanism at the mesoporous surface. The nanodevice was successfully employed for the enzyme-controlled release of doxorubicin in HeLa cancer cells.

El objetivo de este trabajo consiste en realizar un estudio comparativo y un análisis de la adaptación de cuentos populares a los mass media, tan recurrentes hoy en día, usando como fuente la serie televisiva El Cuentacuentos (1987-1988). Para ello, previamente se presenta una aproximación teórica al concepto “cuento popular”, en el que se incluyen desde su descripción, origen y evolución en la literatura oral, hasta la exposición de rasgos y elementos que lo definen (características, estructuras, personajes). Una vez aclarada esta parte más conceptual, se hace uso de dos capítulos del programa, en este caso “Juan Sin Miedo” y “Cenicienta”, para elaborar un análisis que contraste las versiones originales que se han encontrado de ambos cuentos con su correspondiente adaptación. Se aplicará la teoría trabajada para determinar qué grado de acercamiento comparten entre ellas. También se encuentran posibles usos como recurso didáctico, además de interpretaciones que nos aportan diferentes autores. Finalmente, a raíz de los resultados extraídos, se presenta una reflexión acerca de la posible adaptación de cuentos populares a los mass media y los aspectos positivos que esto implica, no sólo a nivel infantil, sino en general., The aim of this work consists in the realization of a comparative study and an analysis of the adaptation of folktales to mass media, so recurrent and popular nowadays, using as a source the television series The Storyteller (1987 – 1988). In order to do this, a theoretical approach of the “folktale” concept is presented, including from its description, origin and evolution in oral literature to the exposition of the features and components that define him (characteristics, structures, characters). Once the most conceptual part has been clarified, two of the chapters of the program (in this case “Fearless John” and “Cinderella”) are used to elaborate an analysis that contrasts the original versions of both folktales which have been found, with their corresponding adaptation. The worked theory will be applied to determine the approaching degree between them. There are also explained the possibilities of using them as an educational resource as well as some interpretations provided by different authors. Finally, using the extracted results, there is a reflection about the possible adaptation of popular folktales to the mass media and the positive aspects that this implies, not only in a child level, but in general., Grado en Educación Infantil

Development of bioinspired nanomachines with an efficient propulsion and cargo-towing has attracted much attention in the last years due to their potential biosensing, diagnostics, and therapeutics applications. In this context, self-propelled synthetic nanomotors are promising carriers for intelligent and controlled release of therapeutic payloads. However, the implementation of this technology in real biomedical applications is still facing several challenges. Herein, we report the design, synthesis, and characterization of innovative multifunctional gated platinum-mesoporous silica nanomotors constituted of a propelling element (platinum nanodendrite face), a drug-loaded nanocontainer (mesoporous silica nanoparticle face), and a disulfide-containing oligo(ethylene glycol) chain (S-S-PEG) as a gating system. These Janus-type nanomotors present an ultrafast self-propelled motion due to the catalytic decomposition of low concentrations of hydrogen peroxide. Likewise, nanomotors exhibit a directional movement, which drives the engines toward biological targets, THP-1 cancer cells, as demonstrated using a microchip device that mimics penetration from capillary to postcapillary vessels. This fast and directional displacement facilitates the rapid cellular internalization and the on-demand specific release of a cytotoxic drug into the cytosol, due to the reduction of the disulfide bonds of the capping ensemble by intracellular glutathione levels. In the microchip device and in the absence of fuel, nanomotors are neither able to move directionally nor reach cancer cells and deliver their cargo, revealing that the fuel is required to get into inaccessible areas and to enhance nanoparticle internalization and drug release. Our proposed nanosystem shows many of the suitable characteristics for ideal biomedical destined nanomotors, such as rapid autonomous motion, versatility, and stimuli-responsive controlled drug release.

This work describes the assembly of a novel enzyme-controlled nanomachine operated through an AND Boolean logic gate for on-command delivery. The nanodevice was constructed on Au-mesoporous silica Janus nanoparticles capped with a thiol-sensitive gate-like molecular ensemble on the mesoporous face and functionalized with glutathione reductase on the gold face. This autonomous nanomachine employed NADPH and glutathione disulfide as input chemical signals, leading to the enzymatic production of reduced glutathione that causes the disruption of the gating mechanism on the mesoporous face and the consequent payload release as an output signal. The nanodevice was successfully used for the autonomous release of doxorubicin in HeLa cancer cells and RAW 264.7 macrophage cells.

[EN] Introduction: Breast cancer has the highest mortality rate among cancers in women. Patients suffering from certain breast cancers, such as triple-negative breast cancer (TNBC), lack effective treatments. This represents a clinical concern due to the associated poor prognosis and high mortality. As an approach to succeed over conventional therapy limitations, we present herein the design and evaluation of a novel nanodevice based on enzyme-functionalized gold nanoparticles to efficiently perform enzyme prodrug therapy (EPT) in breast cancer cells. Results: In particular, the enzyme horseradish peroxidase (HRP) which oxidizes the prodrug indole-3-acetic acid (IAA) to release toxic oxidative species is incorporated on gold nanoconjugates (HRP-AuNCs), obtaining an efficient nanoplatform for EPT. The nanodevice is biocompatible and effectively internalized by breast cancer cell lines. Remarkably, co-treatment with HRP-AuNCs and IAA (HRP-AuNCs/IAA) reduces the viability of breast cancer cells below 5%. Interestingly, 3D tumor models (multicellular tumor spheroid-like cultures) co-treated with HRP-AuNCs/IAA exhibit a 74% reduction of cell viability, whereas the free formulated components (HRP, IAA) have no effect. Conclusion: Altogether, our results demonstrate that the designed HRP-AuNCs nanoformulation shows a remarkable therapeutic performance. These findings might help to bypass the clinical limitations of current tumor enzyme therapies and advance towards the use of nanoformulations for EPT in breast cancer., Gema Vivo-Llorca thanks the Generalitat Valenciana for her fellowship ACIF/2017/072. Paula Díez thanks the Generalitat Valenciana for her contract APOSTD/2020/153 and to the Instituto de Salud Carlos III and the European Social Fund for the financial support "Sara Borrell", CD20/00120. Antoni Llopis-Lorente thanks funding from the European Union¿s Horizon 2020 research and innovation programme under grant agreement No 847675. Images were prepared using Servier Medical Art (https://smart.servier.com/). We thank the Spanish Government (project RTI2018-100910-B-C41 (MCUI/AEI/FEDER, UE); SAF2017-84689-R-B (MCUI/ AEI/FEDER, UE)) and the Generalitat Valenciana (project PROMETEO/2018/024 and PROMETEO/2019/065) for support. A pre-print version of this work was published as a chapter of the PhD thesis of the first author Gema VivoLlorca

Dravet Syndrome (DS) is caused by mutations in the Scn1a gene encoding the α1 subunit of the sodium channel Nav1.1, which results in febrile seizures that progress to severe tonic-clonic seizures and associated comorbidities. Treatment with cannabidiol has been approved for the management of seizures in DS patients, but it appears to be also active against associated comorbidities. In this new study, we have investigated β-caryophyllene (BCP), a cannabinoid with terpene structure that appears to also have a broad-spectrum profile, as a useful therapy against both seizuring activity and progression of associated comorbidities. This has been studied in heterozygous conditional knock-in mice carrying a missense mutation (A1783V) in Scn1a gene expressed exclusively in neurons of the Central Nervous System (Syn-Cre/Scn1aWT/A1783V), using two experimental approaches. In the first approach, an acute treatment with BCP was effective against seizuring activity induced by pentylenetetrazole (PTZ) in wildtype (Scn1aWT/WT) and also in Syn-Cre/Scn1aWT/A1783V mice, with these last animals having a greater susceptibility to PTZ. Such benefits were paralleled by a BCP-induced reduction in PTZ-induced reactive astrogliosis (labelled with GFAP) and microgliosis (labelled with Iba-1) in the prefrontal cortex and the hippocampal dentate gyrus, which were visible in both wildtype (Scn1aWT/WT) and Syn-Cre/Scn1aWT/A1783V mice. In the second approach, both genotypes were treated repeatedly with BCP to investigate its effects on several DS comorbidities. Thus, BCP corrected important behavioural abnormalities of Syn-Cre/Scn1aWT/A1783V mice (e.g. delayed appearance of hindlimb grasp reflex, induction of clasping response, motor hyperactivity, altered social interaction and memory impairment), attenuated weight loss, and slightly delayed premature mortality. Again, these benefits were paralleled by a BCP-induced reduction in reactive astrogliosis and microgliosis in the prefrontal cortex and the hippocampal dentate gyrus typical of Syn-Cre/Scn1aWT/A1783V mice. In conclusion, BCP was active in Syn-Cre/Scn1aWT/A1783V mice against seizuring activity (acute treatment) and against several comorbidities (repeated treatment), in both cases in association with its capability to reduce glial reactivity in areas related to these behavioural abnormalities. This situates BCP in a promising position for further preclinical evaluation towards a close translation to DS patients.

Nowadays, massive genomics and transcriptomics data can be generated at the single-cell level. However, proteomics in this setting is still a big challenge. Despite the great improvements in sensitivity and performance of mass spectrometry instruments and the better knowledge on sample preparation processing, it is widely acknowledged that multistep proteomics workflows may lead to substantial sample loss, especially when working with paucicellular samples. Still, in clinical fields, frequently limited sample amounts are available for downstream analysis, thereby hampering comprehensive characterization at protein level. To aim at better protein and peptide recoveries, we compare existing and novel approaches in the multistep sample preparation protocols for mass spectrometry studies, from sample collection, cell lysis, protein quantification, and electrophoresis/ staining to protein digestion, peptide recovery, and LC-MS/MS instruments. From this critical evaluation, we conclude that the recent innovations and technologies, together with high quality management of samples, make proteomics on paucicellular samples possible, which will have immediate impact for the proteomics community.

[EN] Here we describe the construction of an integrated and pH-sensitive nanomachine with layer-by-layer supramolecular design and enzymatic control for on-command delivery. The nanodevice comprises a first layer of ß-cyclodextrin-coated gold nanoparticles as capping element of benzimidazole functionalized mesoporous silica nanoparticles, and a second control layer based on an adatamantane-modified glucose oxidase derivative. The nanomachine was selectively fuelled by glucose and successfully employed for the autonomous release of doxorubicin in HeLa cancer cells., Financial support from the Spanish Ministry of Economy and Competitiveness (projects CTQ2014-58989-P, CTQ2015-71936-REDT, CTQ2017-87954-P, and MAT2015-64139-C4-1-R) and generalitat Valenciana (project PROMETEO/2018/024) is gratefully acknowledged.

© 2021 by the authors., Human Proteome Project (HPP) presents a systematic characterization of the protein landscape under different conditions using several complementary-omic techniques (LC-MS/MS proteomics, affinity proteomics, transcriptomics, etc.). In the present study, using a B-cell lymphoma cell line as a model, comprehensive integration of RNA-Seq transcriptomics, MS/MS, and antibody-based affinity proteomics (combined with size-exclusion chromatography) (SEC-MAP) were performed to uncover correlations that could provide insights into protein dynamics at the intracellular level. Here, 5672 unique proteins were systematically identified by MS/MS analysis and subcellular protein extraction strategies (neXtProt release 2020-21, MS/MS data are available via ProteomeXchange with identifier PXD003939). Moreover, RNA deep sequencing analysis of this lymphoma B-cell line identified 19,518 expressed genes and 5707 protein coding genes (mapped to neXtProt). Among these data sets, 162 relevant proteins (targeted by 206 antibodies) were systematically analyzed by the SEC-MAP approach, providing information about PTMs, isoforms, protein complexes, and subcellular localization. Finally, a bioinformatic pipeline has been designed and developed for orthogonal integration of these high-content proteomics and transcriptomics datasets, which might be useful for comprehensive and global characterization of intracellular protein profiles., We are financial support from the Spanish Health Institute Carlos III (ISCIII) for the grants: FIS PI14/01538, FIS PI17/01930 and CB16/12/00400.

© 2021 by the authors., Sporadic Colorectal Cancer (sCRC) is the third leading cause of cancer death in the Western world, and the sCRC patients presenting with synchronic metastasis have the poorest prognosis. Genetic alterations accumulated in sCRC tumor cells translate into mutated proteins and/or abnormal protein expression levels, which contribute to the development of sCRC. Then, the tumor-associated proteins (TAAs) might induce the production of auto-antibodies (aAb) via humoral immune response. Here, Nucleic Acid Programmable Protein Arrays (NAPPArray) are employed to identify aAb in plasma samples from a set of 50 sCRC patients compared to seven healthy donors. Our goal was to establish a systematic workflow based on NAPPArray to define differential aAb profiles between healthy individuals and sCRC patients as well as between non-metastatic (n = 38) and metastatic (n = 12) sCRC, in order to gain insight into the role of the humoral immune system in controlling the development and progression of sCRC. Our results showed aAb profile based on 141 TAA including TAAs associated with biological cellular processes altered in genesis and progress of sCRC (e.g., FSCN1, VTI2 and RPS28) that discriminated healthy donors vs. sCRC patients. In addition, the potential capacity of discrimination (between non-metastatic vs. metastatic sCRC) of 7 TAAs (USP5, ML4, MARCKSL1, CKMT1B, HMOX2, VTI2, TP53) have been analyzed individually in an independent cohort of sCRC patients, where two of them (VTI2 and TP53) were validated (AUC ~75%). In turn, these findings provided novel insights into the immunome of sCRC, in combination with transcriptomics profiles and protein antigenicity characterizations, wich might lead to the identification of novel sCRC biomarkers that might be of clinical utility for early diagnosis of the tumor. These results explore the immunomic analysis as potent source for biomarkers with diagnostic and prognostic value in CRC. Additional prospective studies in larger series of patients are required to confirm the clinical utility of these novel sCRC immunomic biomarkers., We gratefully acknowledge financial support from the Spanish Health Institute Carlos III (ISCIII) for the grants: FIS PI14/01538, FIS PI17/01930 and CB16/12/00400. We also acknowledge Fondos FEDER (EU) “Una manera de hacer Europa” and Junta Castilla-León (COVID19 grant COV20EDU/00187). Fundación Solórzano FS/38-2017. The Proteomics Unit belongs to ProteoRed, PRB3-ISCIII, supported by grant PT17/0019/0023, of the PE I + D + I 2017-2020, funded by ISCIII and FEDER. CNPq-National Council for Scientific and Technological Development (Brazil) (306258/2019-6) and FAPERJ-Foundation for Research Support of Rio de Janeiro State for the financial support (E-26/201.670/2017 and 210.379/2018). M. González-González is supported by MINECOPTA2019-017870-I.A. Landeira-Viñuela is supported by VIII Centenario-USAL PhD Program. P.J.-V. is supported by JCYL PhD Program and scholarship JCYL-EDU/601/2020. P.D. and E.B. are supported by a JCYL-EDU/346/2013 Ph.D. scholarship.

© 2020 by the authors., Natural products are the ideal basis for the design of novel efficient molecular entities. Podophyllotoxin, a naturally occurring cyclolignan, is an example of natural product which displays a high versatility from a biological activity point of view. Based on its unique chemical structure, different derivatives have been synthesized presenting the original antitumoral properties associated with the compound, i.e., the tubulin polymerization inhibition and arising anti-topoisomerase II activity from structural modifications on the cyclolignan skeleton. In this report, we present a novel conjugate or hybrid which chemically combines both biological activities in one single molecule. Chemical design has been planned based in our lead compound, podophyllic aldehyde, as an inhibitor of tubulin polymerization, and in etoposide, an approved antitumoral drug targeting topoisomerase II. The cytotoxicity and selectivity of the novel synthetized hybrid has been evaluated in several cell lines of different solid tumors. In addition, these dual functional effects of the novel compound have been also evaluated by molecular docking approaches., Financial support came from Spanish MINECO (CTQ2015-68175-R, AGL2016-79813-C2-2-R), Junta de Castilla y León (BIO/SA07/15), ISCIII-RICET Network (RD16/0027/0018) cofinanced by the Fondo Social Europeo of the European Union (Fondos FEDER–EU) and USAL (Financiación GIR). Fundación Solórzano (FS/23-2015) has also sponsored this work. The Proteomics Unit belongs to ProteoRed, PRB2-ISCIII, supported by Grant No. PT13/0001, of the PE I+D+I 2013–2016, funded by ISCIII and FEDER.

Se presenta el caso de un paciente varón de 41 años con hidradenitis supurativa de 25 años de evolución. Presenta, desde entonces, un transcurso inestable de la enfermedad que le ha impedido llevar sus actividades cotidianas con total bienestar. Las comorbilidades asociadas, las complicaciones, los cuadros clínicos producidos como efecto secundario a los tratamientos recibidos y los ingresos se han convertido en una constante en su vida. Se pretende mostrar, desde un enfoque holístico y práctico, a través de los patrones funcionales de Marjory Gordon, la evaluación y el manejo de un paciente con estas características. [Hidradenitis suppurativa and autoinflammatory syndrome. A case report] ABSTRACT: The case of a 41-year-old male with 25 years of evolution of hidradenitis suppurativa is presented. Since then, he presents an unstable course of the disease that has prevented him from carrying out his daily activities with total well-being. The associated comorbidities, complications, the clinic produced as a side effect of the treatments received and admissions have become a constant in his life. The aim is to show, from a holistic and practical approach, through Marjory Gordon's functional patterns, the evaluation and management of a patient with these characteristics.

[EN] A novel nanomachine for dual and sequential delivery of two different compounds was developed by grafting a thiol group and a pH sensitive beta-cyclodextrin-based gate-like ensemble on acetylcholinesterase-modified Au-mesoporous silica Janus nanoparticles., This work was supported by CTQ2017-87954-P and RTI2018100910-B-C41 grants from the Spanish Ministry of Economy and Competitiveness and PROMETEO2018/024 from the Generalitat Valenciana.

Caso Clínico: Se presenta un caso de un carcinoma verrucoso en el hallux del pie derecho, que inicialmente fue extirpado completamente, evolucionando de forma tórpida hasta una posterior amputación transmetatarsiana sin evidencia de recidiva tumoral, durante su seguimiento en un periodo de 3 años, para evolucionar a otra entidad patológica como es el Pie Diabético.Discusión: Este carcinoma se caracteriza clínicamente por un lento crecimiento de aspecto similar a la verruga plantar, siendo su histología típica la invasión local mínima con algo de displasia y baja incidencia de metástasis y presenta buen pronóstico. Para el diagnóstico correcto, es imprescindible la biopsia quirúrgica. La resección radical es el tratamiento de elección con confirmación histológica de los márgenes de resección libres del tumor.Conclusiones: El carcinoma verrucoso plantar presenta una evidente dificultad diagnóstica, constituyendo la resección quirúrgica completa de la lesión el tratamiento de elección. Posee un buen pronóstico y mejor resolución. Excepcionalmente hay pacientes que presentan complicaciones a este tratamiento. [Clinical case of plantar verrucous carcinoma versus diabetic foot] ABSTRACT: Case report: A case of a verrucous carcinoma in the hallux of the right foot is presented, which was initially completely removed, evolving torpidly until a subsequent transmetatarsal amputation without evidence of tumor recurrence, during its follow-up over a period of 3 years, to evolve to another pathological entity such as the Diabetic Foot.Discussion: Verrucous carcinoma is clinically characterized by a slow growth similar in appearance to the plantar wart. The typical histology is a minimal local invasion with low incidence of metastasis and a good prognosis. Surgical biopsy is essential for correct diagnosis. Radical resection is the treatment of choice with histological confirmation of free resection margins of the tumor.Conclusions: The plantar verrucous carcinoma usually presents diagnostic difficulty. The complete surgical resection of the lesion is the treatment of choice presenting a good prognosis and prompt healing. As an exception, there are patients who present complications to this treatment.

Made available in DSpace on 2018-11-26T15:37:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-15 Spanish Ministry of Economy and Competitiveness Comunidad de Madrid, Programme NANOAVANSENS A novel nanocomposite material consisting of reduced graphene oxide/Rh nanoparticles was prepared by a one-pot reaction process. The strategy involved the simultaneous reduction of RhCl3 and graphene oxide with NaBH4 and the in situ deposition of the metal nanoparticles on the 2D carbon nanomaterial planar sheets. Glassy carbon electrode coated with this nanocomposite was employed as nanostructured support for the cross-linking of the enzyme laccase with glutaraldehyde to construct a voltammperometric biosensor for 17 beta-estradiol in the 0.9-11 pM range. The biosensor showed excellent analytical performance with high sensitivity of 25.7 A mu M-1 cm(-1), a very low detection limit of 0.54 pM and high selectivity. The biosensor was applied to the rapid and successful determination of the hormone in spiked synthetic and real human urine samples. (C) 2016 Elsevier B.V. All rights reserved. Univ Complutense Madrid, Fac Chem, Dept Analyt Chem, E-28040 Madrid, Spain State Univ Sao Paulo, Inst Chem, POB 780, BR-13560970 Sao Carlos, SP, Brazil Mackenzie Presbiterian Univ, Engn Sch, BR-01302907 Sao Paulo, SP, Brazil State Univ Sao Paulo, Inst Chem, POB 780, BR-13560970 Sao Carlos, SP, Brazil Spanish Ministry of Economy and Competitiveness: CTQ2014-58989-P Spanish Ministry of Economy and Competitiveness: CTQ2011-24355 Spanish Ministry of Economy and Competitiveness: CTQ2015-71936-REDT Spanish Ministry of Economy and Competitiveness: CTQ2012-34238 Comunidad de Madrid, Programme NANOAVANSENS: S2013/MIT-3029

We report herein the design of an autonomous glucose-responsive smart nanomachine for insulin (In) delivery based on ultrasound (US)-propelled nanomotors combined with an enzyme-based sensing-effector unit. Gold nanowire (AuNW) motors have been coupled with a mesoporous silica (MS) segment, which was gated with pH-responsive phenylboronic acid (PBA)-glucose oxidase (GOx) supramolecular nanovalves responsible for the insulin release. Glucose-induced protonation of the PBA groups triggers the opening of the pH-driven gate and uncapping of the insulin-loaded nanovalves. The insulin-loaded MS-Au nanomotors displayed an efficient US-driven movement that dramatically accelerates the glucose-triggered insulin release when compared to their static counterparts. Such coupling of the locomotion of nanovehicles with gated insulin-containing nanocontainers and glucose-responsive nanovalves holds great promise for the improved management of diabetes.

[EN] We describe herein the preparation of glucose-sensitive capped mesoporous silica nanoparticles for insulin delivery. The new material consists of an expanded-pore nanometric silica support grafted with 1-propyl-1-H-benzimidazole groups, loaded with fluorescein isothiocyanate-labeled insulin (FITC-Ins) and capped by the formation of inclusion complexes between cyclodextrin-modified glucose oxidase (CD-GOx) and the benzimidazole groups grafted on the mesoporous support. Insulin delivery from the gated material in simulated blood plasma was assessed upon addition of glucose. Glucose is transformed by GOx into gluconic acid, which promoted the dethreading of the benzimidazole-CD-GOx inclusion complexes, allowing cargo release. Small quantities of this support would be needed to release the amount of insulin necessary to decrease diabetic blood glucose concentrations to regular levels., The authors thank the Spanish Government (projects CTQ2011-24355, MAT2015-64139-C4-1-R, CTQ2014-58989-P, and AGL2015-70235-C2-2-R (MINECO/FEDER)) and the Generalitat Valenciana (project PROMETEOII/2014/047) for support. M.O. thanks the Universitat Politecnica de Valencia for her FPI grant. P.D. thanks the Ministerio de Economia y Competitividad for her FPI grant (BES-2012-054066). C.C. thanks the Generalitat Valenciana for her postdoctoral contract VALi+D.

Au nanoparticles modified with 4-mercaptopyridine and 6-mercapto-1-hexanol were used as coordination agents to prepare a novel hybrid nanomaterial with Ag:4,4'-bipyridine nanobelts. This nanohybrid was employed to modify glassy carbon electrodes and to construct a horseradish peroxidase-based mediatorless amperometric biosensor for H2O2. The electrode, poised at -100mV, exhibited a rapid response within 4s and a linear calibration range from 90pM to 6.5nM H2O2. The biosensor showed a high sensitivity of 283A/Mcm(2) and a very low detection limit of 45pM at a signal-to-noise ratio of 3. The enzyme biosensor showed high stability when stored at 4°C under dry conditions, retaining over 96% and 78% of its initial activity after 15 and 30days of storage at 4°C, respectively.

Resumen: Se presenta el caso clínico de una paciente de 38 años que padece una úlcera neuropática infectada en pie derecho (Charcot). Tras 6 semanas de tratamiento combinando antibioterapia, cura en ambiente húmedo y descarga con bota óptima, se logra la epitelización y resolución definitiva de la lesión. [Infected plantar ulcer secondary to Charcot neuro-arthropathy] ABSTRACT: The clinical case of a 38-year-old patient who has an infected neuropathic ulcer with Charcot's right foot, is presented. After 6 weeks of treatment combining antibiotic therapy, cure in a humid environment and discharge with optimal boot, epithelialization and definitive resolution of the lesion is achieved. &nbsp

RESUMEN: Objetivo: Analizar la epidemiología y cuidados de los pacientes con diagnóstico de calcifilaxis tratados en el Servicio de Dermatología y la Unidad de Úlceras del Consorcio Hospital General Universitario de Valencia (CHGUV). Material y método: Estudio observacional descriptivo retrospectivo de casos clínicos de calcifilaxia vistos en el Servicio de Dermatología y la Unidad de Úlceras del CHGUV en el periodo comprendido entre los años 1997 a 2016. Variables principales: factores de riesgo, supervivencia, mortalidad y causas, tratamientos. Estadística descriptiva. Resultados: Se identificaron 11 pacientes, todos del sexo femenino. El 63% de los pacientes estaban en tratamiento en hemodiálisis. Se observó un 81% de mortalidad. La causa prevalente fue: un 9,1% se debió a sepsis y un 54,5% a shock cardíaco. Conclusiones: consideramos que sería conveniente aplicar un enfoque sistemático respecto al tratamiento de las úlceras en pacientes con calcifilaxis, adoptando en ocasiones una visión paliativa, valorando el desbridamiento en cada caso según la evaluación de las comorbilidades, la perfusión vascular y la presencia de infección. CALCIFYLAXIS ULCERS: PALLIATIVE CARE OR DEBRIDEMENT? ABSTRACT: Objective: To analyze the epidemiology and care of patients diagnosed with calciphylaxis treated in the Dermatology Service and the Ulcers Unit of the University General Hospital of Valencia Consortium (CHGUV). Material and method: Retrospective descriptive observational study of clinical cases of calciphylaxis seen in the Dermatology Department and the Ulcers Unit of the CHGUV in the period from 1997 to 2016. Main variables: risk factors, survival, mortality and causes, treatments. Descriptive statistics. Results: Eleven patients were identified, all of them female. 63% of the patients were under treatment in hemodialysis. An 81% mortality was observed. The prevalent cause was: 9.1% was due to sepsis and 54.5% to cardiac shock. Conclusions: we consider that it would be appropriate to apply a systematic approach to the treatment of ulcers in patients with calciphylaxis, adopting at times a palliative view, assessing debridement in each case according to the evaluation of comorbidities, vascular perfusion and the presence of infection.

RESUMEN: Presentamos un caso clínico de paniculitis en muslo que aparentaba ser una calcifilaxia, pero tras realizar dos biopsias se consiguió determinar el diagnóstico correcto y posteriormente instaurar el tratamiento terapéutico adecuado basado en desbridamiento y abordaje de cura en ambiente húmedo hasta su cicatrización. CLINICAL CASE OF PANNICULITIS CHARACTERIZED BY SUBCUTANEOUS TUMOR IN THE THIGH ABSTRACT: We present a clinical case of panniculitis in the thigh that appeared to be a calciphylaxis, but after carrying out two biopsies, it was possible to finish the correct diagnosis and later to establish the adequate therapeutic treatment based on debridement and treatment of cure in a humid environment until it is healing.

We report herein the design of a novel biosensing strategy for the detection of carcinoembryonic antigen (CEA), based on the use of Janus-type nanoparticles having Au and silica opposite faces as integrated electrochemical biorecognition-signaling system. The Janus nanoparticles were properly functionalized with horseradish peroxidase on the silica surface to act as signaling element, and a biotin thiol-modified anti-CEA DNA hairpin aptamer the Au face to assemble the biorecognition element. The sensing approach relies on the first specific recognition of CEA by the bifunctionalized Janus nanoparticles, causing unfolding of the DNA hairpin structure and unmasking the biotin residues at the aptamer chain. This CEA-Janus nanoparticle complex was then captured by avidin-modified Fe3O4@SiO2 NanoCaptors®, allowing further magnetic deposition on carbon screen printed electrodes for the amperometric detection of the cancer biomarker. The Janus nanoparticles-based aptasensor was able to detect CEA in the range from 1 to 5000 ng mL−1 (5.5 pM–28 nM) with a detection limit of 210 pg mL−1 (1.2 pM). The aptasensor also showed high reproducibility and storage stability, and was successfully validated in human serum.

We report the construction of a novel amperometric aptasensor for the highly specific detection of carcinoembryonic antigen (CEA). The sensing interface was assembled by electrodeposition of reduced graphene oxide (rGO) and gold nanoparticles (AuNPs) on a carbon screen-printed electrode (SPE) and further functionalization with a biotin and thiol-modified anti-CEA DNA hairpin aptamer. The sensing approach relies on the specific recognition of CEA by the folded aptamer, causing unfolding of the DNA hairpin structure and unmasking the biotin residues at the aptamer chain. Further incubation with a sptreptavidin-peroxidase conjugate allows the amperometric detection of the cancer biomarker. This aptasensor was able to detect CEA in the broad range from 20 pg mL−1 to 2 μg mL−1 (112 fM–11 μM) with a detection limit of 16 pg mL−1 (90 fM). The aptasensor also showed high reproducibility, specificity and stability, and was successfully validated in diluted human serum samples.

Chronic lymphocytic leukaemia (CLL) is a malignant B cell disorder characterized by its high heterogeneity. Although genomic alterations have been broadly reported, protein studies are still in their early stages. Herein, a 224-antibody microarray has been employed to study the intracellular signalling pathways in a cohort of 14 newly diagnosed B-CLL patients as a preliminary study for further investigations. Several protein profiles were differentially identified across the cytogenetic and molecular alterations presented in the samples (deletion 13q14 and 17p13.1, trisomy 12, and NOTCH1 mutations) by a combination of affinity and MS/MS proteomics approaches. Among others altered cell signalling pathways, PKC family members were identified as down-regulated in nearly 75% of the samples tested with the antibody arrays. This might explain the rapid progression of the disease when showing p53, Rb1, or NOTCH1 mutations due to PKC-proteins family plays a critical role favouring the slowly progressive indolent behaviour of CLL. Additionally, the antibody microarray results were validated by a LC-MS/MS quantification strategy and compared to a transcriptomic CLL database. In summary, this research displays the usefulness of proteomic strategies to globally evaluate the protein alterations in CLL cells and select the possible biomarkers to be further studied with larger sample sizes., We gratefully acknowledge financial support from the Carlos III Health Institute of Spain (ISCIII, FIS PI11/02114 and FIS PI14/01538), Fondos FEDER (EU), Junta Castilla-Leon SA198A12-2 and BIO/SA03/15. The proteomics analysis was performed in the Proteomics facility of Centro de Investigacion del Cancer (Salamanca, Spain) that belongs to ProteoRed, PRB2- ISCIII, supported by grant PT13/0001. P.D. is supported by a JCYL-EDU/346/2013 PhD scholarship.

Background:ALPS patients show impaired generation of humoral memory for T independent antigens whereas they generate memory for self-antigens due to impaired FAS-dependent removal of autoreactive germinal center B cells. It is known that FAS signaling via caspase activation results in cell apoptosis. However, FAS ligation may also initiate or modulate non-apoptotic signaling as shown for example by its ability to activate NF-κB. Recent data implicate a regulatory role of FAS in the modulation of mTOR signaling in ALPS double-negative T cells. Moreover, a recently described C194V FAS mutation disturbs its post-translational modification leading to impaired apoptosis induction while non-apoptotic signalling is still intact. Consequently, C194V FAS protects from the autoimmune phenotype in the murine ALPS system. This supports the view that FAS may prevent autoimmunity with other mechanisms than inducing apoptosis.Objectives:We hypothesize that FAS mutations impair this modulatory signaling, leading to hyper-activation of B cells. Therefore we aim to investigate non apoptotic FAS signaling in B cells derived from healthy individuals and ALPS patients.Methods:We studied resting and activated B cells in ALPS patients in presence or absence of FAS ligand by flow cytometry analysing relevant molecules to the CD40 signaling pathway. We used mass cytometry to perform functional phenotyping of B cells isolated from secondary lymphoid organs. Proteomic studies were performed to identify potential signaling circuits and RNA sequencing to study the consequences of FAS signaling on B cell fate.Results:In CD40L activated B cells, FAS signaling results in specific modulation of the mTOR signaling pathway. This modulation is absent in ALPS derived B cells. In line with these data germinal center B cells and plasmablast from secondary lymphoid organs of ALPS patients show hyperactive mTOR signaling pathway. Proteomic studies identify a circuit that links FAS to the phosphatase PTEN via DAXX and the deubiquitinase USP7.Conclusion:We describe a new role of FAS in the regulation of B cell activation. Defects in FAS signaling in ALPS contribute to dysregulation of the mTOR signaling pathway and disturbed B cell development.Disclosure of Interests:None declared

Monocytes are bone marrow-derived leukocytes that are part of the innate immune system. Monocytes are divided into three subsets: classical, intermediate and non-classical, which can be differentiated by their expression of some surface antigens, mainly CD14 and CD16. These cells are key players in the inflammation process underlying the mechanism of many diseases. Thus, the molecular characterization of these cells may provide very useful information for understanding their biology in health and disease. We performed a multicentric proteomic study with pure classical and non-classical populations derived from 12 healthy donors. The robust workflow used provided reproducible results among the five participating laboratories. Over 5000 proteins were identified, and about half of them were quantified using a spectral counting approach. The results represent the protein abundance catalogue of pure classical and enriched non-classical blood peripheral monocytes, and could serve as a reference dataset of the healthy population. The functional analysis of the differences between cell subsets supports the consensus roles assigned to human monocytes., All participating laboratories are members of the PRBB-ISCIII platform. This work was supported by the Carlos III Health Institute of Spain (ISCIII, FIS PI11/02114 and FIS PI14/01538)-Fondos FEDEREU); grants SAF2014-5478-R from Ministerio de Economía y Competitividad to FJC; grant DPI2015-68982-R from Ministerio de Ciencia e Innovación.

A new family of molecular hybrids, between cyclolignans related to podophyllic aldehyde and several diterpenylnaphthohydroquinones (DNHQ), was prepared and its biological activity evaluated in several human solid tumor cell lines, which are representative of the most prevalent solid tumors in the Western world. Both cyclolignan and quinone fragments were linked through aliphatic or aromatic spacers. The new hybrid family was evaluated for its cytotoxicity, and it was found that the hybrids were several times more potent against the osteosarcoma cell line MG-63 than against MCF-7 and HT-29 cell lines. The presence of an aromatic ring in the linker gave the most potent and selective agent, improving the cytotoxicity of the parent compounds. Cell cycle studies demonstrated that this hybrid induces a strong and rapid apoptotic effect and arrests cells at the G2/M phase of the cell cycle, in the same way that the parent compound podophyllic aldehyde does., Financial support came from Spanish MINECO (CTQ2015-68175-R, AGL2016-79813-C2-2-R), Junta de Castilla y Leon (BIO/SA07/15), and ISCIII-RICET Network (RD06/0022/1004) cofinanced by the Fondo Social Europeo of the European Union (Fondos FEDER − EU. Fundacion Solorzano (FS/23-2015) has also sponsored this work. The Proteomics Unit belongs to ProteoRed, PRB2-ISCIII, supported by Grant No. PT13/0001, of the PE I+D+I 2013-2016, funded by ISCIII and FEDER. Á.P.H. and P.D. are supported by JCYL-EDU/ 346/2013 Ph.D. scholarships.

[EN] Although many nanodevices have been described in recent years, nanoparticles capable of communicating with each other have been barely reported. Traditional communication technologies cannot be applied on the nanoscale, but a potential approach to achieve this goal is to mimic how nature communicates by exchanging information using molecules. Based on these concepts, some examples using DNA, enzymes and small molecules for information processing, nanoparticles capable of modulating chemical communication between cells and nanoparticles that can communicate with each another have been reported. Communication between nanodevices may find applications in different areas and a number of future new results are envisioned in this research field. (C) 2017 Elsevier Ltd. All rights reserved., A. Llopis-Lorente is thankful to the La Caixa Banking Foundation for his PhD grant. The authors are grateful to the Spanish Government (MINECO Projects MAT2015-64139-C4-1, CTQ2014-58989-P, CTQ2015-71936-REDT and AGL2015-70235-C2-2-R) and the Generalitat Valenciana (Project PROMETEOII/2014/047). The authors also thank the Comunidad de Madrid (S2013/MIT-3029, Programme NANOAVANSENS) for support.

Combining biomolecules with materials used in medicine allows for local control of the biological response and can be used for modulating the host immune response, a major challenge in the efficacy of many medical devices. In Subchapter 3.1, we will review different methods used to attach biomolecules to materials, focusing on protein conjugation methods. We will begin by describing noncovalent immobilization strategies, including encapsulation within biomaterials and adsorption to material surfaces. We will then discuss strategies to covalently attach biomolecules to materials via the use of specific functional groups, thus enhancing the stability of the interaction. Finally, we will describe emerging methods to site specifically immobilize biomolecules to materials such that proteins are presented in an oriented manner, improving their overall functionality. Throughout the subchapter, we will emphasize the advantages and disadvantages of each technique, successes achieved, as well as the challenges remaining in this growing field.During last years, increasing development of nanoparticles as targeted drug delivery agents, has led to a wide amount of studies involving their characterization to the application as novel therapeutic agents. Hence, the nanoparticles interact with biological environments when they enter in the human body, and then proteins bind to the nanomaterial surface forming the protein corona. Protein corona has a great relevance in the interaction and function of the nanoparticle-drug conjugates. In fact, its characterization is one of the main challenges for nanoscience development. Herein, it is reviewed the main proteomic methods described for quantify and qualify the protein corona formed around nanoparticles to better understand the process of interaction with the biological media, and to decipher key parameters to control the effects of the protein corona.In Subchapter 3.3, the structure and working principles of coatings for controlled drug release in oral drug administration are presented. The release mechanisms, including diffusion, dissolution, osmotic pumping, and swelling are described. The soft materials used in the majority of controlled drug release formulations are natural and synthetic polymers. They are presented here and examples of specific polymers applied in controlled release formulations are provided. There is also a section containing characterization of soft materials using in situ electron microscopy for studying water transport through coatings at high-spatial resolution. The reason for this is that the detailed properties and release mechanisms of the controlled release depend on the material nanostructure. The in situ characterization gives access not only the information about the nanostructure but also the direct correlation between structure and properties on different length scales. Finally, an overview of the present major challenges and future possibilities concerning controlled drug release formulations is presented.Targeting cancer cells with functional nanoprobes possessing a targeting drug unit and an imaging moiety carries great potential for early detection, accurate diagnosis, and targeted therapy of various diseases. Given their nanoscopic dimensions, ultrasmall particles ( < 100nm) are in general well suited for interactions with the cells; however, the current challenge of the nanomedicine is to transform inorganic nanoparticles of metals (e.g., gold) or metal oxide (e.g., magnetite) into signal-generating vectors. Engineered nanostructures can act as vehicles for a large number of signaling centers and/or targeting units thereby offering unique opportunity to enhance the sensitivity by locally enhancing the density of signal groups. For this purpose, creation of surface groups enabling chemical attachment of antibodies or other targeting biomolecules are essential that will allow the delivery of therapeutic payloads to the diseased sites. Multimodal nanoprobes functionalized with different diagnostic and therapeutic options within a single nanoparticle followed by their functionalization with organic ligands and biomolecules can provide specific uptake and high sensitivity toward anatomical information. However, the vision of making clinical theranostics a routine clinical procedure is encumbered by limited stability of complex nanoparticles in biological milieu and lack of standardization of therapy response. Despite the widely acclaimed advantages of integrating diagnostic imaging, drug delivery, and therapeutic monitoring in a single nanotheranostic probe, the clinical utilization of engineered nanoprobes demands concerted efforts in the domains of nanoparticle and surface chemistry/charge, new chelator ligands, pharmaceutical technology, radioactive labeling of nanovectors, biokinetics, and pharmacodynamics of nanoprobes, and biological tests (cell tests and animal models).

Phage-display technology constitutes a powerful tool for the generation of specific antibodies against a predefined antigen. The main advantages of phage-display technology in comparison to conventional hybridoma-based techniques are: (1) rapid generation time and (2) antibody selection against an unlimited number of molecules (biological or not). However, the main bottleneck with phage-display technology is the validation strategies employed to confirm the greatest number of antibody fragments. The development of new high-throughput (HT) techniques has helped overcome this great limitation. Here, we describe a new method based on an array technology that allows the deposition of hundreds to thousands of phages by micro-contact on a unique nitrocellulose surface. This setup comes in combination with bioinformatic approaches that enables simultaneous affinity screening in a HT format of antibody-displaying phages.