Multi-electrode architecture modeling and optimization for homogeneous electroporation of large volumes of tissue

Electroporation is a phenomenon that consists of increasing the permeability of the cell membrane by means of high-intensity electric field application. Nowadays, its clinical application to cancer treatment is one of the most relevant branches within the many areas of electroporation. In this area, it is essential to apply homogeneous treatments to achieve complete removal of tumors and avoid relapse. It is necessary to apply an optimized transmembrane potential at each point of the tissue by means of a homogenous electric field application and appropriated electric field orientation. Nevertheless, biological tissues are composed of wide variety, heterogeneous and anisotropic structures and, consequently, predicting the applied electric field distribution is complex. Consequently, by applying the parallel-needle electrodes and single-output generators, homogeneous and predictable treatments are difficult to obtain, often requiring several repositioning/application processes that may leave untreated areas. This paper proposes the use of multi-electrode structure to apply a wide range of electric field vectors to enhance the homogeneity of the treatment. To achieve this aim, a new multi-electrode parallel-plate configuration is proposed to improve the treatment in combination with a multiple-output generator. One method for optimizing the electric field pattern application is studied, and simulation and experimental results are presented and discussed, proving the feasibility of the proposed approach

Real-time impedance monitoring during electroporation processes in vegetal tissue using a high-performance generator

Classical application of electroporation is carried out by using fixed protocols that do not clearly assure the complete ablation of the desired tissue. Nowadays, new methods that pursue the control of the treatment by studying the change in impedance during the applied pulses as a function of the electric field are being developed. These types of control seek to carry out the treatment in the fastest way, decreasing undesired effects and treatment time while ensuring the proper tumour ablation. The objective of this research is to determine the state of the treatment by continuously monitoring the impedance by using a novel versatile high-voltage generator and sensor system. To study the impedance dynamics in real time, the use of pulses of reduced voltage, below the threshold of reversible electroporation, is tested to characterise the state-of-the-treatment without interfering with it. With this purpose, a generator that provides both low voltage for sense tissue changes and high voltage for irreversible electroporation (IRE) was developed. In conclusion, the characterisation of the effects of electroporation in vegetal tissue, combined with the real-time monitoring of the state-of-the-treatment, will enable the provision of safer and more effective treatments

Implementación del protocolo DNP3 en una red de sistemas SCADA empotrados para la monitorización de variables y dispositivos

Este Trabajo de Fin de Grado se enmarca dentro de mi trabajo en Pariver S.A. y está compuesto por tres partes diferenciadas. Pariver recibió una oferta para un proyecto en El Salvador en la que se buscaba el uso del protocolo DNP3 en la comunicación entre sistemas SCADA. En este Trabajo de Fin de Grado se ha incluido también la creación del propio sistema SCADA y formar redes de comunicación entre ellos. \\ La primera tarea consistió en el estudio y análisis del protocolo DNP3 buscando toda la documentación posible del protocolo. Para subsanar las lagunas y dudas surgidas de la documentación he recurrido a simuladores del protocolo DNP3 y a un analizador de paquetes de red para realizar ingeniería inversa. La segunda tarea trató la implementación del protocolo DNP3 en sendas aplicaciones cliente y servidor que leen los valores de un fichero, los traducen al formato del protocolo y los envían para que sean leídos por el servidor. El servidor analiza los datos y verifica que son aceptables, si no lo son responde al cliente comunicando los cambios necesarios. La tercera y última tarea abarcó la configuración de las placas ARM GuruPlug sobre las que se desplegaron los clientes y servidores DNP3 y la utilización de sensores que comuniquen valores a los SCADA a través de placas Arduino mediante el protocolo Modbus. \\ El resultado es un sistema capaz de recolectar datos del entorno, enviarselos al servidor de forma encriptada, analizar los datos y corregir posibles problemas en la instalación en la que se ha implantado de forma automatica

Focalización de ablación tumoral basada en sistema de electroporación versátil multi-salida

La ablación tumoral mediante IRE ya es empleada a nivel clínico, mas tiene diversos retos por superar, como la focalización del tratamiento, preservando tejido sano y asegurando su eficacia. Este trabajo propone una metodología basada en un sistema experimental de electroporación para mejorar el control y la focalización del tratamiento

Plataforma de electrónica de potencia versátil para aplicaciones de campo eléctrico pulsado de nanosegundos.

En el campo de la electroporación, las tendencias actuales apuntan a que los pulsos de nanosegundos podrían presentar grandes ventajas en diversas aplicaciones, pero la sintetización de estas señales supone un reto tecnológico. Actualmente existen pocos generadores experimentales y con muchas limitaciones. Este artículo propone un diseño con menos limitaciones

Characterization by Electrical Impedance of an In Vitro Model Based on Tumor Cell Spheroids

The main objective of this work is to assess the loosening effect of blebbistatin, a specific inhibitor of myosin II activity, on glioblastoma spheroids. Electric impedance will be used to quantify the effect of this drug, with therapeutic potential for facilitating the access of other treatments into the tumor

Posicionamiento tumoral mediante multielectrodos y redes neuronales

En este trabajo se ha propuesto un método basado en redes neuronales que permite estimar la posición de un tumor por medio de un generador multisalida, estructuras multi-electrodo y redes neuronales, para mejorar el control, focalización, y homogeneidad de las aplicaciones médicas actuales de la electroporación

Processing additivity in Spanish: incluso vs además

El presente trabajo aporta un análisis experimental sobre el procesamiento de relaciones discursivas aditivas en español. Los datos se obtuvieron a partir de dos experimentos en los que el conector argumentativo "además" y el operador focal "incluso" estaban presentes o no en una serie de enunciados sometidos a lectura controlada por eye-tracking. "Incluso" actúa fundamentalmente en el plano de la estructura informativa, mientras que "además" explicita relaciones argumentativas entre segmentos discursivos. Los resultados evidencian que, a pesar de sus diferencias semánticas y sintácticas, la presencia de ambos marcadores discursivos afecta principalmente el procesamiento de alto nivel. Estos resultados constituyen una prueba de cómo funciona el significado fundamentalmente procedimental que se atribuye a estas unidades discursivas

Control of protein synthesis and memory by GluN3A-NMDA receptors through inhibition of GIT1/mTORC1 assembly

De novo protein synthesis is required for synapse modifications underlying stable memory encoding. Yet neurons are highly compartmentalized cells and how protein synthesis can be regulated at the synapse level is unknown. Here, we characterize neuronal signaling complexes formed by the postsynaptic scaffold GIT1, the mechanistic target of rapamycin (mTOR) kinase, and Raptor that couple synaptic stimuli to mTOR-dependent protein synthesis; and identify NMDA receptors containing GluN3A subunits as key negative regulators of GIT1 binding to mTOR. Disruption of GIT1/mTOR complexes by enhancing GluN3A expression or silencing GIT1 inhibits synaptic mTOR activation and restricts the mTOR-dependent translation of specific activity-regulated mRNAs. Conversely, GluN3A removal enables complex formation, potentiates mTOR-dependent protein synthesis, and facilitates the consolidation of associative and spatial memories in mice. The memory enhancement becomes evident with light or spaced training, can be achieved by selectively deleting GluN3A from excitatory neurons during adulthood, and does not compromise other aspects of cognition such as memory flexibility or extinction. Our findings provide mechanistic insight into synaptic translational control and reveal a potentially selective target for cognitive enhancement

Beneficial Effect of Ursodeoxycholic Acid in Patients with ACOX2 Deficiency-Associated Hypertransaminasemia

Background: A variant (p.Arg225Trp) of peroxisomal acyl-CoA oxidase 2 (ACOX2), involved in bile acid (BA) side-chain shortening, has been associated with unexplained persistent hypertransaminasemia and accumulation of C27-BAs, mainly trihydroxycholestanoic acid (THCA). Aims: To investigate the prevalence of ACOX2 deficiency-associated hypertransaminasemia (ADAH), its response to ursodeoxycholic acid (UDCA), elucidate its pathophysiological mechanism and identify other inborn errors that could cause this alteration. Methods & results: Among 33 patients with unexplained hypertransaminasemia from 11 hospitals, and 13 of their relatives, 7 individuals with abnormally high C27-BA levels (>50% of total BAs) were identified by HPLC-MS/MS. The p.Arg225Trp variant was found in homozygosity (exon amplification/sequencing) in 2 patients and 3 family members. Two additional non-related patients were heterozygous carriers of different alleles: c.673C>T (p.Arg225Trp) and c.456_459del (p.Thr154fs). In ADAH patients, impaired liver expression of ACOX2, but not ACOX3, was found (immunohistochemistry). Treatment with UDCA normalized transaminases levels. Incubation of HuH-7 liver cells with THCA, which was efficiently taken up, but not through BA transporters, increased ROS production (flow cytometry), ER stress biomarkers (GRP78, CHOP and XBP1-S/XBP1-U ratio), and BAX¿ expression (RT-qPCR and immunoblot), whereas cell viability was decreased (MTT). THCA-induced cell toxicity was higher than that of major C24-BAs and was not prevented by UDCA. Fourteen predicted ACOX2 variants were generated (site-directed mutagenesis) and expressed in HuH-7 cells. Functional tests to determine their ability to metabolize THCA identified six with the potential to cause ADAH. Conclusion: Dysfunctional ACOX2 has been found in several patients with unexplained hypertransaminasemia. This condition can be accurately identified by a non-invasive diagnostic strategy based on plasma BA profiling and ACOX2 sequencing. Moreover, UDCA treatment can efficiently attenuate liver damage in these patients.This study was supported by the following grants: CIBERehd (EHD15PI05/2016); Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III, Spain (PI19/00819 and PI20/00189), co-funded by European Regional Development Fund/European Social Fund, “Investing in your future”; “Junta de Castilla y León” (SA074P20); Fundació Marato TV3 (201916–31); AECC Scientific Foundation (2017/2020), Spain; and “Centro Internacional sobre el Envejecimiento” (OLD-HEPAMARKER, 0348_CIE_6_E), Spain. We also acknowledge support from grants PID2019-111669RBI- 100, PID2020-115055RB- I00 from Plan Nacional de I+D funded by the “Agencia Estatal de Investigación” (AEI) and the center grant P50AA011999 Southern California Research Center for ALPD and Cirrhosis funded by NIAAA/NIH, as well as support from AGAUR of the “Generalitat de Catalunya” SGR-2017- 1112, European Cooperation in Science & Technology (COST) ACTION CA17112 Prospective European Drug-Induced Liver Injury Network. Marta Alonso-Peña was the recipient of a predoctoral fellowship from “Ministerio de Educación, Cultura y Deporte” (BOE-A- 2015- 9456; FPU-14/ 00214) and a Mobility Grant for Short Stays from “Ministerio de Ciencia, Innovación y Universidades” (EST17/00186). Ricardo Espinosa-Escudero is the recipient of a predoctoral fellowship from “Junta de Castilla y León” and “Fondo Social Europeo” (EDU/574/2018). The funding sources were not involved in the research design or preparation of the articl