A Statistical Study of Resistive Switching Parameters in Au/Ta/ZrO2(Y)/Ta2O5/TiN/Ti Memristive Devices

The authors acknowledge the Consejería de Conocimiento, Investigación y Universidad, Junta de Andalucía (Spain), European Regional Development Fund (ERDF) under projects A-TIC-117-UGR18, B-TIC-624-UGR20, and IE2017-5414. Support from the Government of the Russian Federation under Megagrant Program (agreement no. 074-02-2018-330 (2)) and the Ministry of Science and Higher Education of the Russian Federation under “Priority-2030” Academic Excellence Program of the Lobachevsky State University of Nizhny Novgorod (N-466-99_2021- 2023) is also acknowledged. Memristive devices were designed in the frame of the scientific program of the National Center for Physics and Mathematics (project “Artificial intelligence and big data in technical, industrial, natural and social systems”).Variability is an inherent property of memristive devices based on the switching of resistance in a simple metal–oxide–metal structure compatible with the standard complementary metal–oxide–semiconductor fabrication process. For each specific structure, the variability should be measured and assessed as both the negative and positive factors for different applications of memristive devices. In this report, it is shown how this variability can be extracted and analyzed for such main parameters of resistive switching as the set and reset voltages/cur- rents and how it depends on the methodology used and experimental conditions. The obtained results should be taken into account in the design and predictive simulation of memristive devices and circuits.Consejería de Conocimiento, Investigación y Universidad, Junta de Andalucía (Spain), European Regional Development Fund (ERDF) under projects A-TIC-117-UGR18, B-TIC-624-UGR20, and IE2017-5414Government of the Russian Federation under Megagrant Program (agreement no. 074-02-2018-330 (2))Ministry of Science and Higher Education of the Russian Federation under “Priority-2030” Academic Excellence Program of the Lobachevsky State University of Nizhny Novgorod (N-466-99_2021- 2023

Specific gene correction of the AGXT gene and direct cell reprogramming for the treatment of Primary Hyperoxaluria Type 1

P428 Primary Hyperoxaluria Type 1 (PH1) is an inherited rare metabolic liver disease caused by the deficiency in the alanine: glyoxylate aminotransferase enzyme (AGXT), involved in the glyoxylate metabolism. The only potentially curative treatment is organ transplantation. Thus, the development of new therapeutic approaches for the treatment of these patients appears as a priority.We propose the combination of site-specific gene correction and direct cell reprogramming for the generation of autologous phenotypically healthy induced hepatocytes (iHeps) from skin-derived fibroblast of PH1 patients. For the correction of AGXT mutations, we have designed specific gene editing tools to address gene correction by two different strategies, assisted by CRISPR/Cas9 system. Accurate specific point mutation correction (c.853T-C) has been achieved by homologydirected repair (HDR) with ssODN harbouring wild-type sequence. In the second strategy, an enhanced version ofAGXTcDNAhas been inserted near the transcription start codon of the endogenous gene, constituting an almost universal correction strategy for PH1 mutations. Direct reprogramming of fibroblasts has been conducted by overexpression of hepatic transcription factors and in vitro culture in defined media. In vitro characterization of healthy induced hepatocytes (iHeps) has demonstrated hepatic function of the reprogrammed cells. PH1 patient fibroblasts and , ,

Disrupted in schizophrenia 1 (DISC1) L100P mutants have impaired activity-dependent plasticity in vivo and in vitro

Major neuropsychiatric disorders are genetically complex but share overlapping etiology. Mice mutant for rare, highly penetrant risk variants can be useful in dissecting the molecular mechanisms involved. The gene disrupted in schizophrenia 1 (DISC1) has been associated with increased risk for neuropsychiatric conditions. Mice mutant for Disc1 display morphological, functional and behavioral deficits that are consistent with impairments observed across these disorders. Here we report that Disc1 L100P mutants are less able to reorganize cortical circuitry in response to stimulation in vivo. Molecular analysis reveals that the mutants have a reduced expression of PSD95 and pCREB in visual cortex and fail to adjust expression of such markers in response to altered stimulation. In vitro analysis shows that mutants have impaired functional reorganization of cortical neurons in response to selected forms of neuronal stimulation, but there is no altered basal expression of synaptic markers. These findings suggest that DISC1 has a critical role in the reorganization of cortical plasticity and that this phenotype becomes evident only under challenge, even at early postnatal stages. This result may represent an important etiological mechanism in the emergence of neuropsychiatric disorders

Growth cartilage expression of growth hormone/insulin-like growth factor I axis in spontaneous and growth hormone induced catch-up growth

Introduction: Catch-up growth following the cessation of a growth inhibiting cause occurs in humans and animals. Although its underlying regulatory mechanisms are not well understood, current hypothesis confer an increasing importance to local factors intrinsic to the long bones' growth plate (GP). Aim: The present study was designed to analyze the growth-hormone (GH)-insulin-like growth factor I (IGF-I) axis in the epiphyseal cartilage of young rats exhibiting catch-up growth as well as to evaluate the effect of GH treatment on this process. Material and methods: Female Sprague-Dawley rats were randomly grouped: controls (group C), 50% diet restriction for 3 days + refeeding (group CR); 50% diet restriction for 3 days + refeeding & GH treatment (group CRGH). Analysis of GH receptor (GHR), IGF-I, IGF-I receptor (IGF-IR) and IGF binding protein 5 (IGFBP5) expressions by real-time PCR was performed in tibial growth plates extracted at the time of catch-up growth, identified by osseous front advance greater than that of C animals. Results: In the absence of GH treatment, catch-up growth was associated with increased IGF-I and IGFBP5 mRNA levels, without changes in GHR or IGF-IR. GH treatment maintained the overexpression of IGF-I mRNA and induced an important increase in IGF-IR expression. Conclusions: Catch-up growth that happens after diet restriction might be related with a dual stimulating local effect of IGF-I in growth plate resulting from overexpression and increased bioavailability of IGF-I. GH treatment further enhanced expression of IGF-IR which likely resulted in a potentiation of local IGF-I actions. These findings point out to an important role of growth cartilage GH/IGF-I axis regulation in a rat model of catch-up growth

Targeting a major HIV-1 vulnerability region : the gp41 membrane proximal external region : balance between neutralizing and non-neutralizing antibodies and implications for vaccine design /

La generació d'anticossos àmpliament neutralitzants contra el VIH-1 es el principal objectiu d'una vacuna que sigui capaç de controlar l'epidèmia causada per aquest virus. La glicoproteïna de l'embolcall del VIH-1 es l'únic antigen viral exposat a la membrana del virus y la principal diana de respostes humorals protectores. Dins d'aquesta proteïna han sigut identificades regions funcionals reconegudes per anticossos àmpliament neutralitzants, fent possible la definició de punts de vulnerabilitat del virus, cap als que una vacuna hauria de dirigir-se. En aquest context, la regió externa pròxima a la membrana (MPER) de la gp41 es un dels llocs de vulnerabilitat més representatius del virus, ja que està altament conservat, té un paper crucial per a la infectivitat del virus i es reconegut per anticossos protectors àmpliament neutralitzants. Tot i així, la seva situació, prop a la membrana, fa que el MPER sigui un domini poc accessible, exposat de manera transitòria, hidrofòbic i altament influenciat pels lípids de la membrana. Per tant, la seva immunogenicitat presenta una alta complexitat que ha de ser explorada en major profunditat. En aquesta tesi, aportem nou coneixement sobre l'immunogenicitat del MPER durant ĺinfecció natural i en models animals d'immunització. S'han avaluat miniproteïnes derivades de la gp41 que sobreexposen el MPER, per ser utilitzades com 1) nous plataformes per a la detecció d'anticossos anti-MPER i 2) prototipus d'immunògens presentats en proteoliposomes de diversa composició. Els resultats van revelar una alta immunogenicitat del MPER en humans, que correlaciona amb la resposta global contra la proteïna de l'embolcall, suggerint que el sistema immune no té cap restricció per a la generació d'aquest tipus d'anticossos. Tot i així, els anticossos detectats van mostrar una funcionalitat heterogènia, tant pel que fa a la capacitat neutralitzant com a la competició per diferents epítops, independentment de l'especificitat per MPER. A més, aquest resultats van ser reproduïts en animals immunitzats. En aquest últims, s'ha aconseguit generar un alt títol d'anticossos específics, que van ser potenciats per la incorporació als proteoliposomes de lípids complexes que mimetitzaven als de la partícula viral. Sorprenentment, es va generar una resposta immunodominant no neutralitzant contra un epítop solapant amb el de l'anticòs neutralitzant 2F5. En resum, la resposta humoral contra el MPER a humans i animals immunitzats no va correlacionar amb la capacitat neutralizant d'aquests i els anticossos generats són principalment no neutralitzants. Aquests resultats suggereixen que el balanç de la resposta neutralitzant i no neutralitzant té una important rellevància en la resposta global contra el MPER. Conseqüentment, sembla necessari un major refinament d'immunògens capaços d'evitar respostes no neutralitzants. Aquest redisseny es veurà altament beneficiat pel coneixement generat a partir de nous anticossos monoclonals contra el MPER que recullin l'heterogeneïtat funcional observada als nostres estudis.La generación de anticuerpos ampliamente neutralizantes contra el VIH-1 es el principal objetivo de una vacuna que sea capaz de controlar la epidemia causada por el virus. La glicoproteína de la envuelta del VIH-1 es el único antígeno viral expuesto en la membrana del virus y la principal diana de respuestas humorales protectoras. Dentro de esta proteína se han identificado regiones funcionales que son reconocidas por anticuerpos ampliamente neutralizantes, lo cual ha permitido establecer puntos de vulnerabilidad del virus hacia los que una vacuna debería dirigirse. En este contexto, la región externa próxima a la membrana (MPER) de gp41 es uno de los sitios de vulnerabilidad del virus más representativos, ya que está altamente conservado, juega un papel crucial en la infectividad del virus y es reconocido por anticuerpos protectores ampliamente neutralizantes. Sin embargo, su localización, contigua a la membrana, hace del MPER un dominio poco accesible, expuesto de forma transitoria, hidrofóbico y altamente influenciado por lípidos de membrana. En consecuencia, su inmunogenicidad presenta una alta complejidad que debe ser explorada en mayor profundidad. En esta tesis, aportamos nuevo conocimiento sobre la inmunogenicidad del MPER a nivel de la infección natural y en modelos animales de inmunización. Hemos evaluado miniproteínas basadas en gp41 que sobreexponen el MPER, para ser utilizadas como 1) nuevas plataformas para la detección de anticuerpos anti-MPER y 2) prototipos de inmunógenos presentados en proteoliposomas de diversa composición. Los resultados han revelado una alta inmunogenicidad del MPER en humanos, que correlaciona con la respuesta global contra la proteína de la envuelta. Esto sugiere que el sistema inmune no tiene una especial restricción para la generación de este tipo de anticuerpos. Sin embargo, los anticuerpos detectados mostraron una funcionalidad heterogénea, en términos de capacidad neutralizante y competición por diferentes epítopos, independientemente de la especificidad por el MPER. Además, los resultados fueron reproducidos en animales inmunizados. En estos últimos, conseguimos generar un alto título de anticuerpos específicos, potenciados por la incorporación en los proteoliposomas de mezclas lipídicas complejas que mimetizaban las de la partícula viral. Sorprendentemente, se generó una respuesta inmunodominante no neutralizante que solapaba con un epítopo reconocido por el anticuerpo neutralizante 2F5. En resumen, la repuesta humoral natural contra el MPER en pacientes infectados por el VIH-1 y la generada en modelos animales mediante inmunización comparten ciertas características como son la especificidad y la escasa capacidad neutralizante. Estos resultados sugieren que el balance entre la respuesta neutralizante y no neutralizante podría tener una importante relevancia en la respuesta global contra el MPER. De este modo, es necesario un mayor refinamiento de inmunógenos que sean capaces de sortear respuestas no neutralizantes. Este rediseño se verá altamente beneficiado del conocimiento generado a partir de nuevos anticuerpos monoclonales contra el MPER que reflejen la heterogeneidad funcional observada en nuestros estudios.The elicitation of broadly neutralizing antibodies (bNAbs) against HIV-1 constitutes the major goal of an effective vaccine able to control the epidemic caused by this virus. The HIV-1 envelope glycoprotein is the only viral antigen exposed on the surface of the virus and the main target of protective humoral responses. The identification within this protein of functional regions recognized by bNAbs has delineated an HIV-1 vulnerability map that has guided efforts for rational immunogen design. In this regard, the gp41 Membrane Proximal External Region (MPER) is a major HIV-1 vulnerability site because is highly conserved, plays a major role in viral infectivity and is targeted by protective bNAbs. However, its localization, next to the viral membrane, results in a hardly accessible, transiently exposed and hydrophobic domain, strongly influenced by membrane lipids. Therefore, the immunogenicity of the MPER offers a high level of complexity that needs to be further explored. In this thesis we provide new knowledge on the MPER immunogenicity in both natural HIV-1 infection and immunization in animal models. We generated gp41-based miniproteins which properly exposed the MPER region that have been used 1) as novel platforms for MPER antibody detection and 2) as immunogen candidates presented in different proteoliposome compositions. In humans, the results revealed a strong immunogenicity of the MPER that correlated with a global response against the envelope glycoprotein suggesting no special constraints for the immune system to target this region. However, the antibodies elicited showed heterogeneous functionality in terms of neutralizing capacity and epitope competition, regardless MPER specificity. Interestingly, these results were reproduced by immunization in animal models. High antibody titers were achieved that were specially enhanced by the addition of lipid mixtures mimicking the viral membrane. Interestingly, a non-neutralizing immunodominant response against an epitope that overlapped the 2F5 neutralizing antibody binding motif was identified. Overall, the anti-MPER response in both humans and animal model settings was not correlated with the neutralizing capacity and antibodies detected or induced by immunization were preferentially non-neutralizing. Our results suggest that the balance between neutralizing and non neutralizing responses may represent an important issue in the global response against MPER. Therefore, further redesign of immunogens able to skip non-neutralizing determinants will benefit from the knowledge derived from new anti-MPER antibodies that reflect the functional heterogeneous profile observed in our studies

Lower Respiratory Tract Infection and Short-Term Outcome in Patients With Acute Respiratory Distress Syndrome

To assess whether ventilator-associated lower respiratory tract infections (VA-LRTIs) are associated with mortality in critically ill patients with acute respiratory distress syndrome (ARDS). Post hoc analysis of prospective cohort study including mechanically ventilated patients from a multicenter prospective observational study (TAVeM study); VA-LRTI was defined as either ventilator-associated tracheobronchitis (VAT) or ventilator-associated pneumonia (VAP) based on clinical criteria and microbiological confirmation. Association between intensive care unit (ICU) mortality in patients having ARDS with and without VA-LRTI was assessed through logistic regression controlling for relevant confounders. Association between VA-LRTI and duration of mechanical ventilation and ICU stay was assessed through competing risk analysis. Contribution of VA-LRTI to a mortality model over time was assessed through sequential random forest models. The cohort included 2960 patients of which 524 fulfilled criteria for ARDS; 21% had VA-LRTI (VAT = 10.3% and VAP = 10.7%). After controlling for illness severity and baseline health status, we could not find an association between VA-LRTI and ICU mortality (odds ratio: 1.07; 95% confidence interval: 0.62-1.83; P =.796); VA-LRTI was also not associated with prolonged ICU length of stay or duration of mechanical ventilation. The relative contribution of VA-LRTI to the random forest mortality model remained constant during time. The attributable VA-LRTI mortality for ARDS was higher than the attributable mortality for VA-LRTI alone. After controlling for relevant confounders, we could not find an association between occurrence of VA-LRTI and ICU mortality in patients with ARDS

Alergia a la proteína de leche de vaca en la infancia: microbiota, hidrolizados y tolerancia

Trabajo presentado al XIII Workshop Sociedad Española de Microbiota, Probióticos y Prebióticos, celebrado en Valencia (España), del 7 la 9 de junio de 2022.Introducción La alergia a proteínas de leche de vaca (APLV) es la alergia alimentaria más frecuente en la infancia, habiéndose descrito posibles relaciones con la microbiota intestinal y con el tipo de alimentación. El objetivo de este trabajo es profundizar en el estudio de la microbiota intestinal en menores de un año con APLV y su relación con la adquisición de tolerancia y dieta, comparando muestras al diagnóstico y a los 6 meses de seguimiento con dieta de exclusión láctea. Metodología Se reclutaron 22 pacientes diagnosticados con APLV (14 mediados por IgE y 8 no mediados) y un grupo control de 25 niños sanos. Se recogieron muestras de heces y se realizó un análisis metataxonómico del ADNr 16S y de las regiones ITS de bifidobacterias por secuenciación. Se evaluaron las características clínico-epidemiológicas de los pacientes y se realizó un seguimiento a los 6 meses para evaluar tolerancia y el uso de distintas fórmulas terapéuticas de sustitución alimentaria. Resultados Se detectó un mayor porcentaje de secuencias pertenecientes al filo Actinobacteria (¿60%) en controles frente a casos (¿30%) al diagnóstico. Además, el patrón de abundancias relativas de bifidobacterias fue diferente entre controles y pacientes no mediados por IgE, con una menor proporción de B. longum en estos últimos. Tras la dieta de exclusión, sólo 3 de los pacientes, que estaban tomando distintos tipos de fórmulas terapéuticas, adquirió tolerancia, de los cuales 2 eran casos no mediados por IgE. Conclusiones En los pacientes APLV no IgE mediada se observaron perfiles microbianos distintos de los lactantes sanos, encontrándose a su vez en este grupo una mayor tolerancia al cabo de 6 meses. En tratamiento y seguimiento de la APLV la determinación de la microbiota intestinal puede ser clave para establecer posibles vínculos con la adquisición de tolerancia y el tipo de hidrolizado

Búsqueda de marcadores intestinales asociados a alergia a la proteína de leche de vaca

Resumen trabajo presentado en el XII Workshop Sociedad Española de Microbiota, Probióticos y Prebióticos (SEMiPyP) y I Congreso Sociedad Iberoamericana de Microbiota, Probióticos y Prebióticos (SIAMPYP), celebrado de forma virtual del 15 al 18 de septiembre de 2021.Introducción. De las reacciones de hipersensibilidad causadas por alimentos, la alergia a la proteína de la leche de vaca (APLV) es la más común en los lactantes. En la mayoría de casos se trata de una alergia mediada por IgE, si bien las formas no mediadas son las más desconocidas y difíciles de diagnosticar, con una afectación retardada y fundamentalmente digestiva. Dentro de estas destaca por su gravedad el síndrome de enterocolitis inducida por proteínas de leche de vaca (FPIES). Objetivo. El objetivo de este estudio es buscar biomarcadores microbianos e inflamatorios en heces asociados con distintas formas de presentación de APLV que pudieran facilitar el diagnóstico de estas patologías en la infancia. Metodología. Se parte de un grupo de pacientes = 1 año con distintas formas de presentación de APLV reclutados a lo largo de un año en tres Hospitales de Asturias. Con fines comparativos, se recluta en centros de atención primaria un grupo control de lactantes sanos sin ninguna enfermedad digestiva ni alérgica. A partir de las heces de los niños se realiza el análisis de parámetros inflamatorios, así como de la composición microbiana y de sus metabolitos. Resultados. Se detectaron distintos perfiles entre el grupo control y los pacientes en función del tipo de hipersensibilidad. Destaca la disminución de la excreción fecal de ácidos grasos de cadena corta (AGCC) en aquellos niños con formas no mediadas por IgE respecto al grupo de niños sanos y a los pacientes con APLV mediada por IgE. Conclusiones. Las poblaciones microbianas que se desarrollan en el intestino del lactante y la producción de metabolitos bacterianos específicos pueden ser claves en la interacción con el sistema inmunitario del lactante en este tipo de alergias. El análisis de muestras de heces podría ayudar al diagnóstico, sobre todo en los casos no mediados por IgE

Accessory gene regulator (Agr) functionality in Staphylococcus aureus derived from lower respiratory tract infections

Altres ajuts: This work has been funded by the project PI13/01418 which is part of "Plan Nacional de I+D+I" and co-funded by ISCIII-Subdirección General de Evaluacioón and "Fondo Europeo de Desarrollo Regional"(FEDER).D. Domínguez-Villanueva is funded by "Plan Nacional de I+D+I" and co-funded by ISCIII-Subdirección General de Evaluación and "Fondo Europeo de Desarrollo Regional"(FEDER). M. Gomes-Fernandes is funded by CAPES Foundation, Ministry of Education of Brazil (Brasılia, Brazil). Maisem Laabei was supported by a joint ERS/SEPAR fellowship (LTRF2015).This work also received a grant from the Spanish Society of Pneumology and Thoracic Surgery (SEPAR054/2011).Objective. Characterization of Staphylococcus aureus clinical isolates derived from lower respiratory tract infections (LRTIs), and correlation between the functionality of the accessory gene regulator (Agr) and genotypic and phenotypic characteristics, clinical variables and clinical outcome. Methods. S aureus isolates derived from LRTIs and control groups (nasal carriage and bacteraemia) were genotyped using StaphyType DNA microarray. Agr activity was evaluated using the CAMP synergistic haemolysis assay and the Vesicle Lysis Test (VLT). Discordant strains were analysed by quantitative reverse- transcriptase real-time PCR (qRT-PCR). Results. Agr was functional in 79.7% and 84.5% of strains according to the CAMP and VLT assays respectively. Higher concordance with RNAIII expression measured by qRT-PCR was observed with the VLT assay (76.2%) compared with the CAMP assay (23.8%). No statistically significant differences were observed in Agr functionality between the study groups, nor the phenotypical/genotypical bacterial characteristics. No association between increased mortality/respiratory complications and Agr function was observed. Conclusions. Agr activity was high (82.2%) in isolates from LRTIs suggesting the importance of this global regulator in lower respiratory tract colonisation and infection. However, equally high Agr activity was observed in isolates derived from nasal carriage and bacteraemia, contradictory to previous observations. Agr functionality measured by the VLT assay was superior to CAMP assay