Engineering self-regulated synthetic gene expression systems and its application on controlled protein expression

One of the limiting factor in industrial biotechnology is the lack of cost-effective and regulated gene expression systems at large-scale. Thus current approaches suffer of suboptimal gene expression levels and/or they require of expensive inducer metabolites. The emergent of synthetic biology including new highthrouput DNA assembly methods and sophisticated biosensors now allow the redesign of complex bacterial trait including automata and full pre-programed gene expression control systems.  This high level of control allows save inducers in bioprocesses and/or reducing the time where the cells and toxic intermediates are in contact by generating an auto-regulated expression system.With the aim to address this important shortcoming in industrial biotechnology and provide a cheap and highly controlled gene expression system at large scale, in this work we firstly have developed a new highthrouput DNA  assembly system based on GoldenGate´s MoClo [1] adapted to the Standard European Vector Architecture (pSEVA) [2]. The new methods was further used to generate a large array of complex circuits capable of regulate genetic expression according to phosphate concentration by taking advantage of an existing collection of synthetic promoter available at SBG group [3].The circuits were generated by assembling synthetic transcriptional units using random assembly of synthetic promoters and ribosome binding sites driving the expression of  well-known reporter  fluorescent proteins such as GFP and RFP, thus resulting on a wide variety of circuits. The circuits obtained where subsequently analyzed by following the fluorescent profile under different phosphate concentrations media. Those that showed a more promissing behavior where selected to be further studied and sequenced to determine those random elements. Ongoing efforts have focus on the construction of a set of well-characterized synthetic circuits driving the a la carte and self-regulated expression of a given gene, even a whole metabolic pathway.In summary, we have obtained a way number of different plasmids, circuits and components that can be used to elaborate new circuits and constructions with all new applications and functions such as the above mentioned genetic expression self-regulation based on externals factors like phosphate concentration and its application to large scale production systems

Rechazo entre iguales: modelos teóricos asociativos

Comunicación presentada en las I Congreso Internacional de Estrategias Didácticas Inclusivas, celebradas en la Universitat Jaume I (Castellón, España) los días 1 y 2 diciembre 2011El rechazo entre iguales es un aspecto de mucha relevancia en el ajuste socio-emocional y cognitivo del niño, lo cual ha motivado que en las tres últimas décadas se hayan generado varias tentativas explicativas tanto del proceso del rechazo per se como de su impacto en el desarrollo del niño. Los primeros modelos teóricos que aparecen tienen una orientación de efectos principales, pero rápidamente son seguidos por modelos más complejos, de efectos aditivos. En el presente trabajo se exponen dichos modelos de forma sintetizada, acompañando cada modelo con su representación esquemática, y con un ejemplo extraído de la literatura científica del ámbito del rechazo entre iguales en la infancia

Non‐branched β‐1,3‐glucan oligosaccharides trigger immune responses in Arabidopsis

[EN] Fungal cell walls, which are essential for environmental adaptation and host colonization by the fungus, have been evolutionarily selected by plants and animals as a source of microbe-associated molecular patterns (MAMPs) that, upon recognition by host pattern recognition receptors (PRRs), trigger immune responses conferring disease resistance. Chito-oligosaccharides [b-1,4-N-acetylglucosamine oligomers, (GlcNAc)n] are the only glycosidic structures from fungal walls that have been well-demonstrated to function as MAMPs in plants. Perception of (GlcNAc)4–8 by Arabidopsis involves CERK1, LYK4 and LYK5, three of the eight members of the LysM PRR family. We found that a glucan-enriched wall fraction from the pathogenic fungus Plectosphaerella cucumerina which was devoid of GlcNAc activated immune responses in Arabidopsis wild-type plants but not in the cerk1 mutant. Using this differential response, we identified the non-branched 1,3-b-D-(Glc) hexasaccharide as a major fungal MAMP. Recognition of 1,3-b-D-(Glc)6 was impaired in cerk1 but not in mutants defective in either each of the LysM PRR family members or in the PRR-co-receptor BAK1. Transcriptomic analyses of Arabidopsis plants treated with 1,3-b-D-(Glc)6 further demonstrated that this fungal MAMP triggers the expression of immunity-associated genes. In silico docking analyses with molecular mechanics and solvation energy calculations corroborated that CERK1 can bind 1,3-b-D-(Glc)6 at effective concentrations similar to those of (GlcNAc)4. These data support that plants, like animals, have selected as MAMPs the linear 1,3-b-D-glucans present in the walls of fungi and oomycetes. Our data also suggest that CERK1 functions as an immune co-receptor for linear 1,3-b-D-glucans in a similar way to its proposed function in the recognition of fungal chito-oligosaccharides and bacterial peptidoglycan MAMPs.S

La imagen que el profesor tiene de los alumnos y la aceptación de los alumnos por sus iguales en primero de primaria

Comunicación presentada en las I Congreso Internacional sobre Ciudadanía Crítica y Mejora Educativa: recursos, estrategias y experiencias, celebradas en la Universitat Jaume I (Castellón, España) los días 13 al 15 de octubre 2011El presente trabajo estudia la relación entre la imagen del alumno que tiene el profesor y la aceptación de los alumnos por sus iguales. La muestra está formada por 247 alumnos escolarizados en 11 aulas de primero de primaria, de cinco colegios públicos de Castellón de la Plana. A nivel metodológico se ha realizado un estudio de correlaciones, el procedimiento Anova de un factor, un análisis de tablas de contingencia r, y un análisis de regresión. Respecto a los resultados, las correlaciones son significativas, el Anova de un factor muestra que el profesor establece claras diferencias entre tipologías sociométricas y el análisis de regresión refleja cierto carácter predictivo de la variable preferencia del maestro sobre la aceptación de los iguales

Potenciación del contexto social escolar que promueve la convivencia, la inclusión, la ciudadanía y el aprendizaje

Comunicación presentada en las I Congreso Internacional sobre Ciudadanía Crítica y Mejora Educativa: recursos, estrategias y experiencias, celebradas en la Universitat Jaume I (Castellón, España) los días 13 al 15 de octubre 2011.El artículo trata sobre la implementación de un programa multinivel, multicomponente y multiagente. Se aplicó a 4 colegios de Castellón en las aulas de 1º de primaria (curso 2010-2011). Se basa en 3 grandes componentes (normas de convivencia y estrategias permanentes, aprendizaje cooperativo y desarrollo socioemocional) a partir de los cuales se capacitó a los profesores durante 6 sesiones de formación (30 horas). Ellos mismos fueron los que implementaron el programa adaptándolo al currículo, por lo que se pudo observar mejoras considerables, por una parte, aprendieron a ser más conscientes en la forma de dirigirse al alumnado; y por otra, en las relaciones interpersonales con su clase y entre los propios alumnos

Gene Expression Patterns of Oxidative Phosphorylation Complex I Subunits Are Organized in Clusters

After the radiation of eukaryotes, the NUO operon, controlling the transcription of the NADH dehydrogenase complex of the oxidative phosphorylation system (OXPHOS complex I), was broken down and genes encoding this protein complex were dispersed across the nuclear genome. Seven genes, however, were retained in the genome of the mitochondrion, the ancient symbiote of eukaryotes. This division, in combination with the three-fold increase in subunit number from bacteria (N = ∼14) to man (N = 45), renders the transcription regulation of OXPHOS complex I a challenge. Recently bioinformatics analysis of the promoter regions of all OXPHOS genes in mammals supported patterns of co-regulation, suggesting that natural selection favored a mechanism facilitating the transcriptional regulatory control of genes encoding subunits of these large protein complexes. Here, using real time PCR of mitochondrial (mtDNA)- and nuclear DNA (nDNA)-encoded transcripts in a panel of 13 different human tissues, we show that the expression pattern of OXPHOS complex I genes is regulated in several clusters. Firstly, all mtDNA-encoded complex I subunits (N = 7) share a similar expression pattern, distinct from all tested nDNA-encoded subunits (N = 10). Secondly, two sub-clusters of nDNA-encoded transcripts with significantly different expression patterns were observed. Thirdly, the expression patterns of two nDNA-encoded genes, NDUFA4 and NDUFA5, notably diverged from the rest of the nDNA-encoded subunits, suggesting a certain degree of tissue specificity. Finally, the expression pattern of the mtDNA-encoded ND4L gene diverged from the rest of the tested mtDNA-encoded transcripts that are regulated by the same promoter, consistent with post-transcriptional regulation. These findings suggest, for the first time, that the regulation of complex I subunits expression in humans is complex rather than reflecting global co-regulation

Quality of Life and Autonomy in Patients with Intermittent Bladder Catheterization Trained by Specialized Nurses

Intermittent bladder catheterization (IBC) involves regular urine draining using a catheter, which is removed immediately after urinary elimination. It allows for the patient's urological health to be managed and their renal function to be preserved, and it promotes autonomy. Compliance with the prescribed number of daily catheterizations, which must be conducted by the patient, and infection prevention measures are crucial. To identify the patients requiring IBC, and to determine their adherence (whether they followed the prescribed guidelines and their difficulty in carrying out the procedure, as well as to assess how the IBC influences their quality of life and state of mind after receiving self-care training from a specialized nurse), we carried out a prospective, multicenter observational study in 24 Spanish hospitals with one month of monitoring and a sample of 99 patients. The sources of information were the patients' clinical records, the King's Health Questionnaire, the Mini-Mental State Examination (MMSE), and the hospital anxiety and depression scale (HADS). Descriptive and bivariate statistics were used to analyses the paired data. After recruitment (n = 99), 79 patients completed the questionnaire at a mean age of 35.2 years (SD = 20.5 years). In total, 53.5% (53) of the sample consisted of men and 32.3% (32) had neurological damage as the reason for prescription; 67% (67.7) performed self-catheterization and 86.7% adhered to the IBC. After one month of monitoring, a statistically significant improvement in quality of life was observed in all criteria, with the exception of personal relationships (p < 0.005), as well as an improvement in anxiety and depression levels (p < 0.001). Patients who require IBC show good adherence to the IBC with a significant percentage of self-catheterization. After one month of IBC, a significant improvement in the patients' quality of life and mood was observed. These results could be attributed to adequate patient training and adequate personalization of the IBC materials by the specialized nurses

Evolution of Quality of Life and Treatment Adherence after One Year of Intermittent Bladder Catheterisation in Functional Urology Unit Patients

Objective: To determine patient difficulties and concerns when performing IBC (Intermittent Bladder Catheterisation), as well as the evolution of adherence, quality of life, and emotional state of patients one year after starting IBC. Method: A prospective, observational, multicentre study conducted in 20 Spanish hospitals with a one-year follow-up. Data sources were patient records and the King's Health Questionnaire on quality of life, the Mini-Mental State Examination (MMSE), and the Hospital Anxiety and Depression Scale (HADS). Perceived adherence was measured using the ICAS (Intermittent Catheterization Adherence Scale) and perceived difficulties with IBC were assessed using the ICDQ (Intermittent Catheterization Difficulty Questionnaire). For data analysis, descriptive and bivariate statistics were performed for paired data at three points in time (T1: one month, T2: three months, T3: one year). Results: A total of 134 subjects initially participated in the study (T0), becoming 104 subjects at T1, 91 at T2, and 88 at T3, with a mean age of 39 years (standard deviation = 22.16 years). Actual IBC adherence ranged from 84.8% at T1 to 84.1% at T3. After one year of follow-up, a statistically significant improvement in quality of life (p <= 0.05) was observed in all dimensions with the exception of personal relationships. However, there were no changes in the levels of anxiety (p = 0.190) or depression (p = 0.682) at T3 compared to T0. Conclusions: Patients requiring IBC exhibit good treatment adherence, with a significant proportion of them performing self-catheterisation. After one year of IBC, a significant improvement in quality of life was noted, albeit with a significant impact on their daily lives and their personal and social relationships. Patient support programmes could be implemented to improve their ability to cope with difficulties and thus enhance both their quality of life and the maintenance of their adherence

Healthcare workers hospitalized due to COVID-19 have no higher risk of death than general population. Data from the Spanish SEMI-COVID-19 Registry

Aim To determine whether healthcare workers (HCW) hospitalized in Spain due to COVID-19 have a worse prognosis than non-healthcare workers (NHCW). Methods Observational cohort study based on the SEMI-COVID-19 Registry, a nationwide registry that collects sociodemographic, clinical, laboratory, and treatment data on patients hospitalised with COVID-19 in Spain. Patients aged 20-65 years were selected. A multivariate logistic regression model was performed to identify factors associated with mortality. Results As of 22 May 2020, 4393 patients were included, of whom 419 (9.5%) were HCW. Median (interquartile range) age of HCW was 52 (15) years and 62.4% were women. Prevalence of comorbidities and severe radiological findings upon admission were less frequent in HCW. There were no difference in need of respiratory support and admission to intensive care unit, but occurrence of sepsis and in-hospital mortality was lower in HCW (1.7% vs. 3.9%; p = 0.024 and 0.7% vs. 4.8%; p<0.001 respectively). Age, male sex and comorbidity, were independently associated with higher in-hospital mortality and healthcare working with lower mortality (OR 0.211, 95%CI 0.067-0.667, p = 0.008). 30-days survival was higher in HCW (0.968 vs. 0.851 p<0.001). Conclusions Hospitalized COVID-19 HCW had fewer comorbidities and a better prognosis than NHCW. Our results suggest that professional exposure to COVID-19 in HCW does not carry more clinical severity nor mortality

Clonación, expresión y caracterización de las penicilina acilasas de Streptomyces lavendulae y Actimnoplanes utahensis

219 p.-83 fig.-25 tab.-2 anexos.La ciencia en muchas ocasiones se nutre de la casualidad y la buena interpretación de las señales observadas puede llegar a grandes descubrimientos. Así la casualidad hizo que en 1928, Alexander Fleming descubriese la capacidad antibacteriana delhon Penicillium notatum, tras observar y analizar la contaminación accidental de las placas de cultivos de Streptococus aureus. Posteriormente, desarrolló el hongo de un medio de cultivo líquido, observando que el exacto filtrado, libre de células, era capaz de inhibir el crecimiento de muchas especies bacterianas. Al principio activo de estos caldos lo denominó "penicilina". Fleming no pudo seguir avanzando en el estudio de la peneicilina, ya que todos los intentos llevados a cabo para su aislamiento y purificación resultaron infroructuosos debido a su inestabilidad. Fue Ernest Chain quien, diez años más tarde, en 1940, consiguió aislar y obtener una preparación estable de penicilina, así como su carazterización físico-química. Esto permitió que el grupo de Horward W. Florey desarrollase los métodos que permitieron el uso terapeútico de la penicilina, al demostrar su baja toxicidad en los animales poniendo de manifiesto su impresionante capacidad como agente bactericida en el tratamiento de enfermedades infecciosas, lo que hizo posible su utilización con gran éxito durante la Segunda Guerra Mundial y supuso el comienzo de la era de los antibióticos.Peer reviewe