Improving carotenoids biosynthesis pathway in the unicelullar microalgae Chlamydomonas reinhardtii

Introduction: Carotenoids are a wide group of isoprenoids synthesized by photosynthetic organisms and some non-photosynthetic yeast and bacteria (1).They are indispensable in light harvesting and energy transference during photosynthesis and in the protection of the photosynthetic apparatus against the photooxidative damage. Mammals cannot synthesize them and must include them in their diet as precursors for essential compounds. The important colorant, antioxidant and provitamin properties of carotenoids, have made of them an important group of high-added value compounds, massively commercialized (1,2).There is an increasing demand of natural carotenoids and microalgae can be an excellent natural source of carotenoids.Results and conclusions: In this work, we describe the subcloning of two genes from the carotenoid biosynthesis pathway in a microalgal expression vector: The PSY gene from Dunaliella salina, encoding phytoene synthase, a key enzyme in the pathway, which catalyzes the formation of phytoene, and CRTI gene encoding for fitoene desaturase from Erwinia Uredowa. The bacterial CRTI gene catalyzes the conversion of phytoene to lycopene, replacing the function of two microalgal enzymes phytoene desaturase (PDS) and chis-carotene desaturase (ZDS) (1,3). PSY and CRTI genes were fused by a short DNA fragment which encodes a self-cleaving peptide and fused to the selective marker gene APHVIII from Streptomyces rimosus, encoding for an aminoglycoside 3'phosphotransferase that confers resistance to the antibiotic paromomycin. All genes were placed under the control of the strong constitutive promoters RBCS2 and HSP70A and terminated by the 3'untranslated region of RBCS2 (plasmid 4-75).C. reinhardtii was nuclear transformated with plasmid 4-75. Obtained transformants were analysed by PCR to check the insertion of the PSY, CRTI and APHVIII genes into the genome of C. reinhardtii and tested for expression at mRNA level. The phenotype of some of the transformants was also analyzed studying their carotenoid composition.The obtained data show that the designed vector allows the insertion of all genes into the genome, although we have detected unexpected DNA cleavage and rearrangements during the integration process, which leads to discontinuous insertion of the genes, as they are placed in the 4-75 plasmid. This has made impossible to isolate any transformant with significant phenotypical changes so far and it is subject of current investigation

Papel de NFkB como posible biomarcador en el desarrollo de cáncer de pulmón en pacientes previamente diagnosticados de EPOC

Comunicaciones a congreso

FGFR1 Cooperates with EGFR in Lung Cancer Oncogenesis, and Their Combined Inhibition Shows Improved Efficacy

Introduction: There is substantial evidence for the onco- genic effects of fi broblast growth factor receptor 1 (FGFR1) in many types of cancer, including lung cancer, but the role of this receptor has not been addressed speci fi cally in lung adenocarcinoma. Methods: We performed FGFR1 and EGFR overexpression and co-overexpression assays in adenocarcinoma and in inmortalized lung cell lines, and we also carried out surrogateandinteractionassays.Weperformedmono- therapy and combination EGFR /FGFR inhibitor sensitivity assays in vitro and in vivo in cell line – and patient- derived xenografts. We determined FGFR1 mRNA expression in a cohort of patients with anti – EGFR ther- apy – treated adenocarcinoma. Results: We have reported a cooperative interaction between FGFR1 and EGFR in this context, resulting in increased EGFR activation and oncogenic signaling. We have provided in vitro and in vivo evidence indicating that FGFR1 expression in- creases tumorigenicity in cells with high EGFR activation in EGFR-mutated and EGFR wild-type models. At the clinical level, we have shown that high FGFR1 expression levels pre- dict higher resistance to erlotinib or ge fi tinib in a cohort of patients with tyrosine kinase inhibitor – treated EGFR-mutated and EGFR wild-type lung adenocarcinoma. Dual EGFR and FGFR inhibition in FGFR1-over expressing, EGFR-activated models shows synergistic effects on tumor growth in vitro and in cell line – and patient-derived xenografts, suggesting that patients with tumors bearing these characteristics may bene fi t from combined EGFR/FGFR inhibition. Conclusion: These results support the extended the use of EGFR inhibitors beyond monotherapy in the EGFR-mutated adenocarcinoma setting in combination with FGFR in- hibitors for selected patients with increased FGFR1 over- expression and EGFR activation.ISCIII PI14/01964 PIE15/00076 PI17/00778 DTS17/00089 PI15/00045 PI17/00033 PI16/01311 FI12/00429CIBERONC CD16/12/00442FEDER CD16/12/00442 PI16/01311Spanish Ministry of Economy and Competitiveness PI15/00045Ministry of Health and Social Welfare of Junta de Andalucía PI-0046-2012 C-0040-2016Ministry of Equality, Health and Social Policies of the Junta de Andalucía PI- 0029-2013Comunidad de Madrid B2017/BMD3884Ministry of Education, Culture and Sports FPU13/0259

Alteraciones de la aorta ascendente en un modelo animal espontáneo de válvula aórtica bicúspide

La válvula aórtica bicúspide (VAB) es la malformación cardiaca congénita más prevalente y se asocia frecuentemente con dilatación de la aorta ascendente (DAA). Existen dos hipótesis etiológicas sobre esta asociación: 1) genética: VAB y alteraciones estructurales de la pared aórtica son el resultado de un mismo defecto embrionario; 2) hemodinámica: alteraciones del flujo causadas por el defecto valvular provocan la aortopatía. El sustrato histopatológico de la DAA es la necrosis cística de la media (NCM): disrupción de las lamelas elásticas, muerte de las células musculares lisas, acúmulo de sustancia basófila en áreas acelulares, ausencia de fibrosis o inflamación. Se desconoce si durante la patogénesis de la DAA, la disrupción de la elástica precede a la muerte celular o si esta última es la causa original. Contamos con el único modelo animal espontáneo de VAB, consistente en una cepa consanguínea de hámsteres con una incidencia de VAB cercana al 40% (CE). Hemos investigado la anatomía e histología de la aorta en 148 animales de la CE y 71 de una cepa control (CC), desde la juventud a la senectud. En los animales de la CC, el diámetro de la aorta ascendente aumentó progresivamente con la edad (incremento del 22% en 1,5 años), mientras que en los de la CE permaneció constante hasta la vejez (15 meses), pero aumentó súbita y significativamente (19%) en los últimos 3 meses. Comparada con la CC, la media aórtica de los hámsteres adultos de la CE presentó mayor grosor, número de lamelas, grado de ondulación de las lamelas y muerte de las células musculares lisas. La muerte celular originó áreas acelulares con acúmulo de sustancia basófila, sin presencia de inflamación ni fibrosis. No se detectó disrupción de las lamelas elásticas. Las diferencias observadas fueron de tipo cuantitativo, no cualitativo, y se detectaron entre animales de la CE y la CC, pero no entre animales de la CE con VAB o con VAT. Presentamos el primer modelo animal espontáneo de alteraciones de la aorta asociadas a la VAB. Nuestros resultados indican que la NCM es una proceso degenerativo de la aorta asociado a la senectud, cuyo factor desencadenante es la muerte de las células musculares lisas. El hecho de que los hámsteres consanguíneos de la CE presenten alteraciones en la aorta independientemente del morfotipo valvular apoya la hipótesis genética sobre la asociación entre VAB y DAA.Universidad de Málaga. Campus de Excelencia. Internacional Andalucía Tech. P10-CTS-6068 (Junta de Andalucía. Red de Investigación Cardiovascular (RIC; RETICs

A novel biocompatible polymer derived from D-mannitol used as a vector in the field of genetic engineering of eukaryotic cells

The design and preparation of new vectors to transport genetic material and increase the transfection efficiency continue being an important research line. Here, a novel biocompatible sugar-based polymer derived from Dmannitol has been synthesized to be used as a gene material nanocarrier in human (gene transfection) and microalga cells (transformation process). Its low toxicity allows its use in processes with both medical and industrial applications. A multidisciplinary study about the formation of polymer/p-DNA polyplexes has been carried out using techniques such as gel electrophoresis, zeta potential, dynamic light scattering, atomic force microscopy, and circular dichroism spectroscopy. The nucleic acids used were the eukaryotic expression plasmid pEGFP-C1 and the microalgal expression plasmid Phyco69, which showed different behaviors. The importance of DNA supercoiling in both transfection and transformation processes was demonstrated. Better results were obtained in microalga cells nuclear transformation than in human cells gene transfection. This was related to the plasmid’s conformational changes, in particular to their superhelical structure. It is noteworthy that the same nanocarrier has been used with eukaryotic cells from both human and microalga.This work was supported by the Consejería de Conocimiento, Innovación y Universidades de la Junta de Andalucía (FQM-206, FQM-274, FQM-135 and P20–01234); VI Plan Propio Universidad de Sevilla (PP2019/00000748), and the European Union (Feder Funds)

Effect of a Graduated Walking Program on the Severity of Obstructive Sleep Apnea Syndrome. A Randomized Clinical Trial

Background: Obstructive sleep apnea syndrome (OSAS) is a common disease. The objective of this research was to determine the effectiveness of a graduated walking program in reducing the apnea–hypopnea index number in patients with obstructive sleep apnea syndrome (OSAS). Methods: A randomized controlled clinical trial with a two-arm parallel in three tertiary hospitals was carried out with seventy sedentary patients with moderate to severe OSAS. Twenty-nine subjects in each arm were analyzed by protocol. The control group received usual care, while usual care and an exercise program based on progressive walks without direct supervision for 6 months were offered to the intervention group. Results: The apnea–hypopnea index decreased by six points in the intervention group, and improvements in oxygen desaturation index, total cholesterol, and Low-Density Lipoprotein of Cholesterol (LDL-c) were observed. A higher decrease in sleep apnea–hypopnea index (45 ± 20.6 vs. 34 ± 26.3/h; p = 0.002) was found in patients with severe vs. moderate OSAS, as well as in oxygen desaturation index from baseline values (43.3 vs. 34.3/h; p = 0.046). Besides, High-Density Lipoprotein of Cholesterol (HDL-c) values showed a higher increase in the intervention group (45.3 vs. 49.5 mg/dL; p = 0.009) and also, a higher decrease in LDL-c was found in this group (141.2 vs. 127.5 mg/dL; p = 0.038). Conclusion: A home physical exercise program is a useful and viable therapeutic measure for the management of OSAS

Dental age estimation based on pulp chamber/crown volume ratio measured on CBCT images in a Spanish population

Dental age estimation in living individuals is one of the most frequent requests undertaken by forensic odontologists. The aim of this study was to estimate the dental age by pulp/tooth volume ratio, as measured on cone beam computed tomography (CBCT) images, in a Spanish population. This study included 313 teeth from 107 adult individuals, 56 females and 51 males with a mean age of 44 ± 14 years. The statistical analysis of the results took account of clustering (multiple teeth in individuals). Linear regression models were constructed on the relationship between pulp/tooth volume ratio and chronological age for each tooth type. The highest coefficient of determination (R2) value was provided by the upper incisors (36.6%), and the difference between chronological and estimated age was less than 5 years in 31.3% of the sample and less than 10 years for 65.7%. CBCT is an accurate imaging technique to measure dental volume with a relatively low radiation dose, and it can be used to assess dental age in living adult individuals. Volumetric changes in the pulp cavity with increasing age proved valuable to estimate dental age in this Spanish populatio

Predicting the distribution of canine leishmaniasis in western Europe based on environmental variables.

The domestic dog is the reservoir host of Leishmania infantum, the causative agent of zoonotic visceral leishmaniasis endemic in Mediterranean Europe. Targeted control requires predictive risk maps of canine leishmaniasis (CanL), which are now explored. We databased 2187 published and unpublished surveys of CanL in southern Europe. A total of 947 western surveys met inclusion criteria for analysis, including serological identification of infection (504, 369 dogs tested 1971-2006). Seroprevalence was 23 2% overall (median 10%). Logistic regression models within a GIS framework identified the main environmental predictors of CanL seroprevalence in Portugal, Spain, France and Italy, or in France alone. A 10-fold cross-validation approach determined model capacity to predict point-values of seroprevalence and the correct seroprevalence class (20%). Both the four-country and France-only models performed reasonably well for predicting correctly the 20% seroprevalence classes (AUC >0 70). However, the France-only model performed much better for France than the four-country model. The four-country model adequately predicted regions of CanL emergence in northern Italy (<5% seroprevalence). Both models poorly predicted intermediate point seroprevalences (5-20%) within regional foci, because surveys were biased towards known rural foci and Mediterranean bioclimates. Our recommendations for standardizing surveys would permit higher-resolution risk mapping

Improvement of carotenoid extraction from a recently isolated, robust microalga, Tetraselmis sp. CTP4 (chlorophyta)

In recent years, there has been increasing consumer interest in carotenoids, particularly of marine sustainable origin with applications in the food, cosmeceutical, nutritional supplement and pharmaceutical industries. For instance, microalgae belonging to the genus Tetraselmis are known for their biotechnologically relevant carotenoid profile. The recently isolated marine microalgal strain Tetraselmis sp. CTP4 is a fast-growing, robust industrial strain, which has successfully been produced in 100-m3 photobioreactors. However, there are no reports on total carotenoid contents from this strain belonging to T. striata/convolutae clade. Although there are several reports on extraction methods targeting chlorophytes, extraction depends on the strength of cell coverings, solvent polarity and the nature of the targeted carotenoids. Therefore, this article evaluates different extraction methods targeting Tetraselmis sp. CTP4, a strain known to contain a mechanically resistant theca. Here, we propose a factorial experimental design to compare extraction of total carotenoids from wet and freeze-dried microalgal biomass using four different solvents (acetone, ethanol, methanol or tetrahydrofuran) in combination with two types of mechanical cell disruption (glass beads or dispersion). The extraction efficiency of the methods was assessed by pigment contents and profiles present in the extracts. Extraction of wet biomass by means of glass bead-assisted cell disruption using tetrahydrofuran yielded the highest amounts of lutein and β-carotene (622 ± 40 and 618 ± 32 µg g-1 DW, respectively). Although acetone was slightly less efficient than tetrahydrofuran, it is preferable due to its lower costs and toxicity.info:eu-repo/semantics/publishedVersio

La Imagen y la Narrativa como Herramientas para el Abordaje Psicosocial en Escenarios de Violencia. Departamento del Cesar

Teniendo en cuenta los referentes bibliográficos y contenidos de las unidades trabajadas a lo largo del curso, logramos evidenciar los diferentes enfoques que abarcan la intervención psicosocial en la problemática del conflicto armado en Colombia, generando profundización de las posibilidades de enfoques psicosocial al momento de abordar a las víctimas de conflicto armado. En el presente trabajo , se encontrará con la propuesta del caso “Camilo”, en el cual se puede dejar a la luz una serie de acontecimientos o sucesos vividos, donde la violencia emerge la vida de los habitantes de un municipio de Colombia, dejando claro que la desigualdad, la injusticia hacen eje central de las vivencias de estas personas, de igual modo, también se refleja como de una manera inhumana, actualmente se tiene que llevar con el peso de la exclusión social, el desalojo de sus tierras, las constantes amenazas, el desarraigo de sus identidad, sobreponiéndolo como una persona empoderada, capaz de cambiar la historia de su vida y la de sus familiares, enfrentando todos los desafíos contantes para salir adelante, haciendo gala de su empoderamiento y fortaleza, mostrándose así mismo como una víctima, pero capaz de sobrepasar los esquemas de dolor, tristeza, terror y victimización. Por otro lado, presentamos el caso Pandurí, en el cual abarcamos teniendo en cuenta los procesos de acción psicosocial en las victimas de conflicto armado. Es importante resaltar que posterior de la irrupción de los grupos armados, se focalizan emergentes psicosociales donde se puede detallar los impactos frente a los hechos de terrorismo. Si nos ponemos de cara a realidad de la moneda, focalizamos que toda incursión de violencia, trae consigo transformaciones que generan impactos en la comunidad y que de ello dependerá el futuro de muchas personas, a corto, mediano y largo plazo, así como lo demuestra el caso Pandurí. Todas estas series de acontecimientos, nos permitirán desde una mirada psicosocial, como se deben intervenir los procesos que emergen del conflicto armado, se debe tomar a los individuos implicados, no como simples víctimas y mediante la intervención psicosocial, postular como resilientes las personas que diariamente viven este conflicto.Taking into account the bibliographic references and contents of the units worked throughout the course, we were able to highlight the different approaches that embrace psychosocial intervention in the problems of the armed conflict in Colombia, generating a deepening of the possibilities of psychosocial approaches when addressing victims of armed conflict. In this work, you will find the proposal of the “Camilo” case, in which a series of events or lived events can be brought to light, where violence emerges the lives of the inhabitants of a municipality in Colombia, leaving Of course, inequality and injustice are the central axis of the experiences of these people, likewise, it is also reflected in an inhuman way, currently it has to be carried with the weight of social exclusion, the eviction of their lands, the constant threats, the uprooting of his identity, overlapping him as an empowered person, capable of changing the history of his life and that of his family, facing all the constant challenges to get ahead, showing off his empowerment and strength, showing himself as a victim, but capable of overcoming the patterns of pain, sadness, terror and victimization. On the other hand, we present the Pandurí case, in which we cover taking into account the processes of psychosocial action in the victims of armed conflict. It is important to highlight that after the irruption of the armed groups, psychosocial emergencies are focused on where the impacts of terrorism can be detailed. If we face the reality of the currency, we focus that any incursion of violence brings about transformations that generate impacts on the community and that the future of many people will depend on it, in the short, medium and long term, as demonstrated the Pandurí case. 5 All these series of events will allow us, from a psychosocial perspective, how the processes that emerge from the armed conflict must be intervened, the individuals involved must be taken, not as simple victims and through psychosocial intervention, postulate as resilient the people who daily they live this conflict