Papel de las dimetilargininas en la hipertensión portal y cirrosis experimental

RESUMEN La alteración del sistema DDAH-ADMA-NOS podría contribuir en la fisiopatogénesis de las alteraciones vasculares que aparecen en la hipertensión portal asociada o no a una cirrosis. Está bien establecido que pacientes con cirrosis hepática tienen niveles plasmáticos elevados de dimetilarginina asimétrica (ADMA). Una de las posibles causas podría deberse a un fallo en su degradación, es decir, una disminución de la actividad de las DDAHs. Por otro lado, el NO intrahepático está disminuido. El objetivo general del trabajo fue estudiar el papel del ADMA en las alteraciones vasculares que aparecen en los modelos de hipertensión portal y cirrosis. En la presente tesis doctoral se comprueba que el ADMA inhibe la relajación dependiente de endotelio, mediada principalmente por NO en la arteria ilíaca. Sin embargo, la arteria mesentérica el ADMA no altera la relajación dependiente de endotelio. Por tanto, el incremento de los niveles plasmáticos de ADMA en la cirrosis no modificaría la relajación dependiente del endotelio en el lecho esplácnico y contribuiría a la excesiva vasodilatación. Existen variaciones importantes en la expresión de las DDAHs inducidas por la hipertensión portal y cirrosis. La elevada expresión de las DDAHs a nivel esplácnico en la hipertensión portal y en la cirrosis contribuiría a una mayor degradación del ADMA que favorecería la síntesis de NO, que tendría especial repercusión en la liberación basal de NO en el territorio esplácnico. Por ultimo, en el hígado cirrótico hay un incremento en la expresión de DDAH-2, que no se acompaña con una mayor síntesis de NO. Por otra parte, el déficit de NO existente en el hígado cirrótico podría ser debido, al menos en parte, a los bajos niveles hepáticos de L-arginina. __________________________________________________________________________________________________Nitric oxide (NO) synthesis is modulated by dimethylarginine dimethylaminohydrolase (DDAH) via metabolizing asymmetric dimethylarginine (ADMA), an endogenous NO synthase (NOS) inhibitor. DDAH-ADMA-NOS pathway could play an important role in the physiopathogenesis of vascular alterations observed in isolated portal hypertension or associated with cirrhosis. The aim of the present study was to examine alterations in DDAH/ADMA/NOS pathway and the ADMA role in portal hypertension (PHT) and cirrhosis. One rat model of cirrhosis (bile-duct-ligation ,BDL), one rat model of PHT without cirrhosis (partial portal vein-ligated, PPVL) and sham-operated control rats were studied. To evaluate endothelial function, concentration-effect curves to acetylcholine were determined in vitro using different arteries preincubated with either vehicle or ADMA/L-NAME (NOS inhibitor). In addition, concentration-effect curves of ADMA and L-NAME were studied to observe NO basal release. The mRNA expression of eNOS gene, DDAH-1 gene and DDAH-2 gene were detected by semi-quantitative RT-PCR in differents arteries and liver. Overexpression of both types of DDAHs in splanchnic territory of PHT and BDL rats could metabolize greater ADMA and could contribute to vasodilatation in this territory. We found an increase in the mRNA expression of DDAH-2 gene not associated with elevated levels of NO in the cirrhotic liver. The effects of decreasing NO synthesis are, at least in part, mediated by lower levels of L-arginine intrahepatic

Ovarian transcriptomic analysis reveals differential expression genes associated with cell death process after selection for ovulation rate in rabbits

[EN] Transcriptomic analysis showed nineteen potential biomarkers in ovarian tissue from females belonged to a rabbit line selected for ovulation rate for 10 generations and the control line. These females differed not only in ovulation rate but also in prenatal survival since similar litter size were observed. Litter size is an essential trait in rabbit meat production but with low heritability. A selection experiment for ovulation rate has been performed for 10 generations to improve litter size in rabbits. The selected line increased two ova more than the control line but nevertheless a negative correlation was observed with prenatal survival. A transcriptomic study was performed, using microarrays, in ovarian tissue from females belonging to the selected line and the control line. Our results showed 1357 differential expressed genes and nineteen potential biomarkers associated with prenatal mortality, which could explain differences between litter size in rabbits. Cell death was the most relevant process.This research was supported by MEC (AGL2014-55921-C2-1-P) and Generalitat Valenciana (Prometeo 2009/125).Serna-García, M.; Peiró Barber, RM.; Serna, E.; Santacreu Jerez, MA. (2020). Ovarian transcriptomic analysis reveals differential expression genes associated with cell death process after selection for ovulation rate in rabbits. Animals. 10(10):1-11. https://doi.org/10.3390/ani10101924S1111010Laborda, P., Mocé, M. L., Blasco, A., & Santacreu, M. A. (2012). Selection for ovulation rate in rabbits: Genetic parameters and correlated responses on survival rates1. Journal of Animal Science, 90(2), 439-446. doi:10.2527/jas.2011-4219Laborda, P., Mocé, M. L., Santacreu, M. A., & Blasco, A. (2011). Selection for ovulation rate in rabbits: Genetic parameters, direct response, and correlated response on litter size1. Journal of Animal Science, 89(10), 2981-2987. doi:10.2527/jas.2011-3906Laborda, P., Santacreu, M. A., Blasco, A., & Mocé, M. L. (2012). 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Gamificación como herramienta docente aplicada a las tutorías de grupo en la Educación Superior

[EN] Our general objective consists in using gamification as a teaching tool in order to get a higher level of students’ involvement in the tutorials, as well as, increasing the motivation to learn Pathophysiology in the Pharmacy Degree. Not all the students groups are uniform, and also the teachers have nonconscious prejudices for each group. Hence, every student group has its own idiosyncrasy. We have the hypothesis that the game can give us really valuable information to let us develop suitable teaching techniques to enhance the motivation for each particular group. Games offers a more relaxed scenario compared to master class or exam, therefore we can ask questions as a team contest that stimulate the comprehension and integration of contents, and at the same time to brainstorm and to learn from mistakes. Games also offers teachers a better interaction with students to reflect on how to motivate each specific group. Our conclusion is, in general terms, that gamification in tutorials let students be more actives in the session. Moreover, for teachers it is an excellent tool to get to know the student’s profile and to find the best way to increase the motivation for learning in each specific group.[ES] El objetivo general del presente trabajo es utilizar la gamificación como herramienta docente para conseguir una mayor implicación del alumnado en las tutorías y aumentar la motivación frente al conocimiento de la asignatura de Fisiopatología en el Grado de Farmacia.No todos los grupos de estudiantes son iguales, y además el profesorado tiene sus prejuicios no conscientes para cada grupo. Nuestra hipótesis de trabajo se basa en que cada grupo tiene su idiosincrasia y a través del juego podemos obtener información muy valiosa que nos permita desarrollar en un futuro técnicas docentes apropiadas para cada grupo de alumnos que hagan resurgir la motivación. Desde el juego, con una perspectiva más relajada comparada con la clase magistral o con el examen, podemos plantear preguntas a modo de concurso por equipos que favorezcan el asentamiento, interiorización e integración de los contenidos, y al mismo tiempo desatar una lluvia de ideas, enseñar a partir del error e interactuar más directamente con los alumnos para reflexionar sobre cómo los docentes podemos motivar y llegar a cada grupo concreto.La conclusión general de este trabajo es que la gamificación en tutorías hace que los alumnos participen activamente en la sesión. Además, desde el punto de vista del profesorado ofrece una herramienta para conocer mejor el perfil de su alumnado y definir la mejor manera en cada caso para aumentar la motivación por la asignatura.Serna García, E.; Pereda, J.; Mauricio, MD.; Pérez, S. (2019). Gamificación como herramienta docente aplicada a las tutorías de grupo en la Educación Superior. En IN-RED 2019. V Congreso de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 750-758. https://doi.org/10.4995/INRED2019.2019.10442OCS75075

Material multimedia para la integración de contenidos en asignaturas y grados del Área de la Salud: Transporte de membrana

[EN] Multimedia material supports both teachers and students in the teaching-learning process, obtaining satisfactory results. The current educational system uses a mechanical and rigid mechanism in which teaching is fragmented and disconnected by subjects. Our work aims to report a new academic proposal where a new integrative multimedia material is developed about membrane transport between different subjects and degrees in the Health Area. The results were very satisfactory because the students' opinion about the video was very positive and well-rated. This work clarified the need for a change in the educational system.[ES] El material multimedia apoya tanto al profesorado como al alumnado en el proceso enseñanza-aprendizaje obteniendo buenos resultados. El sistema educativo actual utiliza un mecanismo mecánico y rígido en el cual la docencia está fragmentada e inconexa por asignaturas. Nuestro trabajo pretende abordar una nueva propuesta educativa en la que se elabora un material multimedia integrativo sobre el transporte de membrana entre diferentes asignaturas y grados del Área de la Salud. Los resultados fueron muy satisfactorios ya que la opinión del alumnado sobre el vídeo fue muy positiva y bien calificada. Este trabajo elucida la necesidad de un cambio en el sistema educativo.Serna García, E.; San-Miguel Diez, T.; Megías Vericat, J.; Mauricio Avñó, M. (2021). Material multimedia para la integración de contenidos en asignaturas y grados del Área de la Salud: Transporte de membrana. En IN-RED 2021: VII Congreso de Innovación Edicativa y Docencia en Red. Editorial Universitat Politècnica de València. 804-811. https://doi.org/10.4995/INRED2021.2021.13703OCS80481

Transcriptomic identification of miR-205 target genes potentially involved in metastasis and survival of cutaneous malignant melanoma

Cutaneous melanoma is an aggressive neoplasm and is responsible for the majority of skin cancer deaths. Several miRNAs are involved in melanoma tumor progression. One of them is miR-205, the loss of which contributes to the development of melanoma metastasis. We evaluated whole-genome mRNA expression profiling associated with different miR-205 expression levels in melanoma cells. Differential expression analysis identified 243 differentially expressed transcripts including inositol polyphosphate 5'-phosphatase-like protein-1 (INPPL1) and BTB/POZ Domain-Containing Protein 3 (BTBD3). INPPL1 and BTBD3 were downregulated when melanoma cells expressed miR-205, indicating that these genes are potential miR-205 targets. Additionally, the target prediction algorithm TargetScan revealed that INPPL1 and BTBD3 genes had predicted target sites of miR-205 in their 3'UTRs and functional analysis demonstrated that these genes were directly linked to miR-205. Interestingly, our clinical data showed that INPPL1 was significantly associated with lymph node metastasis-free survival (LNMFS), distant metastasis-free survival (DMFS) and melanoma specific survival (MSS). This study supports INPPL1 as a miR-205 target gene and, therefore, that the involvement of miR-205 in the metastatic dissemination of malignant melanoma is, at least in part, via INPPL1

Analysis of Exosomal Cargo Provides Accurate Clinical, Histologic and Mutational Information in Non-Small Cell Lung Cancer

Lung cancer is a malignant disease with high mortality and poor prognosis, frequently diagnosed at advanced stages. Nowadays, immense progress in treatment has been achieved. However, the present scenario continues to be critical, and a full comprehension of tumor progression mechanisms is required, with exosomes being potentially relevant players. Exosomes are membranous vesicles that contain biological information, which can be transported cell-to-cell and modulate relevant processes in the hallmarks of cancer. The present research aims to characterize the exosomes' cargo and study their role in NSCLC to identify biomarkers. We analyzed exosomes secreted by primary cultures and cell lines, grown in monolayer and tumorsphere formations. Exosomal DNA content showed molecular alterations, whereas RNA high-throughput analysis resulted in a pattern of differentially expressed genes depending on histology. The most significant differences were found in XAGE1B, CABYR, NKX2-1, SEPP1, CAPRIN1, and RIOK3 genes when samples from two independent cohorts of resected NSCLC patients were analyzed. We identified and validated biomarkers for adenocarcinoma and squamous cell carcinoma. Our results could represent a relevant contribution concerning exosomes in clinical practice, allowing for the identification of biomarkers that provide information regarding tumor features, prognosis and clinical behavior of the disease. Keywords: non-small cell lung cancer; liquid biopsy; exosomes; extracellular vesicles; cell cultures; adenocarcinoma; squamous cell carcinoma; biomarker; tumorsphere

Uso del debate como herramienta metodológica docente en estudios del Grado en Medicina: DEBATMITAL

[EN] The main purpose of the DEBATMITAL project is to increase the participation of students in the practical session on "Myths of Food" that is part of the optional subject “Nutrition and Dietetics" of the degree in Medicine of the University of Valencia. In front of the master class, which was basically the tool used in previous courses, students are organized into discussion groups to debate some food-related statements commonly heard. The assessment that students make of this initiative is very positive. They indicate that they have learned, they have had fun while participating in the debates, and that they prefer this tool to the master class. Therefore, the debate is a teaching tool that can not only be used in Social Sciences area, but can also be very useful for certain topics in Health Sciences, as in the case of the degree in Medicine.[ES] El proyecto DEBATMITAL tiene como objetivo principal aumentar la participación de los alumnos en la sesión práctica sobre “Mitos de la Alimentación” que forma parte de la asignatura optativa “Alimentación y Dietética” del grado en Medicina de la Universidad de Valencia. Frente a la clase magistral, que era básicamente la herramienta utilizada en cursos anteriores, se organiza la clase en grupos de debate para tratar diversas afirmaciones relacionadas con la alimentación que se pueden escuchar comúnmente. La valoración que los estudiantes hacen de esta iniciativa es muy positiva. Indican que han aprendido, que se han divertido mientras participaban en los debates, y que prefieren esta herramienta a la clase magistral. Por tanto, el debate es una herramienta docente que no solo se puede utilizar en el área de las Ciencias Sociales, sino que también puede ser muy útil para determinados temas en grados del área de Ciencias de la Salud, como es el caso del grado en Medicina.Olaso-González, G.; Romá-Mateo, C.; Serna García, E.; Gambini, J.; Correas, ÁG.; Gimeno, L.; Escrivá, C.... (2019). Uso del debate como herramienta metodológica docente en estudios del Grado en Medicina: DEBATMITAL. En IN-RED 2019. V Congreso de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 1022-1033. https://doi.org/10.4995/INRED2019.2019.10471OCS1022103

Intrinsic functional and architectonic heterogeneity of tumor-targeted protein nanoparticles

Altres ajuts: CIBER de Bioingeniería, Biomateriales y Nanomedicina (project NANOPROTHER) (to AV), Marató de TV3 foundation (TV32013-132031) (TV32013-133930). Protein production has been partially performed by the ICTS "NANBIOSIS", more specifically by the Protein Production Platform of CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN)/IBB, at the UAB SepBioES scientific-technical service (http://www.nanbiosis.es/unit/u1-protein-production-platform-ppp/) and DLS measurements have been done at the Biomaterial Processing and Nanostructuring Unit of NANBIOSIS. We are also indebted to Fran Cortés from the Cell Culture and Cytometry Units of the Servei de CultiusCel·lulars, Producciód'AnticossosiCitometria (SCAC), and to the Servei de Microscòpia, both at the UAB. Strain KPM335 was kindly provided by Research Corporation Technologies, Tucson, AZ. AV received an ICREA ACADEMIA award.Self-assembling proteins are gaining attention as building blocks for application-tailored nanoscale materials. This is mostly due to the biocompatibility, biodegradability, and functional versatility of peptide chains. Such a potential for adaptability is particularly high in the case of recombinant proteins, which are produced in living cells and are suitable for genetic engineering. However, how the cell factory itself and the particular protein folding machinery influence the architecture and function of the final material is still poorly explored. In this study we have used diverse analytical approaches, including small-angle X-ray scattering (SAXS) and field emission scanning electron microscopy (FESEM) to determine the fine architecture and geometry of recombinant, tumor-targeted protein nanoparticles of interest as drug carriers, constructed on a GFP-based modular scheme. A set of related oligomers were produced in alternative Escherichia coli strains with variant protein folding networks. This resulted in highly regular populations of morphometric types, ranging from 2.4 to 28 nm and from spherical- to rod-shaped materials. These differential geometric species, whose relative proportions were determined by the features of the producing strain, were found associated with particular fluorescence emission, cell penetrability and receptor specificity profiles. Then, nanoparticles with optimal properties could be analytically identified and further isolated from producing cells for use. The cell's protein folding machinery greatly modulates the final geometry reached by the constructs, which in turn defines the key parameters and biological performance of the material

Human exceptional longevity: transcriptome from centenarians is distinct from septuagenarians and reveals a role of Bcl‐xL in successful aging

Centenarians not only enjoy an extraordinary aging, but also show a compression of morbidity. Using functional transcriptomic analysis of peripheral blood mononuclear cells (PMBC) we identified 1721 mRNAs differentially expressed by centenarians when compared with septuagenarians and young people. Sub‐network analysis led us to identify Bcl ‐ xL as an important gene up‐regulated in centenarians. It is involved in the control of apoptosis, cellular damage protection and also in modulation of immune response, all associated to healthy aging. Indeed, centenarians display lower plasma cytochrome C levels, higher mitochondrial membrane potential and also less cellular damage accumulation than septuagenarians. Leukocyte chemotaxis and NK cell activity are significantly impaired in septuagenarians compared with young people whereas centenarians maintain them. To further ascertain the functional role of Bcl‐ xL in cellular aging, we found that lymphocytes from septuagenarians transduced with Bcl‐xL display a reduction in senescent‐related markers. Finally, to demonstrate the role of BcL‐xL in longevity at the organism level, C. elegans bearing a gain of function mutation in the BcL‐xL ortholog ced‐9, showed a significant increase in mean and maximal life span. These results show that mRNA expression in centenarians is unique and reveals that BcL‐ xL plays an important role in exceptional aging

Lack of replication of interactions between polymorphisms in rheumatoid arthritis susceptibility: case-control study

Introduction: Approximately 100 loci have been definitively associated with rheumatoid arthritis (RA) susceptibility. However, they explain only a fraction of RA heritability. Interactions between polymorphisms could explain part of the remaining heritability. Multiple interactions have been reported, but only the shared epitope (SE) × protein tyrosine phosphatase nonreceptor type 22 (PTPN22) interaction has been replicated convincingly. Two recent studies deserve attention because of their quality, including their replication in a second sample collection. In one of them, researchers identified interactions between PTPN22 and seven single-nucleotide polymorphisms (SNPs). The other showed interactions between the SE and the null genotype of glutathione S-transferase Mu 1 (GSTM1) in the anti-cyclic citrullinated peptide-positive (anti-CCP+) patients. In the present study, we aimed to replicate association with RA susceptibility of interactions described in these two high-quality studies. Methods: A total of 1,744 patients with RA and 1,650 healthy controls of Spanish ancestry were studied. Polymorphisms were genotyped by single-base extension. SE genotypes of 736 patients were available from previous studies. Interaction analysis was done using multiple methods, including those originally reported and the most powerful methods described. Results: Genotypes of one of the SNPs (rs4695888) failed quality control tests. The call rate for the other eight polymorphisms was 99.9%. The frequencies of the polymorphisms were similar in RA patients and controls, except for PTPN22 SNP. None of the interactions between PTPN22 SNPs and the six SNPs that met quality control tests was replicated as a significant interaction term the originally reported finding or with any of the other methods. Nor was the interaction between GSTM1 and the SE replicated as a departure from additivity in anti-CCP+ patients or with any of the other methods. Conclusions: None of the interactions tested were replicated in spite of sufficient power and assessment with different assays. These negative results indicate that whether interactions are significant contributors to RA susceptibility remains unknown and that strict standards need to be applied to claim that an interaction exists