The impact of respiration and oxidative stress response on recombinant ?-amylase production by Saccharomyces cerevisiae

Studying protein production is important for fundamental research on cell biology and applied research for biotechnology. Yeast Saccharomyces cerevisiae is an attractive workhorse for production of recombinant proteins as it does not secrete many endogenous proteins and it is therefore easy to purify a secreted product. However, recombinant production at high rates represents a significant metabolic burden for the yeast cells, which results in oxidative stress and ultimately affects the protein production capacity. Here we describe a method to reduce the overall oxidative stress by overexpressing the endogenous HAP1 gene in a S. cerevisiae strain overproducing recombinant α-amylase. We demonstrate how Hap1p can activate a set of oxidative stress response genes and meanwhile contribute to increase the metabolic rate of the yeast strains, therefore mitigating the negative effect of the ROS accumulation associated to protein folding and hence increasing the production capacity during batch fermentations

Expanded metabolite coverage of Saccharomyces cerevisiae extract through improved chloroform/methanol extraction and tert-butyldimethylsilyl derivatization

AbstractWe present an improved extraction and derivatization protocol for GC–MS analysis of amino/non-amino acids in Saccharomyces cerevisiae. Yeast cells were extracted with chloroform: aqueous-methanol (1:1, v/v) and the resulting non-polar and polar extracts combined and dried for derivatization. Polar and non-polar metabolites were derivatized using tert-butyldimethylsilyl (t-BDMS) dissolved in acetonitrile. Using microwave treatment of the samples, the derivatization process could be completed within 2 h (from >20 h of the conventional method), providing fully derivatized metabolites that contain multiple derivatizable organic functional groups. This results in a single derivative from one metabolite, leading to increased accuracy and precision for identification and quantification of the method. Analysis of combined fractions allowed the method to expand the coverage of detected metabolites from polar metabolites i.e. amino acids, organic acids and non-polar metabolites i.e. fatty alcohols and long-chain fatty acids which are normally non detectable. The recoveries of the extraction method was found at 88 ± 4%, RSD, N = 3 using anthranilic acid as an internal standard. The method promises to be a very useful tool in various aspects of biotechnological applications i.e. development of cell factories, metabolomics profiling, metabolite identification, 13C-labeled flux analysis or semi-quantitative analysis of metabolites in yeast samples

Subsidence and thermal history of an inverted Late Jurassic-Early Cretaceous extensional basin (Cameros, North-central Spain) affected by very low- to low-grade metamorphism.

The Cameros Basin (North Spain) is a Late Jurassic-Early Cretaceous extensional basin, which was inverted during the Cenozoic. It underwent a remarkable thermal evolution, as indicated by the record of anomalous high temperatures in its deposits. In this work the subsidence and thermal history of the basin is reconstructed, using subsidence analysis and 2D thermal modeling. Tectonic subsidence curves provide evidence of the occurrence of two rapid subsidence phases during the syn-extensional stage. In the first phase (Tithonian-Early Berriasian), the largest accommodation space was formed in the central sector of the basin, whereas in the second (Early Barremian-Early Albian), it was formed in the northern sector. These rapid subsidence phases could correspond to relevant tectonic events affecting the Iberian Plate at that time. By distinguishing between the initial and thermal subsidence and defining their relative magnitudes, Royden's (1986) method was used to estimate the heat flow at the end of the extensional stage. A maximum heat flow of 60-65 mW/m2 is estimated, implying only a minor thermal disturbance associated with extension. In contrast with these data, very high vitrinite reflectance, anomalously distributed in some case with respect to the typical depth-vitrinite reflectance relation, was measured in the central-northern sector of the basin. Burial and thermal data are used to construct a 2D thermal basin model, to elucidate the role of the processes involved in sediment heating. Calibration of the thermal model with the vitrinite reflectance (%Ro) and fluid inclusion (FI) data indicates that in the central and northern sectors of the basin, an extra heat source, other than a typical rift, is required to explain the observed thermal anomalies. The distribution of the %Ro and FI values in these sectors suggests that the high temperatures and their distribution are related to the circulation of hot fluids. Hot fluids were attributed to the hydrothermal metamorphic events affecting the area during the early post-extensional and inversion stages of the basin

Il6 gene promoter polymorphism (-174G/C) influences the association between fat mass and cardiovascular risk factors

During the last decades, the prevalence of obesity has increased rapidly among young people. A polymorphism in the promoter region of the IL6 gene (-174G/C), has been previously reported to be involved in obesity and metabolic syndrome development. Therefore, the aim of the study was to examine whether the IL6 -174G/C polymorphism influence the association of body fat with low-grade inflammatory markers and blood lipids and lipoproteins in Spanish adolescents. 504 Spanish adolescents participating in the AVENA study were genotyped for the -174G/C polymorphism of the IL6 gene. Anthropometric and body composition measurements were taken and blood samples were collected for plasma molecules determinations. No differences between genotypes were observed in anthropometric values, body composition measurements and plasma markers concentration. Physical activity level differ between genotypes with subjects carrying the C allele of the polymorphism being significantly (p<0.05) more active than GG subjects. The association between body fat mass and plasma glucose was influenced by the -174G/C polymorphism of the IL6 gene. Subjects carrying the C allele of the mutation seem to have higher values of lipoprotein (a) and C-reactive protein as their percentage of body fat mass increase. Our results suggest that this promoter polymorphism influences the association between adiposity and some plasma markers

Genetic Deletion of NOD1 Prevents Cardiac Ca2+ Mishandling Induced by Experimental Chronic Kidney Disease

© 2020 by the authors.Risk of cardiovascular disease (CVD) increases considerably as renal function declines in chronic kidney disease (CKD). Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) has emerged as a novel innate immune receptor involved in both CVD and CKD. Following activation, NOD1 undergoes a conformational change that allows the activation of the receptor-interacting serine/threonine protein kinase 2 (RIP2), promoting an inflammatory response. We evaluated whether the genetic deficiency of Nod1 or Rip2 in mice could prevent cardiac Ca2+ mishandling induced by sixth nephrectomy (Nx), a model of CKD. We examined intracellular Ca2+ dynamics in cardiomyocytes from Wild-type (Wt), Nod1−/− and Rip2−/− sham-operated or nephrectomized mice. Compared with Wt cardiomyocytes, Wt-Nx cells showed an impairment in the properties and kinetics of the intracellular Ca2+ transients, a reduction in both cell shortening and sarcoplasmic reticulum Ca2+ load, together with an increase in diastolic Ca2+ leak. Cardiomyocytes from Nod1−/−-Nx and Rip2−/−-Nx mice showed a significant amelioration in Ca2+ mishandling without modifying the kidney impairment induced by Nx. In conclusion, Nod1 and Rip2 deficiency prevents the intracellular Ca2+ mishandling induced by experimental CKD, unveiling new innate immune targets for the development of innovative therapeutic strategies to reduce cardiac complications in patients with CKD.This work was supported by Spanish Ministry of Economy and Competitiveness and European Regional Development Fund (SAF-2017-84777R), Institute of Health Carlos III (PI17/01093 and PI17/01344), Sociedad Española de Cardiología, Proyecto Traslacional 2019, Fundación Renal Íñigo Álvarez de Toledo (FRIAT), Fondo Europeo de Desarrollo Regional (FEDER), FSE, and CIBER-CV, a network funded by ISCIII. M.F.-V. is Miguel Servet II researcher of ISCIII (MSII16/00047 Carlos III Health Institute). G.R.-H. is Miguel Servet I researcher of ISCIII (CP15/00129 Carlos III Health Institute). M.T. is a PhD student funded by the FPU program of the Spanish Ministry of Science, Innovation and Universities (FPU17/06135). A.R. was supported by Fondo SEP-Cinvestav project #601410 FIDSC 2018/2; and Fondo SEP-Conacyt Ciencia Básica A1-S-9082

Talento 360: Potenciar la satisfacción de los estudiantes a través de la evaluación continua y la Gestión del Talento de los Grupos Multigeneracionales (GIMM)

Para mejorar los criterios de satisfacción de los estudiantes de la asignatura se ha creado un Grupo de Iniciativa y Mejora Multidisciplinar y Multigeneracional (GIMM) conformado por estudiantes de diferentes cursos y de doctorado, así como por profesores de diferentes asignaturas. Este grupo tiene por finalidad gestionar el talento de diferentes generaciones en la creación de un Now How que mejore la satisfacción de los estudiantes que cursan la asignatura. Para realizar este proyecto se hace una evaluación en 2 momentos: evaluación inicial y final. Cada evaluación será analizada por el GIMM siguiendo una metodología Focus Group y PDCA. Este grupo propondrá medidas innovadoras de la asignatura para mejorar las puntuaciones en cada una de las evaluaciones de satisfacción. Con el análisis de la evaluación final se realizara un informe de buenas prácticas para conocer los aspectos más importantes a tener en cuenta para impartir una asignatura de calidad

Gerentur: Una aplicación transversal de gamificación para la estadística en turismo y ciencias económicas

Depto. de Economía Financiera y Actuarial y EstadísticaFac. de Ciencias Económicas y EmpresarialesFALSEsubmitte

Assessment of Multilocus Sequence Analysis (MLSA) for Identification of Candidatus Liberibacter Solanacearum from Different Host Plants in Spain

[EN] Liberibacteris a bacterial group causing different diseases and disorders in plants. Among liberibacters,CandidatusLiberibacter solanaceraum (CLso) produces disorders in several species mainly within Apiaceae and Solanaceae families. CLso isolates are usually grouped in defined haplotypes according to single nucleotide polymorphisms in genes associated with ribosomal elements. In order to characterize more precisely isolates of CLso identified in potato in Spain, a Multilocus Sequence Analysis (MLSA) was applied. This methodology was validated by a complete analysis of ten housekeeping genes that showed an absence of positive selection and a nearly neutral mechanism for their evolution. Most of the analysis performed with single housekeeping genes, as well as MLSA, grouped together isolates of CLso detected in potato crops in Spain within the haplotype E, undistinguishable from those infecting carrots, parsnips or celery. Moreover, the information from these housekeeping genes was used to estimate the evolutionary divergence among the different CLso by using the concatenated sequences of the genes assayed. Data obtained on the divergence among CLso haplotypes support the hypothesis of evolutionary events connected with different hosts, in different geographic areas, and possibly associated with different vectors. Our results demonstrate the absence in Spain of CLso isolates molecularly classified as haplotypes A and B, traditionally considered causal agents of zebra chip in potato, as well as the uncertain possibility of the present haplotype to produce major disease outbreaks in potato that may depend on many factors that should be further evaluated in future worksThis research was funded by Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA), grant numbers AT2016-007 and RTA2014-00008-C04-03-E, co-financed by FEDER.Ruiz-Padilla, A.; Redondo, C.; Asensio, A.; Garita-Cambronero, J.; Martinez, C.; Perez-Padilla, V.; Marquinez, R.... (2020). Assessment of Multilocus Sequence Analysis (MLSA) for Identification of Candidatus Liberibacter Solanacearum from Different Host Plants in Spain. Microorganisms. 8(9):1-19. https://doi.org/10.3390/microorganisms8091446S11989Haapalainen, M. (2014). Biology and epidemics ofCandidatusLiberibacter species, psyllid-transmitted plant-pathogenic bacteria. Annals of Applied Biology, 165(2), 172-198. doi:10.1111/aab.12149Raddadi, N., Gonella, E., Camerota, C., Pizzinat, A., Tedeschi, R., Crotti, E., … Alma, A. (2010). ‘Candidatus Liberibacter europaeus’ sp. nov. that is associated with and transmitted by the psyllid Cacopsylla pyri apparently behaves as an endophyte rather than a pathogen. Environmental Microbiology, 13(2), 414-426. doi:10.1111/j.1462-2920.2010.02347.xWang, N., Pierson, E. A., Setubal, J. C., Xu, J., Levy, J. G., Zhang, Y., … Martins, J. (2017). The Candidatus Liberibacter–Host Interface: Insights into Pathogenesis Mechanisms and Disease Control. Annual Review of Phytopathology, 55(1), 451-482. doi:10.1146/annurev-phyto-080516-035513Morris, J., Shiller, J., Mann, R., Smith, G., Yen, A., & Rodoni, B. (2017). Novel ‘Candidatus Liberibacter’ species identified in the Australian eggplant psyllid, Acizzia solanicola. Microbial Biotechnology, 10(4), 833-844. doi:10.1111/1751-7915.12707Alfaro-Fernández, A., Hernández-Llopis, D., & Font, M. I. (2017). Haplotypes of ‘Candidatus Liberibacter solanacearum’ identified in Umbeliferous crops in Spain. European Journal of Plant Pathology, 149(1), 127-131. doi:10.1007/s10658-017-1172-2Haapalainen, M., Wang, J., Latvala, S., Lehtonen, M. T., Pirhonen, M., & Nissinen, A. I. (2018). Genetic Variation of ‘Candidatus Liberibacter solanacearum’ Haplotype C and Identification of a Novel Haplotype from Trioza urticae and Stinging Nettle. Phytopathology®, 108(8), 925-934. doi:10.1094/phyto-12-17-0410-rHaapalainen, M., Latvala, S., Wickström, A., Wang, J., Pirhonen, M., & Nissinen, A. I. (2019). A novel haplotype of ‘Candidatus Liberibacter solanacearum’ found in Apiaceae and Polygonaceae family plants. European Journal of Plant Pathology, 156(2), 413-423. doi:10.1007/s10658-019-01890-0Mauck, K. E., Sun, P., Meduri, V. R., & Hansen, A. K. (2019). New Ca. Liberibacter psyllaurous haplotype resurrected from a 49-year-old specimen of Solanum umbelliferum: a native host of the psyllid vector. Scientific Reports, 9(1). doi:10.1038/s41598-019-45975-6Teixeira, D. C., Eveillard, S., Sirand-Pugnet, P., Wulff, A., Saillard, C., Ayres, A. J., & Bove, J. M. (2008). The tufB-secE-nusG-rplKAJL-rpoB gene cluster of the liberibacters: sequence comparisons, phylogeny and speciation. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 58(6), 1414-1421. doi:10.1099/ijs.0.65641-0Glaeser, S. P., & Kämpfer, P. (2015). Multilocus sequence analysis (MLSA) in prokaryotic taxonomy. Systematic and Applied Microbiology, 38(4), 237-245. doi:10.1016/j.syapm.2015.03.007Gevers, D., Cohan, F. M., Lawrence, J. G., Spratt, B. G., Coenye, T., Feil, E. J., … Swings, J. (2005). Re-evaluating prokaryotic species. Nature Reviews Microbiology, 3(9), 733-739. doi:10.1038/nrmicro1236Swisher Grimm, K. D., & Garczynski, S. F. (2019). Identification of a New Haplotype of ‘CandidatusLiberibacter solanacearum’ inSolanum tuberosum. Plant Disease, 103(3), 468-474. doi:10.1094/pdis-06-18-0937-reLin, H., Lou, B., Glynn, J. M., Doddapaneni, H., Civerolo, E. L., Chen, C., … Vahling, C. M. (2011). The Complete Genome Sequence of ‘Candidatus Liberibacter solanacearum’, the Bacterium Associated with Potato Zebra Chip Disease. PLoS ONE, 6(4), e19135. doi:10.1371/journal.pone.0019135Thompson, S. M., Johnson, C. P., Lu, A. Y., Frampton, R. A., Sullivan, K. L., Fiers, M. W. E. J., … Smith, G. R. (2015). Genomes of ‘Candidatus Liberibacter solanacearum’ Haplotype A from New Zealand and the United States Suggest Significant Genome Plasticity in the Species. Phytopathology®, 105(7), 863-871. doi:10.1094/phyto-12-14-0363-fiLin, H., Pietersen, G., Han, C., Read, D. A., Lou, B., Gupta, G., & Civerolo, E. L. (2015). Complete Genome Sequence of « Candidatus Liberibacter africanus,» a Bacterium Associated with Citrus Huanglongbing. Genome Announcements, 3(4). doi:10.1128/genomea.00733-15Wulff, N. A., Zhang, S., Setubal, J. C., Almeida, N. F., Martins, E. C., Harakava, R., … Gabriel, D. W. (2014). The Complete Genome Sequence of ‘Candidatus Liberibacter americanus’, Associated with Citrus Huanglongbing. Molecular Plant-Microbe Interactions®, 27(2), 163-176. doi:10.1094/mpmi-09-13-0292-rDuan, Y., Zhou, L., Hall, D. G., Li, W., Doddapaneni, H., Lin, H., … Gottwald, T. (2009). Complete Genome Sequence of Citrus Huanglongbing Bacterium, ‘CandidatusLiberibacter asiaticus’ Obtained Through Metagenomics. Molecular Plant-Microbe Interactions®, 22(8), 1011-1020. doi:10.1094/mpmi-22-8-1011Katoh, H., Miyata, S., Inoue, H., & Iwanami, T. (2014). Unique Features of a Japanese ‘Candidatus Liberibacter asiaticus’ Strain Revealed by Whole Genome Sequencing. PLoS ONE, 9(9), e106109. doi:10.1371/journal.pone.0106109Leonard, M. T., Fagen, J. R., Davis-Richardson, A. G., Davis, M. J., & Triplett, E. W. (2012). Complete genome sequence of Liberibacter crescens BT-1. Standards in Genomic Sciences, 7(2), 271-283. doi:10.4056/sigs.3326772Teresani, G. R., Bertolini, E., Alfaro-Fernández, A., Martínez, C., Tanaka, F. A. O., Kitajima, E. W., … Font, M. I. (2014). Association of ‘Candidatus Liberibacter solanacearum’ with a Vegetative Disorder of Celery in Spain and Development of a Real-Time PCR Method for Its Detection. Phytopathology®, 104(8), 804-811. doi:10.1094/phyto-07-13-0182-rLi, W., Hartung, J. S., & Levy, L. (2006). Quantitative real-time PCR for detection and identification of Candidatus Liberibacter species associated with citrus huanglongbing. Journal of Microbiological Methods, 66(1), 104-115. doi:10.1016/j.mimet.2005.10.018Munyaneza, J. E., Sengoda, V. G., Crosslin, J. M., De la Rosa-Lozano, G., & Sanchez, A. (2009). First Report of ‘Candidatus Liberibacter psyllaurous’ in Potato Tubers with Zebra Chip Disease in Mexico. Plant Disease, 93(5), 552-552. doi:10.1094/pdis-93-5-0552aPhillips, J. L., & Gnanakaran, S. (2014). A data-driven approach to modeling the tripartite structure of multidrug resistance efflux pumps. Proteins: Structure, Function, and Bioinformatics, 83(1), 46-65. doi:10.1002/prot.24632Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Molecular Biology and Evolution, 35(6), 1547-1549. doi:10.1093/molbev/msy096Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. (1993). Molecular Biology and Evolution. doi:10.1093/oxfordjournals.molbev.a040023Rozas, J., Ferrer-Mata, A., Sánchez-DelBarrio, J. C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S. E., & Sánchez-Gracia, A. (2017). DnaSP 6: DNA Sequence Polymorphism Analysis of Large Data Sets. Molecular Biology and Evolution, 34(12), 3299-3302. doi:10.1093/molbev/msx248Liao, J., Wiedmann, M., & Kovac, J. (2017). Genetic Stability and Evolution of the sigB Allele, Used for Listeria Sensu Stricto Subtyping and Phylogenetic Inference. Applied and Environmental Microbiology, 83(12). doi:10.1128/aem.00306-17Tamura, K., Battistuzzi, F. U., Billing-Ross, P., Murillo, O., Filipski, A., & Kumar, S. (2012). Estimating divergence times in large molecular phylogenies. Proceedings of the National Academy of Sciences, 109(47), 19333-19338. doi:10.1073/pnas.1213199109Tamura, K., Tao, Q., & Kumar, S. (2018). Theoretical Foundation of the RelTime Method for Estimating Divergence Times from Variable Evolutionary Rates. Molecular Biology and Evolution, 35(7), 1770-1782. doi:10.1093/molbev/msy044López-Hermoso, C., de la Haba, R. R., Sánchez-Porro, C., Papke, R. T., & Ventosa, A. (2017). Assessment of MultiLocus Sequence Analysis As a Valuable Tool for the Classification of the Genus Salinivibrio. Frontiers in Microbiology, 8. doi:10.3389/fmicb.2017.01107Hajri, A., Loiseau, M., Cousseau-Suhard, P., Renaudin, I., & Gentit, P. (2017). Genetic Characterization of ‘Candidatus Liberibacter solanacearum’ Haplotypes Associated with Apiaceous Crops in France. Plant Disease, 101(8), 1383-1390. doi:10.1094/pdis-11-16-1686-reFang, Y., Wang, Y., Liu, Z., Dai, H., Cai, H., Li, Z., … Wang, D. (2019). Multilocus Sequence Analysis, a Rapid and Accurate Tool for Taxonomic Classification, Evolutionary Relationship Determination, and Population Biology Studies of the Genus Shewanella. Applied and Environmental Microbiology, 85(11). doi:10.1128/aem.03126-18Konstantinidis, K. T., Ramette, A., & Tiedje, J. M. (2006). Toward a More Robust Assessment of IntraspeciesDiversity, Using Fewer GeneticMarkers. Applied and Environmental Microbiology, 72(11), 7286-7293. doi:10.1128/aem.01398-06Ajene, I. J., Khamis, F., Ballo, S., Pietersen, G., van Asch, B., Seid, N., … Mohamed, S. (2020). Detection of Asian Citrus Psyllid (Hemiptera: Psyllidae) in Ethiopia: A New Haplotype and its Implication to the Proliferation of Huanglongbing. Journal of Economic Entomology, 113(4), 1640-1647. doi:10.1093/jee/toaa113Thapa, S. P., De Francesco, A., Trinh, J., Gurung, F. B., Pang, Z., Vidalakis, G., … Coaker, G. (2020). Genome‐wide analyses of Liberibacter species provides insights into evolution, phylogenetic relationships, and virulence factors. Molecular Plant Pathology, 21(5), 716-731. doi:10.1111/mpp.12925Antolinez, C. A., Fereres, A., & Moreno, A. (2017). Risk assessment of ‘Candidatus Liberibacter solanacearum’ transmission by the psyllids Bactericera trigonica and B. tremblayi from Apiaceae crops to potato. Scientific Reports, 7(1). doi:10.1038/srep45534Antolínez, Moreno, Ontiveros, Pla, Plaza, Sanjuan, … Fereres. (2019). Seasonal Abundance of Psyllid Species on Carrots and Potato Crops in Spain. Insects, 10(9), 287. doi:10.3390/insects10090287Wang, J., Haapalainen, M., Schott, T., Thompson, S. M., Smith, G. R., Nissinen, A. I., & Pirhonen, M. (2017). Genomic sequence of «Candidatus Liberibacter solanacearum» haplotype C and its comparison with haplotype A and B genomes. PLOS ONE, 12(2), e0171531. doi:10.1371/journal.pone.0171531Katsir, L., Zhepu, R., Santos Garcia, D., Piasezky, A., Jiang, J., Sela, N., … Bahar, O. (2018). Genome Analysis of Haplotype D of Candidatus Liberibacter Solanacearum. Frontiers in Microbiology, 9. doi:10.3389/fmicb.2018.02933Quintana-González de Chaves, M., Teresani, G. R., Hernández-Suárez, E., Bertolini, E., Moreno, A., Fereres, A., … Siverio, F. (2020). ‘Candidatus Liberibacter Solanacearum’ Is Unlikely to Be Transmitted Spontaneously from Infected Carrot Plants to Citrus Plants by Trioza Erytreae. Insects, 11(8), 514. doi:10.3390/insects1108051

The confining-Higgs phase transition in U(1)-Higgs LGT

Abstract We simulate the compact U(1)-Higgs model in a four-dimensional lattice. We present a numerical study for the behaviour around the region where the transition between the confining and Higgs phases disappears. The transition line is found to be first order and to end in a second-order point. We measure the critical exponents of the endpoint obtaining the mean field ones within errors