Comprehensive Cell Surface Protein Profiling Identifies Specific Markers of Human Naive and Primed Pluripotent States

Human pluripotent stem cells (PSCs) exist in naive and primed states and provide important models to investigate the earliest stages of human development. Naive cells can be obtained through primed-to-naive resetting, but there are no reliable methods to prospectively isolate unmodified naive cells during this process. Here we report comprehensive profiling of cell surface proteins by flow cytometry in naive and primed human PSCs. Several naive-specific, but not primed-specific, proteins were also expressed by pluripotent cells in the human preimplantation embryo. The upregulation of naive-specific cell surface proteins during primed-to-naive resetting enabled the isolation and characterization of live naive cells and intermediate cell populations. This analysis revealed distinct transcriptional and X chromosome inactivation changes associated with the early and late stages of naive cell formation. Thus, identification of state-specific proteins provides a robust set of molecular markers to define the human PSC state and allows new insights into the molecular events leading to naive cell resetting.Imaging was performed at the Live Cell Imaging Facility/Nikon Center of Excellence, Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden, supported by grants from the Knut and Alice Wallenberg Foundation, the Swedish Research Council, the Centre for Innovative Medicine, and the Jonasson donation to the School of Technology and Health, Royal Institute of Technology, Sweden. We would like to acknowledge the MedH Flow Cytometry facility at Karolinska Institutet, supported by grants from Karolinska Institutet and the Stockholm County Council. We thank Céline Vallot and Claire Rougeulle at the Université Paris Diderot for providing X chromosome SNP coordinates. We are grateful to Rudolph Jaenisch at the Whitehead Institute for Biomedical Research for providing WIBR3 cells and Austin Smith at the WT–MRC Cambridge Stem Cell Institute for providing H9 NK2 and FiPS cells. We thank all couples who donated embryos to this study. S.P., A.P.R., J.P.S., and F.L. are supported by grants from the Swedish Research Council (2013-2570), Ragnar Söderberg Foundation (M67/13), Swedish Foundation for Strategic Research (ICA-5), Knut and Alice Wallenberg Foundation (4-1205/2016 and 4-148/2017), and Centre for Innovative Medicine and by a Lau fellowship. R.W. is an ISAC Shared Resource Laboratory Emerging Leader. A.J.C. is supported by an MRC DTG Studentship (MR/J003808/1). P.J.R.G. is supported by the Wellcome Trust (WT093736) and BBSRC (BBS/ E/B/000C0402)

Development of anatomic models through plastination techniques as an alternative to the use of animals in teaching and as an essential resource in the teaching-learning process

La implantación del principio de las 3Rs (Reducción, Refinamiento y Reemplazo), descritas por Russell y Burch en 1959, fue el punto de partida de numerosos cambios en la utilización de los animales, tanto en el ámbito de la investigación como de la docencia. Por tanto, es necesario ir propiciando el empleo de alternativas y que el profesorado disponga de recursos y de una buena información. Este proyecto se propone como objetivo esencial la elaboración de modelos animales mediante técnicas de plastinación como recurso docente alternativo al uso de animales vivos, y proporcionando al profesorado la disposición de medios y recursos esenciales en el proceso de enseñanza-aprendizaje del alumnado. Los alumnos podrán disponer de una variedad de modelos animales en las sesiones prácticas, y/o teórico/prácticas de las distintas disciplinas en las que sea necesario el uso de animales. Podrán trabajar con un modelo fiel al original sin necesidad de utilizar modelos vivos y podrán desarrollar las distintas técnicas y procedimientos que realizarían in vivo sin necesidad de generar un estrés innecesario a los animales por su manejo por personal aún no capacitado para ello, evitando situaciones que hagan peligrar el bienestar animal (RD 53/2003). El desarrollo de estos modelos plastinados supondrá recurso docente esencial para la adquisición de las habilidades específicas en distintas disciplinas Biomédicas que conlleven el uso de animales, tales como los Grados en Veterinaria, Biología, Medicina, Enfermería, etc.The implementation of the 3Rs principles (Reduction, Refinement and Replacement), described by Russell and Burch in 1959, was the starting point of numerous changes in the use of animals in the fields of research and teaching. Therefore, it is necessary to foster the use of alternative methods and provide teachers with resources and good information. This project aims to develop animal models through the application of plastination techniques as an alternative teaching resource to the use of animals. In this way, the project aims to provide teaching staff with tools and resources that are essential in the teaching-learning process. The students will be able to use a variety of animal models in practical and/or theoretical-practical sessions of the different subject areas, in which the use of animals is necessary. They will be able to work with animal models that are similar to the original, while avoiding the use of animals. Moreover, they will be able to learn and practice different techniques and procedures that they would carry out in vivo, without generating unnecessary stress to the animals caused by their management by untrained personnel. This approach is therefore in full conformity with existing animal welfare regulations (i.e. RD 53/2003). Plastinated models will become a an essential teaching resource for the acquisition of the specific skills in different Biomedical and Science disciplines that involve the use of animals, such as Veterinary Science, Biology, Environmental Sciences, Medicine, Nursing, etc

Microwave sensor system for continuous monitoring of adhesive curing processes

A microwave sensor system has been developed for monitoring adhesive curing processes. The system provides continuous, real-time information about the curing progress without interfering with the reaction. An open-coaxial resonator is used as the sensor head, and measurements of its resonance frequency and quality factor are performed during cure to follow the reaction progress. Additionally, the system provides other interesting parameters such as reaction rate or cure time. The adhesive dielectric properties can also be computed off-line, which gives additional information about the process. The results given by the system correlate very well with conventional measurement techniques such as differential scanning calorimetry, combining accuracy and rate with simplicity and an affordable cost. © 2012 IOP Publishing Ltd.The authors thank Rut Benavente Martinez for her assistance in the DSC experiments. The contract of BG-B is financed by the Ministry of Science and Innovation of Spain, through the 'Torres Quevedo' Sub-programme, which is also co-financed by the European Social Fund (ESF). This work has been financed by the Ministry of Science and Innovation of Spain through the project MONIDIEL (TEC2008-04109).García Baños, B.; Catalá Civera, JM.; Penaranda-Foix, FL.; Canós Marín, AJ.; Sahuquillo Navarro, O. (2012). Microwave sensor system for continuous monitoring of adhesive curing processes. Measurement Science and Technology. 23(3). https://doi.org/10.1088/0957-0233/23/3/035101S233Jost, M., & Sernek, M. (2008). Shear strength development of the phenol–formaldehyde adhesive bond during cure. Wood Science and Technology, 43(1-2), 153-166. doi:10.1007/s00226-008-0217-2Costa, M. L., Botelho, E. C., Paiva, J. M. F. de, & Rezende, M. C. (2005). Characterization of cure of carbon/epoxy prepreg used in aerospace field. Materials Research, 8(3), 317-322. doi:10.1590/s1516-14392005000300016Chen, J., & Hojjati, M. (2007). Microdielectric analysis and curing kinetics of an epoxy resin system. Polymer Engineering & Science, 47(2), 150-158. doi:10.1002/pen.20687Sernek, M., & Kamke, F. A. (2007). Application of dielectric analysis for monitoring the cure process of phenol formaldehyde adhesive. International Journal of Adhesion and Adhesives, 27(7), 562-567. doi:10.1016/j.ijadhadh.2006.10.004Núñez, L., Gómez-Barreiro, S., Gracia-Fernández, C. A., & Núñez, M. R. (2004). Use of the dielectric analysis to complement previous thermoanalytical studies on the system diglycidyl ether of bisphenol A/1,2 diamine cyclohexane. Polymer, 45(4), 1167-1175. doi:10.1016/j.polymer.2003.12.024Lefebvre, D. R., Han, J., Lipari, J. M., Long, M. A., McSwain, R. L., & Wells, H. C. (2006). Dielectric analysis for in-situ monitoring of gelatin renaturation and crosslinking. Journal of Applied Polymer Science, 101(5), 2765-2775. doi:10.1002/app.21631Cordovez, M., Li, Y., & Karbhari, V. M. (2004). Assessment of Dielectrometry for Characterization of Processing and Moisture Absorption in FRP Composites. Journal of Reinforced Plastics and Composites, 23(4), 445-456. doi:10.1177/0731684404031980Das, N. K., Voda, S. M., & Pozar, D. M. (1987). Two Methods for the Measurement of Substrate Dielectric Constant. IEEE Transactions on Microwave Theory and Techniques, 35(7), 636-642. doi:10.1109/tmtt.1987.1133722Fioretto, D., Livi, A., Rolla, P. A., Socino, G., & Verdini, L. (1994). The dynamics of poly(n-butyl acrylate) above the glass transition. Journal of Physics: Condensed Matter, 6(28), 5295-5302. doi:10.1088/0953-8984/6/28/007Givot, B. L., Krupka, J., & Belete, D. Y. (s. f.). Split post dielectric resonator technique for dielectric cure monitoring of structural adhesives. 13th International Conference on Microwaves, Radar and Wireless Communications. MIKON - 2000. Conference Proceedings (IEEE Cat. No.00EX428). doi:10.1109/mikon.2000.913931Canos, A. J., Catala-Civera, J. M., Penaranda-Foix, F. L., & Reyes-Davo, E. (2006). A novel technique for deembedding the unloaded resonance frequency from measurements of microwave cavities. IEEE Transactions on Microwave Theory and Techniques, 54(8), 3407-3416. doi:10.1109/tmtt.2006.877833Marks, R. B., & Williams, D. F. (1992). A general waveguide circuit theory. Journal of Research of the National Institute of Standards and Technology, 97(5), 533. doi:10.6028/jres.097.024Harrington, R. F. (1967). Matrix methods for field problems. Proceedings of the IEEE, 55(2), 136-149. doi:10.1109/proc.1967.5433Baker-Jarvis, J., Janezic, M. D., Domich, P. D., & Geyer, R. G. (1994). Analysis of an open-ended coaxial probe with lift-off for nondestructive testing. IEEE Transactions on Instrumentation and Measurement, 43(5), 711-718. doi:10.1109/19.328897Taylor, B. N. (1994). Guidelines for evaluating and expressing the uncertainty of NIST measurement results. doi:10.6028/nist.tn.1297Casalini, R., Corezzi, S., Livi, A., Levita, G., & Rolla, P. A. (1997). Dielectric parameters to monitor the crosslink of epoxy resins. Journal of Applied Polymer Science, 65(1), 17-25. doi:10.1002/(sici)1097-4628(19970705)65:13.0.co;2-tPreu, H., & Mengel, M. (2007). Experimental and theoretical study of a fast curing adhesive. International Journal of Adhesion and Adhesives, 27(4), 330-337. doi:10.1016/j.ijadhadh.2006.06.004Harper, D. P., Wolcott, M. P., & Rials, T. G. (2001). Evaluation of the cure kinetics of the wood/pMDI bondline. International Journal of Adhesion and Adhesives, 21(2), 137-144. doi:10.1016/s0143-7496(00)00045-2Garcia-Banos, B., Canos, A. J., Penaranda-Foix, F. L., & Catala-Civera, J. M. (2011). Noninvasive Monitoring of Polymer Curing Reactions by Dielectrometry. IEEE Sensors Journal, 11(1), 62-70. doi:10.1109/jsen.2010.2050475He, Y. (2001). DSC and DEA studies of underfill curing kinetics. Thermochimica Acta, 367-368, 101-106. doi:10.1016/s0040-6031(00)00654-7Núñez-Regueira, L., Gracia-Fernández, C. A., & Gómez-Barreiro, S. (2005). Use of rheology, dielectric analysis and differential scanning calorimetry for gel time determination of a thermoset. Polymer, 46(16), 5979-5985. doi:10.1016/j.polymer.2005.05.06

Convergent functional genomics of anxiety disorders: translational identification of genes, biomarkers, pathways and mechanisms

Anxiety disorders are prevalent and disabling yet understudied from a genetic standpoint, compared with other major psychiatric disorders such as bipolar disorder and schizophrenia. The fact that they are more common, diverse and perceived as embedded in normal life may explain this relative oversight. In addition, as for other psychiatric disorders, there are technical challenges related to the identification and validation of candidate genes and peripheral biomarkers. Human studies, particularly genetic ones, are susceptible to the issue of being underpowered, because of genetic heterogeneity, the effect of variable environmental exposure on gene expression, and difficulty of accrual of large, well phenotyped cohorts. Animal model gene expression studies, in a genetically homogeneous and experimentally tractable setting, can avoid artifacts and provide sensitivity of detection. Subsequent translational integration of the animal model datasets with human genetic and gene expression datasets can ensure cross-validatory power and specificity for illness. We have used a pharmacogenomic mouse model (involving treatments with an anxiogenic drug—yohimbine, and an anti-anxiety drug—diazepam) as a discovery engine for identification of anxiety candidate genes as well as potential blood biomarkers. Gene expression changes in key brain regions for anxiety (prefrontal cortex, amygdala and hippocampus) and blood were analyzed using a convergent functional genomics (CFG) approach, which integrates our new data with published human and animal model data, as a translational strategy of cross-matching and prioritizing findings. Our work identifies top candidate genes (such as FOS, GABBR1, NR4A2, DRD1, ADORA2A, QKI, RGS2, PTGDS, HSPA1B, DYNLL2, CCKBR and DBP), brain–blood biomarkers (such as FOS, QKI and HSPA1B), pathways (such as cAMP signaling) and mechanisms for anxiety disorders—notably signal transduction and reactivity to environment, with a prominent role for the hippocampus. Overall, this work complements our previous similar work (on bipolar mood disorders and schizophrenia) conducted over the last decade. It concludes our programmatic first pass mapping of the genomic landscape of the triad of major psychiatric disorder domains using CFG, and permitted us to uncover the significant genetic overlap between anxiety and these other major psychiatric disorders, notably the under-appreciated overlap with schizophrenia. PDE10A, TAC1 and other genes uncovered by our work provide a molecular basis for the frequently observed clinical co-morbidity and interdependence between anxiety and other major psychiatric disorders, and suggest schizo-anxiety as a possible new nosological domain

Canagliflozin and Renal Outcomes in Type 2 Diabetes and Nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to 300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m 2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

SUPLEMENTO DEL XV CONGRESO INTERNACIONAL DE ACTIVIDAD FÍSICA Y DEPORTE. 17,18 y 19 DE OCTUBRE DEL 2018. ENSENADA, BAJA CALIFORNIA, MÉXICO

Compendio, en formato artículo, de los mejores trabajos presentados en el XV CONGRESO INTERNACIONAL DE ACTIVIDAD FÍSICA Y DEPORTE celebrado los días  17,18 y 19 de octubre del 2018 en la Universidad Autónoma de Baja California en la ciudad de Ensenada, Baja California, México:La nutrición en la actividad física y deportiva: alimentos funcionales con nanotecnología, aplicaciones potenciales. González González, K.Y.; Huerta Plaza, B.A.; Amaya Parra, G. (118-130)Perfil antropométrico, físico y hábitos alimentarios en escolares indígenas de Tijuana México. Avendaño Cano, D.L.; Gómez Miranda, L.M.; Aburto Corona, J.A. (131-142)Relación entre el clima de aprendizaje en Educación Física y la percepción de los estudiantes en las competencias del profesorado. Baños, R; Ortiz-Camacho, M.M.; Baena-Extremera, A.; Granero-Gallegos, A.; Machado-Parra, J.P.; Rentería, I.; Acosta, I.; Ramírez, L. (143-153)Valoración de capacidades físicas, composición corporal y consumo de vitaminas en una competencia de Crossfit. Cervantes-Hernández, N.; Hernández Nájera, N.; Carrasco Legleu, C.E.; Candia Lujan, R.; Enríquez Del Castillo, L.A. (154-164)Relación de la actividad física, características antropométricas y VO2máx en jóvenes universitarios: características por género. Enríquez-del Castillo, L.A.; Cervantes-Hernández, N.; Carrasco-Legleu, C.E.; Candia Luján, R. (165-174)Entrenamiento vibratorio de cuerpo completo y sus efectos sobre la composición corporal en jóvenes universitarios. Flores-Chico, B.; Bañuelos-Teres, L.E.; Buendía Lozada, E.R.P. (175-183)Actualización curricular, plan 2016 de la Licenciatura en Cultura Física de la BUAP. Flores-Chico, B.; Flores-Flores, A.; López de La Rosa, LE.; Aguilar-Enríquez, R.I.; Caballero Gómez, JM; Villanueva-Huerta, JA. (184-192)Evaluación psicológica y de la musculatura isquiosural de basquetbolistas universitarias en distintas etapas deportivas. Moranchel-Charros, R.; Martínez-Velázquez, E.S. (193-203)Efecto del ejercicio físico sobre la fuerza, resistencia y riesgo de caída en mujeres adultas. Ortiz Ortiz, M; Espinoza Gutiérrez, R; Gómez Miranda, LM.; Guzmán Gutiérrez, EC.; Calleja Núñez, JJ. (204-212)Desigualdad vs igualdad numérica y su efecto en la técnica de jugadores de fútbol infantil. Vega-Orozco, SI; Gavotto Nogales, OI; Bernal Reyes, F; Horta Gim, MA; Sarabia Sainz, HM. (213-224

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to &lt;90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], &gt;300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of &lt;15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P&lt;0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P&lt;0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years