Electrodeposition of WO3 Nanoparticles for Sensing Applications

The motivation of using metal oxides is mainly due to its charge storage capabilities, and electrocatalytic, electrochromic and photoelectrochemical properties. But comparing with bulk, nanostructured materials present several advantages related with the spatial confinement, large fraction of surface atoms, high surface energy, strong surface adsorption and increased surface to volume ratio, which greatly improves the performances of these materials. The deposition of this materials can be accomplished by a variety of physical and chemical techniques but nowadays, electrodeposited metal oxides are generally used in both laboratories and industries due to the flexibility to control structure and morphology of the oxide electrodes combined with a reduced cost. Tungsten oxide (WO3) is a well-studied semiconductor and is used for several applications as chromogenic material, sensor and catalyst. The major important features is its low cost and availability, improved stability, easy morphologic and structural control of the nanostructures, reversible change of conductivity, high sensitivity, selectivity and biocompatibility. For the electrodeposition of WO3, more than one method can be adopted: electrodeposition from a precursor solution, anodic oxidation, and electrodeposition of already produced nanoparticles; however, in this case the mechanism of the electrodeposition is not fully understood. In this chapter, a review of the latest published work of electrodeposited nanostructured metal oxides is provided to the reader, with a more detailed explanation of WO3 material applied in sensing devices

Evaluación de la Competencia Transversal 2: Aplicación y pensamiento práctico

[ES] En este trabajo se presenta la competencia transversal 2 evaluada en la Universitat Politècnica de València, “Aplicación y Pensamiento Práctico”. En concreto se centra en la evaluación de dicha competencia en 3 asignaturas del Grado en Administración y Dirección de Empresas que se imparte en dicha Universidad. Las asignaturas corresponden tanto al nivel uno (primer y segundo curso) como al dos (tercer y cuarto curso) de la titulación.[EN] En este trabajo se presenta la competencia transversal 2 evaluada en la Universitat Politècnica de València, “Aplicación y Pensamiento Práctico”. En concreto se centra en la evaluación de dicha competencia en 3 asignaturas del Grado en Administración y Dirección de Empresas que se imparte en dicha Universidad. Las asignaturas corresponden tanto al nivel uno (primer y segundo curso) como al dos (tercer y cuarto curso) de la titulaciónVallada Regalado, E.; Martín Marín, J.; Úbeda García, JE.; Crespo Abril, F. (2019). Evaluación de la Competencia Transversal 2: Aplicación y pensamiento práctico. En JIDDO. I Jornada de innovación en docencia universitaria para la dirección de organizaciones públicas y privadas. Editorial Universitat Politècnica de València. 17-23. https://doi.org/10.4995/JIDDO2019.2019.10188OCS172

Advancing manufacture of hiPSC-derived hepatocytes with improved functionality: A nature-inspired protocol

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Metodologías aplicadas en la evaluación de la competencia transversal “Comprensión e Integración” en los estudios universitarios del Grado de Administración y Dirección de Empresas en la Universitat Politècnica de València

[ES] Las metodologías aplicadas en la implementación de la Competencia Transversal “Comprensión e integración” en los estudios de Administración y Dirección de Empresas, son muy diversas predominando el estudio de casos y la resolución de problemas. En el presente trabajo se analiza cómo se aplican y qué ventajas e inconvenientes se observan. Se concluye que al evaluar al alumno desde distintas perspectivas con diferentes metodologías, el resultado global integra los distintos niveles de dominio de la competencia[EN] In this paper we analyze the implementation of the Transversal Competence “Understanding and integration” through the subjects of Business Administration and Management degree. It is kept in mind that the proficiency level of the competence can not be the same for 1st and 2nd years, which would correspond to level 1, that for 3rd and 4th years, to which they correlate a higher level 2 The broad range of methodologies and techniques used in the assessment of this skill, allows us to infer that the overall evaluation is correctMarín Sánchez, MDM.; Crespo Abril, F.; Maroto Álvarez, MC.; Palau Ramírez, F.; Palmer Gato, ME.; Úbeda García, JE. (2019). Metodologías aplicadas en la evaluación de la competencia transversal “Comprensión e Integración” en los estudios universitarios del Grado de Administración y Dirección de Empresas en la Universitat Politècnica de València. En JIDDO. I Jornada de innovación en docencia universitaria para la dirección de organizaciones públicas y privadas. Editorial Universitat Politècnica de València. 8-16. https://doi.org/10.4995/JIDDO2019.2019.10247OCS81

A new wildland fire danger index for a Mediterranean region and some validation aspects

Wildland fires are the main cause of tree mortality in Mediterranean Europe and a major threat to Spanish forests. This paper focuses on the design and validation of a new wildland fire index especially adapted to a Mediterranean Spanish region. The index considers ignition and spread danger components. Indicators of natural and human ignition agents, historical occurrence, fuel conditions and fire spread make up the hierarchical structure of the index. Multi-criteria methods were used to incorporate experts¿ opinion in the process of weighting the indicators and to carry out the aggregation of components into the final index, which is used to map the probability of daily fire occurrence on a 0.5-km grid. Generalised estimating equation models, which account for possible correlated responses, were used to validate the index, accommodating its values onto a larger scale because historical records of daily fire occurrence, which constitute the dependent variable, are referred to cells on a 10-km grid. Validation results showed good index performance, good fit of the logistic model and acceptable discrimination power. Therefore, the index will improve the ability of fire prevention services in daily allocation of resources.The authors acknowledge the support received from the Ministry of Science and Innovation through the research project Modelling and Optimisation Techniques for a Sustainable Development, Ref. EC02008-05895-C02-01/ECON.Vicente López, FJD.; Crespo Abril, F. (2012). A new wildland fire danger index for a Mediterranean region and some validation aspects. International Journal of Wildland Fire. 21(8):1030-1041. https://doi.org/10.1071/WF11046S10301041218Aguado, I., Chuvieco, E., Borén, R., & Nieto, H. (2007). Estimation of dead fuel moisture content from meteorological data in Mediterranean areas. Applications in fire danger assessment. International Journal of Wildland Fire, 16(4), 390. doi:10.1071/wf06136Andrews, P. L., Loftsgaarden, D. O., & Bradshaw, L. S. (2003). Evaluation of fire danger rating indexes using logistic regression and percentile analysis. International Journal of Wildland Fire, 12(2), 213. doi:10.1071/wf02059Bradstock, R. A., Cohn, J. S., Gill, A. M., Bedward, M., & Lucas, C. (2009). Prediction of the probability of large fires in the Sydney region of south-eastern Australia using fire weather. International Journal of Wildland Fire, 18(8), 932. doi:10.1071/wf08133Buizza, R., & Hollingsworth, A. (2002). Storm prediction over Europe using the ECMWF Ensemble Prediction System. Meteorological Applications, 9(3), 289-305. doi:10.1017/s1350482702003031Carmel, Y., Paz, S., Jahashan, F., & Shoshany, M. (2009). Assessing fire risk using Monte Carlo simulations of fire spread. Forest Ecology and Management, 257(1), 370-377. doi:10.1016/j.foreco.2008.09.039Castedo-Dorado, F., Rodríguez-Pérez, J. R., Marcos-Menéndez, J. L., & Álvarez-Taboada, M. F. (2011). Modelling the probability of lightning-induced forest fire occurrence in the province of León (NW Spain). Forest Systems, 20(1), 95. doi:10.5424/fs/2011201-9409Catry, F. X., Rego, F. C., Bação, F. L., & Moreira, F. (2009). Modeling and mapping wildfire ignition risk in Portugal. International Journal of Wildland Fire, 18(8), 921. doi:10.1071/wf07123CHUVIECO, E., & SALAS, J. (1996). Mapping the spatial distribution of forest fire danger using GIS. International journal of geographical information systems, 10(3), 333-345. doi:10.1080/02693799608902082Chuvieco, E., Cocero, D., Riaño, D., Martin, P., Martı́nez-Vega, J., de la Riva, J., & Pérez, F. (2004). Combining NDVI and surface temperature for the estimation of live fuel moisture content in forest fire danger rating. Remote Sensing of Environment, 92(3), 322-331. doi:10.1016/j.rse.2004.01.019Chuvieco, E., Aguado, I., Yebra, M., Nieto, H., Salas, J., Martín, M. P., … Zamora, R. (2010). Development of a framework for fire risk assessment using remote sensing and geographic information system technologies. Ecological Modelling, 221(1), 46-58. doi:10.1016/j.ecolmodel.2008.11.017Danson, F. M., & Bowyer, P. (2004). Estimating live fuel moisture content from remotely sensed reflectance. Remote Sensing of Environment, 92(3), 309-321. doi:10.1016/j.rse.2004.03.017Dasgupta, S., Qu, J. J., & Hao, X. (2006). Design of a Susceptibility Index for Fire Risk Monitoring. IEEE Geoscience and Remote Sensing Letters, 3(1), 140-144. doi:10.1109/lgrs.2005.858484Fairbrother, A., & Turnley, J. G. (2005). Predicting risks of uncharacteristic wildfires: Application of the risk assessment process. Forest Ecology and Management, 211(1-2), 28-35. doi:10.1016/j.foreco.2005.01.026Finney, M. A. (2005). The challenge of quantitative risk analysis for wildland fire. Forest Ecology and Management, 211(1-2), 97-108. doi:10.1016/j.foreco.2005.02.010Gouma, V., & Chronopoulou-Sereli, A. (1998). Wildland Fire Danger Zoning - a Methodology. International Journal of Wildland Fire, 8(1), 37. doi:10.1071/wf9980037Hernandez-Leal, P. A., Arbelo, M., & Gonzalez-Calvo, A. (2006). Fire risk assessment using satellite data. Advances in Space Research, 37(4), 741-746. doi:10.1016/j.asr.2004.12.053Li, L.-M., Song, W.-G., Ma, J., & Satoh, K. (2009). Artificial neural network approach for modeling the impact of population density and weather parameters on forest fire risk. International Journal of Wildland Fire, 18(6), 640. doi:10.1071/wf07136Maingi, J. K., & Henry, M. C. (2007). Factors influencing wildfire occurrence and distribution in eastern Kentucky, USA. International Journal of Wildland Fire, 16(1), 23. doi:10.1071/wf06007Martell, D. L., Otukol, S., & Stocks, B. J. (1987). A logistic model for predicting daily people-caused forest fire occurrence in Ontario. Canadian Journal of Forest Research, 17(5), 394-401. doi:10.1139/x87-068Martínez, J., Vega-Garcia, C., & Chuvieco, E. (2009). Human-caused wildfire risk rating for prevention planning in Spain. Journal of Environmental Management, 90(2), 1241-1252. doi:10.1016/j.jenvman.2008.07.005Moffett, A., Garson, J., & Sarkar, S. (2005). MultCSync: a software package for incorporating multiple criteria in conservation planning. Environmental Modelling & Software, 20(10), 1315-1322. doi:10.1016/j.envsoft.2004.10.001Nieto, H., Aguado, I., Chuvieco, E., & Sandholt, I. (2010). Dead fuel moisture estimation with MSG–SEVIRI data. Retrieval of meteorological data for the calculation of the equilibrium moisture content. Agricultural and Forest Meteorology, 150(7-8), 861-870. doi:10.1016/j.agrformet.2010.02.007Noble, B. F., & Christmas, L. M. (2007). Strategic Environmental Assessment of Greenhouse Gas Mitigation Options in the Canadian Agricultural Sector. Environmental Management, 41(1), 64-78. doi:10.1007/s00267-007-9017-yNúñez-Regueira, L. (1997). Calorific values and flammability of forest species in galicia. Continental high mountainous and humid Atlantic zones. Bioresource Technology, 61(2), 111-119. doi:10.1016/s0960-8524(97)00053-9Padilla, M., & Vega-García, C. (2011). On the comparative importance of fire danger rating indices and their integration with spatial and temporal variables for predicting daily human-caused fire occurrences in Spain. International Journal of Wildland Fire, 20(1), 46. doi:10.1071/wf09139Pendergast, J. F., Gange, S. J., Newton, M. A., Lindstrom, M. J., Palta, M., & Fisher, M. R. (1996). A Survey of Methods for Analyzing Clustered Binary Response Data. International Statistical Review / Revue Internationale de Statistique, 64(1), 89. doi:10.2307/1403425Pew, K. ., & Larsen, C. P. . (2001). GIS analysis of spatial and temporal patterns of human-caused wildfires in the temperate rain forest of Vancouver Island, Canada. Forest Ecology and Management, 140(1), 1-18. doi:10.1016/s0378-1127(00)00271-1Podur, J., Martell, D. L., & Csillag, F. (2003). Spatial patterns of lightning-caused forest fires in Ontario, 1976–1998. Ecological Modelling, 164(1), 1-20. doi:10.1016/s0304-3800(02)00386-1Preisler, H. K., Brillinger, D. R., Burgan, R. E., & Benoit, J. W. (2004). Probability based models for estimation of wildfire risk. International Journal of Wildland Fire, 13(2), 133. doi:10.1071/wf02061Preisler, H. K., Chen, S.-C., Fujioka, F., Benoit, J. W., & Westerling, A. L. (2008). Wildland fire probabilities estimated from weather model-deduced monthly mean fire danger indices. International Journal of Wildland Fire, 17(3), 305. doi:10.1071/wf06162Romero-Calcerrada, R., Novillo, C. J., Millington, J. D. A., & Gomez-Jimenez, I. (2008). GIS analysis of spatial patterns of human-caused wildfire ignition risk in the SW of Madrid (Central Spain). Landscape Ecology, 23(3), 341-354. doi:10.1007/s10980-008-9190-2Saaty, T. L. (1987). RANK GENERATION, PRESERVATION, AND REVERSAL IN THE ANALYTIC HIERARCHY DECISION PROCESS. Decision Sciences, 18(2), 157-177. doi:10.1111/j.1540-5915.1987.tb01514.xSahin, Y. G., & Ince, T. (2009). Early Forest Fire Detection Using Radio-Acoustic Sounding System. Sensors, 9(3), 1485-1498. doi:10.3390/s90301485López, A. S., San-Miguel-Ayanz, J., & Burgan, R. E. (2002). Integration of satellite sensor data, fuel type maps and meteorological observations for evaluation of forest fire risk at the pan-European scale. International Journal of Remote Sensing, 23(13), 2713-2719. doi:10.1080/01431160110107761Sharples, J. J., McRae, R. H. D., Weber, R. O., & Gill, A. M. (2009). A simple index for assessing fire danger rating. Environmental Modelling & Software, 24(6), 764-774. doi:10.1016/j.envsoft.2008.11.004Stocks, B. J., Lynham, T. J., Lawson, B. D., Alexander, M. E., Wagner, C. E. V., McAlpine, R. S., & Dubé, D. E. (1989). The Canadian Forest Fire Danger Rating System: An Overview. The Forestry Chronicle, 65(6), 450-457. doi:10.5558/tfc65450-6Sturtevant, B. R., & Cleland, D. T. (2007). Human and biophysical factors influencing modern fire disturbance in northern Wisconsin. International Journal of Wildland Fire, 16(4), 398. doi:10.1071/wf06023Swets, J. (1988). Measuring the accuracy of diagnostic systems. Science, 240(4857), 1285-1293. doi:10.1126/science.3287615Vadrevu, K. P., Eaturu, A., & Badarinath, K. V. S. (2009). Fire risk evaluation using multicriteria analysis—a case study. Environmental Monitoring and Assessment, 166(1-4), 223-239. doi:10.1007/s10661-009-0997-3Vasilakos, C., Kalabokidis, K., Hatzopoulos, J., Kallos, G., & Matsinos, Y. (2007). Integrating new methods and tools in fire danger rating. International Journal of Wildland Fire, 16(3), 306. doi:10.1071/wf05091Verde, J. C., & Zêzere, J. L. (2010). Assessment and validation of wildfire susceptibility and hazard in Portugal. Natural Hazards and Earth System Science, 10(3), 485-497. doi:10.5194/nhess-10-485-2010Wotton, B. M., & Martell, D. L. (2005). A lightning fire occurrence model for Ontario. Canadian Journal of Forest Research, 35(6), 1389-1401. doi:10.1139/x05-071Yebra, M., Chuvieco, E., & Riaño, D. (2008). Estimation of live fuel moisture content from MODIS images for fire risk assessment. Agricultural and Forest Meteorology, 148(4), 523-536. doi:10.1016/j.agrformet.2007.12.00

Defining the scope of the European Antimicrobial Resistance Surveillance network in Veterinary medicine (EARS-Vet): a bottom-up and One Health approach

Background Building the European Antimicrobial Resistance Surveillance network in Veterinary medicine (EARS-Vet) was proposed to strengthen the European One Health antimicrobial resistance (AMR) surveillance approach. Objectives To define the combinations of animal species/production types/age categories/bacterial species/specimens/antimicrobials to be monitored in EARS-Vet. Methods The EARS-Vet scope was defined by consensus between 26 European experts. Decisions were guided by a survey of the combinations that are relevant and feasible to monitor in diseased animals in 13 European countries (bottom-up approach). Experts also considered the One Health approach and the need for EARS-Vet to complement existing European AMR monitoring systems coordinated by the ECDC and the European Food Safety Authority (EFSA). Results EARS-Vet plans to monitor AMR in six animal species [cattle, swine, chickens (broilers and laying hens), turkeys, cats and dogs], for 11 bacterial species (Escherichia coli, Klebsiella pneumoniae, Mannheimia haemolytica, Pasteurella multocida, Actinobacillus pleuropneumoniae, Staphylococcus aureus, Staphylococcus pseudintermedius, Staphylococcus hyicus, Streptococcus uberis, Streptococcus dysgalactiae and Streptococcus suis). Relevant antimicrobials for their treatment were selected (e.g. tetracyclines) and complemented with antimicrobials of more specific public health interest (e.g. carbapenems). Molecular data detecting the presence of ESBLs, AmpC cephalosporinases and methicillin resistance shall be collected too. Conclusions A preliminary EARS-Vet scope was defined, with the potential to fill important AMR monitoring gaps in the animal sector in Europe. It should be reviewed and expanded as the epidemiology of AMR changes, more countries participate and national monitoring capacities improve.Peer reviewe

Human-based fibrillar nanocomposite hydrogels as bioinstructive matrices to tune stem cell behavior

The extracellular matrix (ECM)-biomimetic fibrillar structure of platelet lysate (PL) gels along with its enriched milieu of biomolecules has drawn significant interest in regenerative medicine applications. However, PL-based gels have poor structural stability which severely limits its performance as a bioinstructive biomaterial. Here, rod-shaped cellulose nanocrystals (CNC) are used as a novel approach to modulate the physical and biochemical microenvironment of PL gels enabling their effective use as injectable human-based cell scaffolds with a level of biomimicry that is difficult to recreate with synthetic biomaterials. The incorporation of CNC (0 to 0.61 wt.%) into the PL fibrillar network during the coagulation cascade leads to decreased fiber branching, increased interfiber porosity (from 66 to 83%) and modulate fiber (from 1.4 ± 0.7 to 27 ± 12 kPa) and bulk hydrogel (from 18 ± 4 to 1256 ± 82 Pa) mechanical properties. As result of these physicochemical alterations, nanocomposite PL hydrogels resist to the typical extensive clot retraction (from 76 ± 1 to 24 ± 3 at Day 7) and show favored retention of PL bioactive molecules. The feedback of these cues on the fate of human adipose-derived stem cells is evaluated, showing how it can be explored to modulate the commitment of encapsulated stem cells toward different genetic phenotypes without the need for additional external biological stimuli. These fibrillar nanocomposite hydrogels allow therefore to explore the outstanding biological properties of human-based PL as an efficient engineered ECM which can be tailored to trigger specific regenerative pathways in minimal invasive strategies.The authors thank the Hospital da Prelada (Porto, Portugal) for providing adipose tissue samples. The authors acknowledge the financial support from project Recognize (UTAP-ICDT/CTM-BIO/0023/2014), project NORTE-01-0145FEDER-000021 supported by the Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), the European Union Framework Programme for Research and Innovation HORIZON 2020, under the TEAMING Grant agreement No. 739572 – The Discoveries CTR EU, Forecast 668983, Marie Skłodowska-Curie grant agreement No. 706996 (PrinTendon) and CHEM2NATURE 692333; FCT/MCTES (Fundação para a Ciência e a Tecnologia/ Ministério da Ciência, Tecnologia, e Ensino Superior) and the Fundo Social Europeu através do Programa Operacional do Capital Humano (FSE/POCH) in the framework of PhD grant PD/59/2013 – PD/BD/113807/2015 for BBM, Post-Doc grant SFRH/BPD/112459/2015 for R.D.info:eu-repo/semantics/publishedVersio

Relevance of Stereotyped B-Cell Receptors in the Context of the Molecular, Cytogenetic and Clinical Features of Chronic Lymphocytic Leukemia

Highly homologous B-cell receptors, characterized by non-random combinations of immunoglobulin heavy-chain variable (IGHV) genes and heavy-chain complementarity determining region-3 (HCDR3), are expressed in a recurrent fraction of patients affected by chronic lymphocytic leukemia (CLL). We investigated the IGHV status of 1131 productive IG rearrangements from a panel of 1126 CLL patients from a multicenter Italian study group, and correlated the presence and class of HCDR3 stereotyped subsets with the major cytogenetic alterations evaluated by FISH, molecular prognostic factors, and the time to first treatment (TTFT) of patients with early stage disease (Binet A). Stereotyped HCDR3 sequences were found in 357 cases (31.7%), 231 of which (64.7%) were unmutated. In addition to the previously described subsets, 31 new putative stereotypes subsets were identified. Significant associations between different stereotyped HCDR3 sequences and molecular prognostic factors, such as CD38 and ZAP-70 expression, IGHV mutational status and genomic abnormalities were found. In particular, deletion of 17p13 was significantly represented in stereotype subset #1. Notably, subset #1 was significantly correlated with a substantially reduced TTFT compared to other CLL groups showing unmutated IGHV, ZAP-70 or CD38 positivity and unfavorable cytogenetic lesions including del(17)(p13). Moreover, subset #2 was strongly associated with deletion of 13q14, subsets #8 and #10 with trisomy 12, whereas subset #4 was characterized by the prevalent absence of the common cytogenetic abnormalities. Our data from a large and representative panel of CLL patients indicate that particular stereotyped HCDR3 sequences are associated with specific cytogenetic lesions and a distinct clinical outcome

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field