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

Transient Porosity in Densely Packed Crystalline Carbazole–(<i>p</i>‑Diethynylphenylene)–Carbazole Rotors: CO₂ and Acetone Sorption Properties

We report for the first time the high sorption properties of a molecular rotor with no permanent voids or channels in its crystal structure. Such crystalline phase originates from THF, DCM, or the irreversible desolvation of entrapped benzene molecules. From these, the benzene in its solvate form acts as rotation stopper, as supported by dynamic characterization using solid-state ²H NMR experiments. In the solvent-free form, the diffusion of small quantities of iodine vapors caused a significant change in the intramolecular rotation, increasing the known activation energy to rotation from 8.5 to 10.6 kcal mol^–1. Notably, those results paved the way for the discovery of the high CO₂ uptake (201.6 cm³ g^–1 at 196 K, under 1 atm) and acetone (5 wt %), a sorption property that was attributed to both, the restriction of the molecular rotation at low temperatures and the flexibility of the molecular axle made of conjugated <i>p</i>-(ethynylphenylene), surrounded by carbazole

Biofabrication of Silk Fibers with Enhanced Conductivity through Silkworm Feeding with Reduced Graphene Oxide: Implications for Smart Textile Innovations

In this study, we present an innovative research within the intersection of biotechnology, materials science, and electronics by presenting an approach to enhance the electrical characteristics of silk fibers. By feeding silkworms with mulberry leaves infused with reduced graphene oxide (rGO), we achieved the integration of the nanomaterial into the biopolymer matrix of silk fibers. Through comprehensive spectroscopic analyses, including Fourier transform infrared (FTIR) and Raman spectroscopy, we confirmed the incorporation of rGO into the silk fiber structure. Key findings emphasize a significant hydrogen interaction between the hydroxyl groups of rGO and the NH groups of silk. This molecular interaction not only bolsters the conductivity of the fibers but also maintains the natural silk’s coloration. Electrical characterization revealed a temperature-dependent conductivity pattern. Significantly, this behavior adheres to the variable range hopping (VRH) formalism, suggesting a σ(T1/4) temperature relationship. Notably, a conductivity of approximately σ =4 × 10–3 S/m was achieved in the modified fibers. This study represents a significant advancement in the innovation of electronic textiles by enhancing the electrical properties of silk fibers. The introduction of nanomaterial opens new opportunities for its implementation in the field of smart textiles, providing improved electronic properties that have the potential to transform the apparel industry

Guía Académica para Estudiantes Internacionales. Departamento de Filología Española, Lingüística General y Teoría de la Literatura de la Universidad de Alicante

[No abstract available] Document in Spanis

Structure–Activity Relationship of a New Series of Reversible Dual Monoacylglycerol Lipase/Fatty Acid Amide Hydrolase Inhibitors

The two endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), play independent and nonredundant roles in the body. This makes the development of both selective and dual inhibitors of their inactivation an important priority. In this work we report a new series of inhibitors of monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH). Among them, (±)-oxiran-2-ylmethyl 6-(1,1′-biphenyl-4-yl)­hexanoate (<b>8</b>) and (2<i>R</i>)-(−)-oxiran-2-ylmethyl­(4-benzylphenyl)­acetate (<b>30</b>) stand out as potent inhibitors of human recombinant MAGL (IC₅₀ (<b>8</b>) = 4.1 μM; IC₅₀ (<b>30</b>) = 2.4 μM), rat brain monoacylglycerol hydrolysis (IC₅₀ (<b>8</b>) = 1.8 μM; IC₅₀ (<b>30</b>) = 0.68 μM), and rat brain FAAH (IC₅₀ (<b>8</b>) = 5.1 μM; IC₅₀ (<b>30</b>) = 0.29 μM). Importantly, and in contrast to the other previously described MAGL inhibitors, these compounds behave as reversible inhibitors either of competitive (<b>8</b>) or noncompetitive nature (<b>30</b>). Hence, they could be useful to explore the therapeutic potential of reversible MAGL inhibitors

Español académico e inmersión universitaria del alumnado extranjero de la Universidad de Alicante

La integración académica de los estudiantes internacionales es un paso imprescindible en la internacionalización del sistema universitario. Para contribuir a ella, el objetivo de esta red ha sido la creación de una Guía académica para estudiantes internacionales, que persigue un mejor rendimiento académico de su estancia. Sus destinatarios son los estudiantes de movilidad internacional de la Facultad de Filosofía y Letras que cursan asignaturas relacionadas con la lengua y la literatura españolas. Metodológicamente, partiendo del diagnóstico de la situación tanto del alumnado extranjero como de su profesorado obtenida como resultado de una red anterior (Pastor, 2014b), hemos trabajado con las asignaturas más demandadas por este colectivo en los últimos tres años, hemos realizado entrevistas personalizadas con el profesorado que las imparte y hemos elaborado la información requerida. Así pues, ofrecemos como resultado la presente Guía, que incluye una introducción sobre la cultura académica universitaria en España, así como las fichas de las asignaturas más solicitadas, diseñadas desde la perspectiva de lo que requiere de ellas un estudiante internacional (requisitos lingüísticos, material adaptado, pautas de evaluación, comentarios de anteriores alumnos, etc.). Confiamos en facilitar con todo ello su interacción con el profesorado y el resto del alumnado e incrementar así su rendimiento y éxito académicos

Total Syntheses of (+)-Vigulariol and (−)-Sclerophytin A

[Image: see text] The total synthesis of (+)-vigulariol and (−)-sclerophytin A are reported in 15 steps and 16 steps, respectively, from a known compound. The flexible, readily scalable synthetic strategy allows for rapid construction of a critical tricyclic intermediate and is demonstrated via the synthesis of these two marine natural products. A key reaction in this synthetic protocol is a combination Wittig/intramolecular Diels-Alder cycloaddition

In-depth search focused on furans, lactones, volatile phenols, and acetals as potential age markers of Madeira wines by comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry combined with solid phase microextraction

The establishment of potential age markers of Madeira wine is of paramount significance as it may contribute to detect frauds and to ensure the authenticity of wine. Considering the chemical groups of furans, lactones, volatile phenols, and acetals, 103 volatile compounds were tentatively identified; among these, 71 have been reported for the first time in Madeira wines. The chemical groups that could be used as potential age markers were predominantly acetals, namely, diethoxymethane, 1,1-diethoxyethane, 1,1-diethoxy-2-methyl-propane, 1-(1-ethoxyethoxy)-pentane, trans-dioxane and 2-propyl-1,3-dioxolane, and from the other chemical groups, 5-methylfurfural and cis-oak-lactone, independently of the variety and the type of wine. GC × GC-ToFMS system offers a more useful approach to identify these compounds compared to previous studies using GC−qMS, due to the orthogonal systems, that reduce coelution, increase peak capacity and mass selectivity, contributing to the establishment of new potential Madeira wine age markers. Remarkable results were also obtained in terms of compound identification based on the organized structure of the peaks of structurally related compounds in the GC × GC peak apex plots. This information represents a valuable approach for future studies, as the ordered-structure principle can considerably help the establishment of the composition of samples. This new approach provides data that can be extended to determine age markers of other types of wines