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

Gene expression in coffee

Coffee is cultivated in more than 70 countries of the intertropical belt where it has important economic, social and environmental impacts. As for many other crops, the development of molecular biology technics allowed to launch research projects for coffee analyzing gene expression. In the 90s decade, the first expression studies were performed by Northern-blot or PCR, and focused on genes coding enzymes of the main compounds (e.g., storage proteins, sugars, complex polysaccharides, caffeine and chlorogenic acids) found in green beans. Few years after, the development of 454 pyrosequencing technics generated expressed sequence tags (ESTs) obviously from beans but also from other organs (e.g., leaves and roots) of the two main cultivated coffee species, Coffea arabica and C. canephora. Together with the use of real-time quantitative PCR, these ESTs significantly raised the number of coffee gene expression studies leading to the identification of (1) key genes of biochemical pathways, (2) candidate genes involved in biotic and abiotic stresses as well as (3) molecular markers essential to assess the genetic diversity of the Coffea genus, for example. The development of more recent Illumina sequencing technology now allows large-scale transcriptome analysis in coffee plants and opens the way to analyze the effects on gene expression of complex biological processes like genotype and environment interactions, heterosis and gene regulation in polypoid context like in C. arabica. The aim of the present review is to make an extensive list of coffee genes studied and also to perform an inventory of large-scale sequencing (RNAseq) projects already done or on-going

Implementing Ultrasound Imaging for the Assessment of Muscle and Tendon Properties in Elite Sports: Practical Aspects, Methodological Considerations and Future Directions.

Ultrasound (US) imaging has been widely used in both research and clinical settings to evaluate the morphological and mechanical properties of muscle and tendon. In elite sports scenarios, a regular assessment of such properties has great potential, namely for testing the response to training, detecting athletes at higher risks of injury, screening athletes for structural abnormalities related to current or future musculoskeletal complaints, and monitoring their return to sport after a musculoskeletal injury. However, several practical and methodological aspects of US techniques should be considered when applying this technology in the elite sports context. Therefore, this narrative review aims to (1) present the principal US measures and field of applications in the context of elite sports; (2) to discuss, from a methodological perspective, the strengths and shortcomings of US imaging for the assessment of muscle and tendon properties; and (3) to provide future directions for research and application