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

Association of the epithelial�mesenchymal transition (EMT) with cisplatin resistance

Therapy resistance is a characteristic of cancer cells that significantly reduces the effectiveness of drugs. Despite the popularity of cisplatin (CP) as a chemotherapeutic agent, which is widely used in the treatment of various types of cancer, resistance of cancer cells to CP chemotherapy has been extensively observed. Among various reported mechanism(s), the epithelial�mesenchymal transition (EMT) process can significantly contribute to chemoresistance by converting the motionless epithelial cells into mobile mesenchymal cells and altering cell�cell adhesion as well as the cellular extracellular matrix, leading to invasion of tumor cells. By analyzing the impact of the different molecular pathways such as microRNAs, long non-coding RNAs, nuclear factor-κB (NF-ĸB), phosphoinositide 3-kinase-related protein kinase (PI3K)/Akt, mammalian target rapamycin (mTOR), and Wnt, which play an important role in resistance exhibited to CP therapy, we first give an introduction about the EMT mechanism and its role in drug resistance. We then focus specifically on the molecular pathways involved in drug resistance and the pharmacological strategies that can be used to mitigate this resistance. Overall, we highlight the various targeted signaling pathways that could be considered in future studies to pave the way for the inhibition of EMT-mediated resistance displayed by tumor cells in response to CP exposure. © 2020 by the authors

Recent advances in natural gum-based biomaterials for tissue engineering and regenerative medicine: A review

The engineering of tissues under a three-dimensional (3D) microenvironment is a great challenge and needs a suitable supporting biomaterial-based scaffold thatmay facilitate cell attachment, spreading, proliferation, migration, and differentiation for proper tissue regeneration or organ reconstruction. Polysaccharides as natural polymers promise great potential in the preparation of a three-dimensional artificial extracellular matrix (ECM) (i.e., hydrogel) via various processing methods and conditions. Natural polymers, especially gums, based upon hydrogel systems, provide similarities largely with the native ECM and excellent biological response. Here, we review the origin and physico-chemical characteristics of potentially used natural gums. In addition, various forms of scaffolds (e.g., nanofibrous, 3D printed-constructs) based on gums and their efficacy in 3D cell culture and various tissue regenerations such as bone, osteoarthritis and cartilage, skin/wound, retinal, neural, and other tissues are discussed. Finally, the advantages and limitations of natural gums are precisely described for future perspectives in tissue engineering and regenerative medicine in the concluding remarks. © 2020 by the authors