1,721,253 research outputs found
Need for Methods to Investigate Endocannabinoid Signaling
No full textEndocannabinoids (eCBs) are endogenous lipids able to bind to cannabinoid receptors, the primary molecular targets of the cannabis (Cannabis sativa) active principle Δ9-tetrahydrocannabinol. During the last 20 years, several N-acylethanolamines and acylesters have been shown to act as eCBs, and a complex array of receptors, metabolic enzymes, and transporters (that altogether form the so-called "eCB system") has been shown to finely tune their manifold biological activities. It appears now urgent to develop methods and protocols that allow to assay in a specific and quantitative manner the distinct components of the eCB system and that can properly localize them within the cell. A brief overview of eCBs and of the proteins that bind, transport, and metabolize these lipids is presented here, in orderto put in a better perspective, the relevance of methodologies that help to disclose molecular details of eCB signaling in health and disease. Proper methodological approaches form also the basis for a more rationale and effective drug design and therapeutic strategy to combat human disorders
A game of networks: Do different lipids interact to orchestrate inflammatory homeostasis?
No full textResolution of inflammation is the physiological process whereby endogenous pro-resolving lipids constrain inflammatory stimuli that would otherwise cause chronic inflammation. In this issue of Cell Chemical Biology, Peltner et al.1 report that the cannabis component cannabidiol induces production of pro-resolving lipids directly activating 15-lipoxygenase and inhibiting 5-lipoxygenase in human macrophages
Endocannabinoids and Skin Barrier Function: Molecular Pathways and Therapeutic Opportunities
No full textAs an interface between the organism and its environment, skin is continuously subjected to a myriad of insults that could impair its structural and functional integrity, ultimately leading to clinical manifestations. Skin reacts to external stresses through a vast array of cellular and molecular components, which form a highly sophisticated and well-organized signaling network. Our knowledge of the molecular pathways underlying this complex interplay and accounting for skin homeostasis , both under normal and pathological conditions, is still in its infancy. Among these pathways, the endocannabinoid system has emerged as a key actor in skin biology , by controlling epidermal barrier formation and maintenance, modulating growth and terminal differentiation of cutaneous cells, and regulating skin inflammatory responses via pleiotropic mechanisms. This chapter summarizes our current knowledge of the manifold effects of endocannabinoids in skin, in order to put in a better perspective their potential as next-generation therapeutics against disorders of this organ, particularly those involving dysregulation of immune system and epidermal barrier, such as allergic and atopic dermatitis, localized scleroderma and psoriasis
Light & biotech application in the study of living organisms.
No full textThis 40-page document sheds light on the outcomes of the European Biotech Week, with short reports for each one of the many events and initiatives which took place in 2015
Radiometric Assay of NAPE-PLD Activity
No full textN-Acyl-phosphatidylethanolamine (NAPE)-hydrolyzing phospholipase D (NAPE-PLD) is a prominent enzyme involved in the biosynthesis of fatty acid amides, a family of bioactive lipids including anandamide as the prototypical member. Here, we describe a NAPE-PLD assay based on radioactive substrates and product separation by thin layer chromatography (TLC)
The endocannabinoid system in ageing: a new target for drug development
No full textEndocannabinoids are a new class of lipids, which include amides, esters andethers of long chain polyunsaturated fatty acids. Anandamide(N-arachidonoylethanolamine; AEA) and 2-arachidonoylglycerol are the mainendogenous agonists of cannabinoid receptors able to mimic severalpharmacological effects of Delta(9)-tetrahydrocannabinol, the active principle ofCannabis sativa preparations like hashish and marijuana. AEA is released "ondemand" from membrane lipids, and its activity at the receptors is limited bycellular uptake followed by intracellular hydrolysis. Together with AEA andcongeners, the proteins which bind, synthesize, transport and hydrolyze AEA form the "endocannabinoid system". Endogenous cannabinoids are present in the central nervous system and in peripheral tissues, suggesting a physiological role asbroad spectrum modulators. This review summarizes the main features of theendocannabinoid system, and the latest advances on its involvement in ageing ofcentral and peripheral cells. In addition, the therapeutic potential of recently developed drugs able to modulate the endocannabinoid tone for the treatment ofageing and age-related human pathologies will be reviewed.[...
Radiometric Assay of ABHD2 Activity
No full textThe α,β-hydrolase fold-containing protein 2 (ABHD2) is a serine hydrolase, responsible for the cleavage of endogenous 2-arachidonoylglycerol (2-AG). ABHD2 is activated by progesterone, thus, it is considered a nonnuclear receptor of this steroid hormone that terminates its biological effects. The products of ABHD2-catalyzed cleavage by the natural substrate 2-AG are glycerol and arachidonic acid; here, instead of 2-AG, the radioactive substrate 2-oleoyl-[3H]glycerol has been used as already done in various acylglycerol lipase activity assays. The amount of [3H]glycerol released allows to measure ABHD2 enzymatic activity
Design and Synthesis of Biotinylated Probes for N-Acyl-Ethanolamines
No full textThe present invention relates to the synthesis and characterization of biotinylated analogue of N-arachidonoylethanolamine (AEA) and its use as a tool to study AEA transport and trafficking through biochemical and morphological techniques. In particular biotinylated AEA (b-AEA, for which we propose the common name MM22) is suitable to design highly sensitive and simple methods for the non-radioactive detection and quantitation of AEA from complex samples, which would offer a useful alternative approach to the routinely used radiometric assays. The invention also relates to the use of b-AEA as a potential therapeutic and diagnostic agent.[...
Fluorimetric Assay of FAAH Activity
No full text: Fatty acid amide hydrolase (FAAH) is the enzyme responsible for the degradation of anandamide (N-arachidonoylethanolamine, AEA) to arachidonic acid (AA) and ethanolamine. The method described here measures FAAH activity through the fluorometric arachidonoyl-7-amino-4-methyl-coumarin amide (AAMCA) substrate, which allows a simple and sensitive assay suitable for high-throughput screening tests. FAAH catalyzes the hydrolysis of AAMCA producing AA and the highly fluorescent compound 7-amino-4-methylcoumarin (AMC)
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