Phase I Hydroxylated Metabolites of the K2 Synthetic Cannabinoid JWH-018 Retain In Vitro and In Vivo Cannabinoid 1 Receptor Affinity and Activity

K2 products are synthetic cannabinoid-laced, marijuana-like drugs of abuse, use of which is often associated with clinical symptoms atypical of marijuana use, including hypertension, agitation, hallucinations, psychosis, seizures and panic attacks. JWH-018, a prevalent K2 synthetic cannabinoid, is structurally distinct from Δ(9)-THC, the main psychoactive ingredient in marijuana. Since even subtle structural differences can lead to differential metabolism, formation of novel, biologically active metabolites may be responsible for the distinct effects associated with K2 use. The present study proposes that K2's high adverse effect occurrence is due, at least in part, to distinct JWH-018 metabolite activity at the cannabinoid 1 receptor (CB1R).JWH-018, five potential monohydroxylated metabolites (M1-M5), and one carboxy metabolite (M6) were examined in mouse brain homogenates containing CB1Rs, first for CB1R affinity using a competition binding assay employing the cannabinoid receptor radioligand [(3)H]CP-55,940, and then for CB1R intrinsic efficacy using an [(35)S]GTPγS binding assay. JWH-018 and M1-M5 bound CB1Rs with high affinity, exhibiting K(i) values that were lower than or equivalent to Δ(9)-THC. These molecules also stimulated G-proteins with equal or greater efficacy relative to Δ(9)-THC, a CB1R partial agonist. Most importantly, JWH-018, M2, M3, and M5 produced full CB1R agonist levels of activation. CB1R-mediated activation was demonstrated by blockade with O-2050, a CB1R-selective neutral antagonist. Similar to Δ(9)-THC, JWH-018 and M1 produced a marked depression of locomotor activity and core body temperature in mice that were both blocked by the CB1R-preferring antagonist/inverse agonist AM251.Unlike metabolites of most drugs, the studied JWH-018 monohydroxylated compounds, but not the carboxy metabolite, retain in vitro and in vivo activity at CB1Rs. These observations, combined with higher CB1R affinity and activity relative to Δ(9)-THC, may contribute to the greater prevalence of adverse effects observed with JWH-018-containing products relative to cannabis

Building a better foundation: improving root-trait measurements to understand and model plant and ecosystem processes

Trait-based approaches provide a useful framework to investigate plant strategies for resource acquisition, growth, and competition, as well as plant impacts on ecosystem processes. Despite significant progress capturing trait variation within and among stems and leaves, identification of trait syndromes within fine-root systems and between fine roots and other plant organs is limited. Here we discuss three underappreciated areas where focused measurements of fine-root traits can make significant contributions to ecosystem science. These include assessment of spatiotemporal variation in fine-root traits, integration of mycorrhizal fungi into fine-root-trait frameworks, and the need for improved scaling of traits measured on individual roots to ecosystem-level processes. Progress in each of these areas is providing opportunities to revisit how below-ground processes are represented in terrestrial biosphere models. Targeted measurements of fine-root traits with clear linkages to ecosystem processes and plant responses to environmental change are strongly needed to reduce empirical and model uncertainties. Further identifying how and when suites of root and whole-plant traits are coordinated or decoupled will ultimately provide a powerful tool for modeling plant form and function at local and global scales

Nitrogen Capture by Grapevine Roots and Arbuscular Mycorrhizal Fungi from Legume Cover-Crop Residues Under Low Rates of Mineral Fertilization

The influence of mineral fertilization on root uptake and arbuscular mycorrhizal fungi-mediated 15N capture from labeled legume (Medicago polymorpha) residue was examined in winegrapes (Vitis vinifera) in the greenhouse, to evaluate compatibility of fertilization with incorporation of cover-crop residue in winegrape production. Plants grown in marginal vineyard soil were either fertilized with 0.25× Hoagland’s solution or not. This low fertilization rate represents the deficit management approach typical of winegrape production. Access to residue in a separate compartment was controlled to allow mycorrhizal roots (roots + hyphae), hyphae (hyphae-intact), or neither (hyphae-rotated) to proliferate in the residue by means of mesh core treatments. Leaves were weekly analyzed for 15N. On day 42, plants were analyzed for 15N and biomass; roots were examined for intraradical colonization; and soils were analyzed for 15N, inorganic N, Olsen-P, X-K, and extraradical colonization. As expected, extraradical colonization of soil outside the cores was unaffected by mesh core treatment, while that inside the cores varied significantly. 15N atom% excess was highest in leaves of roots + hyphae. In comparison, leaf 15N atom% excess in hyphae-intact was consistently intermediate between roots + hyphae and hyphae-rotated, the latter of which remained unchanged over time. Fertilization stimulated host and fungal growth, based on higher biomass and intraradical colonization of fertilized plants. Fertilization did not affect hyphal or root proliferation in residue but did lower %N derived from residue in leaves and stems by 50%. Our results suggest that even low fertilization rates decrease grapevine N uptake from legume crop residue by both extraradical hyphae and roots