Microwave-mediated synthesis of N-methyliminodiacetic acid (MIDA) boronates

A library of over 20, mainly aryl or heteroaryl, N-methyliminodiacetic acid (MIDA) boronates have been synthesised. A rapid microwave-mediated (MW) method (5–10 min) has been developed using polyethylene glycol 300 (PEG 300) as solvent. However, acetonitrile (MeCN) and dimethylformamide (DMF) were found to be alternative solvents, the latter especially for 2-substituted aryl boronic acids

Membrane Microviscosity Modulates Μ-Opioid Receptor Conformational Transitions and Agonist Efficacy

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66282/1/j.1471-4159.1999.0730289.x.pd

Lipophilicity of opioids determined by a novel micromethod

The lipophilicity of various [mu]-selective opioids was determined by measuring their distribution between n-octanol and Tris.HCl buffer, pH 7.4, by a procedure requiring submicromolar concentrations (submilligram amounts) of the compounds. After partitioning at 25[deg]C, the concentration of opioids in the aqueous phase was quantified by their displacement of bound [3H]Tyr-Ala-Gly-(Me)Phe-Gly-ol (DAMGO) from opioid receptor in brain membranes. The obtained distribution coefficients (log Papp) agreed well with respective values determined previously with other, less sensitive or more cumbersome, methods of quantitation. The procedure is precise and versatile, and offers the routine assessment of lipophilicity as part of the in vitro characterization of opioids frequently available in limited quantities. In principle, the method is applicable to any compound whose binding to its receptor is quantifiable.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30114/1/0000489.pd

Synthesis and characterization of 7-nitrobenzo-2-oxa-1,3-diazole (NBD)-labeled fluorescent opioids

Alkylation of sarcosine with 4-chloro-nitrobenzo-2-oxa-1,3-diazole (NBD-chloride) furnished a fluorescent tag that was coupled with a tetrahydrothebaine derivative and [beta]-naltrexamine, respectively, to yield the fluorescent opioids 7[alpha]-(1R)-1-hydroxy-1-methyl-3-(4-hydroxyphenyl)-propyl]-6,14-endoethenotetrahydrothebaine NBD-sarcosinate (ASM-5-10) and N-cyclopropylmethyl-3-hydroxy-14[beta]-hydroxy-6[beta]-(NBD sarcosinyl)-amino-epoxymorphinan (ASM-5-67). The fluorescence intensity of the novel opioids allowed their detection at subnanomolar concentrations, and was dependent on the polarity of the solvent. Maximum quantum yield was obtained in ethyl acetate and ethanol, and minimal fluorescence in heptane and water. Compounds ASM-5-10 and ASM-5-67 displaced the opioid receptor binding of [3H]Tyr--Ala-Gly-(Me)Phe-Gly-ol in monkey brain membranes with IC50 values of 8.4 and 1.5nM, respectively. Whereas ASM-5-67 bound to [mu], [delta], and [kappa] receptors with comparable affinities, ASM-5-10 was [mu]-selective, with selectivity indices (ratio of respective IC50 values) of 0.04 for both [mu]/[delta] and [mu]/[kappa]. The sodium response ratio in binding revealed a pronounced agonist property of ASM-5-10. Both opioids were lipophilic, with octanol-water partition coefficients (log Papp) of 2.8 (ASM-5-10) and 1.0 (ASM-5-67). ASM-5-10 exhibited particularly strong membrane retention that was not reversible by four washes. Their favorable characteristics in fluorescence, receptor binding, and membrane interaction make these newly developed ligands useful molecular probes to study opioid receptor mechanisms.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30242/1/0000637.pd

GDF15 and Growth Control

Growth/differentiation factor-15 (GDF-15) is a distant member of the transforming growth factor β (TGF-β) superfamily and is widely expressed in multiple mammalian tissues. Its expression is highly regulated and is often induced in response to conditions associated with cellular stress. GDF15 serum levels have a strong association with many diseases, including inflammation, cancer, cardiovascular diseases, and obesity, and potentially serve as reliable predictor of disease progression. A functional role for GDF15 has been suggested in cancer, cardiovascular disease, kidney disease and metabolic disease. However, the knowledge of its pathophysiological function at the molecular level is still limited and requires more investigation. Recent identification of the endogenous receptor for GDF15 may provide additional insight in to its’ molecular mechanisms and relationship to disease states

Photoperiodic regulation of FGF21 production in the Siberian hamster

FGF21 is an endocrine member of the fibroblast growth factor superfamily that has been shown to play an important role in the physiological response to nutrient deprivation. Food restriction enhances hepatic FGF21 production, which serves to engage an integrated response to energy deficit. Specifically, elevated FGF21 levels lead to reduced gluconeogenesis and increased hepatic ketogenesis. However, circulating FGF21 concentrations also paradoxically rise in states of metabolic dysfunction such as obesity. Furthermore, multiple peripheral tissues also produce FGF21 in addition to the liver, raising questions as to its endocrine and paracrine roles in the control of energy metabolism. The objectives of this study were to measure plasma FGF21 concentrations in the Siberian hamster, a rodent which undergoes a seasonal cycle of fattening and body weight gain in the long days (LD) of summer, followed by reduction of appetite and fat catabolism in the short days (SD) of winter. Groups of adult male hamsters were raised in long days, and then exposed to SD for up to 12 weeks. Chronic exposure of LD animals to SD led to a significant increase in circulating FGF21 concentrations. This elevation of circulating FGF21 was preceded by an increase in liver FGF21 protein production evident as early as 4 weeks of exposure to SD. FGF21 protein abundance was also increased significantly in interscapular brown adipose tissue, with a positive correlation between plasma levels of FGF21 and BAT protein abundance throughout the experimental period. Epididymal white adipose tissue and skeletal muscle (gastrocnemius) also produced FGF21, but levels did not change in response to a change in photoperiod. In summary, a natural programmed state of fat catabolism was associated with increased FGF21 production in the liver and BAT, consistent with the view that FGF21 has a role in adapting hamsters to the hypophagic winter state

Predicting future stability of ecosystem functioning under climate change

To maintain food security under global change, we need to consider the stability of ecosystem functioning into the future, particularly in resource production landscapes such as agricultural pasture. With ongoing climate change, extreme climatic events are predicted to become more frequent and severe globally, impacting crop production. The whole process of farming will become more uncertain, from choice of crop and crop productivity to the timing of the windows of opportunity for management decisions. Future agricultural policies, therefore, should not only consider changes in grassland production, but also its future stability. We use a case study of agricultural pastures on the island of Ireland to project different components of ecosystem stability (resistance, recovery time and recovery rate) to 2050 and 2080 under different future climate scenarios: a peak and decline scenario; and a continued emissions scenario. We show that future climate change will have substantial effects on both the future resistance and the recovery of ecosystem functioning following environmental disturbances, but the spatial pattern of effect sizes is not the same for these two measures of stability. National level analyses and agricultural policies, therefore, are likely to ignore regional variation in future change. From this, we encourage the translation of stability-based constructs, as well as maximum yield considerations, into future agricultural policy at the regional level

Dual effects of fibroblast growth factor 21 on hepatic energy metabolism

The aim of this study was to investigate the mechanisms by which FGF21 affects hepatic integration of carbohydrate and fat metabolism in Siberian hamsters, a natural model of adiposity. Twelve aged matched adult male Siberian hamsters maintained in their long day fat state since birth were randomly assigned to one of two treatment groups and were continuously infused with either vehicle (saline; n=6) or recombinant human FGF21 protein (1 mg/kg/day; n=6) for 14 days. FGF21 administration caused a 40% suppression (P<0.05) of hepatic pyruvate dehydrogenase complex (PDC), the rate-limiting step in glucose oxidation, a 34% decrease (P<0.05) in hepatic acetylcarnitine accumulation, an index of reduced PDC flux, a 35% increase (P<0.05) in long-chain acylcarnitine content (an index of flux through β-oxidation) and a 47% reduction (P<0.05) in hepatic lipid content. These effects were underpinned by increased protein abundance of PD kinase-4 (a negative regulator of PDC), the phosphorylated (inhibited) form of acetyl-CoA carboxylase (ACC, a negative regulator of delivery of fatty acids into the mitochondria), and the transcriptional co-regulators of energy metabolism peroxisome proliferator activated receptor gamma co-activator alpha (PGC1 and sirtuin-1 (SIRT-1). These findings provide novel mechanistic basis to support the notion that FGF21 exerts profound metabolic benefits in the liver by modulating nutrient flux through both carbohydrate (mediated by a PDK4-mediated suppression of PDC activity) and fat (mediated by deactivation of ACC) metabolism, and therefore may be an attractive target for protection from increased hepatic lipid content and insulin resistance that frequently accompany obesity and diabetes

Anorectic and aversive effects of GLP-1 receptor agonism are mediated by brainstem cholecystokinin neurons, and modulated by GIP receptor activation

This work was funded by an MRC Career Development Award (MR/ P009824/1 and MR/P009824/2) to GD’A, as well as an MRC grant to SML/GD’A (MR/T032669/1), a BBSRC grant to SML (BB/M001067/1), and an additional direct contribution from Eli Lilly. D.J.H. was sup- ported by MRC (MR/N00275X/1 and MR/S025618/1), Diabetes UK (17/ 0005681), and the European Research Council (ERC) under the Eu- ropean Union’s Horizon 2020 research and innovation programme (Starting Grant 715884 to D.J.H.). AC was supported for part of this project by a travel grant from the Italian Society of Pharmacology and a fellowship from the Veronesi Foundation (Italy).Peer reviewedPublisher PD

Ecosystem stability at the landscape scale is primarily associated with climatic history

There is an increasing interest in landscape-scale perspectives of ecosystem functioning to inform policy and conservation decisions. However, we need a better understanding of the stability of ecosystem functioning (e.g. plant productivity) at the landscape scale to inform policy around topics such as global food security. We investigate the role of the ecological and environmental context on landscape-scale stability of plant productivity in agricultural pasture using remotely sensed enhanced vegetation index data. We determine whether four measures of stability (variability, magnitude of extreme anomalies, recovery time and recovery rate) are predicted by (a) species richness of vascular plants, (b) regional land cover heterogeneity and (c) climatic history. Stability of plant productivity was primarily associated with climatic history, particularly a history of extreme events. These effects outweighed any positive effects of species richness in the agricultural landscape. A history of variable and extreme climates both increased and decreased contemporary ecosystem stability, suggesting both cumulative and legacy effects, whereas land cover heterogeneity had no effect on stability. The landscape scale is a relevant spatial scale for the management of an ecosystem's stability. At this scale, we find that past climate is a stronger driver of stability in plant productivity than species richness, differing from results at finer field scales. Management should take an integrated approach by incorporating the environmental context of the landscape, such as its climatic history, and consider multiple components of stability to maintain functioning in landscapes that are particularly vulnerable to environmental change