Monoacylglycerol Lipase (MAGL) Inhibition Attenuates Acute Lung Injury in Mice

Endocannabinoid signaling is terminated by enzymatic hydrolysis, a process that, for 2-Arachidonoylglycerol (2-AG), is mediated by monoacylglycerol lipase (MAGL). The piperidine carbamate, 4-​nitrophenyl- ​4-​(dibenzo[d] [1,3]dioxol-​5-​yl (hydroxy) methyl) piperidine- 1-​carboxylate (JZL184), is a drug that inhibits MAGL and presents high potency and selectivity. Thus, JZL184 increases the levels of 2-AG, an endocannabinoid that acts on the CB₁ and CB₂ cannabinoid receptors. Here, we investigated the effects of MAGL inhibition, with a single dose (16 mg/kg, intraperitoneally (i.p.)) of JZL184, in a murine model of lipopolysaccharide (LPS) -induced acute lung injury (ALI) 6, 24 and 48 hours after the inflammatory insult. Treatment with JZL184 decreased the leukocyte migration into the lungs as well as the vascular permeability measured through the bronchoalveolar lavage fluid (BAL) and histological analysis. JZL184 also reduced the cytokine and chemokine levels in the BAL and adhesion molecule expression in the blood and BAL. The CB₁ and CB₂ receptors were considered involved in the anti-inflammatory effects of JZL184 because the AM281 selective CB₁ receptor antagonist (1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-4-morpholinyl-1H-pyrazole-3-carboxamide) and the AM630 selective CB₂ receptor antagonist ([6-​iodo-​2-​methyl-​1-​[2-​(4-​morpholinyl)ethyl]-​1H-​indol-​3-​yl](4-​methoxyphenyl)-​methanone) blocked the anti-inflammatory effects previously described for JZL184. It was concluded that MAGL inhibition, and consequently the increase in 2-AG levels, produced anti-inflammatory effects in a murine model of LPS-induced ALI, a finding that was considered a consequence of the activation of the CB₁ and CB₂ receptors.     

Nature of the effect of ther locus on the lipid content of embryos of peas (Pisum sativum L.)

The aim of this work was to discover why pea (Pisum sativum L.) embryos recessive at the r locus (rr) have a higher lipid content than embryos dominant at this locus (RR). The r locus is a gene encoding starch-branching enzyme, rr embryos have a much lower activity of this enzyme than RR embryos, and hence a reduced rate of starch synthesis. The higher lipid content of rr embryos must be a consequence of this. We suggest that neither differences in the availability of substrate for lipid synthesis as a consequence of different rates of starch synthesis, nor differences in the capacity of the pathway for malonyl-CoA synthesis, account for the different lipid contents of RR and rr embryos. Lipid contents of the two sorts of embryo first diverge at a much later stage in development than divergence in starch content. Amounts of pyruvate and acetate, and activities of enzymes that convert triose phosphate to malonyl CoA are the same in the two sorts of embryo. Most of the lipid in developing embryos is polar, structural lipid, and polar lipid accounts for a large proportion of the difference in lipid content between the two sorts of embryo. This difference in structural-lipid content reflects considerable structural differences between the two sorts of embryo and is presumably the consequence of differences in rates of lipid turnover.Abbreviations DW dry weight - FW fresh weight - FAME fatty-acid methyl esters This work was supported by a grant-in-aid from the Agricultural and Food Research Council to the John Innes Institute. We are very grateful to Alan Jones for his valuable advice on lipid analysis and to Dr. Kay Denyer (Advanced Technologies, Cambridge, UK) for valuable discussions. We thank Dr. Cliff Hedley for the gift of the seed of the peas used in this work.     

Tracheary-element differentiation in suspension-cultured cells ofZinnia requires uptake of extracellular Ca2+

Tracheary-element (TE) differentiation in suspension cultures ofZinnia elegans L. mesophyll cells was inhibited by blocking calcium uptake in three ways: 1) reducing the [Ca²⁺] of the culture medium, 2) blocking calcium channels with the non-permeant cation La³⁺, and 3) blocking calcium channels with permeant dihydropyridine calcium-channel blockers. Calcium-channel blockers were effective when added at any time between 0 and 48 h after culture initiation; after 48h, calcium sequestration and secondary cell-wall deposition began. In contrast, calmodulin antagonists inhibited TE differentiation when added at the beginning of culture, but not when added after 24h. These results indicate that TE differentiation involves at least two calcium-regulated events: one calmodulin-dependent and occurring shortly after exposure to inductive conditions, and the other calmodulin-independent and occurring just prior to secondary cell-wall deposition.     

Biosynthesis,cDNA and amino acid sequences of a precursor of conglutin δ, a sulphur-rich protein from Lupinus angustifolius

The biosynthesis of conglutin has been studied in developing cotyledons of Lupinus angustifolius L. Precursors of conglutin formed the major sink for [³⁵S]-cysteine incorporated by developing lupin cotyledons, and these precursors were rapidly sequestered into the endoplasmic reticulum. The sequence of a cDNA clone coding for one such precursor of conglutin was determined. The structure of the precursor polypeptide for conglutin predicted from the cDNA sequence contained an N-terminal leader peptide of 22 amino acids directly preceding a subunit polypeptide of M _r 4520, together with a linking region of 13 amino acids and a subunit polypeptide of M _r 9558 at the C-terminus. The amino acid sequence predicted from the cDNA sequence showed minor variations from that established by sequencing of the protein purified from mature dried seeds (Lilley and Inglis, 1986). These were consistent with the existence of a multi-gene family coding for conglutin . Comparison of the sequences of conglutin with those of other 2S storage proteins showed that the cysteines involved in internal disulphide bridges between the mature subunits of conglutin , were maintained throughout this family of proteins but that little else was conserved either at the protein or DNA level.     

The effects of thermal biofeedback and autogenic training of cardiovascular reactivity: The joint USSR-USA behavioral hypertension treatment project

Cardiovascular reactivity (heart rate, systolic, and diastolic BP) to mental arithmetic and cold pressor were measured before and after treatment as part of the cross-cultural (USSR and USA) evaluation of thermal biofeedback and autogenic training (in comparison with self-relaxation) as treatments for mild hypertension in unmedicated males. There were no statistically reliable decreases in cardiovascular reactivity from before to after treatment. However, downward shifts in basal levels of systolic and diastolic BP at post-treatment led treated patients to have lower stress-induced levels of BP.The American portion of this research was supported by a grant from the NHLBI, HL-31189. We express appreciation to officials at the USSR Cardiology Research Center, Academician Igor Shkhvatsabaya, and Professor Vadim Zaitsev for their support, as well as to Dr. Steve Weiss, of NHLBI, for his instigation of this research.     

Major differences in isoforms of starch-branching enzyme between developing embryos of round- and wrinkled-seeded peas (Pisum sativum L.)

In order to determine whether round-and wrinkled-seeded peas (Pisum sativum L.) differ in the activity and properties of starch-branching enzyme (1,4--D-glucan, 1,4--D-glucan-6-glycosyl transferase; EC 2.4.1.18) in their developing embryos, essentially isogenic lines of peas, differing only at the r (rugosus) locus that confers the round (RR, Rr) or wrinkled (rr) phenotype, were studied. Activity of the enzyme rises rapidly from an early stage of development in embryos of round peas, but only at later stages in embryos of wrinkled peas. The purified enzyme from mature embryos of round peas can be resolved into two isoforms that differ in molecular weight and in their ability to branch amylose. The purified enzyme from mature embryos of wrinkled peas is a single protein with the same molecular weight and branching properties as one of the isoforms from embryos of round peas. The difference in activity of starch-branching enzyme between embryos of round and wrinkled peas is likely to be due to the absence from embryos of wrinkled peas of one of the isoforms occurring in embryos of round peas.Abbreviations DEAE diethylaminoethyl - FW fresh weight - kDa kilodalton - SDS sodium dodecyl sulfate     

Extracellular fibril production by freshwater algae and cyanobacteria

In order to study the ability of freshwater algae and cyanobacteria to form extracellular fibrils, a screening test using ruthenium red (RR) staining was carried out on 28 species. Five of these were examined for growth and production of fibrillar material in culture media of different phosphate (P;) contents. RR-staining and uronic acid determinations at various stages of algal growth were complemented by electron microscopy of the cells and of fibrillar material released into the medium. The lower P_i concentrations enhanced growth of Micrasterias radiata, Eremosphaera sp., and Microcystis aeruginosa, and had little or no effect on growth of a Xanthidium sp. and Scenedesmus quadricauda. Extracellular uronic acid production, which was higher in low P_i medium in M. radiata, M. aeruginosa, and Xanthidium sp., could reach levels of 50 mg/liter or more. Algae with high proportions of RR-positive cells (M. radiata, Eremosphaera sp., Xanthidium sp., and M. aeruginosa) produced high levels of slime-like material and distinct fibrils that were often seen attached to the cell surface and only slowly released into the medium. No such material was found in cultures (or supernatants) of Sc. quadricauda, which also produced relatively low amounts of polyuronic acids. Specific types of filaments, often forming fascicles with rectangular arrays of globular particles were observed by negative staining electron microscopy of some algal cultures. RR-positive material was also observed in the cytoplasm and on the cell walls and surfaces of M. radiata and M. aeruginosa.Offprint requests to: D. J. Kushner.     

Paralytic shellfish toxins in Australian Southern Rock Lobster (Jasus edwardsii): Acute human exposure from consumption of hepatopancreas

Paralytic shellfish toxins (PST) were identified in the hepatopancreas of Southern Rock Lobster (Jasus edwardsii) during Alexandrium tamarense blooms in Tasmania, Australia. Human health risk from PST in lobsters was unknown – this study assesses exposure to PST from hepatopancreas consumption. Lobster hepatopancreas samples collected during blooms (n = 181) were mostly positive for PST (>88%), the highest concentration was 4032 μg STX-2HCl eq/kg. Consumer exposure to PST was estimated using a 2-D Monte Carlo model. Mean PST intake (pi) from hepatopancreas consumption (raw and cooked) was below the lowest-observed-adverse-effect-level (LOAEL) for PST (<2.0 μg/kg bw), however the 97.5th percentile pi for raw meals (2.64 μg/kg bw) exceeded the LOAEL. A total of 4.1% of raw hepatopancreas meals were estimated to exceed the LOAEL. Lobster hepatopancreas consumption during A. tamarense blooms may be concerning for a small proportion of consumers, particularly those that eat large meals at the bloom peak. However, when the model was re-run with PST concentration capped at the bivalve regulatory limit (800 μg STX-2HCl eq/kg) pi decreased, with the 97.5th percentile values below the LOAEL. Thus, issuing public health warnings and harvesting restrictions for lobsters when levels exceed 800 μg STX-2HCl eq/kg would reduce the probability of illness occurring.