Distribution and Accumulation of the Alkaloid Peramine in Neotyphodium lolii-Infected Perennial Ryegrass

Through the production of alkaloids and mycotoxins, the endophytic fungus of perennial ryegrass, Neotyphodium lolii, confers resistance to the host grass against several insect pasture pests and causes health problems in livestock. The effects of plant component, plant reproductive development, and plant management (trimming) on the concentration and accumulation of peramine, the alkaloid thought to be most important for conferring resistance against Argentine stem weevil, were investigated. Clones of eight different naturally occurring perennial ryegrass-N. lolii associations were used in the experiment, which was conducted during spring–summer, which corresponded with the height of plant reproductive activity. Clones were either trimmed regularly to maintain them in a vegetative state (V clones) or not trimmed, allowing them to become fully reproductive (R clones). V and R clones of each grass/endophyte association were destructively harvested at times that corresponded roughly with the start, middle, and end of the plant reproductive phase. Harvested material was dissected into a number of plant components that were analyzed for peramine concentration by HPLC. All clippings from the V clones were also analyzed. Peramine concentrations were usually similar in leaf sheaths and leaf blades, but they decreased as leaf age increased. The seed from R clones and younger sheaths and blades of leaves from vegetative tillers contained the highest concentrations, while the root, crown, and dead leaf tissue contained the lowest. During the inflorescence phase, tissues from reproductive tillers generally contained lower concentrations than corresponding tissues from vegetative tillers. Also at this time, mean peramine concentrations were higher in V clones than in R clones. By the end of the plant reproductive phase, at the mature seed stage, mean peramine concentrations in R and V clones were similar. The seed component, which contained approximately 75% of the total peramine present in the plant (at a mean concentration of 63.7 μg/g), probably accounted for much of the recovery in the R clones.     

Distribution and Accumulation of the Mycotoxin Lolitrem B in Neotyphodium lolii- Infected Perennial Ryegrass

The symbiotic relationship between perennial ryegrass and its endophytic fungus, Neotyphodium lolii, is of considerable agronomic significance in New Zealand. Livestock ingesting N. lolii-infected perennial ryegrass may succumb to a neuromuscular condition, ryegrass staggers, the probable cause of which has been identified as the endophyte-produced indole–diterpenoid mycotoxin lolitrem B. The effects of plant component, plant reproductive development, and plant management (clipping) on the concentration and accumulation of lolitrem B were investigated. Eight different naturally occurring perennial ryegrass–N. lolii associations were cloned. They were either trimmed regularly to maintain their vegetative state (V clones) or not trimmed at all to enable full plant reproductive development (R clones). At times that corresponded approximately with the start, middle, and end of the plant reproductive phase, V and R clones of each grass–endophyte association were destructively harvested. After dissection into a number of plant components, harvested material was analyzed for lolitrem B content by HPLC. Analyses were also performed on all clippings taken from the V clones. Highest lolitrem B concentrations in vegetative parts were found in older leaf sheaths and dead leaves. Towards the end of the plant reprodutive phase, dead leaves also contained a large proportion of the total lolitrem B content of the V clones. The highest concentration of lolitrem B was found in the seed, which accounted for nearly 60% of the total quantity in R clones. Leaf blades, young leaf sheaths, roots, and crowns generally contained low to moderate concentrations. In late spring, mean concentrations in R and V clones were similar. By the time mature seed was present (mid-summer), mean concentrations in R clones were three times higher than in V clones. This difference was attributed largely to the contribution by the seed in the R clones. The implications of these findings in relation to ryegrass staggers in livestock are discussed.     

Hybrid Photopatterned Enzymatic Reaction (HyPER) for in Situ Cell Manipulation

The ability to design artificial extracellular matrices as cell‐instructive scaffolds has opened the door to technologies capable of studying the fate of cells in vitro and to guiding tissue repair in vivo. One main component of the design of artificial extracellular matrices is the incorporation of biochemical cues to guide cell phenotype and multicellular organization. The extracellular matrix (ECM) is composed of a heterogeneous mixture of proteins that present a variety of spatially discrete signals to residing cell populations. In contrast, most engineered ECMs do not mimic this heterogeneity. In recent years, photo‐deprotection has been used to spatially immobilize signals. However, this approach has been limited mostly to small peptides. Here we combine photo‐deprotection with enzymatic reaction to achieve spatially controlled immobilization of active bioactive signals that range from small molecules to large proteins. A peptide substrate for transglutaminase factor XIII (FXIIIa) was caged with a photo‐deprotectable group, which was then immobilized to the bulk of a cell‐compatible hydrogel. With focused light, the substrate can be deprotected and used to immobilize patterned bioactive signals. This approach offers an innovative strategy to immobilize delicate bioactive signals, such as growth factors, without loss of activity and enables in situ cell manipulation of encapsulated cells.     

An in situ intercalative polymerization method for preparing UV curable clay–polymer nanocomposites

A novel in situ intercalative polymerization technique was used to disperse clay mineral in a precursor resin for use in UV curing by performing an in situ ion exchange reaction during polyesterification. Unmodified montmorillonite (MMT) was added to a reaction mixture composed of monomers and methyl, tallow, bis‐2‐hydroxyethyl ammonium (MTEtOH) during the synthesis of unsaturated polyesters to create resins containing highly dispersed, organically modified MMT. UV‐curable clay–polymer nanocomposite (CPN) films were then prepared utilizing donor–acceptor chemistry through reactions of the unsaturated polyester resin with triethylene glycol divinyl ether. Functional group conversion improved up to 15% by the incorporation of clay mineral into the polymer matrix through the in situ polymerization method. The CPNs also had improved barrier, mechanical, and thermal properties over a control film containing no clay mineral. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42601.     

Evaluation of Agronomic and Seed Characteristics in Elevated Oleic Acid Soybean Lines in the South-Eastern US

Increasing oleic acid, a monounsaturated fatty acid, is reported to strike the best balance between cold flow properties and oxidative stability in soybean seed oil to enhance biodiesel and produce a better burning fuel. In addition, it is important that elevated oleic acid soybeans have the agronomic traits of local cultivars and maintain oleic acid stability across environments. Research was conducted in 2007–2008 to evaluate six Roundup Ready^® soybean recombinant inbred lines exhibiting enhanced levels of oleic acid. The six elevated oleic lines averaged a 55% increase in oleic acid and a 43% decrease in linolenic acid over the two commercial cultivars (AG3906 and AG4103). Some elevated oleic acid genotypes fulfilled the linear regression definition of a stable genotype. TN03-93RR was the best genotype because of its oleic acid content (397 g kg^?1) and desirable regression estimates for stability. Iodine value (IV), peroxide value (PV), and induction period (IP) were used to evaluate the fuel properties of the two lines with the highest oleic acid content and the two commercial cultivars. The elevated oleic acid lines had significantly better IP, PV and IV for better biodiesel properties and oxidative stability than the two commercial cultivars.     

Termini and main-chain composition of monomethoxy-terminated poly(ethylene glycol) studied by two-dimensional column chromatography

Samples of poly(ethylene glycol) especially those with functionalized ends have dimeric, trimeric, and tetrameric components at twice, three times, and four times as high a molecular weight (MW) as that of the main component. We used two-dimensional column chromatography to evaluate the MW and identify the terminal groups in each component. The first step is high osmotic pressure chromatography to prepare a sufficient amount of fractions that contain different percentages of the multimeric components. In the second step, each of the fractions was characterized by using regular size exclusion chromatography and high-resolution NMR. Decomposition of the chromatogram combined with isolation of the terminal methoxy peak in the NMR spectrum led to unique identification of the terminal groups. We thus found that the monomeric component is monomethoxy-terminated whereas the dimeric and other multimeric components are mostly dihydroxy-terminated.     

(Z,Z)-4,7-tridecadien-(S)-2-yl acetate: sex pheromone of Douglas-fir cone gall midge,Contarinia oregonensis

Our objectives were to identify and field test the sex pheromone of female Douglas-fir cone gall midge, Contarinia oregonensis (Diptera: Cecidomyiidae). Coupled gas chromatographic–electroantennographic detection (GC-EAD) analyses of pheromone extract revealed a single compound (A) that elicited responses from male antennae. Hydrogenation of pheromone extract, followed by renewed GC-EAD analysis, revealed a new EAD-active compound with chromatographic characteristics identical to those of tridecan-2-yl acetate on five fused silica columns (DB-5, DB-210, DB-23, SP-1000, and Cyclodex-B). Syntheses, chromatography, and retention index calculations of all possible tridecen-2-yl acetates suggested that the candidate pheromone A was a tridecadien-2-yl acetate with nonconjugated double bonds. Synthetic candidate pheromone component (Z,Z)-4,7-tridecadien-2-yl acetate (Z4Z7) cochromatographed with A on all analytical columns and elicited comparable antennal activity. In GC-EAD analyses that separated the enantiomers (Z,Z)-4,7-tridecadien-(S)-2-yl acetate (2S-Z4Z7) and (Z,Z)-4,7-tridecadien-(R)-2-yl acetate (2R-Z4Z7) with baseline resolution, only 2S-Z4Z7 as a component in a racemic standard or in pheromone extract elicited antennal responses. In Douglas-fir seed orchards, sticky traps baited with 2S-Z4Z7 captured male C. oregonensis, whereas 2R-Z4Z7 was behaviorally benign. Comparable catches of males in traps baited with racemic Z4Z7 (50 g) or virgin female C. oregonensis suggested that synthetic pheromone baits could be developed for monitoring C. oregonensis populations in commercial Douglas-fir seed orchards.     

Characterization of the adipose tissue atrophy induced by peroxisome proliferators in mice

In the present study, we characterized the effects of peroxisome proliferators (PP) on adipose tissue in mice. Treatment with potent PP, such as perfluorooctanoic acid (PFOA), 2-methyl-2-(p(1,2,3,4-tetrahydroxy-naphthyl)-phenoxy)propionic acid, (4-chloro-6-(2,3-xylidino)2-pyrimidinylthio) acetic acid, and di(2-ethylhexyl)phthalate, caused dramatic decreases in adipose tissue weight, whereas the moderately potent PP, acetylsalicylic acid, had a relatively weak effect. This decrease in weight reflects a loss of fat from adipocytes rather than a loss of cells, as demonstrated by constant DNA content. The dose-dependency and time-course experiments indicate that peroxisome proliferation occurs simultaneously with or prior to adipose tissue atrophy. Thus, hepatic peroxisome proliferation might result in the increased mobilization of lipids and lipid utilization in liver. The enhanced adipose tissue hormonesensitive lipase (HSL) activity and down-regulated lipoprotein lipase (LPL) activity observed upon PP treatment might, at least in part, explain the loss of fat via increase FA release from adipocytes and/or decreased FA uptake from the circulation, respectively. In addition, the possible involvement of the increased tumor necrosis factor α expression found upon PFOA treatment in reducing the insulin sensitivity of adipose tissue and thereby altering LPL and HSL activities is discussed.     

Separation and spectral data of the six isomeric ozonides from methyl oleate

The structures of the products obtained on ozonation of methyl oleate have been re-examined. The assignments for the six isomeric ozonides of methyl oleate have been made by¹H and¹³C nuclear magnetic resonance (NMR), which were consistent with the retention times observed in high-performance liquid chromatography; the assignments were confirmed by mass and infrared spectroscopy. Two triplets for the ozonide ring protons of thecis andtrans isomers in the normal (MOO1) and the two cross ozonides (MOO1 and MOO3) can be resolved by 400 MHz NMR. For MOO1 and MOO3, where the two ring carbons are equivalent, two peaks for the ring carbons of each cross ozonides are resolved in the¹³C NMR spectra, one for thecis and one for thetrans isomer. For MOO2, four peaks for the ring carbons are resolved in the¹³C NMR spectra, two for thecis and two for thetrans isomer.