Crystal structure of the malaria vaccine candidate apical membrane antigen 1

Apical membrane antigen 1 from Plasmodium is a leading malaria vaccine candidate. The protein is essential for host-cell invasion, but its molecular function is unknown. The crystal structure of the three domains comprising the ectoplasmic region of the antigen from P. vivax, solved at 1.8 angstrom resolution, shows that domains I and II belong to the PAN motif, which defines a superfamily of protein folds implicated in receptor binding. We also mapped the epitope of an invasion-inhibitory monoclonal antibody specific for the P. falciparum ortholog and modeled this to the structure. The location of the epitope and current knowledge on structure-function correlations for PAN domains together suggest a receptor-binding role during invasion in which domain II plays a critical part. These results are likely to aid vaccine and drug design.     

Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis

The Golgi enzyme beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is up-regulated in carcinomas and promotes the substitution of N-glycan with poly N-acetyllactosamine, the preferred ligand for galectin-3 (Gal-3). Here, we report that expression of Mgat5 sensitized mouse cells to multiple cytokines. Gal-3 cross-linked Mgat5-modified N-glycans on epidermal growth factor and transforming growth factor-beta receptors at the cell surface and delayed their removal by constitutive endocytosis. Mgat5 expression in mammary carcinoma was rate limiting for cytokine signaling and consequently for epithelial-mesenchymal transition, cell motility, and tumor metastasis. Mgat5 also promoted cytokine-mediated leukocyte signaling, phagocytosis, and extravasation in vivo. Thus, conditional regulation of N-glycan processing drives synchronous modification of cytokine receptors, which balances their surface retention against loss via endocytosis.     

Resolving the urban nest predator paradox: The role of alternative foods for nest predators

Urbanization is a leading cause of species endangerment in the United States; however, certain species thrive in urban habitats. The loss of key predators or the addition of new predators in urban areas could alter the structure of urban communities. A reduction in nest predation is hypothesized to explain the high density of urban birds, yet urban areas typically have increased populations of avian nest predators. The loss of important nest predators in urban habitats, prey switching of urban predators, or successful nest defense against avian nest predators could explain this urban nest predator paradox. To assess these hypotheses I compared nest predation rates of Northern mockingbirds (Mimus polyglottos) in parking lots and residential neighborhoods to populations in pastures and wildlife preserves during 2007–2009 in Florida, USA and placed video cameras on a subset of nests in 2008–2009. Data do not support the hypothesis that urban nest predation rates are consistently lower than non-urban nest predation rates. Of the 56 nest predation events recorded, cats were the dominant urban predator and Cooper’s hawks (Accipiter cooperii) were the dominant non-urban predator. There was no evidence for a loss of important nest predators in urban habitats; however, prey switching by Cooper’s hawks likely occurred. There was also indirect evidence for the importance of nest defense. Furthermore, some of the cats recorded as nest predators in residential neighborhoods were owned cats and all but one cat predation event occurred at night. To reduce nest predation rates, cat owners should keep their cats indoors at night.     

Bioaccumulation of melamine in catfish muscle following continuous, low-dose, oral administration

In this study, catfish muscle was analyzed for melamine (MEL) and cyanuric acid (CYA) residues following experimental feeding with low doses of MEL and MEL and CYA (MEL+CYA) and with the insoluble melamine-cyanurate complex (MEL=CYA). Catfish were daily fed 0.1 mg/kg BW of MEL for 15, 28, or 42 days, 0.1 mg/kg BW of MEL+CYA for 28 days, 2.5 mg/kg BW of MEL+CYA for 14 days, or 400 mg/kg BW of MEL=CYA for 3 days. Residues in the tissue were determined by LC-MS/MS. MEL was extracted with acidic acetonitrile, followed by defatting with dichloromethane, and isolated with cation exchange solid phase extraction (SPE). For CYA analysis, fish were extracted with dilute acetic acid, defatted with hexane, and cleaned up with a graphitic carbon SPE. Catfish fed 0.1 mg/kg BW of MEL reached a maximum muscle residue concentration of 0.33 ± 0.04 mg/kg (ppm) after 28 days of continuous feeding. The same concentration was found for MEL+CYA feeding at the 0.1 mg/kg BW level for 28 days. Feeding at 2.5 mg/kg BW of MEL+CYA yielded muscle concentrations above the 2.5 mg/kg level of concern for most of the study fish. Finally, catfish fed high levels of the MEL=CYA complex (400 mg/kg BW) accumulated relatively little MEL in the muscle (0.14 ± 0.07 mg/kg) and, unlike treatment with MEL+CYA, did not form renal melamine-cyanurate crystals. Appreciable concentrations of CYA were not detected in any of the muscles tested. These studies provide data to model the bioaccumulation of triazine residues into edible fish tissue as a result of the continuous consumption of adulterated feed.     

Denitrification potential of intermittently saturated floodplain soils from a subtropical perennial stream and an ephemeral tributary

Denitrification has the potential to remove excess nitrogen from groundwater passing through riparian buffers, thus improving water quality downstream. In regions with markedly seasonal precipitation, transient stream flow events may be important in saturating adjacent floodplain soils and intermittently providing the anaerobic conditions necessary for denitrification to occur. In two experiments we characterised the denitrification potential of soils from two contrasting floodplains that experience intermittent saturation. We quantified under controlled laboratory conditions: 1) potential rates of denitrification in these soils with depth and over time, for a typical period of saturation; and 2) the influences on rates of nitrate and organic carbon. Treatments differed between experiments, but in each case soil-water slurries were incubated anaerobically with differing amendments of organic carbon and nitrate; denitrification rates were measured at selected time intervals by the acetylene-block technique; and slurry filtrates were analysed for various chemical constituents. In the first experiment (ephemeral tributary), denitrification was evident in soils from both depths (0-0.3 m; 0.3-1.1 m) within hours of saturation. Before Day 2, mean denitrification rates at each depth were generally comparable, irrespective of added substrates; mean rates (Days 0 and 1) were 5.2 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0-0.3 m) and 1.6 ± 0.2 mg N kg dry soil⁻¹ day⁻¹ (0.3-1.1 m). Rates generally peaked on Days 2 or 3. The availability of labile organic carbon was a major constraint on denitrification in these soils. Acetate addition greatly increased rates, reaching a maximum in ephemeral floodplain soils of 17.4 ± 1.8 mg N kg dry soil⁻¹ day⁻¹ on Day 2: in one deep-soil treatment (low nitrate) this overcame differences in rates observed with depth when acetate was not added, although the rate increase in the other deep-soil treatment (high nitrate) was significantly less (P ≤ 0.01). Without acetate, peak denitrification rates in this experiment were 6.9 ± 0.4 and 2.8 ± 0.2 mg N kg dry soil⁻¹ day⁻¹ in surface and deep soils, respectively. Differences in rates were observed with depth on all occasions, despite similar initial concentrations of dissolved organic carbon (DOC) at both depths. Levels of substrate addition in the second experiment (perennial stream) more closely reflected natural conditions at the site. Mean denitrification rates were consistently much higher in surface soil (P ≤ 0.001), while the source of water used in the slurries (surface water or groundwater from the site) had little effect on rates at any depth. Mean rates when all treatments retained nitrate were: 4.5 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0-0.3 m depth); 0.8 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0.3-1.0 m); and 0.6 ± 0.1 mg N kg dry soil⁻¹ day⁻¹ (1.8-3.5 m). For comparable treatments and soil depths, denitrification potentials at both sites were similar, apart from higher initial rates in the ephemeral floodplain soils, probably associated with their higher DOC content and possibly also their history of more frequent saturation. The rapid onset of denitrification and the rates measured in these soils suggest there may be considerable potential for nitrate removal from groundwater in these floodplain environments during relatively short periods of saturation.     

Formation of Maillard reaction products during heat treatment of carrots

As indicators of the early stage of the Maillard reaction in carrots, N-(furoylmethyl) amino acids (FMAAs) formed during acid hydrolysis of the corresponding Amadori products were analyzed using RP-HPLC with UV detection. N(ε)-FM-Lys (furosine), FM-Gly, FM-Ala, FM-Val, FM-Ile, FM-Leu, and FM-GABA were identified using synthesized standard material by means of mass spectrometry. Furthermore, N(ε)-carboxymethyllysine (CML) and pyrraline were analyzed as indicators for advanced stages of glycation. For commercial samples with high water content, the formation of Amadori compounds predominates, whereas the advanced stage of Maillard reaction plays only a minor part. Carrot juices, baby food, and tinned carrots showed quite low rates of amino acid modification up to 5%. For dehydrated carrots, significantly higher values for Amadori products were measured, corresponding to a lysine derivatization of up to 58% and nearly 100% derivatization of GABA. Drying experiments revealed great differences in reactivity between the amino acids studied. Whereas furosine reached constant values quite quickly, some FMAAs showed a continuous increase with heating time, indicating that selected FMAAs can be used as a hallmark for the early Maillard reaction to control processing conditions.     

Silencing of host basal defense response-related gene expression increases susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis

Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato.     

Uncommon associations in target resistance among French populations of Myzus persicae from oilseed rape crops

Within the framework of a molecular exploration of target resistance in populations of Myzus persicae on oilseed rapes in France, (1) the S431F mutation (coding gene ace2), although previously reckoned to be rare, revealed to be frequent, (2) M918L (phenotypically characterised) and L932F (both on para) were found for the first time in M. persicae, and (3) a linkage was revealed between M918L and S431F. While until recently populations developing on French oilseed rapes were dominated by genotypes possessing pyrethroid target resistance and esterase overproduction, to date a different type of dominating genotype, equipped with carbamate and pyrethroid target resistance, seems to be invading such fields.     

Comparison of foliar and soil formulations of neonicotinoid insecticides for control of potato leafhopper, Empoasca fabae (Homoptera: Cicadellidae), in wine grapes

BACKGROUND: The potential of systemic neonicotinoid insecticides to control potato leafhopper, Empoasca fabae (Harris), a damaging pest of wine grapes in the eastern United States, was investigated. Soil or foliar applications were made to potted or field‐grown vines, and the response of leafhoppers was determined in clip cages over the following month on young or mature leaves. RESULTS: Foliar application of imidacloprid caused immediate and long‐lasting reductions in E. fabae survival on both leaf ages, whereas the activity of soil‐applied imidacloprid was delayed. Clothianidin, imidacloprid and thiamethoxam all provided long‐lasting reduction in leafhopper survival on young and mature foliage when applied through either delivery route. However, the percentage of moribund nymphs was significantly greater on foliar‐treated vines and increased over time in mature and immature leaves compared with soil‐treated vines. Residue analysis of foliar‐applied imidacloprid showed an 89% decline in mature leaves from day 1 to day 27, and a 98% decline in immature leaves over the same time period. Comparison of soil‐applied clothianidin, imidacloprid and thiamethoxam in field‐grown vines showed significant reduction in E. fabae only on mature leaves of vines treated with thiamethoxam. CONCLUSIONS: Neonicotinoids can control E. fabae in small vines, even in rapidly expanding foliage where this pest causes greatest injury. Soil application provides superior long‐term vine protection because declining residues on foliar‐treated vines lead to suboptimal activity within 2–3 weeks. Vineyard managers of susceptible cultivars may take advantage of this approach to E. fabae management by using foliar applications of the three neonicotinoids tested here, or by using soil‐applied thiamethoxam. Copyright © 2011 Society of Chemical Industry     

Colluvisols under cultivation in Schleswig-Holstein. 2. Carbon distribution and soil organic matter composition

Soils under intensive cultivation have altered due to water erosion. This study was conducted to determine whether soil organic matter (SOM) composition of the colluvial source (Ap horizons) differs from the colluvial sink (M horizons). The SOM of a sandy Catena with erodic Cambisols and colluvic soils (Colluvisols) in Schleswig-Holstein, Northwest Germany, was investigated. A wet chemical analysis was combined with CPMAS ¹³C-NMR spectroscopy. In one case a significant correlation between the SOM composition of the Ap horizon of the erodic Cambisol and the M horizon of the Colluvisol was high (r² = 0.904^-), whereas the correlation for the other set was much weaker (r² = 0.640*). Two possible paths of pedogenesis are discussed. About 70% of the SOM of the colluvial source is decomposed during translocation or after deposition. A selective preservation or new formation of humins in the M material is probable. These humins contain, obviously, large amounts of polysaccharides, which were not detected by the wet chemical analysis. Further investigations of colluvic and erodic soils are necessary in order to specify the SOM quality and its possible modification due to soil translocation and accumulation.