Kinetic models as a route to control acrylamide formation in food

Reports that heat processing of foods induces the formation of acrylamide heightened interest in the chemistry, biochemistry, and safety of this compound. Acrylamide-induced neurotoxicity, reproductive toxicity, genotoxicity, and carcinogenicity are potential human health risks based on animal studies. Because exposure of humans to acrylamide can come from both external sources and the diet, there exists a need to develop a better understanding of its formation and distribution in food and its role in human health. To contribute to this effort, experts from eight countries have presented data on the chemistry, analysis, metabolism, pharmacology, and toxicology of acrylamide. Specifically covered are the following aspects: exposure from the environment and the diet; biomarkers of exposure; risk assessment; epidemiology; mechanism of formation in food; biological alkylation of amino acids, peptides, proteins, and DNA by acrylamide and its epoxide metabolite glycidamide; neurotoxicity, reproductive toxicity, and carcinogenicity; protection against adverse effects; and possible approaches to reducing levels in food. Cross-fertilization of ideas among several disciplines in which an interest in acrylamide has developed, including food science, pharmacology, toxicology, and medicine, will provide a better understanding of the chemistry and biology of acrylamide in food, and can lead to the development of food processes to decrease the acrylamide content of the diet.

Field evaluation of systemic inducing resistance chemicals at different growth stages for the control of apple (Venturia inaequalis) and pear (Venturia pirina) scab

Two field trials were conducted using established apple (Malus cv. Golden Delicious) and pear (Pyrus communis 'Williams' Bon Chretien') to assess the efficacy of three commercially available systemic inducing resistance (SIR) products, Messenger (a.i. Harpin protein), Phoenix (a.i. Potassium phosphite) and Rigel (a.i. Salicylic acid derivative) applied at four different growth stages of tree development (bud break, green cluster, 90% petal fall, early fruitlet) against the foliar pathogens Venturia inaequalis and Venturia pirina which cause apple and pear scab respectively. A conventional synthetic fungicide (penconazole) used within the UK for apple and pear scab control was included for comparison. Little efficacy as scab protectants was demonstrated when each SIR product and penconazole was applied at only two growth stages (bud break, green cluster). However when the above compounds were applied at three or more growth stages efficacy as scab protectants was confirmed. The synthetic fungicide penconazole provided greatest protection against apple and pear scab in both the 2006 and 2007 field trials. There was little difference in the magnitude of scab protection conferred by each SIR agent. Results suggest application of at least three sprays during bud break to early fruitlet formation with an appropriate SIR agent may provide a useful addition to existing methods of apple and pear scab management under field conditions. (C) 2009 Published by Elsevier Ltd.

Evaluation of film forming polymers to control apple scab (Venturia inaequalis (Cooke) G. Wint.) under laboratory and field conditions

A detached leaf bioassay was used to determine the influence of several film forming polymers and a conventional triazole fungicide on apple scab (Venturia inaequalis (Cooke) G. Wint.) development under laboratory in vitro conditions, supported by two field trials using established apple cv. Golden Delicious to further assess the efficacy of foliar applied film forming polymers as scab protectant compounds. All film forming polymers used in this investigation (Bond, Designer, Nu-Film P, Spray Gard, Moisturin, Companion PCT12) inhibited germination of conidia, subsequent formation of appressoria and reduced leaf scab severity using a detached leaf bioassay. Regardless of treatment, there were no obvious trends in the percentage of conidia with one to four appressoria 5 days after inoculation. The synthetic fungicide penconazole resulted in the greatest levels of germination inhibition, appressorium development and least leaf scab severity. Under field conditions, scab severity on leaves and fruit of apple cv. Golden Delicious treated with a film forming polymer (Bond, Spray Gard, Moisturin) was less than on untreated controls. However, greatest protection in both field trials was provided by the synthetic fungicide penconazole. Higher chlorophyll fluorescence Fv/Fm emissions in polymer and penconazole treated trees indicated less damage to the leaf photosynthetic system as a result of fungal invasion. In addition, higher SPAD values as measures of leaf chlorophyll content were recorded in polymer and penconazole treated trees. Application of a film forming polymer or penconazole resulted in a higher apple yield per tree at harvest in both the 2005 and 2006 field trials compared to untreated controls. Results suggest application of an appropriate film forming polymer may provide a useful addition to existing methods of apple scab management. (C) 2008 Elsevier Ltd. All rights reserved.

Regulated deficit irrigation - a means to control growth in woody ornamentals

Improving plant quality and the uniformity of a crop are major objectives for growers of ornamental nursery stock. The potential to control excess vigour and to improve quality through regulated deficit irrigation (RDI) was investigated using a range of woody ornamental species. RDI regimes reduced vegetative growth consistently across different species and growing seasons. Plants adapted to reduced water supplies primarily via stomatal control, but also by osmotic adjustment when grown under the most severe RDI regimes. Only plants exposed to <= 25% of potential evapo-transpiration demonstrated any evidence of leaf injury, and the extent was slight. Growth inhibition increased as the severity of RDI increased. Improvements in quality were attained through a combination of shorter internodes and final shoot lengths, yet the number of 'formative' primary shoots remained unaffected. Compact, well-branched plants could be formed without a requirement for mid-season pruning. In addition to severity, the timing of RDI also influenced growth responses. Applying 50% ETp for 8 weeks during July-August resulted in the formation of good quality plants, which retained their shape until the following Spring. Re-positioning irrigation drippers within the pots of well-watered plants, in an attempt to induce a partial root drying (PRD) treatment, reduced growth, but not significantly. The adoption of irrigation scheduling, based on 50-100% ETp, has the potential to improve commercial crop quality across a range of ornamental species.

Platelet PECAM-1 inhibits thrombus formation in vivo

Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a cell surface glycoprotein receptor expressed on a range of blood cells, including platelets, and on vascular endothelial cells. PECAM-1 possesses adhesive and signaling properties, the latter being mediated by immunoreceptor tyrosine-based inhibitory motifs present on the cytoplasmic tail of the protein. Recent studies in vitro have demonstrated that PECAM-1 signaling inhibits the aggregation of platelets. In the present study we have used PECAM-1-deficient mice and radiation chimeras to investigate the function of this receptor in the regulation of thrombus formation. Using intravital microscopy and laser-induced injury to cremaster muscle arterioles, we show that thrombi formed in PECAM-1-deficient mice were larger, formed more rapidly than in control mice, and were more stable. Larger thrombi were also formed in control mice that received transplants of PECAM-1-deficient bone marrow, in comparison to mice that received control transplants. A ferric chloride model of thrombosis was used to investigate thrombus formation in carotid arteries. In PECAM-1-deficient mice the time to 75% vessel occlusion was significantly shorter than in control mice. These data provide evidence for the involvement of platelet PECAM-1 in the negative regulation of thrombus formation.

Control of human immunodeficiency virus type-1 protease activity in insect cells expressing Gag-Pol rescues assembly of immature but not mature virus-like particles

Expression of human immunodeficiency virus type 1 (HIV-1) Gag protein in insect cells using baculovirus vectors leads to the abundant production of virus-like particles (VLPs) that represent the immature form of the virus. When Gag-Pol is included, however, VLP production is abolished, a result attributed to premature protease activation degrading the intracellular pool of Gag precursor before particle assembly can occur. As large-scale synthesis of mature noninfectious VLPs would be useful, we have sought to control HIV protease activity in insect cells to give a balance of Gag and Gag-Pol that is compatible with mature particle formation. We show here that intermediate levels of protease activity in insect cells can be attained through site-directed mutagenesis of the protease and through antiprotease drug treatment. However, despite Gag cleavage patterns that mimicked those seen in mammalian cells, VLP synthesis exhibited an essentially all-or-none response in which VLP synthesis occurred but was immature or failed completely. Our data are consistent with a requirement for specific cellular factors in addition to the correct ratio of Gag and Gag-Pol for assembly of mature retrovirus particles in heterologous cell types. (C) 2003 Elsevier Science (USA). All rights reserved.

A novel technique for differentiation of proteins in the development of acid gel structure from control and heat treated milk using confocal scanning laser microscopy

The incorporation of caseins and whey proteins into acid gels produced from unheated and heat treated skimmed milk was studied by confocal scanning laser microscopy (CSLM) using fluorescent labelled proteins. Bovine casein micelles were labelled using Alexa Fluor 594, while whey proteins were labelled using Alexa Fluor 488. Samples of the labelled protein solutions were introduced into aliquots of pasteurised skim milk, and skim milk heated to 90 degrees C for 2 min and 95 degrees C for 8 min. The milk was acidified at 40 degrees C to a final pH of 4.4 using 20 g gluconodelta-lactone/l (GDL). The formation of gels was observed with CSLM at two wavelengths (488 nm and 594 nm), and also by visual and rheological methods. In the control milk, as pH decreased distinct casein aggregates appeared, and as further pH reduction occurred, the whey proteins could be seen to coat the casein aggregates. With the heated milks, the gel structure was formed of continuous strands consisting of both casein and whey protein. The formation of the gel network was correlated with an increase in the elastic modulus for all three treatments, in relation to the severity of heat treatment. This model system allows the separate observation of the caseins and whey proteins, and the study of the interactions between the two protein fractions during the formation of the acid gel structure, on a real-time basis. The system could therefore be a valuable tool in the study of structure formation in yoghurt and other dairy protein systems.

Free amino acids and sugars in rye grain: implications for acrylamide formation

Acrylamide forms from free asparagine and sugars during cooking, and products derived from the grain of cereals, including rye, contribute a large proportion of total dietary intake. In this study, free amino acid and sugar concentrations were measured in the grain of a range of rye varieties grown at locations in Hungary, France, Poland, and the United Kingdom and harvested in 2005, 2006, and 2007. Genetic and environmental (location and harvest year) effects on the levels of acrylamide precursors were assessed. The data showed free asparagine concentration to be the main determinant of acrylamide formation in heated rye flour, as it is in wheat. However, in contrast to wheat, sugar, particularly sucrose, concentration also correlated both with asparagine concentration and with acrylamide formed. Free asparagine concentration was shown to be under genetic (G), environmental (E), and integrated (G × E) control. The same was true for glucose, whereas maltose and fructose were affected mainly by environmental factors and sucrose was largely under genetic control. The ratio of variation due to varieties (genotype) to the total variation (a measure of heritability) for free asparagine concentration in the grain was 23%. Free asparagine concentration was closely associated with bran yield, whereas sugar concentration was associated with low Hagberg falling number. Rye grain was found to contain much higher concentrations of free proline than wheat grain, and less acrylamide formed per unit of asparagine in rye than in wheat flour.

Redox control of light-induced charge separation in a transition metal cluster: photochemistry of a methyl viologen-substituted [Os-3(CO)10(a-diimine)] cluster

Sub)picosecond transient absorption (TA) and time-resolved infrared (TRIR) spectra of the cluster [OS3(CO)(10-) (AcPy-MV)](2+) (the clication AcPy-MV = Acpy-MV2+ = [2-pyridylacetimine-N-(2-(1'-methyl-4,4'-bipyridine-1,1'-diium-1-yl) ethyl)] (PF6)(2)) (1(2+)) reveal that photoinduced electron transfer to the electron-accepting 4,4'-bipyridine-1,1'diium (MV2+) moiety competes with the fast relaxation of the initially populated sigmapi* excited state of the cluster to the ground state and/or cleavage of an Os-Os bond. The TA spectra of cluster 12 in acetone, obtained by irradiation into its lowest-energy absorption band, show the characteristic absorptions of the one-electron-reduced MV*(+) unit at 400 and 615 nm, in accordance with population of a charge-separated (CS) state in which a cluster-core electron has been transferred to the lowest pi* orbital of the remote MV2+ unit. This assignment is confirmed by picosecond TRIR spectra that show a large shift of the pilot highest-frequency nu(CO) band of 1(2+) by ca. +40 cm(-1), reflecting the photooxidation of the cluster core. The CS state is populated via fast (4.2 x 10(11) s(-1)) and efficient (88%) oxidative quenching of the optically populated sigmapi* excited state and decays biexponentially with lifetimes of 38 and 166 ps (1:2:1 ratio) with a complete regeneration of the parent cluster. About 12% of the cluster molecules in the sigmapi* excited state form long-lived open-core biradicals. In strongly coordinating acetonitrile, however, the cluster core-to-MV2+ electron transfer in cluster 12+ results in the irreversible formation of secondary photoproducts with a photooxidized cluster core. The photochemical behavior of the [Os-3(CO)(10)(alpha-diimine-MV)](2+) (donor-acceptor) dyad can be controlled by an externally applied electronic bias. Electrochemical one-electron reduction of the MV2+ moiety prior to the irradiation reduces its electron-accepting character to such an extent that the photoinduced electron transfer to MV*+ is no longer feasible. Instead, the irradiation of reduced cluster 1(.)+ results in the reversible formation of an open-core zwitterion, the ultimate photoproduct also observed upon irradiation of related nonsubstituted clusters [Os-3(CO)(10)(alpha-diimine)] in strongly coordinating solvents such as acetonitrile.

Stress field control of eruption dynamics at a rift volcano: Nyamuragira, D.R.Congo

Using the record of 30 flank eruptions over the last 110 years at Nyamuragira, we have tested the relationship between the eruption dynamics and the local stress field. There are two groups of eruptions based on their duration (< 80days >) that are also clustered in space and time. We find that the eruptions fed by dykes parallel to the East African Rift Valley have longer durations (and larger volumes) than those eruptions fed by dykes with other orientations. This is compatible with a model for compressible magma transported through an elastic-walled dyke in a differential stress field from an over-pressured reservoir (Woods et al., 2006). The observed pattern of eruptive fissures is consistent with a local stress field modified by a northwest-trending, right lateral slip fault that is part of the northern transfer zone of the Kivu Basin rift segment. We have also re-tested with new data the stochastic eruption models for Nyamuragira of Burt et al. (1994). The time-predictable, pressure-threshold model remains the best fit and is consistent with the typically observed declining rate of sulphur dioxide emission during the first few days of eruption with lava emission from a depressurising, closed, crustal reservoir. The 2.4-fold increase in long-term eruption rate that occurred after 1977 is confirmed in the new analysis. Since that change, the record has been dominated by short-duration eruptions fed by dykes perpendicular to the Rift. We suggest that the intrusion of a major dyke during the 1977 volcano-tectonic event at neighbouring Nyiragongo volcano inhibited subsequent dyke formation on the southern flanks of Nyamuragira and this may also have resulted in more dykes reaching the surface elsewhere. Thus that sudden change in output was a result of a changed stress field that forced more of the deep magma supply to the surface. Another volcano-tectonic event in 2002 may also have changed the magma output rate at Nyamuragira.

Control and analysis of oriented thin films of lipid inverse bicontinuous cubic phases using grazing incidence small-angle X‑ray scattering

Lipid cubic phases are complex nanostructures that form naturally in a variety of biological systems, with applications including drug delivery and nanotemplating. Most X-ray scattering studies on lipid cubic phases have used unoriented polydomain samples as either bulk gels or suspensions of micrometer-sized cubosomes. We present a method of investigating cubic phases in a new form, as supported thin films that can be analyzed using grazing incidence small-angle X-ray scattering (GISAXS). We present GISAXS data on three lipid systems: phytantriol and two grades of monoolein (research and industrial). The use of thin films brings a number of advantages. First, the samples exhibit a high degree of uniaxial orientation about the substrate normal. Second, the new morphology allows precise control of the substrate mesophase geometry and lattice parameter using a controlled temperature and humidity environment, and we demonstrate the controllable formation of oriented diamond and gyroid inverse bicontinuous cubic along with lamellar phases. Finally, the thin film morphology allows the induction of reversible phase transitions between these mesophase structures by changes in humidity on subminute time scales, and we present timeresolved GISAXS data monitoring these transformations.

Synoptic versus orographic control on stationary convective banding

Quasi-stationary convective bands can cause large localised rainfall accumulations and are often anchored by topographic features. Here, the predictability of and mechanisms causing one such band are determined using ensembles of the Met Office Unified Model at convection-permitting resolution (1.5 km grid length). The band was stationary over the UK for 3 h and produced rainfall accumulations of up to 34 mm. The amount and location of the predicted rainfall was highly variable despite only small differences between the large-scale conditions of the ensemble members. Only three of 21 members of the control ensemble produced a stationary rain band; these three had the weakest upstream winds and hence lowest Froude number. Band formation was due to the superposition of two processes: lee-side convergence resulting from flow around an upstream obstacle and thermally forced convergence resulting from elevated heating over the upstream terrain. Both mechanisms were enhanced when the Froude number was lower. By increasing the terrain height (thus reducing the Froude number), the band became more predictable. An ensemble approach is required to successfully predict the possible occurrence of such quasi-stationary convective events because the rainfall variability is largely modulated by small variations of the large-scale flow. However, high-resolution models are required to accurately resolve the small-scale interactions of the flow with the topography upon which the band formation depends. Thus, although topography provides some predictability, the quasi-stationary convective bands anchored by it are likely to remain a forecasting challenge for many years to come.

Ecophysiology of seed dormancy and the control of germination in early spring-flowering Galanthus nivalis and Narcissus pseudonarcissus (Amaryllidaceae)

Seed dormancy induction and alleviation in the winter-flowering moist temperate woodland species Galanthus nivalis and Narcissus pseudonarcissus are complex and poorly understood. Temperature, light and desiccation were investigated to elucidate their role in the germination ecophysiology of these species. Outdoor and laboratory experiments simulating different seasonal temperatures, seasonal durations, and temperature fluctuations; the presence of light during different seasons; and intermittent drying (during the summer period) over several ‘years’ investigated the importance of these factors in germination. Warm summer-like temperatures (20°C) were necessary for germination at subsequent cooler autumn-like temperatures (greatest at 15°C in G. nivalis and 10°C in N. pseudonarcissus). As the warm temperature duration increased so did germination at subsequent cooler temperatures; further germination occurred in subsequent ‘years’ at cooler temperatures following a second, and also third, warm period. Germination was significantly greater in darkness, particularly in G. nivalis. Dormancy increased with seed maturation period in G. nivalis, because seeds extracted from green capsules germinated more readily than those from yellow. Desiccation increased dormancy in an increasing proportion of N. pseudonarcissus seeds the later they were dried in ‘summer’. Seed viability was only slightly reduced by desiccation in N. pseudonarcissus but was poor and variable in G. nivalis. Shoot formation occurred both at the temperature at which germination was greatest and also if 5°C cooler. In summary, continuous hydration of seeds of both species during warm summer-like temperatures results in the gradual release of seed dormancy; thereafter, darkness and cooler temperatures promote germination. Cold temperatures, increased seed maturity (G. nivalis), and desiccation (N. pseudonarcissus) increase dormancy while light inhibits germination.

EphB2 regulates contact-dependent and independent signalling to control platelet function

The Eph kinases, EphA4 and EphB1 and their ligand, ephrinB1 have been previously reported to be present in platelets where they contribute to thrombus stability. While thrombus formation allows for Eph-ephrin engagement and bidirectional signalling, the importance specifically of Eph kinase or ephrin signalling in regulating platelet function remained unidentified. In the present study, a genetic approach was used in mice to establish the contribution of signalling orchestrated by the cytoplasmic domain of EphB2 (a newly discovered Eph kinase in platelets) in platelet activation and thrombus formation. We conclude that EphB2 signalling is involved in the regulation of thrombus formation and clot retraction. Furthermore, the cytoplasmic tail of this Eph kinase regulates initial platelet activation in a contact-independent manner in the absence of Eph-ephrin ligation between platelets. Together these data demonstrate that EphB2 signalling not only modulates platelet function within a thrombus but is also involved in the regulation of the function of isolated platelets in a contact-independent manner.

Impacts of uplift of northern Tibetan Plateau and formation of Asian inland deserts on regional climate and environment

Based on the geological evidence that the northern Tibetan Plateau (NTP) had an uplift of a finite magnitude since the Miocene and the major Asian inland deserts formed in the early Pliocene, a regional climate model (RegCM4.1) with a horizontal resolution of 50 km was used to explore the effects of the NTP uplift and the related aridification of inland Asia on regional climate. We designed three numerical experiments including the control experiment representing the present-day condition, the high-mountain experiment representing the early Pliocene condition with uplifted NTP but absence of the Asian inland deserts, and the low-mountain experiment representing the mid-Miocene condition with reduced topography in the NTP (by as much as 2400 m) and also absence of the deserts. Our simulation results indicated that the NTP uplift caused significant reductions in annual precipitation in a broad region of inland Asia north of the Tibetan Plateau (TP) mainly due to the enhanced rain shadow effect of the mountains and changes in the regional circulations. However, four mountainous regions located in the uplift showed significant increases in precipitation, stretching from the Pamir Plateau in the west to the Qilian Mountains in the east. These mountainous areas also experienced different changes in the rainfall seasonality with the greatest increases occurring during the respective rainy seasons, predominantly resulted from the enhanced orographically forced upwind ascents. The appearance of the major deserts in the inland Asia further reduced precipitation in the region and led to increased dust emission and deposition fluxes, while the spatial patterns of dust deposition were also changed, not only in the regions of uplift-impacted topography, but also in the downwind regions. One major contribution from this study is the comparison of the simulation results with 11 existing geological records representing the moisture conditions from Miocene to Pliocene. The comparisons revealed good matches between the simulation results and the published geological records. Therefore, we conclude that the NTP uplift and the related formation of the major deserts played a controlling role in the evolution of regional climatic conditions in a broad region in inland Asia since the Miocene.