Protease-activated receptor 2, dipeptidyl peptidase I, and proteases mediate Clostridium difficile toxin A enteritis

BACKGROUND & AIMS: We studied the role of protease-activated receptor 2 (PAR(2)) and its activating enzymes, trypsins and tryptase, in Clostridium difficile toxin A (TxA)-induced enteritis. METHODS: We injected TxA into ileal loops in PAR(2) or dipeptidyl peptidase I (DPPI) knockout mice or in wild-type mice pretreated with tryptase inhibitors (FUT-175 or MPI-0442352) or soybean trypsin inhibitor. We examined the effect of TxA on expression and activity of PAR(2) and trypsin IV messenger RNA in the ileum and cultured colonocytes. We injected activating peptide (AP), trypsins, tryptase, and p23 in wild-type mice, some pretreated with the neurokinin 1 receptor antagonist SR140333. RESULTS: TxA increased fluid secretion, myeloperoxidase activity in fluid and tissue, and histologic damage. PAR(2) deletion decreased TxA-induced ileitis, reduced luminal fluid secretion by 20%, decreased tissue and fluid myeloperoxidase by 50%, and diminished epithelial damage, edema, and neutrophil infiltration. DPPI deletion reduced secretion by 20% and fluid myeloperoxidase by 55%. In wild-type mice, FUT-175 or MPI-0442352 inhibited secretion by 24%-28% and tissue and fluid myeloperoxidase by 31%-71%. Soybean trypsin inhibitor reduced secretion to background levels and tissue myeloperoxidase by up to 50%. TxA increased expression of PAR(2) and trypsin IV in enterocytes and colonocytes and caused a 2-fold increase in Ca(2+) responses to PAR(2) AP. AP, tryptase, and trypsin isozymes (trypsin I/II, trypsin IV, p23) caused ileitis. SR140333 prevented AP-induced ileitis. CONCLUSIONS: PAR(2) and its activators are proinflammatory in TxA-induced enteritis. TxA stimulates existing PAR(2) and up-regulates PAR(2) and activating proteases, and PAR(2) causes inflammation by neurogenic mechanisms.

Mast cell tryptase and proteinase-activated receptor 2 induce hyperexcitability of guinea-pig submucosal neurons

Mast cells that are in close proximity to autonomic and enteric nerves release several mediators that cause neuronal hyperexcitability. This study examined whether mast cell tryptase evokes acute and long-term hyperexcitability in submucosal neurons from the guinea-pig ileum by activating proteinase-activated receptor 2 (PAR2) on these neurons. We detected the expression of PAR2 in the submucosal plexus using RT-PCR. Most submucosal neurons displayed PAR2 immunoreactivity, including those colocalizing VIP. Brief (minutes) application of selective PAR2 agonists, including trypsin, the activating peptide SL-NH2 and mast cell tryptase, evoked depolarizations of the submucosal neurons, as measured with intracellular recording techniques. The membrane potential returned to resting values following washout of agonists, but most neurons were hyperexcitable for the duration of recordings (> 30 min-hours) and exhibited an increased input resistance and amplitude of fast EPSPs. Trypsin, in the presence of soybean trypsin inhibitor, and the reverse sequence of the activating peptide (LR-NH2) had no effect on neuronal membrane potential or long-term excitability. Degranulation of mast cells in the presence of antagonists of established excitatory mast cell mediators (histamine, 5-HT, prostaglandins) also caused depolarization, and following washout of antigen, long-term excitation was observed. Mast cell degranulation resulted in the release of proteases, which desensitized neurons to other agonists of PAR2. Our results suggest that proteases from degranulated mast cells cleave PAR2 on submucosal neurons to cause acute and long-term hyperexcitability. This signalling pathway between immune cells and neurons is a previously unrecognized mechanism that could contribute to chronic alterations in visceral function.

Seasonality and the valuation of commodity options

Price movements in many commodity markets exhibit significant seasonal patterns. However, given an observed futures price, a deterministic seasonal component at the price level is not relevant for the pricing of commodity options. In contrast, this is not true for the seasonal pattern observed in the volatility of the commodity price. Analyzing an extensive sample of soybean, corn, heating oil and natural gas options, we find that seasonality in volatility is an important aspect to consider when valuing these contracts. The inclusion of an appropriate seasonality adjustment significantly reduces pricing errors in these markets and yields more improvement in valuation accuracy than increasing the number of stochastic factors.

Contrasting arbuscular mycorrhizal communities colonizing different host plants show a similar response to a soil phosphorus concentration gradient

High soil phosphorus (P) concentration is frequently shown to reduce root colonization by arbuscular mycorrhizal (AM) fungi, but the influence of P on the diversity of colonizing AM fungi is uncertain. We used terminal restriction fragment length polymorphism (T-RFLP) of 18S rDNA and cloning to assess diversity of AM fungi colonizing maize (Zea mays), soybean (Glycene max) and field violet (Viola arvensis) at three time points in one season along a P gradient of 10280mgl1 in the field. Percentage AM colonization changed between sampling time points but was not reduced by high soil P except in maize. There was no significant difference in AM diversity between sampling time points. Diversity was reduced at concentrations of P > 25mgl1, particularly in maize and soybean. Both cloning and T-RFLP indicated differences between AM communities in the different host species. Host species was more important than soil P in determining the AM community, except at the highest P concentration. Our results show that the impact of soil P on the diversity of AM fungi colonizing plants was broadly similar, despite the fact that different plants contained different communities. However, subtle differences in the response of the AM community in each host were evident.

Lunasin in cereal seeds: what is the origin?

Lunasin is a peptide from soybean seeds which has been demonstrated to have anticancer properties. It has also been reported in cereal seeds: wheat, rye, barley and Triticale. However, extensive searches of transcriptome and DNA sequence databases for wheat and other cereals have failed to identify sequences encoding either the lunasin peptide or a precursor protein. This raises the question of the origin of the lunasin reported in cereal grain.

Cereal aphids and their natural enemies: responses to landscape composition and agri-environment schemes

There are potential conflicts between food security, biodiversity conservation and ecosystem services. Currently, there are still gaps in our understanding on the links between land use, biodiversity and ecosystem services; all have implications for sustainable agriculture. To improve food productivity in an ecologically friendly manner we should consider adapting current pest control techniques from being reliant on chemical means towards a more integrated approach. However, to do this, farmers and land owners require more information in order to make informed decisions. This brief review explores field level and landscape scale impacts on aphid control by their natural enemies. This will be done by exploring the effects of local field margin flower strips and two key landscape scale factors, winter wheat and lowland calcareous grasslands on aphids and their natural enemies. Research questions which need answering are discussed.

Phospholipid-based microemulsions suitable for use in foods

The preparation of nonaqueous microemulsions using food-acceptable components is reported. The effect of oil on the formation of microemulsions stabilized by lecithin (Epikuron 200) and containing propylene glycol as immiscible solvent was investigated. When the triglycerides were used as oil, three types of phase behavior were noted, namely, a two-phase cloudy region (occurring at low lecithin concentrations), a liquid crystalline (LC) phase (occurring at high surfactant and low oil concentrations), and a clear monophasic microemulsion region. The extent of this clear one-phase region was found to be dependent upon the molecular volume of the oil being solubilized. Large molecular volume oils, such as soybean and sunflower oils, produced a small microemulsion region, whereas the smallest molecular volume triglyceride, tributyrin, produced a large, clear monophasic region. Use of the ethyl ester, ethyl oleate, as oil produced a clear, monophasic region of a size comparable to that seen with tributyrin. Substitution of some of the propylene glycol with water greatly reduced the extent of the clear one-phase region and increased the extent of the liquid crystalline region. In contrast, ethanol enhanced the clear, monophasic region by decreasing the LC phase. Replacement of some of the lecithin with the micelle-forming nonionic surfactant Tween 80 to produce mixed lecithin/Tween 80 mixtures of weight ratios (Km) 1:2 and 1:3 did not significantly alter the phase behavior, although there was a marginal increase in the area of the two-phase, cloudy region of the phase diagram. The use of the lower phosphatidylcholine content lecithin, Epikuron 170, in place of Epikuron 200 resulted in a reduction in the LC region for all of the systems investigated. In conclusion, these studies show that it is possible to prepare one-phase, clear lecithin-based microemulsions over a wide range of compositions using components that are food-acceptable.

Investigations into the formation and characterization of phospholipid microemulsions. IV. Pseudo-ternary phase diagrams of systems containing water-lecithin-alcohol and oil; The influence of oil

Phase studies have been performed for quaternary systems composed of egg lecithin, cosurfactant, water and oil. The lecithin used was the commercially available egg lecithin Ovothin 200 (which comprises ≥ 92% phosphatidylcholine). The cosurfactants employed were propanol and butanol, and these were used at lecithin/cosurfactant mixing ratios (Km) of 1:1 and 1.94:1 (weight basis). Six polar oils were investigated, including the alkanoic acids, octanoic and oleic, their corresponding ethyl esters and the medium and long chain triglycerides, Miglyol 812 and soybean oil. All oils, irrespective of the alcohol and the Km used, gave rise to systems that produced a stable isotropic region along the surfactant/oil axis (designated as a reverse microemulsion system). In addition, the systems incorporating propanol at both Km and butanol at a Km of 1.94: 1, generally gave rise to a liquid crystalline region and, in some cases, a second isotropic non-birefingent area (designated as a normal microemulsion system). The phase behaviour observed was largely dependent upon the alcohol and Km used and the size and the polarity of the oil present.

Characterization of the lipoxygenase (LOX) gene family in the Chinese white pear (Pyrus bretschneideri) and comparison with other members of the Rosaceae

Background Lipoxygenases (LOXs), a type of non-haem iron-containing dioxygenase, are ubiquitous enzymes in plants and participate in the formation of fruit aroma which is a very important aspect of fruit quality. Amongst the various aroma volatiles, saturated and unsaturated alcohols and aldehydes provide the characteristic aroma of the fruit. These compounds are formed from unsaturated fatty acids through oxidation, pyrolysis and reduction steps. This biosynthetic pathway involves at least four enzymes, including LOX, the enzyme responsible for lipid oxidation. Although some studies have been conducted on the LOX gene family in several species including Arabidopsis, soybean, cucumber and apple, there is no information from pear; and the evolutionary history of this gene family in the Rosaceae is still not resolved. Results In this study we identified 107 LOX homologous genes from five Rosaceous species (Pyrus bretschneideri, Malus × domestica, Fragaria vesca, Prunus mume and Prunus persica); 23 of these sequences were from pear. By using structure analysis, phylogenic analysis and collinearity analysis, we identified variation in gene structure and revealed the phylogenetic evolutionary relationship of this gene family. Expression of certain pear LOX genes during fruit development was verified by analysis of transcriptome data. Conclusions 23 LOX genes were identified in pear and these genes were found to have undergone a duplication 30–45 MYA; most of these 23 genes are functional. Specific gene duplication was found on chromosome4 in the pear genome. Useful information was provided for future research on the evolutionary history and transgenic research on LOX genes.

Antibiosis in wheat interacts with crowding stress to affect Metopolophiumdirhodum development and susceptibility to malathion

We used a laboratory study to compare the performance of rose-grain aphid, Metopolophium dirhodum(Walker)(Hemiptera:Aphididae),onthewheatcultivars‘Huntsman’(susceptible)and‘Rapier’ (partiallyresistant)inbothlowdensity(uncrowded)andhighdensity(crowded)coloniesandexamined the consequences for aphid susceptibility to malathion. Adult apterae that developed on Rapier wheat had their mean relative growth rate (MRGR) reduced by 6 and 9% under uncrowded and crowded conditions, respectively, whereas the crowding treatment reduced MRGR by 3%, but only in Rapier aphids. Rapier resistance also reduced adult dry weight by 13 and 14% under crowded and uncrowded conditions, respectively, whereas crowding reduced it by 34 and 35% in Rapier and Huntsman aphids, respectively. Development on Rapier substantially reduced the topical LC50 of malathion by 37.8 and 34.8% under crowded and uncrowded conditions, suggesting that plant antibiosis increased malathion susceptibility. By comparison, crowding only reduced the LC50 by 29.5 and 26.0% on Huntsman and Rapier. The LD50 data showed that reductions on aphid body size on Rapier and through crowding did not fully explain the differences in LC50. This was particularly in the values for crowded aphids that were actually 80% higher than for uncrowded ones. Thi sapparent tolerance of crowded aphids, however, may partly be due to loss of insecticide from small aphids at dosing. Evidence of synergy between plant resistance and insecticide susceptibility raisest he possibility of using reduced concentrations of pesticides to control aphids on resistant crop cultivars, with diminished impacts on non-target and beneficial species important in integrated pest management(IPM)program

Behavioural responses of the seven-spot ladybird Coccinella septempunctata to plant headspace chemicals collected from four crop Brassicas and Arabidopsis thaliana, infested with Myzus persicae

1 Insects using olfactory stimuli to forage for prey/hosts are proposed to encounter a ‘reliability–detectability problem’, where the usability of a stimulus depends on its reliability as an indicator of herbivore presence and its detectability. 2 We investigated this theory using the responses of female seven-spot ladybirds Coccinella septempunctata (Coleoptera: Coccinellidae) to plant headspace chemicals collected from the peach-potato aphid Myzus persicae and four commercially available Brassica cultivars; Brassica rapa L. cultivar ‘turnip purple top’, Brassica juncea L. cultivar ‘red giant mustard’, Brassica napus L. cultivar ‘Apex’, Brassica napus L. cultivar ‘Courage’ and Arabidopsis thaliana. For each cultivar/species, responses to plants that were undamaged, previously infested by M. persicae and infested with M. persicae, were investigated using dual-choice Petri dish bioassays and circular arenas. 3 There was no evidence that ladybirds responded to headspace chemicals from aphids alone. Ladybirds significantly preferred headspace chemicals from B. napus cv. Apex that were undamaged compared with those from plants infested with aphids. For the other four species/cultivars, there was a consistent trend of the predators being recorded more often in the half of the Petri dish containing plant headspace chemicals from previously damaged and infested plants compared with those from undamaged ones. Furthermore, the mean distance ladybirds walked to reach aphid-infested A. thaliana was significantly shorter than to reach undamaged plants. These results suggest that aphid-induced plant chemicals could act as an arrestment or possibly an attractant stimulus to C. septempunctata. However, it is also possible that C. septempunctata could have been responding to aphid products, such as honeydew, transferred to the previously damaged and infested plants. 4 The results provide evidence to support the ‘reliability–detectability’ theory and suggest that the effectiveness of C. septempunctata as a natural enemy of aphids may be strongly affected by which species and cultivar of Brassica are being grown.

Do turning biases by the 7-spot ladybird, Coccinella septempunctata, increase their foraging efficiency?

The hypothesis that foraging male and female Coccinella septempunctata L. would exhibit a turning bias when walking along a branched linear wire in a Y-maze was tested. Individuals were placed repeatedly in the maze. Approximately 45% of all individuals tested displayed significant turning biases, with a similar number of individuals biased to the left and right. In the maze right-handed individuals turned right at 84.4% of turns and the left-handed individuals turned left at 80.2% of turns. A model of the searching efficiency of C. septempunctata in dichotomous branched environments showed that model coccinellids with greater turning biases discovered a higher proportion of the plant for a given number of searches than those with no bias. A modification of the model to investigate foraging efficiency, by calculating the mean time taken by individuals to find randomly distributed aphid patches, suggested that on four different sizes of plants, with a variety of aphid patch densities, implementing a turning bias was a significantly more efficient foraging strategy than no bias. In general the benefits to foraging of implementing a turning bias increased with the degree of the bias. It may be beneficial for individuals in highly complex branched environments to have a turning bias slightly lower than 100% in order to benefit from increased foraging efficiency without walking in circles. Foraging bias benefits increased with increasing plant size and decreasing aphid density. In comparisons of two different plant morphologies, one with a straight stem and side branches and one with a symmetrically branched morphology, there were few significant differences in the effects of turning biases on foraging efficiency between morphologies

JULES-crop: a parametrisation of crops in the Joint UK Land Environment Simulator

Studies of climate change impacts on the terrestrial biosphere have been completed without recognition of the integrated nature of the biosphere. Improved assessment of the impacts of climate change on food and water security requires the development and use of models not only representing each component but also their interactions. To meet this requirement the Joint UK Land Environment Simulator (JULES) land surface model has been modified to include a generic parametrisation of annual crops. The new model, JULES-crop, is described and evaluation at global and site levels for the four globally important crops; wheat, soybean, maize and rice. JULES-crop demonstrates skill in simulating the inter-annual variations of yield for maize and soybean at the global and country levels, and for wheat for major spring wheat producing countries. The impact of the new parametrisation, compared to the standard configuration, on the simulation of surface heat fluxes is largely an alteration of the partitioning between latent and sensible heat fluxes during the later part of the growing season. Further evaluation at the site level shows the model captures the seasonality of leaf area index, gross primary production and canopy height better than in the standard JULES. However, this does not lead to an improvement in the simulation of sensible and latent heat fluxes. The performance of JULES-crop from both an Earth system and crop yield model perspective is encouraging. However, more effort is needed to develop the parametrisation of the model for specific applications. Key future model developments identified include the introduction of processes such as irrigation and nitrogen limitation which will enable better representation of the spatial variability in yield.

Factors affecting the concentration in seven-spotted ladybirds (Coccinella septempunctata L.) of Cd and Zn transferred through the food chain

The transfer of Cd and Zn from soils amended with sewage sludge was followed through a food chain consisting of wheat, aphids and the predator Coccinella septempunctata. Multiple regression models were generated to predict the concentrations of Cd and Zn in C. septempunctata. No significant model could be generated for Cd, indicting that the concentration of this metal was maintained within relatively narrow limits. A model predicting 64% of the variability in the Zn concentration of C. septempunctata was generated from of the concentration of Zn in the diet, time and rate of Zn consumption. The results suggest that decreasing the rate of food consumption is an effective mechanism to prevent the accumulation of Zn and that the availability of Zn in the aphid prey increased with the concentration in the aphids. The results emphasise the importance of using ecologically relevant food chains and exposure pathways during ecotoxicological studies.

Tales of the unexpected: research discoveries that occurred by accident

Data from four experimental research projects are presented which have in common that unexpected results caused a change in direction of the research. A plant growth accelerator caused the appearance of white black bean aphids, a synthetic pyrethroid suspected of enhancing aphid reproduction proved to enhance plant growth, a chance conversation with a colleague initiated a search for fungal DNA in aphids, and the accidental invasion of aphid cultures by a parasitoid reversed the aphid population ranking of two Brussels sprout cultivars. This last result led to a whole series of studies on the plant odour preferences of emerging parasitoids which in turn revealed the unexpected phenomenon that chemical cues to the maternal host plant are left with the eggs at oviposition. It is pointed out that, too often, researchers fail to follow up unexpected results because they resist accepting flaws in their hypotheses; also that current application criteria for research funding make it hard to accommodate unexpected findings.