Nanostructural organization of naturally occurring composites - part II: silica-chitin-based biocomposites

Investigations of the micro- and nanostructures and chemical composition of the sponge skeletons as examples for natural structural biocomposites are of fundamental scientific relevance. Recently, we show that some demosponges (Verongula gigantea, Aplysina sp.) and glass sponges (Farrea occa, Euplectella aspergillum) possess chitin as a component of their skeletons. The main practical approach we used for chitin isolation was based on alkali treatment of corresponding external layers of spicules sponge material with the aim of obtaining alkali-resistant compounds for detailed analysis. Here, we present a detailed study of the structural and physicochemical properties of spicules of the glass sponge Rossella fibulata. The structural similarity of chitin derived from this sponge to invertebrate alpha chitin has been confirmed by us unambiguously using physicochemical and biochemical methods. This is the first report of a silica-chitin composite biomaterial found in Rossella species. Finally, the present work includes a discussion related to strategies for the practical application of silica-chitin-based composites as biomaterials

Reciprocal Inhibition of Adiponectin and Innate Lung Immune Responses to Chitin and Aspergillus fumigatus

Chitin is a structural biopolymer found in numerous organisms, including pathogenic fungi, and recognized as an immune-stimulating pathogen associated molecular pattern by pattern recognition molecules of the host immune system. However, programming and regulation of lung innate immunity to chitin inhalation in the context of inhalation of fungal pathogens such as Aspergillus fumigatus is complex and our understanding incomplete. Here we report that the systemic metabolism-regulating cytokine adiponectin is decreased in the lungs and serum of mice after chitin inhalation, with a concomitant decrease in surface expression of the adiponectin receptor AdipoR1 on lung leukocytes. Constitutive lung expression of acidic mammalian chitinase resulted in decreased inflammatory cytokine gene expression and neutrophil recruitment, but did not significantly affect lung adiponectin transcription. Exogenous recombinant adiponectin specifically dampened airway chitin-mediated eosinophil recruitment, while adiponectin deficiency resulted in increased airway eosinophils. The presence of adiponectin also resulted in decreased CCL11-mediated migration of bone marrow-derived eosinophils. In contrast to purified chitin, aspiration of viable conidia from the high chitin-expressing A. fumigatus isolate Af5517 resulted in increased neutrophil recruitment and inflammatory cytokine gene expression in adiponectin-deficient mice, while no significant changes were observed in response to the isolate Af293. Our results identify a novel role for the adiponectin pathway in inhibition of lung inflammatory responses to chitin and A. fumigatus inhalation

Functional Compost

The aim of the research program Functional Compost is to develop and test compost, which have been enriched with chitin, for plant growth promoting properties and to recognise specific mechanisms. Two types of compost were included in the program: source separated biodegradable municipal solid waste compost (DM = 62 %) and garden and park waste compost (DM = 66 %). Chitin was added in trace amounts during the maturity phase, combined with two levels of trace amounts immediately before adding the compost to the growth medium. The research program includes several parallel experiments. In experiment I, compost (20 vol. %) was added to soil (no plants) and incubated at 15 C for 5 month, under regular determination of microbial respiration and gross and net N mineralization. There was a significant increase in respiration due to chitin enrichment, which could not be explained by the amount of C derived from the chitin, which therefore suggest a priming effect. The N analyses are still being processed in the laboratory, but data are expected to be available at the conference. In experiment II, compost was mixed with sand, put into pots in a climate chamber, and spring barley seeds infected with Fusarium culmorum were sown in the pots. After 3 weeks of growth, the health of the plants was determined, and the chitinase activity in the sand was measured. The health of the plants and the chitinase activity was significantly higher in the treatments receiving municipal waste compared to the treatments receiving garden waste compost. However, there was no clear effect of the chitin enrichment. Additionally, the microbial community structure of the two types of compost, with and without early chitin, was determined by Denaturing Gradient Gel Electrophoresis (DGGE). There was a clear separation between compost types, and with or without early chitin amendment. Experiment III is a regular growth experiment, and is running right now. Compost has been incorporated into soil, put into pots in the greenhouse, and spring barley is grown for 2 month before determination for wet and dry weight and N uptake. Data from experiment III is expected to be available at the conference

Chitin-Induced Airway Epithelial Cell Innate Immune Responses Are Inhibited by Carvacrol/Thymol.

Chitin is produced in large amounts by fungi, insects, and other organisms and has been implicated in the pathogenesis of asthma. Airway epithelial cells are in direct contact with environmental particles and serve as the first line of defense against inhaled allergens and pathogens. The potential contributions of airway epithelial cells to chitin-induced asthma remain poorly understood. We hypothesized that chitin directly stimulates airway epithelial cells to release cytokines that promote type 2 immune responses and to induce expression of molecules which are important in innate immune responses. We found that chitin exposure rapidly induced the expression of three key type 2-promoting cytokines, IL-25, IL-33 and TSLP, in BEAS-2B transformed human bronchial epithelial cells and in A549 and H292 lung carcinoma cells. Chitin also induced the expression of the key pattern recognition receptors TLR2 and TLR4. Chitin induced the expression of miR-155, miR-146a and miR-21, each of which is known to up-regulate the expression of pro-inflammatory cytokines. Also the expression of SOCS1 and SHIP1 which are known targets of miR-155 was repressed by chitin treatment. The monoterpene phenol carvacrol (Car) and its isomer thymol (Thy) are found in herbal essential oils and have been shown to inhibit allergic inflammation in asthma models. We found that Car/Thy inhibited the effects of chitin on type 2-promoting cytokine release and on the expression of TLRs, SOCS1, SHIP1, and miRNAs. Car/Thy could also efficiently reduce the protein levels of TLR4, inhibit the increase in TLR2 protein levels in chitin plus Car/Thy-treated cells and increase the protein levels of SHIP1 and SOCS1, which are negative regulators of TLR-mediated inflammatory responses. We conclude that direct effects of chitin on airway epithelial cells are likely to contribute to allergic airway diseases like asthma, and that Car/Thy directly inhibits epithelial cell pro-inflammatory responses to chitin

Exploiting prokaryotic chitin-binding proteins for glycan recognition

• The cloning, expression and characterisation of prokaryotic chitin-binding proteins from Serratia marcescens, Pseudomonas aeruginosa, Photorhabdus luminescens Microfluidics and Photorhabdus asymbiotica • Development of an assay to assess the activity of chitin-binding proteins • Mutagenesis of chitin-binding proteins to alter glycan recognition pattern

The PKC, HOG and Ca2+ signalling pathways co-ordinately regulate chitin synthesis in Candida albicans

Open Access via PMC2649417Peer reviewedPublisher PD

A structural and biochemical model of processive chitin synthesis

Chitin synthases (CHS) produce chitin, an essential component of the fungal cell wall. The molecular mechanism of processive chitin synthesis is not understood, limiting the discovery of new inhibitors of this enzyme class. We identified the bacterial glycosyltransferase NodC as an appropriate model system to study the general structure and reaction mechanism of CHS. A high throughput screening-compatible novel assay demonstrates that a known inhibitor of fungal CHS also inhibit NodC. A structural model of NodC, on the basis of the recently published BcsA cellulose synthase structure, enabled probing of the catalytic mechanism by mutagenesis, demonstrating the essential roles of the DD and QXXRW catalytic motifs. The NodC membrane topology was mapped, validating the structural model. Together, these approaches give insight into the CHS structure and mechanism and provide a platform for the discovery of inhibitors for this antifungal target

The Fungal Cell Wall : Structure, Biosynthesis, and Function

N.G. is funded by the Wellcome Trust via a senior investigator award and a strategic award and by the MRC Centre for Medical Mycology. C.M. acknowledges the support of the Wellcome Trust and the MRC. N.G. and C.M. are part of the MRC Centre for Medical Mycology. J.P.L. acknowledges support from ANR, Aviesan, and FRM.Peer reviewedPublisher PD

Integration of fertility management, cultivar selection and alternative spray treatments to optimize control of foliar diseases of greenhouse grown tomatoes

Powdery mildew caused by Leveillula taurica (Lev.) Arn. is one of the most serious foliar diseases of greenhouse and open field tomato. The disease is currently controlled with the use of organic fungicides and sulphur, the latter being the only product permitted in organic crops. The aim of this study was to investigate the potential of controlling the disease by integrating: a) hybrids of low susceptibility to the disease, b) organic fertilisers (chitin) and c) alternative spray treatments. Some of the combinations of the above factors were highly effective in decreasing the percentage of disease severity. Specifically the combination of the hybrid of low susceptibility with the addition of chitin in the substrate and the spray treatment Milsana®+chitosan, was equally effective to sulphur. These results indicate that the combination of the above factors could probably be used as an alternative to sulphur for control of L. taurica in the greenhouse