IL‐23 plays a significant role in the augmentation of particulate matter‐mediated allergic airway inflammation

It has been recently that particulate matter (PM) exposure increases the risk and exacerbation of allergic asthma. However, the underlying mechanisms and factors associated with increased allergic responses remain elusive. We evaluated IL‐23 and IL‐23R (receptor) expression, as well as changes in the asthmatic phenotype in mice administered PM and a low dose of house dust mite (HDM). Next, changes in the phenotype and immune responses were evaluated after intranasal administration of anti‐IL‐23 antibody during co‐exposure to PM and low‐dose HDM. We also performed in vitro experiments to investigate the effect of IL‐23. IL‐23 expression was significantly increased in Epcam+CD45− and CD11c+ cells, while that of IL‐23R was increased in Epcam+CD45− cells only in mice administered PM and low‐dose HDM. Administration of anti‐IL‐23 antibody led to decreased airway hyperresponsiveness, eosinophils, and activation of dendritic cells, reduced populations of Th2 Th17, ILC2, the level of IL‐33 and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Inhibition of IL‐23 in PM and low‐dose HDM stimulated airway epithelial cell line resulted in decreased IL‐33, GM‐CSF and affected ILC2 and the activation of BMDCs. PM augmented the phenotypes and immunologic responses of asthma even at low doses of HDM. Interestingly, IL‐23 affected immunological changes in airway epithelial cells.     

Identification of lucerne stem cell wall traits related to in vitro neutral detergent fibre digestibility

Genetic improvement of forage digestibility, especially utilizing marker assisted selection and recombinant DNA techniques, requires identification of specific biochemical traits and associated genes that impact digestibility. We undertook a study to identify cell wall (CW) traits of lucerne (Medicago sativa L.) stems that were consistently and strongly correlated with in vitro neutral detergent fibre (NDF) digestibility, a measurement that has been shown to correlate with animal performance. Spring and summer harvested lucerne stem material, for 2 years, from 24 individual plants in each of two germplasm sources were analyzed for 16 and 96 h in vitro NDF digestibility, and cell wall concentration and composition (monosaccharide constituents of cellulose, hemicellulose, and pectin; and Klason lignin (KL)) by the Uppsala dietary fibre method using near-infrared reflectance spectroscopy (NIRS). Pearson correlation coefficients were calculated for the relationships among these cell wall traits and with in vitro NDF digestibility. Concentrations of the pectin monosaccharide components were all negatively correlated (r=−0.73 to −0.94) with total cell wall concentration. In contrast, the three most abundant cell wall components glucose (Glc), xylose (Xyl) and Klason lignin were not correlated, or only weakly positively correlated (r<0.35), with cell wall concentration. Cell wall concentration was consistently negatively correlated (r=−0.60 to −0.94) with both 16 and 96 h in vitro NDF digestibility. In contrast, Klason lignin concentration was only marginally correlated (r<0.30) with 16 h in vitro NDF digestibility, but strongly negatively correlated (r=−0.71 to −0.74) with 96 h in vitro NDF digestibility. This is consistent with previous reports which show that lignin affects potential extent of digestion, but not rate. Cell wall glucose and xylose concentrations were inconsistently correlated with fibre digestibility. The monosaccharide components of pectin were consistently positively correlated (r=0.54–0.90) with in vitro NDF digestibility, except for 96 h in vitro NDF digestibility of spring harvested stems. Growth environment (year) and germplasm source had only minor impacts on the preceding correlation patterns, whereas spring versus summer harvests accounted for the inconsistencies observed among correlations for cell wall traits. The results of this study indicate that genetic improvement of fibre digestibility of lucerne stems should target genes that reduce total cell wall concentration, perhaps by reducing the rate of xylem tissue deposition during maturation, and reduce Klason lignin and increase pectin concentrations in the cell wall to improve potential extent and rate of fibre digestibility, respectively.     

Kinetic Monte Carlo simulation for the striation distribution of void defects in Czochralski silicon growth

Unique crystal-originated pit (COP) distribution, similar to a striation pattern, is well matched with the oxygen profile in experimental analysis. It shows the strong relationship between oxygen concentration and COP distribution. In this paper, we study the generation of void defects and the relationship between interstitial oxygen and vacancy using the kinetic lattice Monte Carlo (KLMC) method. The KLMC method has been applied extensively in various forms to the study of micro-defects in silicon wafers. It explained well the formation of void defects such as vacancy–oxygen complex and vacancy–vacancy complex. The formation of clusters is strongly affected by oxygen concentration, which showed the relationship between COP distribution and oxygen concentration. The unique COP distribution could be correctly explained with KLMC results, and this kind of meso-scale results has not yet been reported.     

Staphyloferrin A: a structurally new siderophore from staphylococci

Two ferric ion-binding compounds, designated staphyloferrin A and B, were detected in the culture filtrates of staphylococci grown under iron-deficient conditions. Staphyloferrin A was isolated from cultures of Staphylococcus hyicus DSM 20459. The structural elucidation of this highly hydrophilic, acid-labile compound revealed a novel siderophore, N2,N5-di-(1-oxo-3-hydroxy-3,4-dicarboxybutyl)-D-ornithine, which consists of one ornithine and two citric acid residues linked by two amide bonds. The two citric acid components of staphyloferrin A provide two tridentate pendant ligands, comprising of a beta-hydroxy, beta-carboxy-substituted carboxylic acid derivative, for octahedral metal chelation. The CD spectrum of the staphyloferrin A ferric complex indicates a predominant A configuration about the ferric ion center. The uptake of ferric staphyloferrin A by S. hyicus obeys Michaelis-Menten kinetics (Km = 0.246 microM; vmax = 82 pmol.mg-1.min-1), indicating active transport of this siderophore. The staphyloferrin A transport system is different from that of the ferrioxamines as shown by an antagonism test. Production of staphyloferrin A is strongly iron-dependent and is stimulated by supplementation of the medium with either D- or L-ornithine. DL-[5-14C]ornithine was incorporated into staphyloferrin A, demonstrating that ornithine is an intermediate in staphyloferrin A biosynthesis.     

Solid cultures of thrips-pathogenic fungi Isaria javanica strains for enhanced conidial productivity and thermotolerance

Entomopathogenic fungi have great potential to control agricultural and horticultural insect pests, however optimizing conidial production systems to demonstrate high productivity and stability still needs additional efforts for successful field application and industrialization. Although many virulent entomopathogenic fungal isolates have been viewed as potential candidates in a laboratory environment, very few of the isolates are being used in practice for application in agricultural fields as commercial products. I. javanicus is an entomopathogenic fungus that is parasitic to various diverse coleopteran and lepidopteran insects and thought good candidate as biopesticdes. In this work, the basic characteristics of two entomopathogenic fungi, I. javanica FG340 and Pf04, were investigated in morphological examinations, genetic identification, and virulence against Thrips palmi, and then the feasibility of various grains substrates for conidial production was assessed, particularly focusing on conidial productivity and thermotolerance. Isaria javanica FG340 and Pf04 conidia were solid-cultured on 12 grains for 14?days in a Petri dish. Of the tested Italian millet, perilla seed, millet and barley-based cultures showed high conidial production. The four-grain media yielded >1?×?10⁹ conidia/g of I. javanica FG340 and Pf04. Pf04 strain had enhanced thermotolerance up to 45?°C when cultured on Italian millet. In application, it was easy to make a conidial suspension using the cultured grains, and several surfactants were tested to release the conidia. This work suggests several possible inexpensive grain substrates by which to promote conidial production combined with enhanced stability against exposure to high temperature.     

Optimization of algal photobioreactors using flashing lights

It has been reported that flashing light enhances microalgal biomass productivity and overall photosynthetic efficiency. The algal growth kinetics and oxygen production rates under flashing light with various flashing frequencies (5 Hz-37 kHz) were compared with those under equivalent continuous light in photobioreactors. A positive flashing light effect was observed with flashing frequencies over 1 kHz. The oxygen production rate under conditions of flashing light was slightly higher than that under continuous light. The cells under the high frequency flashing light were also observed to be healthier than those under continuous light, particularly at higher cell concentrations. When 37 kHz flashing light was applied to an LED-based photobioreactor, the cell concentration was higher than that obtained under continuous light by about 20%. Flashing light may be a reasonable solution to overcome mutual shading, particularly in high-density algal cultures.     

Species-crossreactive scFv against the tumor stroma marker "fibroblast activation protein" selected by phage display from an immunized FAP-/- knock-out mouse.

BACKGROUND: Fibroblast activation protein (FAP) is a type II membrane protein expressed on tumor stroma fibroblasts in more than 90% of all carcinomas. FAP serves as a diagnostic marker and is potential therapeutic target for treatment of a wide variety of FAP+ carcinomas. Murine tumor stroma models and FAP-specific antibodies are required to investigate the functional role of FAP in tumor biology and its usefulness for drug targeting. We here describe the development of antibodies with crossreactivity for human (hFAP) and murine FAP (mFAP), which share 89% amino acid identity. MATERIAL AND METHODS: An FAP-/- mouse was sequentially immunized with recombinant murine and human FAP-CD8 fusion proteins. Immunoglobulin cDNA derived from hyperimmune spleen cells was used for the construction of a combinatorial single chain Fv (scFv) library. Phage display selection of FAP-specific scFv was performed on immobilized hFAP followed by selection on cells expressing murine FAP. RESULTS: High-affinity, species-crossreactive, FAP-specific scFv were isolated upon sequential phage display selection. A bivalent derivative (minibody M036) constructed thereof was applied for immunohistochemical analyses and allowed detection of FAP expression on stroma cells of different human carcinomas as well as on murine host stroma in a tumor xenograft model. CONCLUSIONS: MB M036, derived from phage display selected species crossreactive scFv, is suitable for tumor stroma targeting and will be a valuable tool in the analyses of the functional role of FAP in tumor biology as well as in the evaluation of the suitability of FAP for drug targeting.     

Eicosanoids in insect immunity: bacterial infection stimulates hemocytic phospholipase A2 activity in tobacco hornworms

Intracellular phospholipase A(2) (PLA(2)) is responsible for releasing arachidonic acid from cellular phospholipids, and is thought to be the first step in eicosanoid biosynthesis. Intracellular PLA(2)s have been characterized in fat body and hemocytes from tobacco hornworms, Manduca sexta. Here we show that bacterial challenge stimulated increased PLA(2) activity in isolated hemocyte preparations, relative to control hemocyte preparations that were challenged with water. The increased activity was detected as early as 15 s post-challenge and lasted for at least 1 h. The increased activity depended on a minimum bacterial challenge dose, and was inhibited in reactions conducted in the presence of oleyoxyethylphosphorylcholine, a site-specific PLA(2) inhibitor. In independent experiments with serum prepared from whole hemolymph, we found no PLA(2) activity was secreted into serum during the first 24 h following bacterial infection. We infer that a hemocytic intracellular PLA(2) activity is increased immediately an infection is detected. The significance of this enzyme lies in its role in launching the biosynthesis of eicosanoids, which mediate cellular immune reactions to bacterial infection.     

Lineage-specific evolution and gene flow in Listeria monocytogenes are independent of bacteriophages

Listeria monocytogenes is a foodborne pathogen causing systemic infection with high mortality. To allow efficient tracking of outbreaks a clear definition of the genomic signature of a cluster of related isolates is required, but lineage-specific characteristics call for a more detailed understanding of evolution. In our work, we used core genome MLST (cgMLST) to identify new outbreaks combined to core genome SNP analysis to characterize the population structure and gene flow between lineages. Whilst analysing differences between the four lineages of L. monocytogenes we have detected differences in the recombination rate, and interestingly also divergence in the SNP differences between sub-lineages. In addition, the exchange of core genome variation between the lineages exhibited a distinct pattern, with lineage III being the best donor for horizontal gene transfer. Whilst attempting to link bacteriophage-mediated transduction to observed gene transfer, we found an inverse correlation between phage presence in a lineage and the extent of recombination. Irrespective of the profound differences in recombination rates observed between sub-lineages and lineages, we found that the previously proposed cut-off of 10 allelic differences in cgMLST can be still considered valid for the definition of a foodborne outbreak cluster of L. monocytogenes.