Genome-Wide Transcriptional Response of Silkworm (Bombyx mori) to Infection by the Microsporidian Nosema bombycis

Microsporidia have attracted much attention because they infect a variety of species ranging from protists to mammals, including immunocompromised patients with AIDS or cancer. Aside from the study on Nosema ceranae, few works have focused on elucidating the mechanism in host response to microsporidia infection. Nosema bombycis is a pathogen of silkworm pébrine that causes great economic losses to the silkworm industry. Detailed understanding of the host (Bombyx mori) response to infection by N. bombycis is helpful for prevention of this disease. A genome-wide survey of the gene expression profile at 2, 4, 6 and 8 days post-infection by N. bombycis was performed and results showed that 64, 244, 1,328, 1,887 genes were induced, respectively. Up to 124 genes, which are involved in basal metabolism pathways, were modulated. Notably, B. mori genes that play a role in juvenile hormone synthesis and metabolism pathways were induced, suggesting that the host may accumulate JH as a response to infection. Interestingly, N. bombycis can inhibit the silkworm serine protease cascade melanization pathway in hemolymph, which may be due to the secretion of serpins in the microsporidia. N. bombycis also induced up-regulation of several cellular immune factors, in which CTL11 has been suggested to be involved in both spore recognition and immune signal transduction. Microarray and real-time PCR analysis indicated the activation of silkworm Toll and JAK/STAT pathways. The notable up-regulation of antimicrobial peptides, including gloverins, lebocins and moricins, strongly indicated that antimicrobial peptide defense mechanisms were triggered to resist the invasive microsporidia. An analysis of N. bombycis-specific response factors suggested their important roles in anti-microsporidia defense. Overall, this study primarily provides insight into the potential molecular mechanisms for the host-parasite interaction between B. mori and N. bombycis and may provide a foundation for further work on host-parasite interaction between insects and microsporidia.     

Characterization of Lignins Isolated with Alkaline Ethanol from the Hydrothermal Pretreated Tamarix ramosissima

The objective of this study was to characterize the changes in lignin structure during hydrothermal pretreatment of shrub Tamarix ramosissima. Lignins in residual wood meal were isolated with alkaline ethanol solution and recovered with acid precipitation. A comparison between the recovered lignin fractions with milled wood lignin has been made in terms of yield, purity, gel permeation chromatography, Fourier transform infrared spectroscopy, pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), 1D ¹³C and 2D heteronuclear single quantum coherence nuclear magnetic resonance (HSQC NMR) spectroscopic techniques. Semiquantitative HSQC NMR showed that the relative amounts of β-O-4′ (around 76 % side chains) and resinol type substructures (16 %) of lignins were significantly modified during hydrothermal pretreatment. Py-GC/MS analyses brought direct evidences of these lignin samples with high S/G ratios ranging from 1.7 to 2.6. Moreover, the results indicated that an increase in the severity of the hydrothermal pretreatment enhanced the degradation of lignin unit side chains and the condensation of lignin and decreased the molecular weight of the recovered lignin fractions. This study demonstrated that the combination of autohydrolysis and alkaline ethanol process could potentially turn the recovered lignin fractions into value added products being in accordance with the “biorefinery” concept.     

Developability studies before initiation of process development

Monoclonal antibodies constitute a robust class of therapeutic proteins. Their stability, resistance to stress conditions and high solubility have allowed the successful development and commercialization of over 40 antibody-based drugs. Although mAbs enjoy a relatively high probability of success compared with other therapeutic proteins, examples of projects that are suspended due to the instability of the molecule are not uncommon. Developability assessment studies have therefore been devised to identify early during process development problems associated with stability, solubility that is insufficient to meet expected dosing or sensitivity to stress. This set of experiments includes short-term stability studies at 2−8 þC, 25 þC and 40 þC, freeze-thaw studies, limited forced degradation studies and determination of the viscosity of high concentration samples. We present here three case studies reflecting three typical outcomes: (1) no major or unexpected degradation is found and the study results are used to inform early identification of degradation pathways and potential critical quality attributes within the Quality by Design framework defined by US Food and Drug Administration guidance documents; (2) identification of specific degradation pathway(s) that do not affect potency of the molecule, with subsequent definition of proper process control and formulation strategies; and (3) identification of degradation that affects potency, resulting in program termination and reallocation of resources.     

Evaluation of genetic diversity of Chinese Pleurotus ostreatus cultivars using DNA sequencing technology

Pleurotus spp. are well-known and economically important cultivated mushrooms in China. Knowledge of the genetic relationship between the Chinese cultivars is essential to the improvement of P. ostreatus strains. Sequence analysis of the internal transcribed spacers (ITS), translation elongation factor (EF1α) and the second largest subunit of RNA polymerase II (RPB2) was performed to assess the genetic diversity of Pleurotus ostreatus strains cultivated in China. The phylogenetic tree constructed using the combined results of the ITS, EF1α and RPB2 sequence analyses showed the genetic relationships between the studied strains. Our phylogenetic analyses therefore provided valuable information on the relationships among the P. ostreatus strains used in this study and that was useful for examining genetic diversity among these strains.     

The effect of polycyclic aromatic hydrocarbon contamination on distribution of the Sphingomonas community in the Shenfu irrigation area of Northeast China

Information about the diversity and community structure of indigenous Sphingomonas communities in natural environments is lacking. In this study, denaturing gradient gel electrophoresis (DGGE) was used to investigate Sphingomonas communities at nine selected sites from the up-, mid- and downstream regions of a wastewater channel, which once flowed with sewage containing high concentrations of polycyclic aromatic hydrocarbons (PAHs). From each region, three samples from channel sediment, rice soil and corn soil were collected. Sediment sites had significantly higher PAH contamination, followed by rice sites and corn sites. In addition, upstream sites had higher PAH accumulation, followed by mid- and downstream sites. For each sample type (sediment, rice and corn soils), the Shannon diversity indices of the Sphingomonas community increased slightly with increasing PAH contamination. Upstream sites had obviously higher diversity than mid- and downstream sites. Both cluster analysis and canonical correspondence analysis indicated that the Sphingomonas community was clearly different among sediment, rice and corn soils. Besides, the Sphingomonas community was affected by different PAH compounds in sediment, rice and corn sites. The Sphingomonas community might degrade mainly benzo[b]fluoranthene, fluorene and fluoranthene in sediment sites by co-metabolism, but degraded mainly pyrene and phenanthrene in corn and rice sites, which provides some suggestions for pollution remediation.     

Eph/ephrin recognition and the role of Eph/ephrin clusters in signaling initiation

The Eph receptors and their ephrin ligands play crucial roles in a large number of cell–cell interaction events, including those associated with axon pathfinding, neuronal cell migration and vasculogenesis. They are also involved in the patterning of most tissues and overall cell positioning in the development of the vertebrate body plan. The Eph/ephrin signaling system manifests several unique features that differentiate it from other receptor tyrosine kinases, including initiation of bi-directional signaling cascades and the existence of ligand and receptor subclasses displaying promiscuous intra-subclass interactions, but very rare inter-subclass interactions. In this review we briefly discuss these features and focus on recent studies of the unique and expansive high-affinity Eph/ephrin assemblies that form at the sites of cell–cell contact and are required for Eph signaling initiation. This article is part of a Special Issue entitled: Emerging recognition and activation mechanisms of receptor tyrosine kinases.     

Display of heterologous protein on the surface of Lactobacillus plantarum by using the CspI anchor protein

The C-terminus of the putative cell surface protein CspI which contains one putative LPxTG motif region and a signal peptides fragment were amplified from L. plantarum CICC6024, and the green fluorescent protein gene gfp was amplified from the plasmid pACGFP. The three genes were ligated and the fusion gene was named SgfpL. The fusion gene SgfpL was then cloned into shuttle expression vector pMG36e and transformed into L. plantarum. SDS-PAGE identified that the fusion protein was expressed and the band of fusion protein was observed at the predicated molecular size. Fluorescence assay, western blot against GFP antibody, protease accessibility and SDS sensitivity assays were performed to determine that the GFP was successfully displayed on the surfaces of L. plantarum cells and the maximum display capacity of the GFP fusion protein was ca. 65 μg?ml^?1. The fermentation condition experiments determined that the amounts of GFP fusion protein were increased at a higher temperature and reached the peak at 2.5 h. Then, the β-galactosidase from Bifidobacterium bifidum was functionally displayed on the surface of L. plantarum cells via CspI to demonstrate the applicability of the CspI-mediated surface display system.