Cytolethal distending toxin (CDT) of the Haemophilus parasuis SC096 strain contributes to serum resistance and adherence to and invasion of PK-15 and PUVEC cells

Cytolethal distending toxin (CDT) is proposed to be an important virulence determinant of many pathogens. Although two cdt gene cluster loci have been identified in Haemophilus parasuis strain SH0165, the characteristics of CDTs associated with pathogenesis remain unclear. In this study, three CDT-deficient mutants, cdt-1, cdt-2 and the double-knockout cdt-1cdt-2 (Δcdt-1, Δcdt-2 and Δcdt-1Δcdt-2, respectively), were obtained in the H. parasuis serovar 4 clinical strain SC096 using a natural transformation method. Compared to the wild-type SC096 strain, the Δcdt-1, Δcdt-2 and Δcdt-1Δcdt-2 mutants showed subtle growth defects and clearly exhibited an increased sensitivity to the bactericidal action of porcine and rabbit sera. Additionally, these mutants had a significantly reduced ability to adhere to and invade porcine umbilicus vein endothelial cells (PUVEC) and porcine kidney epithelial cells (PK-15). These findings suggest that both CDTs in the H. parasuis SC096 strain are involved in serum resistance and adherence and invasion of host cells.     

Denoising technique based on cascaded filtering of particle filter and ANFIS

For the practical application of nonlinear, non-Gaussian noise system state estimation,this paper develops the ANFIS- Particle filter cascaded filtering model based on the adaptive neuro-fuzzy inference system(ANFIS) nonlinear approximation function and particle filter’s obvious advantages for non-linear state estimation. ANFIS is used to eliminate the bias in the colored noise of the signal, then the filtered signal is processed by the particle filter to realize the optimal state estimation. The simulation results demonstrate that with the cascade filter model the mean and variance are reduced by 65% and 74% respectively, ANFIS-particle filter model has significant noise cancellation effect for strongly nonlinear systems, and the state estimation accuracy has been greatly enhanced, which verifies the effectiveness of the proposed model.     

Direct imaging of the diacetylene solid-state monomer-polymer phase transformation

The solid-state phase transformation from 1,6-di(N-carbazolyl)-2-4-hexadiyne (DCHD) diacetylene monomer to polymer has been studied dynamically by low-dose selected area electron diffraction and high-resolution electron microscopy. The total exposure required to induce polymerization is five orders of magnitude smaller than the critical dose for electron beam damage. The phase transformation is quasi-homogeneous, with the lattice parameters changing continuously as a function of beam dose. Characteristic streaking that develops in the selected area electron diffraction patterns in the [200] reciprocal directions during the intermediate stages of the transformation provides information about the defect-mediated mechanisms of this reaction.     

Prevalence of plasmid-mediated quinolone resistance qnr genes in poultry and swine clinical isolates of Escherichia coli

The prevalence of qnr genes was investigated in veterinary clinical isolates of Escherichia coli in Guangdong province, China, and the aac (6')-Ib gene and the mutations in QRDRs of gyrase and topoisomerase IV were examined in qnr-positive strains. A total of 232 E. coli strains isolated from pig and poultry were screened for the presence of the qnrA, qnrB and qnrS genes by PCR and sequencing. The aac (6')-Ib gene was detected in qnr-bearing strains by PCR and sequencing. For all strains carrying qnr, MICs for six quinolones were determined. Mutations within the gyrase and topoisomerase were analyzed by PCR and sequencing for all the QRDRs of gyrA, gyrB, parC and parE. Among 232 E. coli isolates, 14 (6%) isolates were positive for the qnr gene, including one for qnrB, 13 for qnrS, but no qnrA was identified in this population. Detection of the aac (6')-Ib gene showed that one qnrS-positive isolate from pig and one qnrB-positive isolate from duck carried aac (6')-Ib gene, and both were the cr variant allele of aac (6')-Ib. All of the 14 isolates had MICs of ciprofloxacin more than 0.25 mg/L. Mutations in the QRDR of gyrA mutations were observed in 5 (35.7%) of the 14 strains. Three fluoroquinolone-resisting strains showed one mutation S83L of gyrA, while one S83I. One high-level resistance strains harboured gyrA S83L and A87N of gyrA. A singe mutation in site 58 of parC was detected in 3 (21.4%) strains. None mutations were found in QRDRs of gyrB and parE. The emergence of qnr genes in veterinary clinical E. coli isolates is described for the first time. This is also the first report of aac (6')-Ib-cr gene in E. coli isolates from food-producing animals.     

The Influence of the Arrangements of Pipes on Gas solid Flow Characteristics in the Dense Section of the Stand Pipe

The influence of the arrangements of pipes on gas solid flow characteristics in the dense section of standpipe in the circulating fluidized bed is studied on a cold state experiment platform.The influence relationships of the percentage of area of pipes in the standpipe cross section and section shape of pipes and the circulation rate,adjusting range and distribution percentage of loose air are studied emphasizedly.     

The Finite Element Simulation of Pipe Bend Forming

A dynamic explicit finite element method is used to pipe bend forming processes on bender in this paper.The degenerated shell element and co-rotational coordinate system are employed.The material of pipe is assumed to satisfy Hill's anisotropic elasto-plastic yield condition.The bend die and press die are assumed as rigid surfaces and the contact force is computed by penalty method.Several numerical examples are given.They show that this method is effective.     

Profiling of miRNAs in porcine Sertoli cells

Background: Sertoli cells(SCs) create a specialized environment to support and dictate spermatogenesis.MicroRNAs(miRNAs), a kind of ~ 22 nt small noncoding RNAs, have been reported to be highly abundant in mouse SCs and play critical roles in spermatogenesis. However, the miRNAs of porcine SCs remain largely unknown.Methods: We isolated porcine SCs and conducted small RNA sequencing. By comparing miRNAs in germ cells, we systematically analyzed the miRNA expression pattern of porcine SCs. We screened the highly enriched SC miRNAs and predicted their functions by Gene Ontology analysis. The dual luciferase assay was used to elucidate the regulation of tumor necrosis factor receptor(TNFR)-associated factor 3(TRAF3) by ssc-miR-149.Results: The analysis showed that 18 miRNAs were highly expressed in SCs and 15 miRNAs were highly expressed in germ cells. These miRNAs were predicted to mediate SC and germ cell functions. In addition, ssc-miR-149 played critical roles in SCs by targeting TRAF3.Conclusion: Our findings provide novel insights into the miRNA expression pattern and their regulatory roles of porcine SCs.     

Heat shock proteins in canine transmissible venereal tumor

SDS-PAGE, Western blot analysis and immunohistochemical staining were used to detect heat shock proteins (HSPs) 60, 70 and 90 in canine transmissible venereal tumor (CTVT). Tissues tested for HSPs included: (1) tissues from different growth phases of CTVT tumors artificially induced in dogs; (2) tissues from other canine tumors; (3) normal dog tissues. Our results indicate that HSP 60 was consistently higher in CTVT cells in regressing phase than those in progressing phase. However, no detectable antibody response specific to the tested HSPs was found in the sera from CTVT-laden dogs in different growth phases. Although levels of the HSPs were all detectable in CTVT cells, only 60 and 70 were higher in CTVT cells than in normal tissues. In addition, none of the HSPs were detected in cells from five other canine tumors. These data suggest that canine HSP 60 and 70 are potential markers for CTVT and HSP 60 is appear to be involved in CTVT regression.PCR was used to confirm the existence of CTVT cells using primers designed to cover the sequence between the 5' end of c-myc near the first exon and the 3' end outside the LINE gene. Only CTVT samples were positive for this sequence; samples from other tumors and normal tissues were negative. The sequenced PCR products indicated that CTVT from Taiwan and other countries exhibited over 98% sequence homology. This reconfirms that, worldwide, all CTVT cells are very similar.