Cytolysin A-mediated protein exportation efficiency and its role in enhancing the fitness of live recombinant Salmonella Typhi vaccine strain.

The genetic fusion of cytolysin A (clyA) to heterologous antigen expressed in live Salmonella vector demonstrated efficient translocation into periplasmic space and extracellular medium. Accumulating evidence has shown that clyA-mediated antigen delivery improved growth fitness and enhanced immunogenicity of live vector vaccine, but the factors influencing this protein exportation has not been investigated. In this study, Toxoplasma gondii antigen fused at C-terminal of clyA protein was expressed in live S. Typhi vector via both plasmid and chromosomal-based expressions. The bivalent strains showed comparable growth rates as monovalent strains, but in varies antigen exportation efficiency. ClyA-fusion antigen with positive charges was translocated to the extracellular spaces, whereas those with negative charges were retained in the cytoplasm. Furthermore, excessive cellular resources expenditure on antigen expression, especially antigen with larger size, could limit the clyA-fusion antigen exportation, resulting in undesirable metabolic burden that eventually affects the growth fitness. Altogether, the present work indicates potential linkage of factors mainly on antigen properties and expression platforms that may affect clyA-mediated antigen delivery to enhance the growth fitness of live vector strain.

Stability, geometry and electronic properties of BH n (n = 0 to 3) radicals on the Si{0 0 1}3 × 1:H surface from first-principles.

A new generation of radiation detectors relies on the crystalline Si and amorphous B (c-Si/a-B) junctions that are prepared through chemical vapor deposition of diborane (B2H6) on Si at low temperature (~400 °C). The Si wafer surface is dominated by the Si{0 0 1}3 × 1 domains that consist of two different Si species at low temperature. Here we investigate the geometry, stability and electronic properties of the hydrogen passivated Si{0 0 1}3 × 1 surfaces with deposited BH n (n = 0 to 3) radicals using parameter-free first-principles approaches. Ab initio molecular dynamics simulations using the density functional theory (DFT) including van der Waals interaction reveal that in the initial stage the BH3 molecules/radicals deposit on the Si(-H), forming (-Si)BH4 radicals which then decompose into (-Si)BH2 with release of H2 molecules. Structural optimizations provide strong local relaxation and reconstructions at the deposited Si surface. Electronic structure calculations reveal the formation of various defect states in the forbidden gap. This indicates limitations of the presently used rigid electron-counting and band-filling models. The attained information enhances our understanding of the initial stage of the PureB process and the electric properties of the products.

Structure and stability of hcp iron carbide precipitates: A first-principles study.

Hexagonal close-packed (hcp) iron carbides play an important role in steel processing and in steel products. The recent discovery of novel ultrafine (2-5 nm) iron carbide (ε'-Fe2+xC) precipitates in TRIP steel sheds a new light on the hcp family of carbides. Here we present a first-principles study on the relative stability, and the electronic, magnetic properties of the ε'-Fe2C phases. Different stackings of Fe-sheets and orderings of C atoms were investigated and compared with experimental data and with Jack's model. We find very favorable formation enthalpies for these new members of the hcp family, and we present a first-principles-refined model for the crystal structure of the ultrafine Fe(C) precipitates. These findings are useful for the characterization of nano-sized iron carbide precipitates, for understanding their role in the microstructure of steels, and for the design of novel steels having even more desirable properties.

Stability and geometry of silica nano-ribbons (SNRs): a first-principles study.

Silica based materials are attractive because of their versatility and their unique structures and properties, which have led to numerous applications of silica in a range of fields. Recently, various low-dimensional silica materials have been synthesized experimentally. Here we present a first-principles study on the geometry and stability of novel low-dimensional silica nano-ribbons (SNRs) using density-functional theory (DFT) with van der Waals interactions (optB88-vdW). SNRs of various widths with different surface groups, and with the geometry of hexagonal rings and squares, were taken into consideration. An atomically flat ribbon with mixing squares and rings is also included. The calculations showed high stability for the single layer and bilayer silica ribbons, both containing hexagonal rings. The calculations also revealed a high flexibility of silica chains. The local structure and chemical bonding were carefully analyzed. Electronic band structure calculations showed an insulating nature of the SNRs with energy gaps of about 5.0 to 6.0 eV, which are determined by nonbonding and anti-bonding O 2p states.

A multi-nuclear magnetic resonance and density functional theory investigation of epitaxially grown InGaP2.

In this paper the short and long range order in In0.483Ga0.517P thin films is investigated by solid-state Nuclear Magnetic Resonance (NMR) spectroscopy. To this end two samples were grown on a GaAs substrate by metal-organic vapor phase epitaxy at two different growth-pressures. From band gap energy measurements, CuPt long range order parameters of SCuPt = 0.22 and 0.39 were deduced, respectively. In the (31)P spectrum five resonances are observed corresponding to the five possible P(GanIn4-n), n = 0-4, coordinations whose relative intensities correspond to the order in the material, but the intensity variations for order parameters between 0 and 0.5 are minimal. (69)Ga, (71)Ga and (115)In (MQ)MAS spectra were acquired to analyze the quadrupolar and chemical shift distributions related to the (dis)order in these materials in more detail. All these spectra clearly reflect the disorder in the sample and do not show the presence of highly ordered domains. The difference in the order parameter in the sample is not clearly reflected in the spectra. (31)P chemical shifts were calculated using Density Functional Theory. The experimentally observed shifts are well reproduced with a simple random model of the disorder, thus confirming the assignment of the resonances. The (31)P chemical shifts are very sensitive to changes in the lattice parameter and chemical surroundings. These effects nearly compensate and explain why the (31)P chemical shifts in pure InP and GaP are nearly identical whereas a large difference would be expected based on the observed shift difference for the P[In4] and P[Ga4] coordinations in In0.483Ga0.517P.

Unique contribution of IRF-5-Ikaros axis to the B-cell IgG2a response.

IRF-5 is a transcription factor activated by toll like receptor (TLR)7 and TLR9 during innate immune responses. IRF-5 activates not only Type I IFN, but also inflammatory cytokines. Most importantly, a genetic variation in the IRF-5 gene shows a strong association with autoimmune diseases such as Lupus. Here, we report that IRF5-deficient mice have attenuated IgG2a/c responses to T-cell-dependent and -independent antigens and to polyoma virus infection. This defect is due to the intrinsic deletion of IRF-5 in B cells, as SCID mice reconstituted with Irf5-/- B cells show a decrease in IgG2a/c expression after viral infection compared with mice that received wild-type B cells. Irf5-/-B cells in vitro have diminished TLR and cytokine-induced class switching to IgG2a/c. Addressing the molecular mechanism, we show that IRF-5 regulates IgG2a/c expression by decreasing Ikaros expression; reconstitution of IRF-5 in Irf5-/- B cells downregulates Ikaros levels and increases switching to IgG2a/c. The IRF site in ikzf1 promoter binds IRF-5, IRF-4 and IRF-8. We show that IRF-8 but not IRF-4 activates the ikzf1 promoter, and IRF-5 inhibits the transcriptional activity of IRF-8. Collectively, these results identify the IRF-5-Ikaros axis as a critical modulator of IgG2a/c class switching.

Origin of predominance of cementite among iron carbides in steel at elevated temperature.

A long-standing challenge in physics is to understand why cementite is the predominant carbide in steel. Here we show that the prevalent formation of cementite can be explained only by considering its stability at elevated temperature. A systematic highly accurate quantum mechanical study was conducted on the stability of binary iron carbides. The calculations show that all the iron carbides are unstable relative to the elemental solids, α-Fe and graphite. Apart from a cubic Fe23C6 phase, the energetically most favorable carbides exhibit hexagonal close-packed Fe sublattices. Finite-temperature analysis showed that contributions from lattice vibration and anomalous Curie-Weis magnetic ordering, rather than from the conventional lattice mismatch with the matrix, are the origin of the predominance of cementite during steel fabrication processes.

Down-regulated expression of atypical PKC-binding domain deleted asip isoforms in human hepatocellular carcinomas.

Asip is a mammalian homologue of polarity protein Par-3 of Caenorhabditis elegans and Bazooka of Drosophila melanogaster. Asip/Par-3/Bazooka are PDZ-motif containing proteins that localize asymmetrically to the cell periphery and play a pivotal role in cell polarity and asymmetric cell division. In the present study, we have cloned human asip cDNA and its splicing variants by 5'-RACE and RT-PCR using candidate human EST clones which have a high homology to rat asip cDNA. The full-length cDNA of human asip encodes a 1,353 aa protein exhibiting 88% similarity to the rat one. Human asip is a single copy gene consisting of at least 26 exons and localizing in human chromosome 10, band p11.2, with some extraordinarily long introns. All exon/intron boundary nucleotides conform to the "gt-ag" rule. Three main transcripts were detected by Northern blot analysis, and at least five variants, from alternative splicing and polyadenylation, have been identified by RT-PCR and liver cDNA library screening. Exon 17b deleted asip mRNAs expressed ubiquitously in normal human tissues, including liver, on RT-PCR analysis. However, they were absent from most human liver cancer cell lines examined. More interestingly, the expression of exon 17b deleted variants was down regulated in 52.6% (10/19) clinic specimens of human hepatocellular carcinomas (HCCs), compared with the surrounding nontumorous liver tissues from the same patients. The presence of various splicing transcripts, the variation of their distribution among different tissues and cells, and their differential expressions in human HCCs suggest that human Asip isoforms may function in different context.

Deregulated c-myc expression in quiescent CHO cells induces target gene transcription and subsequent apoptotic phenotype.

Human c-myc cDNA was fused with the hormone-binding domain (HBD) cDNA of murine estrogen receptor gene and the chimeric gene was introduced into the CHO cells. The fusion protein, c-MycER, becomes activated when the synthetic steroid, 4-hydroxy-tamoxifen (OHT), binds HBD. Activated c-MycER, likely c-Myc, can induce quiescent CHO cells reentry into S phase and subsequent cell death under serum-free condition. In addition, the expression of some proposed c-myc target genes such as ODC, MrDb, cad, rcc1 and rcl were found to increase upon OHT induction before S phase entry and apoptosis, indicating that these target genes are involved in cell cycle regulation and/or apoptosis control. However, the mutant D106-143c-MycER protein does not have above activities.

Reverse pedicled lateral arm flap for reconstruction of posterior soft-tissue defects of the elbow.

Posterior soft-tissue defects of the elbow are difficult to reconstruct by conventional techniques such as closure by approximation or skin graft. An ideal technique should be an easy and reliable one-stage procedure that provides predictable surgical results with regard to elbow function and cosmesis. This report details our experience in 7 patients who underwent a one-stage procedure for coverage of the posterior elbow employing the reverse pedicled lateral arm flap. All flaps survived and all patients were able to resume full range of motion of the elbow joint at the 6-month follow-up. Complications included forearm paraesthesia in 3 patients and conspicuous scarring in a young female patient. We emphasize two valuable refinements in surgical technique including measuring posterior elbow defect in full flexion and postoperative elbow extension splinting. In trauma-related defects of the posterior elbow, a preoperative angiogram is important before raising this flap.

Pyomyositis of the calf muscles mimicking distal deep venous thrombosis: a case report.

Pyomyositis is a pyogenic infection of skeletal muscle. It is relatively rare in temperate climates. Unfamiliarity with the lesion may lead to a delay in diagnosis. This report describes pyomyositis of the calf muscles in a patient with chronic hepatitis whose clinical symptoms simulated those of distal deep venous thrombosis. The correct diagnosis was not made until computed tomography revealed a local abscess in the calf muscles.

Intradural lumbar disc herniation--a case report.

One case of intradural lumbar disc herniation at the L3-L4 disc level with cauda equina syndrome is reported. Myelo-CT demonstrated an intradural tumor-like lesion with complete block. An intradural fragment of sequestrated disc material was found intraoperatively. Accurate preoperative diagnosis of the intradural nature of the disease and prompt surgical treatment resulted in a smooth recovery.

[Measurements of the lumbar spinal canal of Chinese by computed tomography].

The study of the lumbar canal sizes in Chinese has not been reported before. Thirty adult patients, aged 20 to 40 years, who sustained thoracolumbar or lumbar fracture-dislocation and needed further computed tomographic (CT) scanning were included in this study. There was no significant back pain or sciatica on these patients before. Measurements of the med-sagittal anteroposterior (A-P) and interpedicular (I-P) diameters of the spinal canals L1 to L5 were requested at the same time during CT scanning. The results show that the lowest normal limits of the A-P and I-P diameters of the lumbar canals in Chinese are 11 mm and 19 mm respectively. The I-P diameters increase progressively from the first to the fifth vertebrae. The A-P diameters, however, are more variable and shortest at L2, L3 and L4 vertebrae.