Regioselective Syntheses of 1,4- and 1,6-Dicarbonyl Compounds via Photoredox-Based Oxidative Heterocoupling of Enolsilanes with Oxygen as an Oxidant.

A photoredox-based oxidative heterocoupling of enolsilanes to the corresponding 1,4- and 1,6-dicarbonyl compounds was developed by using Mes-Acr+BF4- as the photocatalyst, and oxygen was used as the oxidant. This newly developed chemistry adheres to the principles of atom economy, step economy, and redox economy, making it a concise and efficient method.

Non-Contact Current Sensing System Based on the Giant Magnetoimpedance Effect of CoFeNiSiB Amorphous Ribbon Meanders.

A sensitive non-contact sensing system based on the CoFeNiSiB amorphous ribbon giant magnetoimpedance (GMI) effect is proposed for current testing. The sensing system consists of a GMI probe, a sinusoidal current generator, a voltage follower, a preamplifier, a low-pass filter, and a peak detector. Four different GMI probes derived from amorphous ribbon meanders are designed and fabricated through MEMS processes. GMI probes were driven by a 10 MHz, 5 mA AC current. A permanent magnet was used to provide a bias magnetic field for the probe. The effect of the bias magnetic field on the output DC voltage was investigated. This non-contact current sensing system exhibits good sensitivity and linearity at a bias magnetic field Hbias = 15 Oe. The sensitivity can reach up to 24.2 mV/A in the ±1.5 A range.

Comparison of magnetoimpedance behaviors in meander CoFeNiSiB amorphous ribbon for deformation sensing applications.

In this work, micro-ribbon strips and meanders based on CoFeNiSiB amorphous ribbons were fabricated by using the lithography technique and chemical etching. Flat and curved holders with different radius of curvature were obtained via 3D printing techniques for GMI testing. Longitudinal and transverse GMI (LGMI and TGMI) behaviors of micro-ribbon sensors in different bending directions and degrees were systematically investigated. The results show the LGMI and TGMI effects of micro-ribbon meanders with one turn is most sensitive to bending. It can be used in the development of deformation sensors. In addition, there is a linear range of field in the LGMI and TGMI curves of micro ribbons under different bending conditions, and the sensitivity of micro-ribbon sensors shows no significant change in the range. In particular, the micro-ribbon meanders with three turns are the least sensitive to bending deformation and can be used to develop stable and flexible GMI sensors for wearable electronics devices.

Giant Magnetoimpedance Effect of Multilayered Thin Film Meanders Formed on Flexible Substrates.

The giant magnetoimpedance effect of multilayered thin films under stress has great application prospects in magnetic sensing, but related studies are rarely reported. Therefore, the giant magnetoimpedance effects in multilayered thin film meanders under different stresses were thoroughly investigated. Firstly, multilayered FeNi/Cu/FeNi thin film meanders with the same thickness were manufactured on polyimide (PI) and polyester (PET) substrates by DC magnetron sputtering and MEMS technology. The characterization of meanders was analyzed by SEM, AFM, XRD, and VSM. The results show that multilayered thin film meanders on flexible substrates also have the advantages of good density, high crystallinity, and excellent soft magnetic properties. Then, we observed the giant magnetoimpedance effect under tensile and compressive stresses. The results show that the application of longitudinal compressive stress increases the transverse anisotropy and enhances the GMI effect of multilayered thin film meanders, while the application of longitudinal tensile stress yields the opposite result. The results provide novel solutions for the fabrication of more stable and flexible giant magnetoimpedance sensors, as well as for the development of stress sensors.

Analysis of Prognosis and Risk Factors for Postoperative Hepatic Dysfunction in Patients with Acute Type A Aortic Dissection.

BACKGROUND: To explore the prognosis and risk factors of postoperative hepatic dysfunction in patients with acute type A aortic dissection. METHODS: A total of 156 patients who underwent surgery for acute type A aortic dissection in our hospital from May 2014 to May 2018 were retrospectively enrolled. The patients were divided into 2 groups based on postoperative liver function. The postop-erative model for end-stage liver disease score was used to define hepatic dysfunction. There were 35 patients with postoperative hepatic dysfunction (group hepatic dysfunction, model for end-stage liver disease score ≥ 15) and 121 patients without postoperative hepatic dysfunction (group non-hepatic dysfunction, model for end-stage liver disease score < 15). Univariate and multiple analyses (logistic regression) were used to identify the predictive risk factors. RESULTS: In-hospital mortality rate was 8.3%. Multiple logistic analysis showed that preoperative alanine aminotransferase (P <.001), cardiopulmonary bypass time (P <.001), and red blood cell transfusion (P <.001) were independent determinants for postoperative hepatic dysfunction. The patients were followed up for 2 years, with an average follow-up of 22.9 ± 3.2 months, and the lost follow-up rate was 9.1%. The short- and medium-term mortality in hepatic dysfunction group was higher than that in non-hepatic dysfunction group (log-rank P =.009). CONCLUSIONS: The incidence of postoperative hepatic dysfunction is high in patients with acute type A aortic dissection. Preoperative alanine aminotransferase, cardiopulmonary bypass time, and red blood cell transfusion were independent risk factors for those patients. The short- and medium-term mortality in hepatic dysfunction group was higher than that in non-hepatic dysfunction group.

Construction of a high-resolution genetic map and identification of single nucleotide polymorphism markers relevant to flower stalk height in onion.

Introduction: Onion (Allium cepa L., 2n=16) is an economically and nutritionally important vegetable crop worldwide. Construction of a high-resolution genetic map and map-based gene mining in onion have lagged behind other vegetable crops such as tomato and pepper. Methods: In this study, we constructed a high-resolution genetic map of onion using 321 F2 individuals from a cross between two double haploid lines DH-1×DH-17 and employing specific length amplified fragment (SLAF)-seq technology. The genetic map containing 10,584 polymorphic SLAFs with 21,250 single nucleotide polymorphism (SNP) markers and 8 linkage groups was developed for onion, which spanned 928.32 cM, with an average distance of 0.09 cM between adjacent markers. Results: Using this map, we carried out QTL mapping of Ms locus related to the male-fertile trait and reproduced previous mapping results, which proved that this map was of good quality. Then, four QTLs (located on LG2, LG5, and LG8) were detected for flower stalk height, explaining 26.60% of the phenotypic variance. Among them, we proposed that 20 SLAF markers (in three QTLs) of flower stalk height trait were effective favorable allelic variant markers associated with heterosis. Discussion: Overall, the genetic map was structured using SLAF-seq based on DH lines, and it is the highest-quality and highest-resolution linkage map of onion to date. It lays a foundation for the fine mapping and candidate gene identification of flower stalk height, and provides new insights into the developmental genetic mechanisms in onion breeding.

Tumor microenvironment characterization in stage IV gastric cancer.

Immunotherapy is remarkably affected by the immune environment of the principal tumor. Nonetheless, the immune environment's clinical relevance in stage IV gastric cancer (GC) is largely unknown. The gene expression profiles of 403 stage IV GC patients in the three cohorts: GEO (Gene Expression Omnibus, GSE84437 (n=292) and GSE62254 (n=77), and TCGA (The Cancer Genome Atlas, n=34) were used in the present study. Using four publicly available stage IV GC expression datasets, 29 immune signatures were expression profiled, and on this basis, we classified stage IV GC. The classification was conducted using the hierarchical clustering method. Three stage IV GC subtypes L, M, and H were identified representing low, medium, and high immunity, respectively. Immune correlation analysis of these three types revealed that Immune H exhibited a better prognostic outcome as well as a higher immune score compared with other subtypes. There was a noticeable difference in the three subgroups of HLA genes. Further, on comparing with other subtypes, CD86, CD80, CD274, CTLA4, PDCD1, and PDCD1LG2 had higher expression in the Immunity H subtype. In stage IV GC, potentially positive associations between immune and pathway activities were displayed, due to the enrichment of pathways including TNF signaling, Th-17 cell differentiation, and JAK-STAT signaling pathways in Immunity H vs Immunity L subtypes. External cohorts from TCGA cohort ratified these results. The identification of stage IV GC subtypes has potential clinical implications in stage IV GC treatment.

Crystallization of SLA-2*04:02:02 complexed with a CTL epitope derived from FMDV.

Presentation of viral epitopes by swine MHC I (termed leukocyte antigen class I, SLA I) to cytotoxic T lymphocytes (CTLs) is crucial for swine immunity. The SLA-2 structure, however, remains largely unknown. To illustrate the structural basis of swine CTL epitope presentation, the crystal structure of SLA-2*04:02:02 complexed with one peptide, derived from foot-and-mouth disease virus (FMDV), was analyzed in this study. SLA-2*04:02:02 and swine ß2-microglobulin (sß2m) were refolded in vitro in the presence of peptides. X-ray diffraction data of SLA-2*04:02:02-peptide-sß2m (referred to as p/SLA-2*04:02:02) were collected. The diffraction dataset was 2.3 Šin resolution and the space group was P3(2)21. Relevant data included a = 101.8 Å, b = 101.8 Å, c = 73.455 Å,α = 90.00°, ß = 90.00°, γ = 120.00°. The structure of p/SLA-2*04:02:02 was analyzed. The results revealed that Glu24, Met68, Gly76, and Gln173 in PBG of SLA-2*04:02:02 are different from other MHC I. Furthermore, Asn63 is different from other SLA I. Gln57, Met174 and Gln180 in PBG of SLA I are different from other species' MHC I. All of these features are different from known mammalian peptide-MHC class I complexes (referred to as p/MHC I). In addition, P4-His, P6-Val, and P8-Pro in the peptide were exposed, and these residues as epitopes can be presented by SLA-2*04:02:02. This study not only provides a structural basis for peptide presentation by SLA-2, but also screens one potential FMDV CTL epitope. The results may be of interest in future vaccine development.

Structural insights into the evolution feature of a bony fish CD8αα homodimer.

The CD8αα homodimer structures of endotherms demonstrate that despite distinct diversity at the amino acid sequence level, a few conserved key amino acids ensure common structural features. The structure of CD8αα in ancient ectotherms, such as lower bony fish, remains unclear. In this study, the high-resolution structure of the grass carp (Ctenopharyngodon idella) CD8αα (Ctid-CD8αα) homodimer was determined using the single-wavelength anomalous diffraction (SAD) method. The structure of Ctid-CD8αα shows distinct differences from the known CD8αα structures of endotherms, including a distinct topological structure with shorter back ß sheets. The configuration and distribution of the hydrophobic core are different from those in endotherms. Interestingly, mutation of the key amino acid F32S, which is very common in fish and lies in the CDR loop region, leads to the absence of the typical cavity that binds to an epitope-MHC I (p/MHC I) in endotherms, yet Ctid-CD8αα can still specifically bind the grass carp peptide-Ctid-UAA-ß2m (p/UAA-ß2m). Our results indicate that during the evolutionary process, CD8αα has undergone dramatic changes that affect its dimeric structure and may use a new strategy to interact with p/MHC I.

Discovery and Analysis of Invertebrate IgVJ-C2 Structure from Amphioxus Provides Insight into the Evolution of the Ig Superfamily.

The emergence of adaptive immunity in jawed vertebrates depended on the appearance of variable immune receptors, BCRs and TCRs, which exhibit variable-J-constant (VJ-C)-type Ig superfamily folds. Hitherto, however, the structures of IgV-J-IgC-type molecules had never been characterized in invertebrates, leaving the origin of BCR/TCR-type molecules unknown. Using x-ray crystallography, the structure of a VJ-C2 molecule, named AmpIgVJ-C2, was determined in amphioxus (Branchiostoma floridae). The first domain shows typical V folding, including the hydrophobic core, CDR analogs, and eight conserved residues. The second domain is a C2-type Ig superfamily domain, as defined by its short length and the absence of ß-strand D- and C1-typical motifs. AmpIgVJ-C2 molecules form homodimers, using "three-layer packing dimerization," as described for TCRs and BCRs. The AmpIgVJ-C2 V domain harbors a diglycine motif in ß-strand G and forms a ß-bulge structure participating in V-V intermolecular interaction. By immunohistochemistry, AmpIgVJ-C2 molecules were primarily found in mucosal tissues, whereas PCR and sequence analysis indicated considerable genetic variation at the single-gene level; these findings would be consistent with an immune function and a basic ability to adapt to binding different immune targets. Our results show a BCR/TCR-ancestral like molecule in amphioxus and help us to understand the evolution of the adaptive immune system.

Conserved peptides enhance immune efficiency of inactive vaccines against emerging avian influenza viruses in chicken.

Avian influenza viruses (AIVs) such as H5N1 and H7N9 are a great threat to poultry economics and public health. Vaccination can effectively inhibit the spread of AIV in poultry, which is also a viable strategy for controlling virus transmission from poultry to human. Adjuvants that are commonly used in current inactivated vaccines to provide stronger anti-AIV immune responses are often limited in their capacity to quantitatively induce both humoral and cellular immune responses. Herein, we assessed the levels of immune responses generated by a vaccine formulation comprising inactivated H5N1 antigen and synthetic peptides covering conserved CD4+, CD8+ T cell, and B cell epitopes. We found that the synthetic peptides enhanced the antibody responses against conserved influenza virus antigen M2e. Notably, the hemagglutination inhibition test results indicated that the peptides significantly augmented the antibody responses of inactivated H5N1 antigen even in the 1/10 or 1/5 dose group, in the identical antibody level as antigen alone used at the full dose. This indicates that the peptide can significantly reduce the use of inactivated virus, lowering the cost of the vaccine. Moreover, the peptides increased the transcript levels of interleukin-4 and interferon-γ cytokines in chicken peripheral blood mononuclear cells, which may facilitate both humoral and cellular immune responses. Our data suggest that this peptide combined with inactivated H5N1 antigen enhances both the humoral and cellular immune responses, which may benefit the prediction and design of synthetic peptide-based adjuvants for vaccines in chicken.

The Structure of the MHC Class I Molecule of Bony Fishes Provides Insights into the Conserved Nature of the Antigen-Presenting System.

MHC molecules evolved with the descent of jawed fishes some 350-400 million years ago. However, very little is known about the structural features of primitive MHC molecules. To gain insight into these features, we focused on the MHC class I Ctid-UAA of the evolutionarily distant grass carp (Ctenopharyngodon idella). The Ctid-UAA H chain and ß2-microglobulin (Ctid-ß2m) were refolded in vitro in the presence of peptides from viruses that infect carp. The resulting peptide-Ctid-UAA (p/Ctid-UAA) structures revealed the classical MHC class I topology with structural variations. In comparison with known mammalian and chicken peptide-MHC class I (p/MHC I) complexes, p/Ctid-UAA structure revealed several distinct features. Notably, 1) although the peptide ligand conventionally occupied all six pockets (A-F) of the Ag-binding site, the binding mode of the P3 side chain to pocket D was not observed in other p/MHC I structures; 2) the AB loop between ß strands of the α1 domain of p/Ctid-UAA complex comes into contact with Ctid-ß2m, an interaction observed only in chicken p/BF2*2101-ß2m complex; and 3) the CD loop of the α3 domain, which in mammals forms a contact with CD8, has a unique position in p/Ctid-UAA that does not superimpose with the structures of any known p/MHC I complexes, suggesting that the p/Ctid-UAA to Ctid-CD8 binding mode may be distinct. This demonstration of the structure of a bony fish MHC class I molecule provides a foundation for understanding the evolution of primitive class I molecules, how they present peptide Ags, and how they might control T cell responses.

Structural and Biochemical Analyses of Swine Major Histocompatibility Complex Class I Complexes and Prediction of the Epitope Map of Important Influenza A Virus Strains.

UNLABELLED: The lack of a peptide-swine leukocyte antigen class I (pSLA I) complex structure presents difficulties for the study of swine cytotoxic T lymphocyte (CTL) immunity and molecule vaccine development to eliminate important swine viral diseases, such as influenza A virus (IAV). Here, after cloning and comparing 28 SLA I allelic genes from Chinese Heishan pigs, pSLA-3*hs0202 was crystalized and solved. SLA-3*hs0202 binding with sß2m and a KMNTQFTAV (hemagglutinin [HA]-KMN9) peptide from the 2009 pandemic swine H1N1 strain clearly displayed two distinct conformations with HA-KMN9 peptides in the structures, which are believed to be beneficial to stimulate a broad spectrum of CTL immune responses. Notably, we found that different HA-KMN9 conformations are caused, not only by the flexibility of the side chains of residues in the peptide-binding groove (PBG), but also by the skewing of α1 and α2 helixes forming the PBG. In addition, alanine scanning and circular-dichroism (CD) spectra confirmed that the B, D, and F pockets play critical biochemical roles in determining the peptide-binding motif of SLA-3*hs0202. Based on biochemical parameters and comparisons to similar pockets in other known major histocompatibility complex class I (MHC-I) structures, the fundamental motif for SLA-3*hs0202 was determined to be X-(M/A/R)-(N/Q/R/F)-X-X-X-X-X-(V/I) by refolding in vitro and multiple mutant peptides. Finally, 28 SLA-3*hs0202-restricted epitope candidates were identified from important IAV strains, and two of them have been found in humans as HLA-A*0201-specific IAV epitopes. Structural and biochemical illumination of pSLA-3*hs0202 can benefit vaccine development to control IAV in swine. IMPORTANCE: We crystalized and solved the first SLA-3 structure, SLA-3*hs0202, and found that it could present the same IAV peptide with two distinct conformations. Unlike previous findings showing that variable peptide conformations are caused only by the flexibility of the side chains in the groove, the skewing of the α1 and α2 helixes is important in the different peptide conformations in SLA-3*hs0202. We also determined the fundamental motif for SLA-3*hs0202 to be X-(M/A/R)-(N/Q/R/F)-X-X-X-X-X-(V/I) based on a series of structural and biochemical analyses, and 28 SLA-3*hs0202-restricted epitope candidates were identified from important IAV strains. We believe our structure and analyses of pSLA-3*hs0202 can benefit vaccine development to control IAV in swine.

Super-flat supercontinuum generation from a Tm-doped fiber amplifier.

Super-flat supercontinua are generated from a double clad Tm-doped fiber amplifier. Two different laser configurations are investigated and compared. In the direct-output configuration, the long-wavelength edge of the supercontinuum spectra is extended to beyond 2.65 µm with a 10 dB bandwidth of 740 nm. In the passive pigtail configuration, the generated supercontinuum features excellent flatness with an intensity difference smaller than 1 dB in the wide central spectral range from 1.98 µm to 2.41 µm.

[A Duplex PCR Method for Detection of Babesia caballi and Theileria equi].

OBJECTIVE: To develop a duplex PCR assay for detection of Babesia caballi and Theileria equi. METHODS: Two pairs of primers were designed according to the BC48 gene of B. caballi and 18 s rRNA gene of T. equi, and a duplex PCR assay was developed by the optimization of reaction conditions. The specificity, sensitivity and reliability of the method were tested. The horse blood samples of suspected cases were collected from Yili region, and detected by the duplex PCR, microspopy, conventional PCR, and fluorescence quantitative PCR, and the results were compared. RESULTS: Using the duplex PCR assay, the specific fragments of 155 bp and 280 bp were amplified from DNA samples of B. caballi and T. equi, respectively. No specific fragment was amplified from DNA samples of B. bigemina, Theilerdia annulata, Theilerdia sergenti, Toxoplasma gondii, Neospora caninum, and Trypanosoma evansi. The limit of detection was 4.85 x 10(5) copies/L for B. caballi DNA and 4.85 x 10(4) copies/µl for T. equi DNA, respectively. Among the 24 blood samples, 11 were found B. caballi-positive by the duplex PCR assay, and 18 were T. equi-positive. The coincidence rate of microscopy, conventional PCR, and fluorescence quantitative PCR with duplex PCR was 91.7% (22/24), 95.8% (23/24), and 95.8% (23/24), respectively. CONCLUSION: A duplex PCR assay for simultaneous detection of B. caballi and T. equi is established.

Crystallization and preliminary X-ray diffraction analysis of a single variable domain of the immunoglobulin superfamily in amphioxus, Amphi-IgSF-V.

Amphioxus is regarded as an essential animal model for the study of immune evolution. Discovery of new molecules with the immunoglobulin superfamily (IgSF) variable (V) domain in amphioxus would help in studying the evolution of IgSF V molecules in the immune system. A protein was found which just contains only one IgSF V domain in amphioxus, termed Amphi-IgSF-V; it has over 30% sequence identity to the V domains of human immunoglobulins and mammalian T-cell receptors. In order to clarify the three-dimensional structure of this new molecule in amphioxus, Amphi-IgSF-V was expressed, purified and crystallized, and diffraction data were collected to a resolution of 1.95 Å. The crystal belonged to space group P3221, with unit-cell parameters a = b = 53.9, c = 135.5 Å. The Matthews coefficient and solvent content were calculated to be 2.58 Å(3) Da(-1) and 52.38%, respectively. The results will provide structural information to study the evolution of IgSF V molecules in the immune system.

Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the bovine CD8αα-BoLA-2*02201 complex.

In order to clarify the structural characteristics of the bovine MHC class I molecule (BoLA-I) complexed with CD8αα (CD8αα-BoLA-I), bovine CD8αα, BoLA-I (BoLA-2*02201) and ß2m were expressed and purified, and were then assembled with a peptide derived from Foot-and-mouth disease virus (FMDV-VP1YY9) and crystallized. The crystal diffracted to 1.7 Šresolution and belonged to space group P21, with unit-cell parameters a=53.9, b=103.8, c=61.8 Å, α=γ=90, ß=96°. The asymmetric unit contained one complex, with a Matthews coefficient of 2.41 Å3 Da(-1) and a solvent content of 48.9%. The rotation-function Z-score and translation-function Z-score for molecular replacement were 3.4 and 8.9, respectively. In addition, SDS-PAGE analysis of CD8αα-BoLA-I crystals showed three bands corresponding to the molecular weights of BoLA-I heavy chain, ß2m and CD8α. The structure of the CD8αα-BoLA-I complex should be helpful in obtaining insight into the interaction between bovine CD8αα and MHC class I molecules. Structure determination of BoLA-2*02201-FMDV-VP1YY9 will be useful in the design of vaccines for foot-and-mouth disease.

Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the chicken MHC class I molecule BF2*1501.

The chicken major histocompatibility complex (MHC) class I molecules named BF are strongly associated with Marek's disease (MD). A single structure, that of chicken BF2*2101 from the B21 haplotype, which might provide resistance to MD, has been determined. However, little is known about other structures apart from BF2*2101. In order to provide further structures of chicken MHC class I molecules, BF2*1501 and chicken ß(2)-microglobulin complexed with a nonapeptide (MDV-MEQ(RRR9)) derived from Marek's disease virus MEQ protein (MDV EcoRI Q fragment, residues 72-80) were assembled and crystallized. Diffraction data from the crystal were collected to 2.6 Å resolution; the crystal belonged to space group P3(1)21, with unit-cell parameters a = 125.1, b = 125.1, c = 80.9 Å and two molecules in the asymmetric unit. The Matthews coefficient V(M) was 2.08 Å(3) Da(-1), with a calculated solvent content of 40.78%. These data will be helpful in obtaining insight into the structural basis of the involvement of BF2*1501 in MD.

Recombinant human endostatin endostar inhibits tumor growth and metastasis in a mouse xenograft model of colon cancer.

To investigate the effects of recombinant human endostatin Endostar on metastasis and angiogenesis and lymphangiogenesis of colorectal cancer cells in a mouse xenograft model. Colon cancer cells SW620 were injected subcutaneously into the left hind flank of nude mice to establish mouse xenograft models. The mice were treated with normal saline or Endostar subcutaneously every other day. The growth and lymph node metastasis of tumor cells, angiogenesis and lymphangiogenesis in tumor tissue were detected. Apoptosis and cell cycle distribution were studied by flow cytometry. The expression of VEGF-A, -C, or -D in SW620 cells was determined by immunoblotting assays. Endostar inhibited tumor growth and the rate of lymph node metastasis (P < 0.01). The density of blood vessels in or around the tumor area was 12.27 ± 1.21 and 22.25 ± 2.69 per field in Endostar-treated mice and controls (P < 0.05), respectively. Endostar also decreased the density of lymphatic vessels in tumor tissues (7.84 ± 0.81 vs. 13.83 ± 1.08, P < 0.05). Endostar suppresses angiogenesis and lymphangiogenesis in the lymph nodes with metastases, simultaneously. The expression of VEGF-A, -C and -D in SW620 cells treated with Endostar was substantially lower than that of controls. Endostar inhibited growth and lymph node metastasis of colon cancer cells by inhibiting angiogenesis and lymphangiogenesis in a mouse xenograft model of colon cancer.