•A novel truncated protein HPV58 L1ΔN4C was highly expressed in baculovirus system.•The yield of L1ΔN4C VLP ranged from 55 mg/L to 60 mg/L by two-step chromatography.•The higher mRNA level with ...improved stability of L1ΔN4C was observed.•L1ΔN4C VLP retained good immunogenicity to induce high level of neutralizing antibodies.
Human papillomavirus (HPV) major capsid protein L1 virus-like particles (VLPs) produced in baculovirus system are highly immunogenic, but the relatively high production cost limits its application in the development of broad-spectrum vaccines. Here we report a novel method for enhancing VLP production in this system. We incorporated respectively 4, 8 or 13 residues truncation mutations in the N-terminus of L1ΔC, a C-terminal 25-residue-deleted L1 of HPV58, to construct three mutants. After expression in Sf9 cells, L1ΔN4C exhibited 2.3-fold higher protein production, 2.0-fold mRNA expression and lower rate of mRNA decay, compared to L1ΔC. More importantly, L1ΔN4C protein was easily purified by two-step chromatography with a VLP yield of up to 60 mg/L (purity > 99 %), 5-fold that of L1ΔC, whereas L1ΔN8C and L1ΔN13C behaved similarly to L1ΔC either in protein or mRNA expression. Moreover, L1ΔN4C VLPs showed similar binding activities with six HPV58 neutralizing monoclonal antibodies and induced comparable level of neutralizing antibody in mice to that of L1ΔC VLPs. Our results demonstrate that certain N- and C-terminal truncations of HPV58 L1 can enhance VLP yield. This method may be used to reduce production costs of other L1VLPs or chimeric VLPs to developing pan-HPV vaccines using baculovirus system.
C2H2 zinc-finger (C2H2-ZF) proteins are a large gene family in plants that participate in various aspects of normal plant growth and development, as well as in biotic and abiotic stress responses. To ...date, no overall analysis incorporating evolutionary history and expression profiling of the C2H2-ZF gene family in model tree species poplar (Populus trichocarpa) has been reported.
Here, we identified 109 full-length C2H2-ZF genes in P. trichocarpa, and classified them into four groups, based on phylogenetic analysis. The 109 C2H2-ZF genes were distributed unequally on 19 P. trichocarpa linkage groups (LGs), with 39 segmental duplication events, indicating that segmental duplication has been important in the expansion of the C2H2-ZF gene family. Promoter cis-element analysis indicated that most of the C2H2-ZF genes contain phytohormone or abiotic stress-related cis-elements. The expression patterns of C2H2-ZF genes, based on heatmap analysis, suggested that C2H2-ZF genes are involved in tissue and organ development, especially root and floral development. Expression analysis based on quantitative real-time reverse transcription polymerase chain reaction indicated that C2H2-ZF genes are significantly involved in drought, heat and salt response, possibly via different mechanisms.
This study provides a thorough overview of the P. trichocarpa C2H2-ZF gene family and presents a new perspective on the evolution of this gene family. In particular, some C2H2-ZF genes may be involved in environmental stress tolerance regulation. PtrZFP2, 19 and 95 showed high expression levels in leaves and/or roots under environmental stresses. Additionally, this study provided a solid foundation for studying the biological roles of C2H2-ZF genes in Populus growth and development. These results form the basis for further investigation of the roles of these candidate genes and for future genetic engineering and gene functional studies in Populus.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
In this paper, we introduce a novel three-dimension chaotic system with strange characteristic by applying construction of a 3D chaotic circuit method. Multiple equilibria and abundant coexisting ...attractors exist in this system. A mathematical model is developed and detailed stability analyses for equilibrium points are executed with obtaining significant results of the period-doubling bifurcation patterns confirmed by phase plane plots and Lyapunov exponent spectra. By varying the initial value and unique controlled parameter, the double-scroll chaotic attractor is broken up into a pair of symmetric singular attractors. Then, the local basins of attraction are investigated concerning the initial condition. Next, the circuit synthesis results generated by Multisim simulation tool validate the self-excitation characteristics of this system. Finally, the feedback control technique is used to study difference synchronization of this system. Main conclusions prove the validity and reliability of difference synchronization.
Degradation of coevaporated CH3NH3PbI3 thin films were investigated with X-ray photoelectron spectroscopy and X-ray diffraction as the films were subjected to exposure of oxygen, low pressure ...atmospheric air, atmospheric air, or H2O. The coevaporated thin films have consistent stoichiometry and crystallinity suitable for detailed surface analysis. The results indicate that CH3NH3PbI3 is not sensitive to oxygen. Even after 1013 Langmuir (L, one L equals 10–6 Torr s) oxygen exposure, no O atoms could be found on the surface. The film is not sensitive to dry air as well. A reaction threshold of about 2 × 1010 L is found for H2O exposure, below which no CH3NH3PbI3 degradation takes place, and the H2O acts as an n-dopant. Above the threshold, the film begins to decompose, and the amount of N and I decrease quickly, leaving the surface with PbI2, hydrocarbon complex, and O contamination.
A series of binuclear cyclometalated Pt(ii) complexes with the bridge ligand of binaphthyl quinoline were synthesized and characterized. Owing to the restriction of intramolecular motions in chiral ...binaphthyl linkers, the complexes exhibited two unusual metal-induced chiral components and aggregation-induced red phosphorescence. The (R,R,R)/(S,S,S) enantiopure complexes that were confirmed by X-ray diffraction, circular dichroism spectra, time-dependent density functional theory calculations, and circularly polarized luminescence did not require purification by chiral high-performance liquid chromatography and could yield unique 1D M or P hierarchical helices of helices via intermolecular noncovalent interactions. Moreover, it is the first time that racemization-induced phosphorescence enhancement has been observed.
As a biomarker for early cancer diagnosis, telomerase are one of the promising targets for cancer therapeutics. Inspired by the fluorescent emission principle of aggregation-induced emission ...fluorogens, we creatively designed an AIE-based turn-on method to detect telomerase activity from cell extracts. A positively charged fluorogen (TPE-Z) is not fluorescent when freely diffused in solution. The fluorescence of TPE-Z is enhanced with the elongation of the DNA strand which could light up telomere elongation process. By exploitation of it, we can detect telomerase activity from different cell lines (E-J, HeLa, MCF-7, and HLF) with high sensitivity and specificity. Moreover, our method is successfully employed to demonstrate the applications in bladder cancer diagnosis (41 urine specimens from bladder cancer patients and 15 urine specimens from normal people are detected). The AIE-based method provides a simple one-pot technique for quantification and monitoring of the telomerase activity and shows great potential for future use in clinical tests.
The receptor-binding domain (RBD) of SARS-CoV-2 S protein is proved to be the major target of neutralizing antibodies. However, on the S protein, only a portion of epitopes in RBD can be effectively ...displayed with dynamic changes in spatial conformations. Using RBD fragment as antigen can better expose the neutralizing epitopes, but the immunogenicity of RBD monomer is suboptimal. Multimeric display of RBD molecules is a feasible strategy to optimize RBD-based vaccines. In this study, RBD single-chain dimer derived from Wuhan-Hu-1 was fused with a trimerization motif, and a cysteine was also introduced at the C-terminus. The resultant recombinant protein 2RBDpLC was expressed in Sf9 cells using a baculovirus expression system. Reducing/non-reducing PAGE, size-exclusion chromatography and in silico structure prediction indicated that 2RBDpLC polymerized and possibly formed RBD dodecamers through trimerization motif and intermolecular disulfide bonds. In mice, 2RBDpLC induced higher levels of RBD-specific and neutralizing antibody responses than RBD dimer, RBD trimer and prefusion-stabilized S protein (S2P). In addition, cross-neutralizing antibodies against Delta and Omicron VOC were also detected in the immune sera. Our results demonstrate that 2RBDpLC is a promising vaccine candidate, and the method of constructing dodecamers may be an effective strategy for designing RBD-based vaccines.
A localization algorithm, which combines Hough transform and a modified high-order ambiguous function (MHAF) process, is proposed to precisely track multiple moving targets by dual-frequency ...continuous-wave through-wall radar (DF-CW TWR). To avoid the cross-term interference, a Hough-based separation method, which uses a novel progressive geometrical model, is first applied to decompose the echo. Then, an MHAF method is applied to real-time estimate the target instantaneous frequency. Finally, the target trajectories are synthesized based on the estimation results.
Myocardial contractile dysfunction in sepsis is associated with the increased morbidity and mortality. Although the underlying mechanisms of the cardiac depression have not been fully elucidated, an ...exaggerated inflammatory response is believed to be responsible. Nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasome is an intracellular platform that is involved in the maturation and release of interleukin (IL)-1β. The aim of the present study is to evaluate whether sepsis activates NLRP3 inflammasome/caspase-1/IL-1β pathway in cardiac fibroblasts (CFs) and whether this cytokine can subsequently impact the function of cardiomyocytes (cardiac fibroblast-myocyte cross-talk). We show that treatment of CFs with lipopolysaccharide (LPS) induces upregulation of NLRP3, activation of caspase-1, as well as the maturation (activation) and release of IL-1β. In addition, the genetic (small interfering ribonucleic acid siRNA) and pharmacological (glyburide) inhibition of the NLRP3 inflammasome in CFs can block this signaling pathway. Furthermore, the inhibition of the NLRP3 inflammasome in cardiac fibroblasts ameliorated the ability of LPS-challenged CFs to impact cardiomyocyte function as assessed by intracellular cyclic adenosine monophosphate (cAMP) responses in cardiomyocytes. Salient features of this the NLP3 inflammasome/ caspase-1 pathway were confirmed in in vivo models of endotoxemia/sepsis. We found that inhibition of the NLRP3 inflammasome attenuated myocardial dysfunction in mice with LPS and increased the survival rate in mice with feces-induced peritonitis. Our results indicate that the activation of the NLRP3 inflammasome in cardiac fibroblasts is pivotal in the induction of myocardial dysfunction in sepsis.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The co-evaporated perovskite thin film has a cubic crystal structure.From XRD, CH3NH3PbI3 converted to PbI2 completely after air exposure for 22h (76.4°F, 25% humidity level).The atomic ratio of ...co-evaporated perovskite observed by XPS is very close to the ideal stoichiometric ratio of CH3NH3PbI3.The degradation process revealed the complete removal of N and substantial reduction of I.The aggregation of PbI2 crystals roughened the film and resulted in a partial exposure of the Au substrate, confirmed by AFM and XPS.
Methylammonium lead halide perovskites as highly promising photovoltaic materials have been found unstable in air. We investigated the degradation of CH3NH3PbI3 by air exposure using X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). The stoichiometric sample was grown with co-evaporation of PbI2 and CH3NH3I on an Au coated Si wafer. It was found that the perovskite thin film gradually turned to PbI2 in air, accompanied with complete removal of N and substantial reduction of I. It was also observed that PbI2 crystallization roughened the film and resulted in a partial exposure of the Au substrate.
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