Construction of a camelid VHH yeast two-hybrid library and the selection of VHH against haemagglutinin-neuraminidase protein of the Newcastle disease virus

Humoral immune response after immunization. Sera from IIama was collected, two-fold diluted and tested by HI using LaSota as antigen. Figure S1 Amplification of VHH through a nested PCR. (A) First round PCR to separate VH from VHH. The upper 900 bp bands represent the VH-CH1-Hinge-CH2 of conventional Abs (lane 1–8). The lower 600 bp bands represent the VHH-Hinge-CH2 of HCAbs (lane 1–8). (B) VHH amplified through nested PCR using 600 bp fragment recovered from first round PCR as template (lane 1–4). M in A and B was the DL2000 DNA marker. C in A and B represent the negative control. Figure S2 PCR identification of inserted VHH. 47 clones were randomly picked to determine the library functional diversity by PCR using universal primers T7 and 3’AD (Table 1). Meanwhile, Sterile water was used as negative controls. 45 clones have amplified the 500 bp VHH fragments (lane 1–47), while negative templates control haven’t amplified any bands (lane C). M indicated the DL2000 DNA marker. Figure S3 Detection of library capacity and library titer. (A) 10-3 dilution plating of the transformed cells calculated a library capacity of 1.25 × 107 independent clones. (B) 10-5 dilution plating of the cultured library indicated a library titer of 3.45 × 108 cfu/mL. Figure S4 Deduced amino acid aligment of 10 random picked VHH. Deduced amino acid sequences were analyzed according to the Kabat numbering. Differences in the sequences are pinked, and the dash represent the missing sequences. Two hallmark Cys residues are labeled by the thick-line boxes. The four conservative hallmark residues of VHH in FR2 are labeled by the dotted line boxes. Figure S5 pGBKT7-HN bait plasmid construction. (A) PCR was carried out to amplify a truncate HN gene (without transmembrane region) from La Sota strain. M, 5000 DNA marker. 1, Truncate HN. C, Negative control. (B) A truncate HN was cloned into pGBKT7 through BamH I and Sal I. M, 5000 DNA marker. 1, Double restriction enzyme digestion of pGBKT7-HN. Figure S6 pHSIE-VHH plasmid construction. (A) 7 positive VHH fragment were amplified from recovered positive clones containing pGADT7-VHH by PCR. M, 5000 DNA marker. 1–7, VHH 1–7. C, Negative control. (B) Double restriction enzyme digestion of pHSIE-VHHs. M, 5000 DNA marker. 1–7, pHSIE-VHH 1–7. Figure S7 Western blot analysis of bait protein expression. 2 mL of Y2HGold(pGBKT7-HN) culture liquid was extracted using yeast protein extraction reagent (Takara). c-Myc tag monoclonal antibody (1:4000 dilution) was used as first antibody and HRP-labeled goat anti-mouse antibody (1:5000) was used as second antibody. The immunoreactive was visualized with cECL Plus Western blotting detection reagent (CWBIO). (DOC 1129 kb

An Ultra-Short-Term Electrical Load Forecasting Method Based on Temperature-Factor-Weight and LSTM Model

Ultra-short-term electrical load forecasting is an important guarantee for the safety and efficiency of energy system operation. Temperature is also an important factor affecting the changes in electric load. However, in different cases, the impact of temperature on load forecasting will vary greatly, and sometimes even lead to the decrease of forecasting accuracy. This often brings great difficulties to researchers’ work. In order to make more scientific use of temperature factor for ultra-short-term electrical load forecasting, especially to avoid the negative influence of temperature on load forecasting, in this paper we propose an ultra-short-term electrical load forecasting method based on temperature factor weight and long short-term memory model. The proposed method evaluates the importance of the current prediction task’s temperature based on the change magnitude of the recent load and the correlation between temperature and load, and therefore the negative impacts of the temperature model can be avoided. The mean absolute percentage error of proposed method is decreased by 1.24%, 1.86%, and 6.21% compared with traditional long short-term memory model, back-propagation neural network, and gray model on average, respectively. The experimental results demonstrate that this method has obvious advantages in prediction accuracy and generalization ability

Magnetic Field-driven Insulator–Metal Transition and Colossal Magnetoresistance of Metamagnetic Semiconductor Mercury Thiodichromite Crystals

A facile method is developed to efficiently prepare metamagnetic mercury thiodichromite (HgCr2S4, HCS) polycrystals and single crystals, and their transport properties are studied. The resistivity of the as-prepared HCS polycrystal shows a semiconducting behavior and no magnetic field dependence in the whole temperature range. In contrast, the annealing treatment of the HCS polycrystal induces gigantic changes: an insulator–metal transition is driven by a magnetic field of 5 T, leading to colossal magnetoresistance (CMR) as high as ∼104. The HCS single crystal grown by a newly developed facile method displays similar properties with a larger CMR up to 106–107. First-principles calculation demonstrates a large spin splitting of band structures, providing the possibility of magnetic polaron existence, which is further evidenced by electron spin resonance spectra. Thus, the insulator–metal transition and CMR can be explained in a magnetic polaronic scenario. This work opens a new window for CMR-based spintronics

Anomaly Internet Network Traffic Detection by Kernel Principle Component Classifier

As a crucial issue in computer network security, anomaly detection is receiving more and more attention from both application and theoretical point of view. In this paper, a novel anomaly detection scheme is proposed. It can detect anomaly network traffic which has extreme large value on some original feature by the major component, or does not follow the correlation structure of normal traffic by the minor component. By introducing kernel trick, the nonlinearity of network traffic can be well addressed. To save the processing time, a simplified version is also proposed, where only major component is adopted. Experimental results validate the effectiveness of the proposed scheme

Modulation of perovskite degradation with multiple-barrier for light-heat stable perovskite solar cells

Abstract The long-term stability of perovskite solar cells remains one of the most important challenges for the commercialization of this emerging photovoltaic technology. Here, we adopt a non-noble metal/metal oxide/polymer multiple-barrier to suppress the halide consumption and gaseous perovskite decomposition products release with the chemically inert bismuth electrode and Al2O3/parylene thin-film encapsulation, as well as the tightly closed system created by the multiple-barrier to jointly suppress the degradation of perovskite solar cells, allowing the corresponding decomposition reactions to reach benign equilibria. The resulting encapsulated formamidinium cesium-based perovskite solar cells with multiple-barrier maintain 90% of their initial efficiencies after continuous operation at 45 °C for 5200 h and 93% of their initial efficiency after continuous operation at 75 °C for 1000 h under 1 sun equivalent white-light LED illumination