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Additional file 8: Supplementary Table 2. Titration of blocking activities of purified LILRB2 antibodies against PS-LILRB2 interactions.

Additional file 5: Supplementary Figure 5. Ab29 increases microglial responses to amyloid plaques in vivo. a. Representative amyloid plaque-microglia co-localization immunofluorescence staining of 5-month-old 5XFAD mice cortex as treated in Fig. 6a. Scale bar = 20 μm. IBA1, microglia marker; 6E10, amyloid plaque marker. b. Quantification of IBA1 area within 30 μm of amyloid plaques in the cortex of mice treated as described in Fig. 6a. n = 5 independent mice. c. Representative amyloid plaque-CD68 co-localization immunofluorescence staining of the cortex of 5XFAD mice treated as described in Fig. 6a. CD68, microglia phagocytic marker. Scale bar = 20 μm. d. Quantification of CD68 area within 30 μm of amyloid plaques in the cortex of mice treated as described in Fig. 6a. n = 5 independent mice. e. Quantification of Aβ co-localized with CD68 per plaque in the cortex of mice treated as described in Fig. 6a. n = 5 independent mice. For all the data presented, bar graphs with error bars represent mean ± SD. For the statistical analysis, *** P < 0.001, two-tailed Student t-test.

Additional file 4: Supplementary Figure 4. Screening of LILRB2 antibodies for rescuing LILRB2-mediated TREM2 signaling inhibition. a-b. LILRB2 antibodies rescue oAβ or PS-LILRB2-mediated inhibition of TREM2 signaling. Plate-coated oAβ (a) or PS (b) was incubated with LILRB2/TREM2 reporter cells in the presence of 10 μg/mL purified LILRB2 antibodies. The activation of LILRB2/TREM2 reporter cells was observed as percentage GFP+ cells. TREM2 signaling in the treatment groups was normalized based on the percentage of GFP+ reporter cells expressing only TREM2 (set to 100%). Data are presented as mean ± SD (n = 4 independent experiments). c. Surface expression of TREM2 and LILRB2 on HMC cells. Indicated antibodies were used to stain surface receptors on HMC cells after Fc blocking by human Fc fragment. The antibody was detected by Alexa Fluor 488-streptavidin, and the fluorescent signals (x-axis) were plotted as a histogram with normalized cell percentage on the y-axis. d. MFI of HMC3 cells stained by indicated antibodies (x-axis) as shown in c. Data are presented as mean ± SD (n = 4 independent experiments). e. Immunoblot of phosphorylated SHP1 (pSHP1), SHP1 of HMC3 upon incubation with oAβ-lipoprotein complex with indicated treatments for 1 hour. Results are from total input cell lysate with β-actin as the loading control. f-g. oAβ or PS activation of TREM2 GFP reporter cells. Plate-coated oAβ (f) or PS (g) was incubated with reporter cells expressing either human or mouse TREM2, and the percentage of GFP+ cells is presented. Data are presented as mean ± SD (n = 4 independent experiments). h-i. Blocking of oAβ or PS-TREM2 signaling by a TREM2 antagonist antibody. Plate-coated oAβ (h) or PS (i) was incubated with reporter cells expressing either human or mouse TREM2 in the presence of TREM2 antagonist antibody or control IgG, the percentage of GFP+ cells are shown with an antibody treatment. Data are presented as mean ± SD (n = 4 independent experiments). j. The LILRB2 antibody clone 42D1 was validated for not blocking either oAβ- or PS-induced LILRB2 signaling. Plate-coated oAβ or PS was incubated with LILRB2-GFP reporter cells in the presence of soluble 42D1 or control IgG, the percentage of GFP+ cells is shown with an antibody treatment. Data are presented as mean ± SD (n = 4 independent experiments). k. Plate-coated 42D1 activates LILRB2 signals. Plate-coated anti-LILRB2 clone 42D1 or control IgG was incubated with LILRB2-GFP reporter cells. The percentages of GFP+ cells are shown. Data are presented at mean ± SD (n = 4 independent experiments). l. Antibody 42D1 showed no recuse of LILRB2-mediated TREM2 signaling inhibition in reporter cells. LILRB2-TREM2 2B4 reporter cells were stimulated with either oAβ or PS co-incubated with Ctrl Ig, 42D1, or Ab29. The TREM2 signaling was detected as increased GFP expression. Data are presented at mean ± SD (n = 3 independent experiments). m. Validation of negative controls and quenching method used in the hMGL phagocytosis assay. hMGLs were incubated with oAβ-lipid (fluorescence-labeled) under the conditions labeled on the x-axis. The temperatures are incubation temperature. CytoD means co-incubated with CytoD during phagocytosis. TB means trypan blue, which was added after phagocytosis to quench cell surface FAM signals as described in the method section. Data are presented at mean ± SD (n = 3 independent experiments).

Additional file 1: Supplementary Figure 1. Immunofluorescence staining of human brain tissue with astrocyte and neuron markers. a. Immunofluorescence staining of human brain tissue of normal subjects showed co-localization of LILRB2 and TREM2 with microglial marker IBA1 (top row). The bottom row showed no co-localization of LILRB2 and TREM2 with astrocyte marker GFAP. Scale bar = 20 μm. b-c. Binding kinetics profiles between oAβ and LILRB2 or TREM2. In the association stage, protein A sensor-captured LILRB2-Fc (b) or TREM2-Fc (c) protein was incubated with oAβ at indicated concentrations The amount of oAβ bound onto the sensors was presented as wavelength shift in nanometers (nm). The red dotted vertical line marks the transit from the association stage to the dissociation stage, where the sensors were dipped into kinetics buffer without oAβ allowing free dissociation. The binding kinetics parameters were calculated using a 1:1 binding model with global fitting.

Additional file 11: Supplementary Table 5. Percentage of similarity of ECDs of LILRB and LILRA family of receptors to the ECD of LILRB2.

Additional file 2: Supplementary Figure 2. Establishment of reporter cell lines for studying LILRB2 and TREM2 a. Schematic diagram showing the cell line LILRB2_muITIM-TREM2. b. Schematic diagram showing oAβ or PS activates LILRB2 chimeric reporter cells. The chimeric LILRB2-ITAM signaling triggers the NFAT pathway upon binding and crosslinking by oAβ or PS, leading to induced GFP expression. The Ig-like domains of LILRB2 are depicted as spheres. The ITAM motifs of the chimeric LILRB2 are from helical and intracellular regions of TREM2, which mediates activation functions after associating with DAP12. c-d. Activation of LILRB2-chimeric GFP reporter cells by oAβ and lipids. Plate-coated oAβ (c) or lipids (d) were incubated with LILRB2-chimeric reporter cells, and the percentage of GFP+ cells is shown in the y-axis. Data are presented as mean ± SD (n = 4 independent experiments). e. Schematic diagram showing oAβ or PS activates TREM2-DAP12 reporter cells. TREM2-DAP12 binding and crosslinking by oAβ or PS trigger the NFAT pathway leading to induced GFP expression. ITAM motifs in the TREM2 figure are from DAP12. f-g. Activation of TREM2 GFP reporter cells by oAβ or lipids. Plate-coated oAβ (f) or lipids (g) were incubated with TREM2-DAP12 reporter cells. The percentage of GFP+ cells is shown on the y-axis. Data are presented as mean ± SD (n = 4 independent experiments). h. Comparison of monomer oAβ versus oligomer oAβ in the activation of LILRB2 or TREM2 reporter cells. i. Schematic diagram showing the reporter cell line muLILRB2-TREM2. j. Surface expression of TREM2 and LILRB2 on BV2-LILRB2 cells. Indicated antibodies were used to stain surface receptors on BV2-LILRB2 cells after Fc blocking. The antibody was detected by Alexa Fluor 488-streptavidin, and the fluorescent signals (x-axis) were plotted as a histogram with normalized cell percentage on the y-axis. k. MFI of LILRB2-BV2 or LILRB2_muITIM-BV2 cells stained by indicated antibodies (x-axis) as shown in j. Data are presented as mean ± SD (n = 3 independent experiments).

Additional file 3: Supplementary Figure 3. LILRB2 targeting antibodies showed potent blocking activity, high affinity, and specificity. a-b. Screening of antibodies inhibiting oAβ or PS-induced activation of LILRB2-chimeric reporter. Plate-coated oAβ (a) or PS (b) was incubated with LILRB2-chimeric reporter cells under the presence of unpurified antibody supernatant (1:20 dilution, antibody name is shown in x-axis). The activation of LILRB2-chimeric reporter cells was observed as percentages of GFP+ cells. Data are presented as mean ± SD (n = 3 independent experiments). c-d. Titration of blocking activities of purified LILRB2 antibodies against oAβ- or PS-LILRB2 interactions. Plate-coated oAβ (c) or PS (d) was incubated with LILRB2-chimeric reporter cells under the presence of increasing concentrations of purified LILRB2 antibodies (antibody names are shown in the figure legend). The activation of LILRB2-chimeric reporter cells was observed as percentages of GFP+ cells. Data are presented as mean ± SD (n = 3 independent experiments). e-f. Maximum wavelength shifts of the oAβ- (e) or PS (f)-LILRB2 binding curve in association stage presented in g and h, respectively. g-h. Antibodies blocking LILRB2 binding with oAβ or PS as measured by BLI. LILRB2 was loaded onto protein A sensors via binding with sensor-captured LILRB2 antibodies. The LILRB2-loaded sensors were then incubated with biotinylated oAβ (1 μM, g) or PS (1 mM, h). The amount of oAβ (g) or PS (h) bound onto the sensors is presented as wavelength shift in nanometers (nm). i. Binding kinetics profile between Ab29 and LILRB2 as measured by BLI. In the association stage, protein A sensor-captured Ab29 was incubated with LILRB2-His at indicated concentrations for the designated time presented on the x-axis. The amount of LILRB2-His bound onto the sensors is presented as a wavelength shift in nanometers (nm). The red dotted vertical line marks the transit from association stage to dissociation stage, where the sensors were dipped into kinetics buffer without LILRB2-His allowing free dissociation. j. Titration profiles of purified antibodies binding to plate-coated LILRB2 as measured by ELISA. Data are presented as mean ± SD (n = 3 independent experiments). k. Binding of purified LILRB2 antibodies to LILRB2 expressed on HEK293T cells. The percentage of positive staining was gated based on control IgG-treated cells. Data are presented as mean ± SD (n = 3 independent experiments). l. Cross-reactivity of all LILRB2 blocking antibodies against other LILRB and LILRA family receptors as measured by ELISA. LILRB2-Fc was included as the positive control. Data are presented as mean ± SD (n = 3 independent experiments). m. Cross-reactivity of Ab29 to LILRB and LILRA family receptors as measured by BLI. In the association stage, protein A sensor-captured Ab29 was incubated with Fc fusion proteins of LILRB and LILRA family receptors. The amount of Fc fusion bound onto the sensors was presented as wavelength shift in nanometers (nm). The red dotted vertical line marks the transit from association stage to dissociation stage, where the sensors were dipped into kinetics buffer without Fc fusion receptors allowing free dissociation. LILRB2-Fc was included as the positive control. n. Maximum wavelength shifts of the Ab29-LILR binding curve in association stage as presented in m.

Additional file 10: Supplementary Table 4. Titration profiles of purified LILRB2 antibodies binding to plate-coated LILRB2 as measured by ELISA.

Additional file 6: Supplementary Figure 6. Identification of key domains and amino acid residues of LILRB2 for ligands and antibody binding. a. HA Ab binding to LILRB2 mutants as measured in chimeric reporter cells. Chimeric NFAT-GFP reporter cells expressing individual mutants of LILRB2 (listed in x-axis) were incubated with plate-coated HA antibody 12CA5. The activation of reporter cells is shown as percentages of GFP+ cells. b-d. oAβ, PS, or Ab29 binding to LILRB2 mutants as measured on chimeric reporter cells. Chimeric NFAT-GFP reporter cells expressing individual mutants of LILRB2 (listed in x-axis) were incubated with plate-coated oAβ (b), PS (c), or Ab29 (d). The activation of reporter cells is shown as percentages of GFP+ cells. Data are presented as mean ± SD (n = 4 independent experiments).

Additional file 9: Supplementary Table 3. Binding kinetics parameters between LILRB2 antibodies and LILRB2.

Additional file 7: Supplementary Table 1. Titration of blocking activities of purified LILRB2 antibodies against oAβ-LILRB2 interactions.

High moisture herbaceous residues (HR) cause a series of environment problems. Torrefaction has proven to be a promising technology for upgrading the fuel properties of biomass. In this study, a typical HR, Licorice residue (LR), was torrefied in a tubular reactor under nitrogen at 210, 240 and 280 °C. The effect of temperature on the fuel properties and the decomposition characteristic of the torrefied samples, as well as the pyrolysis products, were investigated. The mass and energy yield were altered in the range of 91.8–52.3% and 99.3–72.9%, respectively. The higher heating value (HHV) value of the torrefied sample increased by 8.1–39.5% from light to severe torrefaction, compared to untorrefied LR, and reached up to 23.3 MJ/kg at 280 °C. Torrefaction decreased the content of oxygenated organic compounds and the acidity of bio-oil from pyrolysis. The increase in alkanes from torrefied sample pyrolysis implies that torrefied LR has the potential for use in liquid fuel production. The present findings revealed that torrefaction turns industrial LR with poor energy content into a more easily exploitable solid fuel.

Digital elevation models (DEMs) are becoming increasingly important components in national and regional spatial data infrastructure. High-quality DEMs can now be derived directly from airborne light detection and ranging (LiDAR) point-cloud data of high spatial density if the derivation process can be verified. However, LiDAR is relatively new compared with other technologies for terrain data collection, and, although offering the potential for providing better spatial resolution than those that have been routinely available before, will not diffuse among DEM users until the results of meeting the verification challenge are favourable enough to inspire re-organisation of spatial data in decision support for catchment management and other third-tier-of-government authorities. By way of exemplification, the research presented in this thesis concerned ways of improving the processing of the airborne LiDAR data for high-quality DEM generation in terms of both accuracy and efficiency, and explored the applications of LiDAR-derived DEMs in the region of the Corangamite Catchment Management Authority, Victoria, Australia. This thesis begins with a review of the traditional technologies for terrain data collection and DEM generation and compares them with the LiDAR technology. Accordingly, a review of the recently-reported advances in LiDAR data deployment for DEM generation is followed by reports of experiments designed to improve selection and deployment of LiDAR data filtering, modelling methods and data reduction, and the achievement of vertical accuracy for different land covers. Also reported are results of deployment of LiDAR data for ground truthing, and application of LiDAR data for the extraction of drainage networks on an area of deranged drainage: the Victorian Volcanic Plain. The show that: (a) the issues of filtering, modelling techniques, interpolation methods, DEM resolution, and data reduction are critical and must be considered carefully when using LiDAR data for a high-quality DEM generation; (b) it is efficient to use survey marks for the accuracy assessment of LiDAR data. Normal distribution must be tested in order to select a suitable measure for the accuracy assessment of LiDAR data over different land covers; (c) LiDAR data reduction can improve the terrain production efficiency without compromising the product quality. The deployment of breaklines made a significant contribution to improving the accuracy of terrain models while allowing for data reduction; (d) it demonstrated the practical feasibility of applying ground control points from LiDAR intensity image and LiDAR-derived DEM in image orthorectification. The resultant orthoimage accuracy was shown to be superior to that achieved by using (lower accuracy) data sources such as those from Vicmap data; and (e) the LiDAR-derived DEM offers the capability of extracting and delineating the drainage networks in much more detail in low¬relief terrain, including areas in which drainage is barely coherent; The advantages of using LiDAR-derived DEM over the lower-accuracy DEM emerge in terms of stream order, stream number and stream length.

To explore the feasibility of using herbal medicine waste (HMW) as solid fuel, HMW was torrefied under different temperatures and atmospheres. The physicochemical properties and combustion behaviors of the torrefied HMW were investigated. Temperature was found to be the most influential factor affecting the torrefaction. Torrefaction improved the hydrophobicity of HMW and decreased the equilibrated moisture uptake from 24.48(0.083) % to 15.22(0.054) %. The HMW samples torrefied under different conditions are easy to ignite. The comprehensive combustibility index (S) of the torrefied HMW increased by 3–5 folds compared to that of the raw sample. In general, the HMW torrefied under lower temperatures and under CO2 and O2 have better flammability. The present results revealed that the torrefied HMW exhibited good combustion characteristics and can thus be used for solid fuel production, such as fuels for co-combustion or raw materials for pelletization.

Gastric cancer (GC) is a malignant tumor with an adverse health effect worldwide, whereas the underlying mechanism of GC development remains controversial. Identification of biomarkers is critical for the treatment of GC. Increasing evidence demonstrates that protein modification plays a pivotal role in carcinogenesis. USP38 is a member of the ubiquitin-specific protease (USP) family, which promotes protein stability by deubiquitinating the target proteins. In this study, we focused on the effect of USP38 on the GC and explored its underlying mechanism. The Cancer Genome Atlas (TCGA) database was used to evaluate the expression of USP38. AGS and HGC27 cells were treated with siRNA targeting USP38 or plasmids overexpressing USP38 to disturb levels of USP38. Immumohistochemical staining was performed to detect the level of USP38 and FASN. RT-qPCR and Western blotting (WB) were used to analyze the expression of mRNA and protein respectively. CCK8 assay, colony formation, cell migration assay, and cell apoptosis and cell cycle were performed to assess cell proliferation and migration ability. A subcutaneous tumor mice model was carried to verify the effect of USP38 on the GC in vivo. In this research, we found that USP38 was overexpressed in GC tissues, and USP38 contributed to GC cell proliferation, migration and tumorigenesis. Cell cycle and apoptosis were also regulated by USP38. Mechanistically, USP38 interacted with FASN, which resulted in enhanced protein stability of FASN and increased triglyceride production. Furthermore, FASN was critical for GC cell growth, migration and tumor development triggered by USP38 overexpression because its inhibitor orilistat reversed phenotypes in USP38 overexpressed GC cells. Collectively, USP38 overexpression is critical for GC cell growth, migration and tumorigenesis. Targeting FASN with inhibitors could be used as a potential treatment for GC patients with highly expressed USP38.

Anatase (TiO₂) has been widely used in photocatalysis. However, it can only absorb near-ultraviolet light with a wavelength below approximately 388 nm due to a wide band gap. Therefore a modification should be made for anatase to increase its capability in utilizing more abundant visible light. We investigated the doped anatase with the most promising 3d transition metal elements, and the results showed that the visible light absorption intensity was increased significantly due to the reduced band gap and the cavitation effects. As compared to other 3d transition metals, Cu was found to be the most effective one in improving anatase photocatalytic effects. In addition, greater Cu concentration doped in the anatase increased the photocatalysis effects but reduced the anatase stability, therefore, an optimized Cu concentration should be considered to optimize the anatase photocatalysis activity. Refereed/Peer-reviewed