High Potency of a Bivalent Human VH Domain in SARS-CoV-2 Animal Models.

Novel COVID-19 therapeutics are urgently needed. We generated a phage-displayed human antibody VH domain library from which we identified a high-affinity VH binder ab8. Bivalent VH, VH-Fc ab8, bound with high avidity to membrane-associated S glycoprotein and to mutants found in patients. It potently neutralized mouse-adapted SARS-CoV-2 in wild-type mice at a dose as low as 2 mg/kg and exhibited high prophylactic and therapeutic efficacy in a hamster model of SARS-CoV-2 infection, possibly enhanced by its relatively small size. Electron microscopy combined with scanning mutagenesis identified ab8 interactions with all three S protomers and showed how ab8 neutralized the virus by directly interfering with ACE2 binding. VH-Fc ab8 did not aggregate and did not bind to 5,300 human membrane-associated proteins. The potent neutralization activity of VH-Fc ab8 combined with good developability properties and cross-reactivity to SARS-CoV-2 mutants provide a strong rationale for its evaluation as a COVID-19 therapeutic.

A tumor-conditional IL-15 safely synergizes with immunotherapy to enhance antitumor immune responses.

It is a challenge to invigorate tumor-infiltrating lymphocytes without causing immune-related adverse events, which also stands as a primary factor contributing to resistance against cancer immunotherapies. IL-15 can potently promote expansion and activation of T cells, but its clinical use has been limited by dose-limiting toxicities. In this study, we develop a tumor-conditional IL-15 (pro-IL-15), which masks IL-15 with steric hindrance caused by Fc fragment and IL-15Rα-sushi domain. Upon reaching the tumor site, it can be cleaved by tumor-associated proteases to release an IL-15 superagonist, resulting in potent antitumor activities. Systemic delivery of pro-IL-15 demonstrates significantly reduced toxicity but uncompromised antitumor efficacy. Pro-IL-15 can yield better effectors and vitalize terminally exhausted CD8+ T cells to overcome checkpoint blockade resistance. Moreover, pro-IL-15 promotes chemotaxis and activation of adoptive T cells, leading to eradication of advanced solid tumors and durable cures. Furthermore, pro-IL-15 shows promise for synergizing with other immunotherapies like IL-12 and oncolytic virus by improving CD8/Treg ratio and IFN-γ levels, resulting in substantial regression of both local and metastatic cold tumors. Collectively, our results suggest that pro-IL-15 represents a compelling strategy for overcoming resistance to current immunotherapies while avoiding toxicities.

Rapid identification of a human antibody with high prophylactic and therapeutic efficacy in three animal models of SARS-CoV-2 infection.

Effective therapies are urgently needed for the SARS-CoV-2/COVID-19 pandemic. We identified panels of fully human monoclonal antibodies (mAbs) from large phage-displayed Fab, scFv, and VH libraries by panning against the receptor binding domain (RBD) of the SARS-CoV-2 spike (S) glycoprotein. A high-affinity Fab was selected from one of the libraries and converted to a full-size antibody, IgG1 ab1, which competed with human ACE2 for binding to RBD. It potently neutralized replication-competent SARS-CoV-2 but not SARS-CoV, as measured by two different tissue culture assays, as well as a replication-competent mouse ACE2-adapted SARS-CoV-2 in BALB/c mice and native virus in hACE2-expressing transgenic mice showing activity at the lowest tested dose of 2 mg/kg. IgG1 ab1 also exhibited high prophylactic and therapeutic efficacy in a hamster model of SARS-CoV-2 infection. The mechanism of neutralization is by competition with ACE2 but could involve antibody-dependent cellular cytotoxicity (ADCC) as IgG1 ab1 had ADCC activity in vitro. The ab1 sequence has a relatively low number of somatic mutations, indicating that ab1-like antibodies could be quickly elicited during natural SARS-CoV-2 infection or by RBD-based vaccines. IgG1 ab1 did not aggregate, did not exhibit other developability liabilities, and did not bind to any of the 5,300 human membrane-associated proteins tested. These results suggest that IgG1 ab1 has potential for therapy and prophylaxis of SARS-CoV-2 infections. The rapid identification (within 6 d of availability of antigen for panning) of potent mAbs shows the value of large antibody libraries for response to public health threats from emerging microbes.

Coseismic Deformation Field and Fault Slip Distribution Inversion of the 2020 Jiashi Ms 6.4 Earthquake: Considering the Atmospheric Effect with Sentinel-1 Data Interferometry.

Due to some limitations associated with the atmospheric residual phase in Sentinel-1 data interferometry during the Jiashi earthquake, the detailed spatial distribution of the line-of-sight (LOS) surface deformation field is still not fully understood. This study, therefore, proposes an inversion method of coseismic deformation field and fault slip distribution, taking atmospheric effect into account to address this issue. First, an improved inverse distance weighted (IDW) interpolation tropospheric decomposition model is utilised to accurately estimate the turbulence component in tropospheric delay. Using the joint constraints of the corrected deformation fields, the geometric parameters of the seismogenic fault and the distribution of coseismic slip are then inverted. The findings show that the coseismic deformation field (long axis strike was nearly east-west) was distributed along the Kalpingtag fault and the Ozgertaou fault, and the earthquake was found to occur in the low dip thrust nappe structural belt at the subduction interface of the block. Correspondingly, the slip model further revealed that the slips were concentrated at depths between 10 and 20 km, with a maximum slip of 0.34 m. Accordingly, the seismic magnitude of the earthquake was estimated to be Ms 6.06. Considering the geological structure in the earthquake region and the fault source parameters, we infer that the Kepingtag reverse fault is responsible for the earthquake, and the improved IDW interpolation tropospheric decomposition model can perform atmospheric correction more effectively, which is also beneficial for the source parameter inversion of the Jiashi earthquake.

Structural details of monoclonal antibody m971 recognition of the membrane-proximal domain of CD22.

Cluster of differentiation-22 (CD22) belongs to the sialic acid-binding immunoglobulin (Ig)-like lectin family of receptors that is expressed on the surface of B cells. It has been classified as an inhibitory coreceptor for the B-cell receptor because of its function in establishing a baseline level of B-cell inhibition. The restricted expression of CD22 on B cells and its inhibitory function make it an attractive target for B-cell depletion in cases of B-cell malignancies. Genetically modified T cells with chimeric antigen receptors (CARs) derived from the m971 antibody have shown promise when used as an immunotherapeutic agent against B-cell acute lymphoblastic leukemia. A key aspect of the efficacy of this CAR-T was its ability to target a membrane-proximal epitope on the CD22 extracellular domain; however, the molecular details of m971 recognition of CD22 have thus far remained elusive. Here, we report the crystal structure of the m971 fragment antigen-binding in complex with the two most membrane-proximal Ig-like domains of CD22 (CD22d6-d7). The m971 epitope on CD22 resides at the most proximal Ig domain (d7) to the membrane, and the antibody paratope contains electrostatic surfaces compatible with interactions with phospholipid head groups. Together, our data identify molecular details underlying the successful transformation of an antibody epitope on CD22 into an effective CAR immunotherapeutic target.

What Role Does the Incident Light Intensity Play in Photocatalytic Conversion of CO2 : Attenuation or Intensification?

Artificial photoreduction of CO2 is vital for the sustainable development of human beings via solar energy storage in stable chemicals. This process involves intricate light-matter interactions, but the role of incident light intensity in photocatalysis remains obscure. Herein, the influence of excitation intensity on charge kinetics and photocatalytic activity is investigated. Model photocatalysts include the pure graphitic carbon nitride (g-C3 N4 ) and g-C3 N4 loaded with noble/non-noble-metal cocatalysts (Ag, TiN, and CuO). It is found that the increase of light intensity does not always improve the electron utilization. Overly high excitation intensities cause charge carrier congestion and changes the recombination mechanism, which is called the light congestion effect. The electron transport channels can be established to mitigate the light-induced effect via the addition of cocatalyst, leading to a nonlinear growth in the reaction rate with increasing light intensity. From experiments and simulations, it is found that the light intensity and active site density should be collectively optimized for increasing the energy conversion efficiency. This work elucidates the effect of light intensity on photocatalytic CO2 reduction and emphasizes the synergistic relationship of matching the light intensity and the photocatalyst category. The study provides guidance for the design of efficient photocatalysts and the operation of photocatalytic systems.

Developability Assessment of an Isolated CH2 Immunoglobulin Domain.

The IgG CH2 domain continues to hold promise for the development of new therapeutic entities because of its bifunctional role as a biomarker and effector protein. The need for further understanding of molecular stability and aggregation in therapeutic proteins has led to the development of a breakthrough quantum cascade laser microscope to allow for real-time comparability assessment of an array of related proteins in solution upon thermal perturbation. Our objective was to perform a comprehensive developability assessment of three similar monoclonal antibody (mAb) fragments: CH2, CH2s, and m01s. The CH2 construct consists of residues Pro238 to Lys340 of the IgG1 heavy chain sequence. CH2s has a 7-residue deletion at the N-terminus and a 16-residue C-terminal extension containing a histidine tag. The m01s construct is identical to CH2s, except for two cysteines introduced at positions 242 and 334. A series of hyperspectral images was acquired during thermal perturbation from 28 to 60 °C for all three proteins in an array. Co-distribution and two-dimensional infrared correlation spectroscopies yielded the mechanism of aggregation and stability for these three proteins. The level of detail is unprecedented, identifying the regions within CH2 and CH2s that are prone to self-association and establishing the differences in stability. Furthermore, CH2 helical segments, ß-sheets, ß-turns, and random coil regions were less stable than in CH2s and m01s because of the presence of the N-terminal 310-helix and ß-turn type III. The engineered disulfide bridge in m01s eliminated the self-association process and rendered this mAb fragment the most stable.

Compressed ultrafast tomographic imaging by passive spatiotemporal projections.

Existing streak-camera-based two-dimensional (2D) ultrafast imaging techniques are limited by long acquisition time, the trade-off between spatial and temporal resolutions, and a reduced field of view. They also require additional components, customization, or active illumination. Here we develop compressed ultrafast tomographic imaging (CUTI), which passively records 2D transient events with a standard streak camera. By grafting the concept of computed tomography to the spatiotemporal domain, the operations of temporal shearing and spatiotemporal integration in a streak camera's data acquisition can be equivalently expressed as the spatiotemporal projection of an (x,y,t) datacube from a certain angle. Aided by a new, to the best of our knowledge, compressed-sensing reconstruction algorithm, the 2D transient event can be accurately recovered in a few measurements. CUTI is exhibited as a new imaging mode universally adaptable to most streak cameras. Implemented in an image-converter streak camera, CUTI captures the sequential arrival of two spatially modulated ultrashort ultraviolet laser pulses at 0.5 trillion frames per second. Applied to a rotating-mirror streak camera, CUTI records an amination of fast-bouncing balls at 5,000 frames per second.

Extraction of a Weak Flow Field for a Multi-Rotor Unmanned Aerial Vehicle (UAV) Using High-Speed Background-Oriented Schlieren (BOS) Technology.

To optically capture and analyze the structure and changes of the flow field of a weak airflow object with high accuracy, this study proposes novel weak flow field extraction methods based on background-oriented schlieren. First, a fine background pattern texture and a sensor network layout were designed to satisfy the requirement of weak flow field extraction. Second, the image displacement was extracted by calculating the correlation matrix in the frequency domain for a particle image velocimetry algorithm, and further calculations were performed for the density field using Poisson's equation. Finally, the time series baseline stacking method was proposed to obtain the flow field changes of weak airflow structures. A combustion experiment was conducted to validate the feasibility and accuracy of the proposed method. The results of a quad-rotor unmanned aerial vehicle experiment showed that the clear, uneven, and continuous quantitative laminar flow field could be obtained directly, which overcame the interference of the weak airflow, large field of view, and asymmetrical steady flow.

Real-time high-speed three-dimensional surface imaging using band-limited illumination profilometry with a CoaXPress interface.

High-speed three-dimensional (3D) surface imaging by structured-light profilometry is currently driven by numerous applications. However, the limited speeds in fringe pattern projection, image acquisition, and data transmission have strained the existing methods from reaching kilohertz-level acquisition, processing, and display of 3D information during the occurrence of dynamic events (i.e., in real time). To overcome these limitations, we have developed band-limited illumination profilometry (BLIP) with a CoaXPress interface (CI), which enables real-time high-speed 3D surface imaging. We have demonstrated the system's performance by imaging various static and fast-moving 3D objects in real time. We have also applied this system in fluid mechanics by imaging dynamics of a flag, which allowed observation of the wave propagation, gravity-induced phase mismatch, and asymmetric flapping motion. We expect CI-BLIP to find diverse scientific and industrial applications.

Single-shot compressed optical-streaking ultra-high-speed photography.

Single-shot ultra-high-speed imaging is of great significance to capture transient phenomena in physics, biology, and chemistry in real time. Existing techniques, however, have a restricted application scope, a low sequence depth, or a limited pixel count. To overcome these limitations, we developed single-shot compressed optical-streaking ultra-high-speed photography (COSUP) with an imaging speed of 1.5 million frames per second, a sequence depth of 500 frames, and an (x,y) pixel count of 0.5 megapixels per frame. COSUP's single-shot ultra-high-speed imaging ability was demonstrated by recording single laser pulses illuminating through transmissive targets and by tracing a fast-moving object. As a universal imaging platform, COSUP is capable of increasing imaging speeds of a wide range of CCD and complementary metal-oxide-semiconductor cameras by four orders of magnitude. We envision COSUP to be applied in widespread applications in biomedicine and materials science.

Deformation Activity Analysis of a Ground Fissure Based on Instantaneous Total Energy.

This study proposes a novel instantaneous total energy method to perform an activity analysis of ground fissures deformation, which is calculated by integrating the extreme-point symmetric mode decomposition (ESMD) method and kinetic energy based on the time-series displacement acquired by shape acceleration array (SAA) sensors. The proposed method is tested on the Xiwang Road fissure in Beijing, China. First, to fully monitor the hanging wall and footwall of the monitored ground fissure, a 4 m-long SAA in the vertical direction and an 8 m-long SAA in the horizontal direction were embedded in a ground fissure to obtain an accurate time-series displacement with an accuracy of ±1.5 mm/32 m and a displacement acquisition frequency of once an hour. Second, to improve the accuracy of the activity analysis, the ESMD method and Spearman's rho are applied to perform signal denoising of the original time-series displacement obtained by the SAA sensors. Finally, the instantaneous total energy is obtained to analyze the activity of the monitored ground fissure. The results demonstrate that the proposed method is more reliable to reflect the activity of a monitored ground fissure compared to the time-series displacement.

Palladium-Catalyzed Heck-type Domino Cyclization and Carboxylation to Synthesize Carboxylic Acids by Utilizing Chloroform as the Carbon Monoxide Source.

A palladium-catalyzed domino cyclization and carboxylation reaction for synthesis of a variety of carboxylic acids was developed, where chloroform was used as "carbon monoxide" source. The in situ generated neopentylpalladium species by Heck cyclization was efficiently trapped by dichlorocarbene to form a series of carboxylic acids. It was found that in this type of domino reaction CHCl3 is a convenient and safe alternation for CO gas.

Palladium/copper-catalyzed oxidative arylation of terminal alkenes with aroyl hydrazides.

An unprecedented oxidative arylation reaction of terminal alkenes with simple aroyl hydrazides has been developed under aerobic conditions for the stereoselective synthesis of 1,2-disubstituted alkenes. A range of aroyl hydrazides underwent palladium/copper-catalyzed oxidative Mizoroki-Heck reaction with terminal alkenes open to air in a 1:1 mixture of dimethyl sulfoxide and acetonitrile to give structurally diverse 1,2-disubstituted alkenes in moderate to excellent yields with excellent regio- and E-selectivity. The reaction tolerated a wide variety of functional groups, such as alkoxy, hydroxy, amino, fluoro, chloro, bromo, cyano, nitro, ester, amide, imide, phosphine oxide, and sulfone groups, and, moreover, molecular oxygen and dimethyl sulfoxide were demonstrated to serve as terminal oxidants. This study provides a useful method for the stereoselective synthesis of 1,2-disubstituted alkenes through direct transformation of the vinylic CH bonds in terminal alkenes.

Metabolic engineering of Escherichia coli to enhance shikimic acid production from sorbitol.

Shikimic acid (SA) is the key synthetic material of Oseltamivir, which is an effective drug for the prevention and treatment of influenza. In this study, to block the downstream metabolic pathway of SA, the shikimate kinase isoenzyme genes aroK and aroL were deleted by Red recombination. Moreover, the key enzyme genes aroG, aroB, tktA and aroE of SA pathway were co-expressed by constructing the recombinant vector pETDuet-GBAE. As a result, SA production of E. coli BW25113 (∆aroL/aroK, DE3)/pETDuet-GBAE reached 1,077.6 mg/l when low amounts of sorbitol (5 g/l) were fed in shake flasks. The yield was 3.7 times that when glucose was used (P < 0.05). The results showed that sorbitol was an optimized carbon source for the high efficient accumulation of SA for the first time, which was applicable to use in the industry for high yields and low consumption.

Ultrastable halide perovskite CsPbBr3 photoanodes achieved with electrocatalytic glassy-carbon and boron-doped diamond sheets.

Halide perovskites exhibit exceptional optoelectronic properties for photoelectrochemical production of solar fuels and chemicals but their instability in aqueous electrolytes hampers their application. Here we present ultrastable perovskite CsPbBr3-based photoanodes achieved with both multifunctional glassy carbon and boron-doped diamond sheets coated with Ni nanopyramids and NiFeOOH. These perovskite photoanodes achieve record operational stability in aqueous electrolytes, preserving 95% of their initial photocurrent density for 168 h of continuous operation with the glassy carbon sheets and 97% for 210 h with the boron-doped diamond sheets, due to the excellent mechanical and chemical stability of glassy carbon, boron-doped diamond, and nickel metal. Moreover, these photoanodes reach a low water-oxidation onset potential close to +0.4 VRHE and photocurrent densities close to 8 mA cm-2 at 1.23 VRHE, owing to the high conductivity of glassy carbon and boron-doped diamond and the catalytic activity of NiFeOOH. The applied catalytic, protective sheets employ only earth-abundant elements and straightforward fabrication methods, engineering a solution for the success of halide perovskites in stable photoelectrochemical cells.

Efficient solar-driven CO2-to-fuel conversion via Ni/MgAlO x @SiO2 nanocomposites at low temperature.

Solar-driven CO2-to-fuel conversion assisted by another major greenhouse gas CH4 is promising to concurrently tackle energy shortage and global warming problems. However, current techniques still suffer from drawbacks of low efficiency, poor stability, and low selectivity. Here, a novel nanocomposite composed of interconnected Ni/MgAlO x nanoflakes grown on SiO2 particles with excellent spatial confinement of active sites is proposed for direct solar-driven CO2-to-fuel conversion. An ultrahigh light-to-fuel efficiency up to 35.7%, high production rates of H2 (136.6 mmol min-1g- 1) and CO (148.2 mmol min-1g-1), excellent selectivity (H2/CO ratio of 0.92), and good stability are reported simultaneously. These outstanding performances are attributed to strong metal-support interactions, improved CO2 absorption and activation, and decreased apparent activation energy under direct light illumination. MgAlO x @SiO2 support helps to lower the activation energy of CH* oxidation to CHO* and improve the dissociation of CH4 to CH3* as confirmed by DFT calculations. Moreover, the lattice oxygen of MgAlO x participates in the reaction and contributes to the removal of carbon deposition. This work provides promising routes for the conversion of greenhouse gasses into industrially valuable syngas with high efficiency, high selectivity, and benign sustainability.

Swept coded aperture real-time femtophotography.

Single-shot real-time femtophotography is indispensable for imaging ultrafast dynamics during their times of occurrence. Despite their advantages over conventional multi-shot approaches, existing techniques confront restricted imaging speed or degraded data quality by the deployed optoelectronic devices and face challenges in the application scope and acquisition accuracy. They are also hindered by the limitations in the acquirable information imposed by the sensing models. Here, we overcome these challenges by developing swept coded aperture real-time femtophotography (SCARF). This computational imaging modality enables all-optical ultrafast sweeping of a static coded aperture during the recording of an ultrafast event, bringing full-sequence encoding of up to 156.3 THz to every pixel on a CCD camera. We demonstrate SCARF's single-shot ultrafast imaging ability at tunable frame rates and spatial scales in both reflection and transmission modes. Using SCARF, we image ultrafast absorption in a semiconductor and ultrafast demagnetization of a metal alloy.

Discovery of a novel highly specific, fully human PSCA antibody and its application as an antibody-drug conjugate in prostate cancer.

Prostate stem cell antigen (PSCA) is expressed in all stages of prostate cancer, including in advanced androgen-independent tumors and bone metastasis. PSCA may associate with prostate carcinogenesis and lineage plasticity in prostate cancer. PSCA is also a promising theranostic marker for a variety of other solid tumors, including pancreatic adenocarcinoma and renal cell carcinoma. Here, we identified a novel fully human PSCA antibody using phage display methodology. The structure-based affinity maturation yielded a high-affinity binder, F12, which is highly specific and does not bind to 6,000 human membrane proteins based on a membrane proteome array assay. F12 targets PSCA amino acids 63-69 as tested by the peptide scanning microarray, and it cross-reacts with the murine PSCA. IgG1 F12 efficiently internalizes into PSCA-expressing tumor cells. The antimitotic reagent monomethyl auristatin E (MMAE)-conjugated IgG1 F12 (ADC, F12-MMAE) exhibits dose-dependent efficacy and specificity in a human prostate cancer PC-3-PSCA xenograft NSG mouse model. This is a first reported ADC based on a fully human PSCA antibody and MMAE that is characterized in a xenograft murine model, which warrants further optimizations and investigations in additional preclinical tumor models, including prostate and other solid tumors.

Revealing the stochastic kinetics evolution of photocatalytic CO2 reduction.

Investigating kinetic mechanisms to design efficient photocatalysts is critical for improving photocatalytic CO2 reduction, but the stochastic photo-physical/chemical properties of kinetics remain unclear. Herein, we propose a statistical study to discuss the stochastic feature evolution of photocatalytic systems. The uncertainties of light absorption, charge carrier migration, and surface reaction are described by nonparametric estimation methods in the proposed model, which includes the effect of operational and material parameters. The density distribution of surface electrons shifts from a skewed distribution to an approximate uniform distribution as incident photon density increases. The system temperature rising induces the rate-determining step of surface reactions to change from charge carrier kinetics to reactant activation processes. Benefiting from the synergistic optimization between the operational parameter and active site density, the electron-capturing probability of active sites is boosted from 0.06 to 0.17. The modified reaction kinetic equation is constructed based on the distribution function of charge carrier kinetics. Furthermore, the experimental photoactivity results are consistent with the statistical analysis, which proves the feasibility of the established model. The characterization tests show that the gap between testing activities and theoretical efficiency is caused by a mismatch between charge carrier supply and mass transfer. Our work unveils the stochastic features in photocatalytic CO2 reduction, offering a comprehensive analytical framework for photocatalytic system optimization.