A potent Henipavirus cross-neutralizing antibody reveals a dynamic fusion-triggering pattern of the G-tetramer.

The Hendra and Nipah viruses (HNVs) are highly pathogenic pathogens without approved interventions for human use. In addition, the interaction pattern between the attachment (G) and fusion (F) glycoproteins required for virus entry remains unclear. Here, we isolate a panel of Macaca-derived G-specific antibodies that cross-neutralize HNVs via multiple mechanisms. The most potent antibody, 1E5, confers adequate protection against the Nipah virus challenge in female hamsters. Crystallography demonstrates that 1E5 has a highly similar binding pattern to the receptor. In cryo-electron microscopy studies, the tendency of 1E5 to bind to the upper or lower heads results in two distinct quaternary structures of G. Furthermore, we identify the extended outer loop ß1S2-ß1S3 of G and two pockets on the apical region of fusion (F) glycoprotein as the essential sites for G-F interactions. This work highlights promising drug candidates against HNVs and contributes deeper insights into the viruses.

Broadly neutralizing antibodies against Omicron-included SARS-CoV-2 variants induced by vaccination.

The SARS-CoV-2 Omicron variant shows substantial resistance to neutralization by infection- and vaccination-induced antibodies, highlighting the demands for research on the continuing discovery of broadly neutralizing antibodies (bnAbs). Here, we developed a panel of bnAbs against Omicron and other variants of concern (VOCs) elicited by vaccination of adenovirus-vectored COVID-19 vaccine (Ad5-nCoV). We also investigated the human longitudinal antibody responses following vaccination and demonstrated how the bnAbs evolved over time. A monoclonal antibody (mAb), named ZWD12, exhibited potent and broad neutralization against SARS-CoV-2 variants Alpha, Beta, Gamma, Kappa, Delta, and Omicron by blocking the spike protein binding to the angiotensin-converting enzyme 2 (ACE2) and provided complete protection in the challenged prophylactic and therapeutic K18-hACE2 transgenic mouse model. We defined the ZWD12 epitope by determining its structure in complex with the spike (S) protein via cryo-electron microscopy. This study affords the potential to develop broadly therapeutic mAb drugs and suggests that the RBD epitope bound by ZWD12 is a rational target for the design of a broad spectrum of vaccines.

Upregulated circRNA_102231 promotes gastric cancer progression and its clinical significance.

Circular RNAs (circRNAs) are a type of endogenous non-coding RNAs implicated in cancer progression. This study explored the expression levels, clinical implication and possible molecular mechanism of circRNA_102231 in gastric cancer (GC). Gene Expression Omnibus (GEO) was used to analyze differentially expressed circRNAs. CircRNA_102231 expression was verified by qRT-PCR in GC tissues and plasma. The effects of circRNA_102231 was tested by CCK-8, colony formation, EdU and Transwell assays and xenograft tumor model. RNA pull-down and immunoprecipitation (RIP) assays were used to analyze the interaction between circRNA_102231 and IRTKS. CircRNA_102231 expression was significantly upregulated in GC tissue and plasma samples, which can be used as a biomarker for GC diagnosis and prognosis. The function assays showed that circRNA_102231 knockdown inhibited GC cell proliferation and invasion both in vitro and in vivo. CircRNA_102231 was able to bind to IRTKS, increasing IRTKS protein stability, leading to GC progression. Overexpression of IRTKS effectively rescued the reduced cell viability and invasion caused by silencing of circRNA_102231. In sum, our data demonstrate that circRNA_102231 is a novel oncogene in GC and acts as a potential biomarker and therapeutic target for GC patients.AbbreviationscircRNAs: circular RNAs; GC: gastric cancer; GEO: Gene Expression Omnibus; RIP: RNA immunoprecipitation; DEGs: differentially expressed genes.

Bioprinting of Complex Vascularized Tissues.

Functional vasculature is crucial for the maintenance of living tissues via the transport of oxygen, nutrients, and metabolic waste products. As a result, insufficient vascularization in thick engineered tissues will lead to cell death and necrosis due to mass transport and diffusional constraints. To circumvent these limitations, we describe the development of a microscale continuous optical bioprinting (µCOB) platform for 3D printing complex vascularized tissues with superior resolution and speed. By using the µCOB system, endothelial cells and other supportive cells can be printed directly into hydrogels with precisely controlled distribution and subsequent formation of lumen-like structures in vitro.

Three-dimensional bioprinted glioblastoma microenvironments model cellular dependencies and immune interactions.

Brain tumors are dynamic complex ecosystems with multiple cell types. To model the brain tumor microenvironment in a reproducible and scalable system, we developed a rapid three-dimensional (3D) bioprinting method to construct clinically relevant biomimetic tissue models. In recurrent glioblastoma, macrophages/microglia prominently contribute to the tumor mass. To parse the function of macrophages in 3D, we compared the growth of glioblastoma stem cells (GSCs) alone or with astrocytes and neural precursor cells in a hyaluronic acid-rich hydrogel, with or without macrophage. Bioprinted constructs integrating macrophage recapitulate patient-derived transcriptional profiles predictive of patient survival, maintenance of stemness, invasion, and drug resistance. Whole-genome CRISPR screening with bioprinted complex systems identified unique molecular dependencies in GSCs, relative to sphere culture. Multicellular bioprinted models serve as a scalable and physiologic platform to interrogate drug sensitivity, cellular crosstalk, invasion, context-specific functional dependencies, as well as immunologic interactions in a species-matched neural environment.

Photopolymerizable Biomaterials and Light-Based 3D Printing Strategies for Biomedical Applications.

Since the advent of additive manufacturing, known commonly as 3D printing, this technology has revolutionized the biofabrication landscape and driven numerous pivotal advancements in tissue engineering and regenerative medicine. Many 3D printing methods were developed in short course after Charles Hull first introduced the power of stereolithography to the world. However, materials development was not met with the same enthusiasm and remained the bottleneck in the field for some time. Only in the past decade has there been deliberate development to expand the materials toolbox for 3D printing applications to meet the true potential of 3D printing technologies. Herein, we review the development of biomaterials suited for light-based 3D printing modalities with an emphasis on bioprinting applications. We discuss the chemical mechanisms that govern photopolymerization and highlight the application of natural, synthetic, and composite biomaterials as 3D printed hydrogels. Because the quality of a 3D printed construct is highly dependent on both the material properties and processing technique, we included a final section on the theoretical and practical aspects behind light-based 3D printing as well as ways to employ that knowledge to troubleshoot and standardize the optimization of printing parameters.

Modulating physical, chemical, and biological properties in 3D printing for tissue engineering applications.

Over the years, 3D printing technologies have transformed the field of tissue engineering and regenerative medicine by providing a tool that enables unprecedented flexibility, speed, control, and precision over conventional manufacturing methods. As a result, there has been a growing body of research focused on the development of complex biomimetic tissues and organs produced via 3D printing to serve in various applications ranging from models for drug development to translational research and biological studies. With the eventual goal to produce functional tissues, an important feature in 3D printing is the ability to tune and modulate the microenvironment to better mimic in vivo conditions to improve tissue maturation and performance. This paper reviews various strategies and techniques employed in 3D printing from the perspective of achieving control over physical, chemical, and biological properties to provide a conducive microenvironment for the development of physiologically relevant tissues. We will also highlight the current limitations associated with attaining each of these properties in addition to introducing challenges that need to be addressed for advancing future 3D printing approaches.

Porous double network gels with high toughness, high stretchability and fast solvent-absorption.

Using the freeze-drying method, we fabricated porous double network gels with high toughness, high stretchability and fast solvent-absorption. When the freezing temperature was -20 °C and the freezing time was 24 hours, pores with diameters around 300 µm could form in the gel. When the freezing temperature was lowered to -196 °C and the freezing time was reduced to 10 minutes, monodisperse pores with diameters around 15 µm could form in the gel. We found out that both porous gels fabricated under different conditions could absorb solvent much more and much faster than a nonporous gel. Furthermore, we found that the rupturing strain, stiffness and strength of the porous double network gels were all comparable to the nonporous double network gel when containing the same amount of solvent. The unique combination of the mechanical properties of the porous double network gels might motivate new explorations of gels in practical applications.

[Priming Effects of Soil Moisture on Soil Respiration Under Different Tillage Practices].

In the early stage of an incubation experiment, soil respiration has a sensitive response to different levels of soil moisture. To investigate the effects of soil moisture on soil respiration under different tillage practices, we designed an incubation trial using air-dried soil samples collected from tillage experiment station established on black soils in 2001. The tillage experiment consisted of no-tillage (NT), ridge tillage (RT), and conventional tillage (CT). According to field capacity (water-holding capacity, WHC), we set nine moisture levels including 30%, 60%, 90%, 120%, 150%, 180%, 210%, 240%, 270% WHC. During the 22-day short-term incubation, soil CO2 emission was measured. In the early stage of incubation, the priming effects occurred under all tillage practices. There were positive correlations between soil respiration and soil moisture. In addition to drought and flood conditions, soil CO2 fluxes followed the order of NT > RT > CT. We fitted the relationship between soil moisture and soil CO2 fluxes under different tillage practices. In the range of 30%-270% WHC, soil CO2 fluxes and soil moisture fitted a quadratic regression equation under NT, and linear regression equations under RT and CT. Under the conditions of 30%-210% WHC of both NT and RT, soil CO2 fluxes and soil moisture were well fitted by the logarithmic equation with fitting coefficient R² = 0.966 and 0.956, respectively.

No tillage combined with crop rotation improves soil microbial community composition and metabolic activity.

Soil microbial community can vary with different agricultural managements, which in turn can affect soil quality. The objective of this work was to evaluate the effects of long-term tillage practice (no tillage (NT) and conventional tillage (CT)) and crop rotation (maize-soybean (MS) rotation and monoculture maize (MM)) on soil microbial community composition and metabolic capacity in different soil layers. Long-term NT increased the soil organic carbon (SOC) and total nitrogen (TN) mainly at the 0-5 cm depth which was accompanied with a greater microbial abundance. The greater fungi-to-bacteria (F/B) ratio was found in NTMS at the 0-5 cm depth. Both tillage and crop rotation had a significant effect on the metabolic activity, with the greatest average well color development (AWCD) value in NTMS soil at all three soil depths. Redundancy analysis (RDA) showed that the shift in microbial community composition was accompanied with the changes in capacity of utilizing different carbon substrates. Therefore, no tillage combined with crop rotation could improve soil biological quality and make agricultural systems more sustainable.

[Effects of corn and soybean straws returning on CO2 efflux at initial stage in black soil].

In this study, the CO2 emission characteristics and its relationships with C and N concentration in soil amended with different types of residues were studied by thermostatic incubation method to investigate the decomposition characteristics of different types of residues after adding to the soil and the effect of C, N concentration in residues on carbon sequestration. The results showed that during 61 days incubation, the CO2 efflux rates in the soils added with the different residues changed over time and exhibited an initial decrease, followed by a stable low plateau, and then an increase to a high plateau and finally followed by a decrease. The characteristics of CO2 emissions varied with residues, with the differences mainly occurring in the starting and duration of the high plateau CO2 emission period. The cumulative CO2-C emission was significantly affected by residue type. The cumulative CO2-C emissions from soils amended with corn roots, bottom corn stalks, corn leaves, and soybean leaves (about 160 µmol · g(-1) of soil and residue) were significantly greater than those from soils amended with other residues for the initial 21 days. Except for soybean leaves, the cumulative soil CO2 emissions over the 61 day incubation period from soils amended with soybean residues were higher than that from soil amended with corn residues. There were significant linear relationships between the ratio of cumulative CO2-C emission to residue carbon concentration (CR), and both C/N and nitrogen concentration of residues in the initial 21 days incubation, but not for the entire 61 days incubation. Our study suggested that soil CO2 emission was closely dependent upon the type of residue. Soybean residues decomposed more easily than corn residues. However, the decay rate of soybean residues was slower than that of corn residues at the initial stage of incubation. Soil CO2 emission was significantly affected by the C/N ratios and nitrogen concentrations of crop residues only at the early phase of incubation.

[Effects of Different Residue Part Inputs of Corn Straws on CO2 Efflux and Microbial Biomass in Clay Loam and Sandy Loam Black Soils].

The decomposed rate of crop residues is a major determinant for carbon balance and nutrient cycling in agroecosystem. In this study, a constant temperature incubation study was conducted to evaluate CO2 emission and microbial biomass based on four different parts of corn straw (roots, lower stem, upper stem and leaves) and two soils with different textures (sandy loam and clay loam) from the black soil region. The relationships between soil CO2 emission, microbial biomass and the ratio of carbon (C) to nitrogen (N) and lignin of corn residues were analyzed by the linear regression. Results showed that the production of CO2 was increased with the addition of different parts of corn straw to soil, with the value of priming effect (PE) ranged from 215. 53 µmol . g-1 to 335. 17 µmol . g -1. Except for corn leaves, the cumulative CO2 production and PE of clay loam soil were significantly higher than those in sandy loam soil. The correlation of PE with lignin/N was obviously more significant than that with lignin concentration, nitrogen concentration and C/N of corn residue. The addition of corn straw to soil increased the contents of MBC and MBN and decreased MBC/MBN, which suggested that more nitrogen rather than carbon was conserved in microbial community. The augmenter of microbial biomass in sandy loam soil was greater than that in clay loam soil, but the total dissolved nitrogen was lower. Our results indicated that the differences in CO2 emission with the addition of residues to soils were primarily ascribe to the different lignin/N ratio in different corn parts; and the corn residues added into the sandy loam soil could enhance carbon sequestration, microbial biomass and nitrogen holding ability relative to clay loam soil.

[Impact of tillage practices on microbial biomass carbon in top layer of black soils].

A study was conducted on a long-term (13 years) tillage and rotation experiment on black soil in northeast China to determine the effects of tillage, time and soil depth on soil microbial biomass carbon (MBC). Tillage systems included no tillage (NT), ridge tillage (RT) and mould-board plough (MP). Soil sampling was done at 0-5, 5-10 and 10-20 cm depths in June, August and September, 2013, and April, 2014 in the corn phase of corn-soybean rotation plots. MBC content was measured by the chloroform fumigation extraction (CFE) method. The results showed that the MBC content varied with sampling time and soil depth. Soil MBC content was the lowest in April for all three tillage systems, and was highest in June for MP, and highest in August for NT and RT. At each sampling time, tillage system had a significant effect on soil MBC content only in the top 0-5 cm layer. The MBC content showed obvious stratification under NT and RT with a higher MBC content in the top 0-5 cm layer than under MP. The stratification ratios under NT and RT were greatest in September when they were respectively 67.8% and 95.5% greater than under MP. Our results showed that soil MBC contents were greatly affected by the time and soil depth, and were more apparently accumulated in the top layer under NT and RT.

25th anniversary article: double emulsion templated solid microcapsules: mechanics and controlled release.

How droplet microfluidics can be used to fabricate solid-shelled microcapsules having precisely controlled release behavior is described. Glass capillary devices enable the production of monodisperse double emulsion drops, which can then be used as templates for microcapsule formation. The exquisite control afforded by microfluidics can be used to tune the compositions and geometrical characteristics of the microcapsules with exceptional precision. The use of this approach to fabricate microcapsules that only release their contents when exposed to a specific stimulus--such as a change in temperature, exposure to light, a change in the chemical environment, or an external stress--only after a prescribed time delay, and at a prescribed rate is reviewed.

Enhanced encapsulation of actives in self-sealing microcapsules by precipitation in capsule shells.

Microcapsules with core-shell structures are excellent vehicles for the encapsulation of active ingredients; however, the actives often leak out of these structures over time, without observable damage to them. We present a novel approach to enhancing the encapsulation of active ingredients inside microcapsules. We use two components that can form solid precipitates upon mixing and add one each to the microcapsule core and to the continuous phase. The components diffuse through the shell in the same manner as the actives, but upon meeting, they precipitate to form solid particles within the shell; this significantly reduces leakage through the shell of the microcapsules. We show that the reduction in the leakage of actives is due to the blockage of channels or pores that exist in the shell of the capsules by the solid precipitates.

Integrated microdynamics mechanism of the thermal-induced phase separation behavior of poly(vinyl methyl ether) aqueous solution.

The thermal behavior of a poly(vinyl methyl ether) (PVME) aqueous solution (30 wt %) during a heating-and-cooling cycle is studied using FTIR spectroscopy in combination with 2D correlation analysis. The FTIR spectral data of O-H, CH(3)-O, and C-H stretching vibration regions provide detailed changes of hydrophilic and hydrophobic groups of PVME. Hydrogen bonds between hydrophilic groups and water and hydration interactions between hydrophobic groups and water are confirmed to be completely reversible in the heating-and-cooling cycle. Two-dimensional correlation method helps us to understand the microdynamics mechanism of phase separation behavior of PVME 30 wt % aqueous solution. During the heating process, the initially hydrated CH(3) groups start to dehydrate as the first action of phase separation, and the initially hydrated CH(2) groups follow to start their dehydration; interestingly, water molecules leave CH(2) groups very fast, and the whole dehydration process of CH(2) groups finishes even earlier than that of CH(3). After hydrophobic groups finish their dehydrations, hydrogen bonds between hydrophilic group and water start to dissociate. 1:2 adducts formed between PVME and water dissociate first and transfer to the 1:1 adducts, whereas with further heating, 1:1 adducts eventually dissociate and release free water and free CH(3)-O. PCMW method is used as supplement to determine changing conditions of various chemical structures. During the phase separation, O-H hydrogen bond in 1:2 adduct is found to dissociate between 35.5 and 39 °C in a [Formula: see text] style, whereas the 1:1 adduct (also considered as free water) increases between 35.5 and 39 °C in a [Formula: see text] style. Moreover, dehydration conditions of hydrophobic groups are also found. Both of the dehydrated states CH(3) and CH(2) increase like [Formula: see text].

Plasticity of marrow mesenchymal stem cells from human first-trimester fetus: from single-cell clone to neuronal differentiation.

Recent results have shown that bone marrow mesenchymal stem cells (BMSCs) from human first-trimester abortus (hfBMSCs) are closer to embryonic stem cells and perform greater telomerase activity and faster propagation than mid- and late-prophase fetal and adult BMSCs. However, no research has been done on the plasticity of hfBMSCs into neuronal cells using single-cell cloned strains without cell contamination. In this study, we isolated five single cells from hfBMSCs and obtained five single-cell cloned strains, and investigated their biological property and neuronal differentiation potential. We found that four of the five strains showed similar expression profile of surface antigen markers to hfBMSCs, and most of them differentiated into neuron-like cells expressing Nestin, Pax6, Sox1, ß-III Tubulin, NF-L, and NSE under induction. One strain showed different expression profile of surface antigen markers from the four strains and hfBMSCs, and did not differentiate toward neuronal cells. We demonstrated for the first time that some of single-cell cloned strains from hfBMSCs can differentiate into nerve tissue-like cell clusters under induction in vitro, and that the plasticity of each single-cell cloned strain into neuronal cells is different.

Trace of the thermally induced evolution mechanism of interactions between water and ionic liquids.

The thermally induced evolution mechanisms of various interactions in ionic liquids (1-butyl-3-methylimidazolium tetrafluoroborate, bmimBF(4)) and water mixtures have been investigated in this paper. In the near-infrared (NIR) spectroscopy, we focus mainly on nu(OH) and nu(CH) overtone regions. During heating of bmimBF(4) and water mixtures, the nu(OH) overtone peak shows a significant blue shift, and the area of this peak shows different changes in three heating regions. By using the perturbation correlation moving window (PCMW) method, we have ascertained the critical temperatures of these three regions: 25-100, 105-160, and 165-190 degrees C, and we have accordingly performed 2D correlation NIR analysis in three parts. On the basis of 2D study results, we find several types of O-H involved hydrogen bonds (H-B's) in a bmimBF(4) and water (15 mol %) mixture and arrive at their evolution mechanisms in each heating region. During heating at 25-100 degrees C, strong H-B's, such as BF(4)(-)...water...BF(4)(-) and BF(4)(-)...cyclic water dimer...BF(4)(-), transform into weaker H-B's with simpler structures; at 105-160 degrees C, the remaining BF(4)(-)...water...BF(4)(-) continues to dissociate, and cation...water H-B's start to dissociate, and a large amount of released free water evaporates; whereas in the final heating region of 165-190 degrees C, BF(4)(-)...water...BF(4)(-) still exists and continues to dissociate, and in the study of the nu(CH) overtone region, we have found that the concentration of water in bmimBF(4) affects interactions between cations and anions. In the mixture of bmimBF(4) with more water (15 mol %), H-B's between water and bmimBF(4) cannot be completely destroyed, even at very high temperature; therefore, only limited new electrostatic interactions would be formed between cations and anions during heating, but in a mixture of bmimBF(4) with less water (15 mol %), cations and anions are able to form new electrostatic interactions during the heating process. However, the intensity of these interactions is smaller than that in the 80 degrees C isothermal process due to the low contacting possibilities among ions at high temperatures.

Trace of the interesting "V"-shaped dynamic mechanism of interactions between water and ionic liquids.

The mixture of ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate, bmimBF4) and water (2.5%, molar fraction) under isothermal conditions at 80 degrees C was investigated by FTIR spectroscopy and two-dimensional correlation infrared spectroscopy (2D-IR) methods. Three regions were focused: the OH stretching band of water (3755-3300 cm (-1)), the stretching band of CH on the imidazole ring (3300-3020 cm (-1)), and the BF stretching band of anions (1310-1260 cm (-1)). During this process, water was gradually evaporated as time passed, which produced influences on the interactions among cations, anions, and water molecules. In the FTIR analysis, we found an interesting "V"-shaped changing trend in peak areas of the C-H on the imidazole ring and the B-F stretching band; the inflection of the system was 913 s, gained through the "moving window" method. A two-step variation was accordingly found during this process. Hydrogen bonds formed by water molecules with cations or water molecules with anions were destroyed by the reduction of water, making a fall in the former period of "V" process, while electrostatic interactions newly formed between anions and cations leading to a rise during the latter period of this course. In this paper, various conformations formed among cations, anions, and water molecules were clearly assigned, and we managed to trace the whole dynamic mechanism of this isothermal process by 2D-IR techniques.

Phase separation of poly(vinyl methyl ether) aqueous solution: a near-infrared spectroscopic study.

The thermosensitive phase separation of poly(vinyl methyl ether) (PVME) aqueous solutions has been investigated using near-infrared spectroscopy in combination with two-dimensional correlation analysis, and a two-step phase separation mechanism during gradual heating has been established. Two-dimensional near-infrared (2D NIR) analysis results indicate that during this two-step process the dehydration of CH 2 groups occurs earlier than that of CH 3 groups. This result suggests that it is the change of the hydrophobic hydrocarbon chain conformation induced by heating that indirectly leads to the dehydration of the hydrophilic ether oxygen side groups.