Silk Fibroin Improves the Biological Properties of Egg White-Based Bioink for the Bioprinting of Tissue Engineering Materials.

Egg white (EW) is a common nutritious food with excellent heat gelation and biocompatibility, but its application in biomaterials is considerably limited. Silk fibroin (SF) is a protein-based fiber with both excellent mechanical properties and biocompatibility, and its application in biomaterials has attracted much attention. Here, the EW/SF composite scaffold was first synthesized with GMA-modified EW/SF composite bioink (G-EW/SF). When homogenized EW and SF were individually grafted with glycidyl methacrylate (GMA), the grafted EW (G-EW) and SF (G-SF) were mixed in different proportions and then added to I2959. The resulting G-EW/SF composite bioink could be bioprinted into various EW/SF composite scaffolds. Among them, the compressive modulus of EW/SF (50%) composite scaffolds incorporating 50% G-SF was significantly improved. It had a three-dimensional (3D) polypore structure with an average pore size of 61 µm and was mainly composed of ß-sheet structures. Compared with the EW scaffold alone, the thermal decomposition temperature of the EW/SF scaffold was 10 °C higher, and the residual rate after 9 days of enzymatic hydrolysis had increased by about 18%. The scaffold prolonged the sustained release of insulin and promoted the adhesion, growth, and proliferation of the L-929 cells. Therefore, the EW/SF composite scaffolds with good cell proliferation ability and certain mechanical properties can be used in different applications including cells, drugs, and tissues. These results provide new prospects for the application of the EW protein to medical tissue engineering materials.

Annotation of cell types (ACT): a convenient web server for cell type annotation.

BACKGROUND: The advancement of single-cell sequencing has progressed our ability to solve biological questions. Cell type annotation is of vital importance to this process, allowing for the analysis and interpretation of enormous single-cell datasets. At present, however, manual cell annotation which is the predominant approach remains limited by both speed and the requirement of expert knowledge. METHODS: To address these challenges, we constructed a hierarchically organized marker map through manually curating over 26,000 cell marker entries from about 7000 publications. We then developed WISE, a weighted and integrated gene set enrichment method, to integrate the prevalence of canonical markers and ordered differentially expressed genes of specific cell types in the marker map. Benchmarking analysis suggested that our method outperformed state-of-the-art methods. RESULTS: By integrating the marker map and WISE, we developed a user-friendly and convenient web server, ACT ( http://xteam.xbio.top/ACT/ or http://biocc.hrbmu.edu.cn/ACT/ ), which only takes a simple list of upregulated genes as input and provides interactive hierarchy maps, together with well-designed charts and statistical information, to accelerate the assignment of cell identities and made the results comparable to expert manual annotation. Besides, a pan-tissue marker map was constructed to assist in cell assignments in less-studied tissues. Applying ACT to three case studies showed that all cell clusters were quickly and accurately annotated, and multi-level and more refined cell types were identified. CONCLUSIONS: We developed a knowledge-based resource and a corresponding method, together with an intuitive graphical web interface, for cell type annotation. We believe that ACT, emerging as a powerful tool for cell type annotation, would be widely used in single-cell research and considerably accelerate the process of cell type identification.

Encephalitis and poor neuronal death-mediated control of herpes simplex virus in human inherited RIPK3 deficiency.

Inborn errors of TLR3-dependent type I IFN immunity in cortical neurons underlie forebrain herpes simplex virus-1 (HSV-1) encephalitis (HSE) due to uncontrolled viral growth and subsequent cell death. We report an otherwise healthy patient with HSE who was compound heterozygous for nonsense (R422*) and frameshift (P493fs9*) RIPK3 variants. Receptor-interacting protein kinase 3 (RIPK3) is a ubiquitous cytoplasmic kinase regulating cell death outcomes, including apoptosis and necroptosis. In vitro, the R422* and P493fs9* RIPK3 proteins impaired cellular apoptosis and necroptosis upon TLR3, TLR4, or TNFR1 stimulation and ZBP1/DAI-mediated necroptotic cell death after HSV-1 infection. The patient's fibroblasts displayed no detectable RIPK3 expression. After TNFR1 or TLR3 stimulation, the patient's cells did not undergo apoptosis or necroptosis. After HSV-1 infection, the cells supported excessive viral growth despite normal induction of antiviral IFN-ß and IFN-stimulated genes (ISGs). This phenotype was, nevertheless, rescued by application of exogenous type I IFN. The patient's human pluripotent stem cell (hPSC)-derived cortical neurons displayed impaired cell death and enhanced viral growth after HSV-1 infection, as did isogenic RIPK3-knockout hPSC-derived cortical neurons. Inherited RIPK3 deficiency therefore confers a predisposition to HSE by impairing the cell death-dependent control of HSV-1 in cortical neurons but not their production of or response to type I IFNs.

Human IRF1 governs macrophagic IFN-γ immunity to mycobacteria.

Inborn errors of human IFN-γ-dependent macrophagic immunity underlie mycobacterial diseases, whereas inborn errors of IFN-α/ß-dependent intrinsic immunity underlie viral diseases. Both types of IFNs induce the transcription factor IRF1. We describe unrelated children with inherited complete IRF1 deficiency and early-onset, multiple, life-threatening diseases caused by weakly virulent mycobacteria and related intramacrophagic pathogens. These children have no history of severe viral disease, despite exposure to many viruses, including SARS-CoV-2, which is life-threatening in individuals with impaired IFN-α/ß immunity. In leukocytes or fibroblasts stimulated in vitro, IRF1-dependent responses to IFN-γ are, both quantitatively and qualitatively, much stronger than those to IFN-α/ß. Moreover, IRF1-deficient mononuclear phagocytes do not control mycobacteria and related pathogens normally when stimulated with IFN-γ. By contrast, IFN-α/ß-dependent intrinsic immunity to nine viruses, including SARS-CoV-2, is almost normal in IRF1-deficient fibroblasts. Human IRF1 is essential for IFN-γ-dependent macrophagic immunity to mycobacteria, but largely redundant for IFN-α/ß-dependent antiviral immunity.

Functional movement screen dataset collected with two Azure Kinect depth sensors.

This paper presents a dataset for vision-based autonomous Functional Movement Screen (FMS) collected from 45 human subjects of different ages (18-59 years old) executing the following movements: deep squat, hurdle step, in-line lunge, shoulder mobility, active straight raise, trunk stability push-up and rotary stability. Specifically, shoulder mobility was performed only once by different subjects, while the other movements were repeated for three episodes each. Each episode was saved as one record and was annotated from 0 to 3 by three FMS experts. The main strength of our database is twofold. One is the multimodal data provided, including color images, depth images, quaternions, 3D human skeleton joints and 2D pixel trajectories of 32 joints. The other is the multiview data collected from the two synchronized Azure Kinect sensors in front of and on the side of the subjects. Finally, our dataset contains a total of 1812 recordings, with 3624 episodes. The size of the dataset is 190 GB. This dataset provides the opportunity for automatic action quality evaluation of FMS.

Silk Protein Composite Bioinks and Their 3D Scaffolds and In Vitro Characterization.

This paper describes the use of silk protein, including fibroin and sericin, from an alkaline solution of Ca(OH)2 for the clean degumming of silk, which is neutralized by sulfuric acid to create calcium salt precipitation. The whole sericin (WS) can not only be recycled, but completely degummed silk fibroin (SF) is also obtained in this process. The inner layers of sericin (ILS) were also prepared from the degummed silk in boiling water by 120 °C water treatment. When the three silk proteins (SPs) were individually grafted with glycidyl methacrylate (GMA), three grafted silk proteins (G-SF, G-WS, G-ILS) were obtained. After adding I2959 (a photoinitiator), the SP bioinks were prepared with phosphate buffer (PBS) and subsequently bioprinted into various SP scaffolds with a 3D network structure. The compressive strength of the SF/ILS (20%) scaffold added to G-ILS was 45% higher than that of the SF scaffold alone. The thermal decomposition temperatures of the SF/WS (10%) and SF/ILS (20%) scaffolds, mainly composed of a ß-sheet structures, were 3 °C and 2 °C higher than that of the SF scaffold alone, respectively. The swelling properties and resistance to protease hydrolysis of the SP scaffolds containing sericin were improved. The bovine insulin release rates reached 61% and 56% after 5 days. The L929 cells adhered, stretched, and proliferated well on the SP composite scaffold. Thus, the SP bioinks obtained could be used to print different types of SP composite scaffolds adapted to a variety of applications, including cells, drugs, tissues, etc. The techniques described here provide potential new applications for the recycling and utilization of sericin, which is a waste product of silk processing.

Ultrashort-Pulse-Microwave Excited Whole-Breast Thermoacoustic Imaging With Uniform Field of Large Size Aperture Antenna for Tumor Screening.

Microwave-induced thermoacoustic imaging (MTAI) has been widely used in biomedical science, and has the potential as an auxiliary measure for clinical diagnosis and treatment. Recently, there are increasing interests in using ultrashort microwave-pumped thermoacoustic imaging techniques to obtain high-efficiency, high-resolution images. However, the traditional imaging system can only provide uniform radiation in a relatively small area, which limits their large field of view in clinical applications (such as whole-breast imaging, brain imaging). To address this problem, we propose an ultrashort pulse microwave thermoacoustic imaging device with a large size aperture antenna. The system can provide a microwave radiation area of 40 cm × 27 cm and a uniform imaging view of 14 cm × 14 cm. With 7 cm imaging depth and a 290 µm resolution. The practical feasibility of the system for breast tumor screening is tested in phantoms with different shapes and in an ex vivo human breast tumor which is embedded in the excised breast of an ewe (π × 5 cm × 5 cm). The tumor can be identified with a contrast of about 1:2. The results demonstrate that the dedicated MTAI system with the uniform large field of view, high imaging resolution, and large imaging depth have the potential for clinical routine breast screening.

A stimulated liquid-gas phase transition nanoprobe dedicated to enhance the microwave thermoacoustic imaging contrast of breast tumors.

Microwave-induced thermoacoustic imaging (MTAI), combining the advantages of the high contrast of microwave imaging and the high resolution of ultrasonic imaging, is a potential candidate for breast tumor detection. MTAI probes have been used to extend thermoacoustic imaging to molecular imaging. However, due to the high content of water molecules in tissues, the thermoelastic expansion-based probes used in conventional MTAI are not capable of adequate enhancement. Herein, an MTAI nanoprobe for amplification of thermoacoustic (TA) signals by the stimulated liquid-gas phase transition mechanism has been developed, providing significantly higher signal amplitude than that from the conventional mechanism of thermoelastic expansion. The nanoprobe consists of liquid perfluorohexane (PFH) and tungsten disulfide (WS2) nanoparticles rich in defect electric dipoles. When irradiated with pulsed microwaves, the defect electric dipoles in WS2 were repeatedly polarized by gigahertz. This results in localized transient heating and an acoustic shockwave, which destroys the van der Waals forces between PFH molecules. Ultimately, liquid PFH droplets undergo a liquid-gas phase transition, generating dramatically enhanced TA signals. The practical feasibility was tested in vitro and in a breast tumor animal model. The results show that the proposed nanoprobe can greatly improve the contrast of tumor imaging. It will be a new generation probe for MTAI.

Sodium N-lauryl amino acids derived from silk protein can form catanionic aggregates with cytarabine as novel anti-tumor drug delivery systems.

A sodium N-lauryl amino acids (shortly silk sericin surfactant, SSS) is synthesized with lauryl chloride and sericin amino acids recovered from silk industrial waste. The purpose of this study is to explore whether the sericin surfactant can be used as a potential drug delivery carrier. By controlling the proportion of cationic drugs, cytarabine hydrochloride (CH) and anionic SSS, the aggregation behavior, slow release capability and toxicological effects of catanionic aggregates or vesicles, formed through CH and SSS, have been investigated in detail. Dynamic light scattering (DLS), transmission electron microscopy (TEM), and zeta potential analysis showed that the aggregate solution could form a stable vesicle structure when the mass fraction of CH is less than or equal to 0.3. The drug release results showed that the cumulative release rate of the catanionic aggregation solution with CH mass fraction of 0.2 reached a maximum at 18 h, being approximately 9 times greater than that of pure cytarabine. The CH/SSS aggregates had a significant sustained release effect compared with the control group. At the same time, vesicles formed by SSS and CH have better anti-tumor effects compared with the pure drug group. In summary, sericin surfactant from silk industrial waste has a potential use as a drug delivery carrier.

Silk sericin has significantly hypoglycaemic effect in type 2 diabetic mice via anti-oxidation and anti-inflammation.

The sericin protein from silk-processing waste added to the normal diet at 0.8% (g%) level was administered orally to type 2 diabetic (T2D) mice to investigate its hypoglycaemic effects and mechanism. The oral protein is in the form of silk sericin hydrolysate, obtained from a boiling treatment of 0.025% calcium hydroxide solution. The protein significantly decreased fasting blood glucose, fasting plasma insulin, and glycosylated serum protein levels; improved oral glucose tolerance and insulin tolerance, and enhanced antioxidative activities. The protein could ameliorate the pathological damage in pancreatic ß-cells and the liver tissue. It enhanced the expression of key proteins and enzymes, including insulin receptor, insulin receptor substrate, PI3K, phosphorylated-AKT, hepatic kinase, GLUT4, glycogen synthase, GSK3ß, GLK, PFK1, PKM2, and AMPKα, which are related to insulin metabolism and glycolysis. The protein also reduced the expression of G6Pase, PCK, and ACC, which are related to gluconeogenesis and lipid metabolism in the liver, and decreased the expression of TNF-α, IL-6, P65, and IKKß related to inflammation. In general, sericin could maintain normal glucose levels and regulate insulin secretion, insulin and lipid metabolism, and inhibition of inflammation. Therefore, sericin protein could be developed into a novel functional health food with significantly hypoglycaemic effect.

Gynura divaricata ameliorates hepatic insulin resistance by modulating insulin signalling, maintaining glycolipid homeostasis and reducing inflammation in type 2 diabetic mice.

Diabetes mellitus, one of the fastest growing epidemics worldwide, has become a serious health problem in modern society. Gynura divaricata (GD), an edible medicinal plant, has been shown to have hypoglycaemic effects. The molecular mechanisms by which GD improves hepatic insulin resistance (IR) in mice with type 2 diabetes (T2D) remain largely unknown. The aerial parts of GD were prepared in a lyophilized powder, which was added into the diet of T2D mice for 4 weeks. GD could result in an obvious decrease in fasting blood glucose and insulin levels in T2D mice. Meanwhile, the underlying mechanisms involved in the insulin-signalling pathway, glucose metabolism, lipid metabolism and inflammatory reaction in the liver tissue were also investigated by western blot, which indicated that GD further ameliorated hepatic IR by activating the PI3K/p-AKT pathway, decreasing the levels of hepatic phosphoenolpyruvate carboxykinase and glucose-6-phosphatase and increasing the levels of glucokinase and peroxisome proliferator-activated receptor-γ in the livers of T2D mice. GD has the potential to alleviate both hyperglycaemia and hepatic IR in T2D mice. Therefore, GD might be a promising functional food or medicine for T2D treatment.