Virus detection and analysis are of critical importance in biological fields and medicine. Surface‐enhanced Raman scattering (SERS) has shown great promise in small molecule and even single molecule ...detection, and can provide fingerprint signals of molecules. Despite the powerful detection capabilities of SERS, the size discrepancy between the SERS “hot spots” (generally, <10 nm) and viruses (usually, sub‐100 nm) yields poor detection reliability of viruses. Inspired by the concept of molecular imprinting, a volume‐enhanced Raman scattering (VERS) substrate composed of hollow nanocones at the bottom of microbowls (HNCMB) is developed. The hollow nanocones of the resulting VERS substrates serve a twofold purpose: 1) extending the region of Raman signal enhancement from the nanocone surface (e.g., surface “hot spots”) to the hollow area within the cone (e.g., volume “hot spots”)—a novel method of Raman signal enhancement, and 2) directing analyte such as viruses of a wide range of sizes to those VERS “hot spots” while simultaneously increasing the surface area contributing to SERS. Using HNCMB VERS substrates, greatly improved Raman signals of single viruses are demonstrated, an achievement with important implications in disease diagnostics and monitoring, biomedical fields, as well as in clinical treatment.
The size discrepency between surface‐enhanced Raman scattering (SERS) “hot spots” (generally, <10 nm) and viruses (usually, sub‐100 nm) makes reliable SERS detection of viruses very challenging. Inspired by the “molecular imprinting” method, a volume‐enhanced Raman scattering (VERS) technique capable of reliably detecting viruses is developed. The VERS technique has promsing applications in virus‐detection‐related biological and biomedical fields.
Dendritic cells (DCs) are central for the initiation and regulation of innate and adaptive immunity in the tumor microenvironment. As such, many kinds of DC-targeted vaccines have been developed to ...improve cancer immunotherapy in numerous clinical trials. Targeted delivery of antigens and adjuvants to DCs in vivo represents an important approach for the development of DC vaccines. However, nonspecific activation of systemic DCs and the preparation of optimal immunodominant tumor antigens still represent major challenges.
We loaded the immunogenic cell death (ICD) inducers human neutrophil elastase (ELANE) and Hiltonol (TLR3 agonist) into α-lactalbumin (α-LA)-engineered breast cancer-derived exosomes to form an in situ DC vaccine (HELA-Exos). HELA-Exos were identified by transmission electron microscopy, nanoscale flow cytometry, and Western blot analysis. The targeting, killing, and immune activation effects of HELA-Exos were evaluated in vitro. The tumor suppressor and immune-activating effects of HELA-Exos were explored in immunocompetent mice and patient-derived organoids.
HELA-Exos possessed a profound ability to specifically induce ICD in breast cancer cells. Adequate exposure to tumor antigens and Hiltonol following HELA-Exo-induced ICD of cancer cells activated type one conventional DCs (cDC1s) in situ and cross-primed tumor-reactive CD8
T cell responses, leading to potent tumor inhibition in a poorly immunogenic triple negative breast cancer (TNBC) mouse xenograft model and patient-derived tumor organoids.
HELA-Exos exhibit potent antitumor activity in both a mouse model and human breast cancer organoids by promoting the activation of cDC1s in situ and thus improving the subsequent tumor-reactive CD8
T cell responses. The strategy proposed here is promising for generating an in situ DC-primed vaccine and can be extended to various types of cancers. Scheme 1. Schematic illustration of HELA-Exos as an in situ DC-primed vaccine for breast cancer. (A) Allogenic breast cancer-derived exosomes isolated from MDA-MB-231 cells were genetically engineered to overexpress α-LA and simultaneously loaded with the ICD inducers ELANE and Hiltonol (TLR3 agonist) to generate HELA-Exos. (B) Mechanism by which HELA-Exos activate DCs in situ in a mouse xenograft model ofTNBC. HELA-Exos specifically homed to the TME and induced ICD in cancer cells, which resulted in the increased release of tumor antigens, Hiltonol, and DAMPs, as well as the uptake of dying tumor cells by cDC1s. The activated cDC1s then cross-primed tumor-reactive CD8+ T cell responses. (C) HELA-Exos activated DCs in situ in the breast cancer patient PBMC-autologous tumor organoid coculture system.
DCs: dendritic cells; α-LA: α-lactalbumin; HELA-Exos: Hiltonol-ELANE-α-LA-engineered exosomes; ICD: immunogenic cell death; ELANE: human neutrophil elastase; TLR3: Toll-like receptor 3; TNBC: triple-negative breast cancer; TME: tumor microenvironment; DAMPs: damage-associated molecular patterns; cDC1s: type 1 conventional dendritic cells; PBMCs: peripheral blood mononuclear cells.
The protein phosphorylation status of exosomes can regulate the activity and function of proteins related to cancer development, and it is highly possible to diagnose cancers through analyzing the ...protein phosphorylation status. However, monitoring the protein phosphorylation status with a simple and label-free method is still clinically challenging. Here, inspired by beehives, we developed an Au-coated TiO2 macroporous inverse opal (MIO) structure with an engineered “slow light effect” and thus with outstanding surface-enhanced Raman scattering (SERS) performance. The MIO structure can capture and analyze the exosomes from plasma of cancer patients without any labeling processes. It was found that the SERS intensity of exosomes at 1087 cm–1 arising from the P–O bond within the phosphoproteins can be used as a criterion for tumor liquid biopsies. The intensity of the 1087 cm–1 SERS peak from exosomes extracted from the plasma of cancer patients (prostate, lung, liver, and colon) is at least two times of that from healthy people. This indicates the simplicity and versatility of this method in cancer diagnostics. Our method has obvious advantages (noninvasive and time-saving) over currently clinically used tumor liquid biopsy techniques (such as western blot), which has great potentials to make vitro cancer diagnostics/monitoring as simple as diagnostics/monitoring of common diseases.
Pathogenic biofilms are up to 1000-fold more drug-resistant than planktonic pathogens and cause about 80% of all chronic infections worldwide. The lack of prompt and reliable biofilm identification ...methods seriously prohibits the diagnosis and treatment of biofilm infections. Here, we developed a machine-learning-aided cocktail assay for prompt and reliable biofilm detection. Lanthanide nanoparticles with different emissions, surface charges, and hydrophilicity are formulated into the cocktail kits. The lanthanide nanoparticles in the cocktail kits can offer competitive interactions with the biofilm and further maximize the charge and hydrophilicity differences between biofilms. The physicochemical heterogeneities of biofilms were transformed into luminescence intensity at different wavelengths by the cocktail kits. The luminescence signals were used as learning data to train the random forest algorithm, and the algorithm could identify the unknown biofilms within minutes after training. Electrostatic attractions and hydrophobic–hydrophobic interactions were demonstrated to dominate the binding of the cocktail kits to the biofilms. By rationally designing the charge and hydrophilicity of the cocktail kit, unknown biofilms of pathogenic clinical isolates were identified with an overall accuracy of over 80% based on the random forest algorithm. Moreover, the antibiotic-loaded cocktail nanoprobes efficiently eradicated biofilms since the nanoprobes could penetrate deep into the biofilms. This work can serve as a reliable technique for the diagnosis of biofilm infections and it can also provide instructions for the design of multiplex assays for detecting biochemical compounds beyond biofilms.
As the blood glucose concentration is an important clinical parameter of diabetes, the rapid and effective detection of blood glucose is very significant for monitoring and managing diabetes. Here, a ...facile method to prepare Rox-DNA functionalized CdZnTeS quantum dots (QDs) was developed. The Rox-DNA functionalized CdZnTeS QDs were prepared by a one-pot hydrothermal method through phosphorothioate DNA bound to QDs, which were employed as a ratiometric fluorescent probe for the rapid and sensitive detection of H2O2 and glucose. Compared with the traditional multistep construction of ratiometric fluorescent probes, this presented approach is simpler and more effective without chemical modification and complicated separation. The CdZnTeS QDs with green fluorescence is specifically sensitive to H2O2, while the red fluorescence of Rox is invariable. H2O2 is the product from the oxidation of glucose catalyzed by glucose oxidase (GOx). Therefore, a facile method to detect H2O2 and glucose with a detection limit of 0.075 μM for H2O2 and 0.042 μM for glucose was developed. In addition, this proposed probe has been employed for the detection of glucose in human serum with a satisfactory result. Moreover, this probe has been used for visual detection, and the health and diabetics can be distinguished by the naked eye. Meanwhile, this nanoprobe is also generalizable and can be extended to the detection of many other H2O2-mediated analytes.
Glucose metabolism and innate immunity evolved side-by-side. It is unclear if and how the two systems interact with each other during hepatitis B virus (HBV) infections and, if so, which mechanisms ...are involved. Here, we report that HBV activates glycolysis to impede retinoic acid-inducible gene I (RIG-I)-induced interferon production. We demonstrate that HBV sequesters MAVS from RIG-I by forming a ternary complex including hexokinase (HK). Using a series of pharmacological and genetic approaches, we provide in vitro and in vivo evidence indicating that HBV suppresses RLR signaling via lactate dehydrogenase-A-dependent lactate production. Lactate directly binds MAVS preventing its aggregation and mitochondrial localization during HBV infection. Therefore, we show that HK2 and glycolysis-derived lactate have important functions in the immune escape of HBV and that energy metabolism regulates innate immunity during HBV infection.
Cholesterol ester (CE) biosynthesis and homeostasis play critical roles in many cancers, including HCC, but their exact mechanistic contributions to HCC disease development require further study.
...Here, we report on a proposed role of tumor suppressor P53 in its repressing ubiquitin-specific peptidase 19 (USP19) and sterol O-acyltransferase (SOAT) 1, which maintains CE homeostasis. USP19 enhances cholesterol esterification and contributes to hepatocarcinogenesis (HCG) by deubiquitinating and stabilizing SOAT1. Loss of either SOAT1 or USP19 dramatically attenuates cholesterol esterification and HCG in P53-deficient mice fed with either a normal chow diet or a high-cholesterol, high-fat diet (HCHFD). SOAT1 inhibitor avasimibe has more inhibitory effect on HCC progression in HCHFD-maintained P53-deficient mice when compared to the inhibitors of de novo cholesterol synthesis. Consistent with our findings in the mouse model, the P53-USP19-SOAT1 signaling axis is also dysregulated in human HCCs.
Collectively, our findings demonstrate that SOAT1 participates in HCG by increasing cholesterol esterification, thus indicating that SOAT1 is a potential biomarker and therapeutic target in P53-deficient HCC.
Environmental models are sensitive to inaccuracies in their approximation algorithms, which can bias model simulations and even lead to incorrect concepts. We present two alternatives to the ...two‐source evapotranspiration (ET) model, which utilize a more accurate approximation of the Clausius‐Clapeyron relation. Model performance was evaluated through a comparison with observed half‐hourly eddy covariance fluxes. Modeled representations of sensible heat dynamics and the limiting behaviors were also evaluated to identify the causes of model inaccuracies. Our analysis shows that the new exponential approximation used here significantly reduces errors that stem from solutions to the energy balance equations. The proposed parallel and series models are both more accurate than the Shuttleworth‐Wallace (SW) model under the conditions evaluated here, but we cannot conclude that the new models are consistently more accurate over a broader range of conditions. The new models can correctly reproduce several theoretical limiting behaviors, whereas the SW model generates conceptually incorrect results for several important limiting cases in which some key conductances approach both zero and infinity. Importantly, these models seriously underestimate the observed latent heat fluxes at night; the major causes of the nighttime model inadequacies are thought to be uncertainties of the estimated conductances and forcing data errors. The central concepts and processes related to new models are described via plots of temperature versus specific humidity to explore the theory behind an ET model. Through these efforts, we anticipate making more reliable ET predictions that will lead to a complete understanding of vegetation–atmosphere interactions.
Key Points
We propose two new two‐source energy balance evapotranspiration models that incorporate a new approximation algorithm
Model performance is evaluated by comparison with observed fluxes, and model behavior is analyzed
The new models are expressed graphically using a plot of temperature versus specific humidity
The galectin 3 binding protein (LGALS3BP, also known as 90K) is a ubiquitous multifunctional secreted glycoprotein originally identified in cancer progression. It remains unclear how 90K functions in ...innate immunity during viral infections. In this study, we found that viral infections resulted in elevated levels of 90K. Further studies demonstrated that 90K expression suppressed virus replication by inducing IFN and pro-inflammatory cytokine production. Upon investigating the mechanisms behind this event, we found that 90K functions as a scaffold/adaptor protein to interact with TRAF6, TRAF3, TAK1 and TBK1. Furthermore, 90K enhanced TRAF6 and TRAF3 ubiquitination and served as a specific ubiquitination substrate of TRAF6, leading to transcription factor NF-κB, IRF3 and IRF7 translocation from the cytoplasm to the nucleus. Conclusions: 90K is a virus-induced protein capable of binding with the TRAF6 and TRAF3 complex, leading to IFN and pro-inflammatory production.
Although immunotherapy is effective in improving the clinical outcomes of patients with bladder cancer (BC), it is only effective in a small percentage of patients. Intercellular crosstalk in the ...tumor microenvironment strongly influences patient response to immunotherapy, while the crosstalk patterns of plasma cells (PCs) as endogenous antibody-producing cells remain unknown. Here, we aimed to explore the heterogeneity of PCs and their potential crosstalk patterns with BC tumor cells.
Crosstalk patterns between PCs and tumor cells were revealed by performing integrated bulk and single-cell RNA sequencing (RNA-seq) and spatial transcriptome data analysis. A risk model was constructed based on ligand/receptor to quantify crosstalk patterns by stepwise regression Cox analysis.
Based on cell infiltration scores inferred from bulk RNA-seq data (n = 728), we found that high infiltration of PCs was associated with better overall survival (OS) and response to immunotherapy in BC. Further single-cell transcriptome analysis (n = 8; 41,894 filtered cells) identified two dominant types of PCs, IgG1 and IgA1 PCs. Signal transduction from tumor cells of specific states (stress-like and hypoxia-like tumor cells) to PCs, for example, via the LAMB3/CD44 and ANGPTL4/SDC1 ligand/receptor pairs, was validated by spatial transcriptome analysis and associated with poorer OS as well as nonresponse to immunotherapy. More importantly, a ligand/receptor pair-based risk model was constructed and showed excellent performance in predicting patient survival and immunotherapy response.
PCs are an important component of the tumor microenvironment, and their crosstalk with tumor cells influences clinical outcomes and response to immunotherapies in BC patients.
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