Integrated Antibody with Catalytic Metal–Organic Framework for Colorimetric Immunoassay

Enzyme-linked immunosorbent assay has been widely used as a gold standard in biomedical field, but some inevitable drawbacks still exist in its practical applications, especially the laborious preparation of enzyme–antibody conjugates by a covalent linkage. In this work, we proposed a new strategy to prepare enzyme–antibody conjugate by integrating antibody with catalytic metal–organic framework (MOF) to form dual-functional MOF/antibody composite. As models, rabbit antimouse immunoglobulin G antibody (RIgG) and Cu-MOF with peroxidase-like activity were used to fabricate RIgG@Cu-MOF composite for colorimetric immunoassay. It was found that Cu-MOF as a host not only has no influence on the original capture ability of RIgG to its corresponding antigen (mIgG), but also can shield RIgG against long-term storage, high temperature, and biological degradation. More importantly, upon the formation of sandwiched immunocomplex between RIgG@Cu-MOF and capture antibody, Cu-MOF can serve as a signal amplification unit to perform colorimetric immunoassay. The detection limit of RIgG@Cu-MOF toward mIgG was obtained at 0.34 ng/mL, which is 3-fold lower than that of horseradish peroxidase labeled RIgG. Furthermore, the successful determination of mIgG in serum sample demonstrates the applicability of RIgG@Cu-MOF in detecting real sample. Therefore, it is highly anticipated that this study can offer a new way to prepare enzyme–antibody conjugates, facilitating the exploration of MOF composites in biomedical field

DataSheet_1_Extracellular Hsp90α Supports the ePKM2-GRP78-AKT Axis to Promote Tumor Metastasis.docx

Tumor-secreted proteins can provide numerous molecular targets for cancer diagnosis and treatment. Of note, pyruvate kinase M2 (PKM2) is secreted by tumor cells to promote malignant progression, while its regulatory mechanism or the interacting network remains uncovered. In the present study, we identified extracellular heat shock protein 90 alpha (eHsp90α) as one potential interacting protein of ePKM2 by mass spectrometry (MS), which was further verified by pull-down and co-immunoprecipitation analysis. Later, we found that eHsp90α enhanced the effect of ePKM2 on migration and invasion of lung cancer cells. Blocking of Hsp90α activity, on the other hand, attenuated tumor migration or invasion induced by ePKM2. Eventually, the in vivo role of Hsp90α in regulating ePKM2 activity was validated by the mouse xenograft tumor model. Mechanistically, we found that eHsp90α binds to and stabilizes ePKM2 to protect it from degradation in the extracellular environment. Besides, eHsp90α promoted the interaction of ePKM2 with cell surface receptor GRP78, which leads to the activation of the ePKM2/GRP78/AKT axis. Collectively, we unraveled the novel molecular mechanism of eHsp90α in regulating ePKM2 activity during tumor progression, which is beneficial for the development of new treatments against lung cancer.</p

Biocompatible, Functional Spheres Based on Oxidative Coupling Assembly of Green Tea Polyphenols

Green luminescent, monodisperse, smooth, porous and hollow spheres were simply prepared by Cu²⁺ and temperature mediated oxidative coupling assembly of green tea polyphenols in water. These polymeric tea polyphenol spheres are GSH responsive, acid resistant but alkali-responsive, ideally used as platform for controlled delivery of functional guests

DataSheet_1_Exogenous Brassinolide Alleviates Salt Stress in Malus hupehensis Rehd. by Regulating the Transcription of NHX-Type Na+(K+)/H+ Antiporters.pdf

Brassinolide (BL) mediates various physiological processes and improves plant tolerance to abiotic stresses. However, the effects and mechanism of exogenous BL on the salt tolerance of apple seedlings remain unclear. Herein, we investigated the role of BL in the salt stress response of Malus hupehensis Rehd., a widely grown apple rootstock. Salt-stressed apple seedlings showed significant decline in chlorophyll content and photosynthetic rate, and the application of 0.2 mg/L exogenous BL alleviated salt stress and maintained photosynthetic capacity. Exogenous BL application can strengthen the activities of superoxide dismutase and catalase and thereby eliminates reactive oxygen species (ROS) production induced by salt stress and promote the accumulation of proline and soluble sugar, thus maintaining osmotic balance. Furthermore, exogenous BL application decreased Na+ accumulation and increased K+ content in shoots and roots under salt stress by regulating the expression levels of Na+(K+)/H+ antiporter genes (MhNHXs). MhBZR1 and MhBZR2, which are the key transcription factors in the BR signal transduction pathway, can directly bind to the promoter of MhSOS1 and MhNHX4-1, respectively, and inhibit their expression. Our findings would provide a theoretical basis for analyzing the mechanism of exogenous BL application on the salt tolerance of apples.</p

Biomimetic Crystallization of Toplike Calcite Single Crystals with an Extensive (00.1) Face in the Presence of Sodium Hyaluronate

In this paper, we report the controlled crystallization of calcite by using hyaluronan, a primary constituent of the extracellular substance. Unusual toplike asymmetric calcite crystals with a single extensive (00.1) exposed face were harvested, in contrast to the {10.4} rhombohedral calcite obtained without any additives. The size of the crystal together with peculiar cap structural modifications could be modulated easily by altering the Hya concentration, possibly due to three-dimensional Hya templating crystallization control. SEM, XRPD, FT-IR, thermogravimetric analyses, and staining experiments with charged dyes were adopted for characterizing the morphology, predominant crystallographic orientations, phase, and the (001) face of the crystal and describing the breaking of the morphological symmetry

Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Method for Profiling Ketolic and Phenolic Sex Steroids Using an Automated Injection Program Combined with Diverter Valve Switch and Step Analysis

Sex steroids are involved in many physiological and pathological processes. The determination of sex steroids is essential to understand the mechanisms of human health and diseases. Derivatization techniques could specifically enhance the sensitivities for sex steroids with a given functional group. However, no derivatization reagents are available for profiling multifunctional sex steroids, including phenolic estrogens, ketolic androgens, and ketolic progestogens, in a single analytical run. In the present study, a novel method involving ultraperformance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC–MS/MS) was developed for profiling both ketolic and phenolic sex steroids in human serum using an automated injection program combined with diverter valve switch and step analysis (AIDSA). The human serum, prepared through liquid–liquid extraction and subsequently derivatized using Girard P offline, was automatically injected twice under the automated injection program. For the first injection, Girard P-derivatized ketolic sex steroids were loaded onto the column, and subsequently, the second injection and online derivatization of the same sample using dansyl chloride were performed in the injector needle for 15 min. The dansyl-labeled phenolic sex steroids were then loaded onto the column. The diverter valve worked in coordination with the injection program to import the derivatized sex steroids and remove excess derivatization reagents. The two types of derivatives were individually analyzed in a step-by-step manner. In addition, online dansyl derivatization and Girard P derivative analyses were simultaneously implemented to shorten the total analysis time. This method was well validated and applied to determine the sex steroid levels in human serum

Data_Sheet_1_Perturbed Lipidomic Profiles in Rats With Chronic Cerebral Ischemia Are Regulated by Xiao-Xu-Ming Decoction.pdf

Chronic cerebral ischemia (CCI) is a serious human health condition with lacking therapeutic agents. Moreover, its mechanism of action remains elusive, and thus novel treatment options are required. Lipid metabolism disorder are closely related to CCI. In this study, a CCI-rats model was established by the permanent occlusion of rat bilateral common carotid arteries, and then the rats were treated with a Xiao-Xu-Ming decoction (XXMD). Lipidomic profiling was conducted in both plasma and brain o determine the effects of the injury and therapy on lipid metabolism. Sphingolipid (particularly long acyl chain and total ceramides), glyceryl phosphatide, and glyceride profiles significantly changed in the brain after model induction and again after dosing. A total of 35 potential biomarkers were found in the brain and four were found in the plasma, representing both CCI injury and XXMD action. Correlations between endogenous lipids and exogenous XXMD compounds were analyzed using linear regression. Two exogenous compounds (cimifugin and 5-O-methylvisamminol) in the brain and 17 exogenous compounds in the plasma, which may represent the active constituents in XXMD, were significantly associated with lipid metabolism. This study provides a new perspective on the potential mechanism of CCI and its treatment with XXMD, as well as on discovering effective components in traditional Chinese medicines.</p

Table_1_Alteration of resting-state network dynamics in autism spectrum disorder based on leading eigenvector dynamics analysis.docx

BackgroundNeurobiological models to explain the vulnerability of autism spectrum disorders (ASDs) are scarce, and previous resting-state functional magnetic resonance imaging (rs-fMRI) studies mostly examined static functional connectivity (FC). Given that FC constantly evolves, it is critical to probe FC dynamic differences in ASD patients.MethodsWe characterized recurring phase-locking (PL) states during rest in 45 ASD patients and 47 age- and sex-matched healthy controls (HCs) using Leading Eigenvector Dynamics Analysis (LEiDA) and probed the organization of PL states across different fine grain sizes.ResultsOur results identified five different groups of discrete resting-state functional networks, which can be defined as recurrent PL state overtimes. Specifically, ASD patients showed an increased probability of three PL states, consisting of the visual network (VIS), frontoparietal control network (FPN), default mode network (DMN), and ventral attention network (VAN). Correspondingly, ASD patients also showed a decreased probability of two PL states, consisting of the subcortical network (SUB), somatomotor network (SMN), FPN, and VAN.ConclusionOur findings suggested that the temporal reorganization of brain discrete networks was closely linked to sensory to cognitive systems of the brain. Our study provides new insights into the dynamics of brain networks and contributes to a deeper understanding of the neurological mechanisms of ASD.</p

Table_2_Alteration of resting-state network dynamics in autism spectrum disorder based on leading eigenvector dynamics analysis.docx

BackgroundNeurobiological models to explain the vulnerability of autism spectrum disorders (ASDs) are scarce, and previous resting-state functional magnetic resonance imaging (rs-fMRI) studies mostly examined static functional connectivity (FC). Given that FC constantly evolves, it is critical to probe FC dynamic differences in ASD patients.MethodsWe characterized recurring phase-locking (PL) states during rest in 45 ASD patients and 47 age- and sex-matched healthy controls (HCs) using Leading Eigenvector Dynamics Analysis (LEiDA) and probed the organization of PL states across different fine grain sizes.ResultsOur results identified five different groups of discrete resting-state functional networks, which can be defined as recurrent PL state overtimes. Specifically, ASD patients showed an increased probability of three PL states, consisting of the visual network (VIS), frontoparietal control network (FPN), default mode network (DMN), and ventral attention network (VAN). Correspondingly, ASD patients also showed a decreased probability of two PL states, consisting of the subcortical network (SUB), somatomotor network (SMN), FPN, and VAN.ConclusionOur findings suggested that the temporal reorganization of brain discrete networks was closely linked to sensory to cognitive systems of the brain. Our study provides new insights into the dynamics of brain networks and contributes to a deeper understanding of the neurological mechanisms of ASD.</p

A Colorimetric Immunoassay Based on Coordination Polymer Composite for the Detection of Carcinoembryonic Antigen

Coordination polymers (CPs) as fascinating materials have been explored in a number of fields due to their diverse properties. In this work, we demonstrate the feasibility of CPs in the facile fabrication of multifunctional composites for establishing an immunoassay. To this end, a zinc­(II)-based CP (ZnCP) with adenine as a bridge ligand was employed to integrate with alkaline phosphatase (ALP) and anticarcinoembryonic antigen (anti-CEA) antibody, which produces ALP/anti-CEA@ZnCPs. Benefiting from the adaptive inclusion property of ZnCPs, the integrated ALP and anti-CEA can maintain their original catalytic activity and capture ability to target antigen, respectively. This allows the ALP/anti-CEA@ZnCPs to be a detection antibody for performing an immunoassay. Meanwhile, ZnCP as a host can effectively protect the loaded ALP and anti-CEA against harsh environments. On this basis, by using iron­(II)-phenanthroline complex as a signal amplifier, a colorimetric immunoassay for CEA detection was developed, and a low detection limit of 21.1 pg/mL has been achieved. This immunoassay was successfully applied to determine CEA levels in serum samples with good recovery and precision. We believe that this study can not only provide a new method for CEA detection but also open up a new way for the rational design and fabrication of multifunctional composites