Recombinase polymerase amplification assay for rapid detection of Seneca Valley Virus.

Seneca Valley virus (SVV) is related to vesicular disease in pigs, and its clinical symptoms are indistinguishable from other notifiable clinical symptoms of vesicular disease such as foot-and-mouth disease. The rapid and accurate detection of SVV is essential to confirm the pathogenic factors and initiate the implementation of control measures. The development of a rapid, simple, convenient, and low-cost molecular (nucleic acid amplification) test that can be used at the sample collection point has been identified as a key component for controlling SVV. This study describes the development and demonstration of recombinase polymerase amplification (RPA) test targeting the conserved regions of SVV for detection of SVV. The Primers and probes designed by us have shown good sensitivity and specificity in RPA test, which is helpful for RPA to be an effective tool for rapid diagnosis of SVV.

Proteome and transcriptome analyses reveal key molecular differences between quality parameters of commercial-ripe and tree-ripe fig (Ficus carica L.).

BACKGROUND: Fig fruit are highly perishable at the tree-ripe (TR) stage. Commercial-ripe (CR) fruit, which are harvested before the TR stage for their postharvest transportability and shelf-life advantage, are inferior to TR fruit in size, color and sugar content. The succulent urn-shaped receptacle, serving as the protective structure and edible part of the fruit, determines fruit quality. Quantitative iTRAQ and RNA-Seq were performed to reveal the differential proteomic and transcriptomic traits of the receptacle at the two harvest stages. RESULTS: We identified 1226 proteins, of which 84 differentially abundant proteins (DAPs) were recruited by criteria of abundance fold-change (FC) ≥1.3 and p < 0.05 in the TR/CR receptacle proteomic analysis. In addition, 2087 differentially expressed genes (DEGs) were screened by ≥2-fold expression change: 1274 were upregulated and 813 were downregulated in the TR vs. CR transcriptomic analysis. Ficin was the most abundant soluble protein in the fig receptacle. Sucrose synthase, sucrose-phosphate synthase and hexokinase were all actively upregulated at both the protein and transcriptional levels. Endoglucanase, expansin, beta-galactosidase, pectin esterase and aquaporins were upregulated from the CR to TR stage at the protein level. In hormonal synthesis and signaling pathways, high protein and transcriptional levels of aminocyclopropane-1-carboxylate oxidase were identified, together with a few diversely expressed ethylene-response factors, indicating the potential leading role of ethylene in the ripening process of fig receptacle, which has been recently reported as a non-climacteric tissue. CONCLUSIONS: We present the first delineation of intra- and inter-omic changes in the expression of specific proteins and genes of TR vs. CR fig receptacle, providing valuable candidates for further study of fruit-quality formation control and fig cultivar innovation to accommodate market demand.

In Situ Endothelialization Promoted by SEMA4D and CXCL12 for Titanium-Based Biomaterials.

In situ endothelialization, aiming to create implantation surfaces capable of self-endothelialization, seems to be an extremely promising solution, particularly on those blood-contacting surfaces. In this research study, we immobilized the soluble semaphorin 4D (SEMA4D) and C-X-C motif chemokine ligand 12 (CXCL12) biomolecules together with heparin onto the metal-based biomaterial surfaces to achieve in situ endothelialization of modified samples both by stimulating the neighboring endothelial cells (ECs) migration and by capturing the circulating endothelial progenitor cells (EPCs) directly from the blood circulation. X-ray photoelectron spectroscopy data demonstrate the successful immobilization of SEMA4D and CXCL12. Due to the presence of heparin, hemocompatibility was also improved after modification. Although EC migration was mainly mediated by SEMA4D with the coordination of CXCL12, EC proliferation and haptotaxis property were also enhanced, while EC chemotaxis was slightly suppressed because the further immobilization of CXCL12 influences the release of SEMA4D. The results of the ex vivo EPC capturing assay indicated the mobilization of CXCL12 promotes EPC adhesion. In vivo implantation further demonstrated that CXCL12 cooperates with SEMA4D to promote a process of in situ endothelialization.

Deep learning-assisted smartphone-based molecularly imprinted electrochemiluminescence detection sensing platform: Protable device and visual monitoring furosemide.

A novel, portable, and smartphone-based molecularly imprinted polymer electrochemiluminescence (MIP-ECL) sensing platform was constructed for sensitive and selective determination of furosemide (FSM). In this platform, MoSe2 nanoparticles/starch-derived biomass carbon (MoSe2/BC) nanocomposites as imprinted material, lucigenin (Luc) as the energy donor, CdS quantum dots (CdS QDs) were used as the luminophore (energy acceptor), and molecularly imprinted polymer (MIP) as the specificity recognition element to construct a MIP-ECL sensing system based on electroluminescence resonance energy transfer (ECL-RET) mechanism, which enhanced the sensitivity and the specificity of this system. Imprinted materials were characterized by SEM, TEM, XRD, FT-IR, etc. and the recognition performance of MIP was characterized using CV, EIS, and ECL methods. The elution and re-sorption of template molecules can be used as a switch to control ECL based on the signal that can be quenched by FSM. Interestingly, deep learning based on convolutional neural networks realizes batch processing of ECL signals. Additionally, this developed MIP-ECL method was established by using the traditional ECL analyzer detector for the assay of FSM with a detection limit of 4 nM in the range of 0.010 µM-100 µM. Besides, the consumer smartphone sensing platform based on deep learning showed an outstanding linear response between the R-value of the picture and the concentration of furosemide in the range of 1-70 µM with a detection limit of 0.25 µΜ, which is much lower than that the reported for other detection methods. More importantly, due to the transferability of deep learning, the smartphone-based MIP-ECL systems can facilitate the real-time monitoring of biochemical analytes in multiple fields.

Fabrication of magnetic polydopamine@naphthyl microporous organic network nanosphere for efficient extraction of hydroxylated polycyclic aromatic hydrocarbons and p-nitrophenol from wastewater samples.

Herein, we report the fabrication of a novel, well-defined core-double-shell-structured magnetic Fe3O4@polydopamine@naphthyl microporous organic network (MON), Fe3O4@PDA@NMON, for the efficient magnetic extraction of hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) and p-nitrophenol (p-Npn) from wastewater samples. The hierarchical nanospheres were designed and constructed with the Fe3O4 nanoparticle core, the inner shell of a polydopamine (PDA) layer, and the outer shell of a porous naphthyl MON (NMON) coating, allowing efficient and synergistic extraction of OH-PAHs and p-Npn via hydrophobic, hydrogen bonding, and π-π interactions. The Fe3O4@PDA@NMON nanospheres were well characterized and employed as an efficient sorbent for magnetic solid-phase extraction (MSPE) coupled with high performance liquid chromatography (HPLC) for analyzing of OH-PAHs and p-Npn. Under optimal conditions, the Fe3O4@PDA@NMON-based-MSPE-HPLC-UV method afforded wide linear range (0.18-500 µg L-1), low limits of detection (0.070 µg L-1 for p-Npn, 0.090 µg L-1 for 2-OH-Nap, 0.090 µg L-1 for 9-OH-Fluo and 0.055 µg L-1 for 9-OH-Phe, respectively), large enrichment factors (92.6-98.4), good precisions (intra-day and inter-day relative standard deviations (RSDs); <6.4%, n=6) and less consumption of the adsorbent. Furthermore, trace OH-PAHs and p-Npn with concentrations of 0.29-0.80 µg L-1 were successfully detected in various wastewater samples. Fe3O4@PDA@NMON also functioned as a good adsorbent to enrich a wide scope of trace contaminants containing hydrogen bonding sites and aromatic structures, highlighting the potential of functional MONs in sample pretreatment.

Fabrication of polyethyleneimine modified magnetic microporous organic network nanosphere for efficient enrichment of non-steroidal anti-inflammatory drugs from wastewater samples prior to HPLC-UV analysis.

Development of novel functionalized adsorbents for efficient magnetic solid phase extraction (MSPE) is essential for promoting their versatile applications in sample pretreatment. Herein, we report the fabrication of a new polyethyleneimine-600 decorated magnetic microporous organic network nanosphere (Fe3O4@MON-PEI600) for effective MSPE of trace non-steroidal anti-inflammatory drugs (NSAIDs) from different water samples. The core-shelled Fe3O4@MON-PEI600 integrates the synergistic effects of Fe3O4, MON and PEI600, providing facile and effective extraction to NSAIDs via multiple hydrogen bonding, π-π and hydrophobic interactions. The inner MON shell employs π-π and hydrophobic interaction sites and the outer PEI-600 coat acts as the hydrogen bonding doner/receptor, which affords good extraction performance for NSAIDs. Under optimal conditions, the Fe3O4@MON-PEI600-MSPE-HPLC-UV method gives wide linear range (0.14-400 µg L-1), low limits of detection (0.042-0.149 µg L-1), good precisions (intra-day and inter-day RSDs < 4.5%, n = 6), and large enrichment factors (97.0-98.2). Extraction mechanisms and selectivity of Fe3O4@MON-PEI600 are evaluated in detail. Moreover, Fe3O4@MON-PEI600 is successfully applied to enrich the trace NSAIDs in different water samples with the concentrations of 0.7 and 0.8 µg L-1 for 1-naphthylacetic acid, 0.5 and 0.1 µg L-1 for naproxen as well as 0.7 µg L-1 for ibuprofen, respectively. The developed method not only affords a novel and efficient magnetic adsorbent for NSAIDs in aqueous media at trace level, but also provides a new strategy for the rational design and synthesis of multiple functionalized MON composites in sample pretreatment.

Heparin/poly-l-lysine nanoplatform with growth factor delivery for surface modification of cardiovascular stents: The influence of vascular endothelial growth factor loading.

The rapid re-endothelialization of the vascular stent surface is desirable for preventing thrombosis or reducing restenosis. Many biological factors that promote the biological behavior of endothelial cells have been used for the surface modification of stents. Vascular endothelial growth factor (VEGF), which plays an important role in angiogenesis, induces strong vascular growth. In this study, we investigated different VEGF concentrations (50 to 500 ng/ml) to determine the optimum concentration for biocompatibility. First, VEGF-loaded heparin/poly-l-lysine (Hep-PLL) nanoparticles were created by electrostatic interactions. Then, the VEGF-loaded nanoparticles were immobilized on dopamine-coated 316 L stainless steel (SS) surfaces. The physical and chemical properties of the modified surface were characterized and the biocompatibility was evaluated in vitro. The results indicated that the VEGF-loaded nanoparticles were immobilized successfully on the 316LSS surface, as evidenced by the results of Alcian Blue staining and water contact angle (WCA) measurements. The low platelet adhesion and activation indicated that the modified surfaces had good blood compatibility. The modified surfaces showed a good inhibitory effect on smooth muscle cells, indicating that they inhibited tissue hyperplasia. In addition, the modified surfaces significantly promoted endothelial cell adhesion, proliferation, migration, and biological activity, especially VEGF concentration was 350 ng/ml (NPV350). The optical VEGF concentration of the surface modified Hep-PLL nanoparticles was 350 ng/ml. The proposed method shows promise for potential applications for cardiovascular devices.

In vivo cancer targeting and imaging with semiconductor quantum dots.

We describe the development of multifunctional nanoparticle probes based on semiconductor quantum dots (QDs) for cancer targeting and imaging in living animals. The structural design involves encapsulating luminescent QDs with an ABC triblock copolymer and linking this amphiphilic polymer to tumor-targeting ligands and drug-delivery functionalities. In vivo targeting studies of human prostate cancer growing in nude mice indicate that the QD probes accumulate at tumors both by the enhanced permeability and retention of tumor sites and by antibody binding to cancer-specific cell surface biomarkers. Using both subcutaneous injection of QD-tagged cancer cells and systemic injection of multifunctional QD probes, we have achieved sensitive and multicolor fluorescence imaging of cancer cells under in vivo conditions. We have also integrated a whole-body macro-illumination system with wavelength-resolved spectral imaging for efficient background removal and precise delineation of weak spectral signatures. These results raise new possibilities for ultrasensitive and multiplexed imaging of molecular targets in vivo.