3D flexible compositing resonant cavity system for high-performance SERS sensing.

Arrayed resonant cavity with outstanding optical trapping ability have received increasing attention in surface-enhanced Raman spectroscopy (SERS). Here, a three-dimensional (3D) composite AgNPs-Al2O3/Au/inverted patterned sapphire substrate PMMA (IPSSPMMA) flexible resonant cavity system is theoretically and experimentally investigated as a flexible SERS sensor. With the help of an effective plasma coupling (localized surface plasmons (LSPs) and surface plasmon polaritons (SPPs)), as shown by the Finite Element Method, a resonant cavity between IPSSPMMA and a particle-film nanostructure is created. Moreover, the proposed fabrication scheme can be easily used for large-scale fabrication. To measure the performance of IPSSPMMA, Rhodamine 6 G (R6G) and Crystalline violet (CV) were used as probe molecules with limit of detection (LOD) of 6.01 × 10-12 M and 5.36 × 10-10 M, respectively, and enhancement factors (EF) of R6G up to 8.6 × 109. Besides, in-situ detection of CV on the surface of aquatic products with a LOD of 3.96 × 10-5 M, enables highly sensitive in-situ detection of surface analytes. The Raman performance and in-situ detection results demonstrate that the proposed flexible compositing resonant cavity system has the advantages of ultra-sensitivity, stability, uniformity, and reproducibility, and has great potential for applications in the food safety field.

Preparation and SERS applications of Ta2O5 composite nanostructures.

Noble metal and semiconductor composite substrates possess high sensitivity, excellent stability, good biocompatibility, and selective enhancement, making them an important research direction in the field of surface-enhanced Raman scattering (SERS). Ta2O5, as a semiconductor material with high thermal stability, corrosion resistance, outstanding optical properties, and catalytic performance, has great potential in SERS research. This study aims to design and fabricate a composite SERS substrate based on Ta2O5 nanostructures, achieving optimal detection performance by combining the urchin-like structure of Ta2O5 with silver nanoparticles (Ag NPs). The urchin-like Ta2O5 nanostructures were prepared using a hydrothermal reaction method. The bandgap was modulated through structure design and the self-doping technique, the charge transfer efficiency and surface plasmon resonance effects were improved, thereby achieving better SERS performance. The composite substrate enables highly sensitive quantitative detection. This composite SERS substrate combines the electromagnetic enhancement mechanism (EM) and chemical enhancement mechanism (CM), achieving ultra-low detection limits of 10-13 M for R6G. Within the concentration range above 10-12 M, there is a good linear relationship between concentration and peak intensity, demonstrating excellent quantitative analysis capabilities. Furthermore, this composite SERS substrate is capable of precise detection of analytes such as crystal violet (CV) and methylene blue (MB), holding broad application prospects in areas such as food safety and environmental monitoring.

Structure and Biochemical Characteristic of the Methyltransferase (MTase) Domain of RNA Capping Enzyme from African Swine Fever Virus.

African swine fever virus (ASFV) is a complex nucleocytoplasmic large DNA virus (NCLDV) that causes a devastating swine disease and it is urgently needed to develop effective anti-ASFV vaccines and drugs. The process of mRNA 5'-end capping is a common characteristic in eukaryotes and many viruses, and the cap structure is required for mRNA stability and efficient translation. The ASFV protein pNP868R was found to have guanylyltransferase (GTase) activity involved in mRNA capping. Here we report the crystal structure of pNP868R methyltransferase (MTase) domain (referred as pNP868RMT) in complex with S-adenosyl-L-methionine (AdoMet). The structure shows the characteristic core fold of the class I MTase family and the AdoMet is bound in a negative-deep groove. Remarkably, the N-terminal extension of pNP868RMT is ordered and keeps away from the AdoMet-binding site, distinct from the close conformation over the active site of poxvirus RNA capping D1 subunit or the largely disordered conformation in most cellular RNA capping MTases. Structure-based mutagenesis studies based on the pNP868RMT-cap analog complex model revealed essential residues involved in substrate recognition and binding. Functional studies suggest the N-terminal extension may play an essential role in substrate recognition instead of AdoMet-binding. A positively charged path stretching from the N-terminal extension to the region around the active site was suggested to provide a favorable electrostatic environment for the binding and approaching of substrate RNA into the active site. Our structure and biochemical studies provide novel insights into the methyltransfer process of mRNA cap catalyzed by pNP868R.IMPORTANCE African swine fever (ASF) is a highly contagious hemorrhagic viral disease in pigs that is caused by African swine fever virus (ASFV). There are no effective drugs or vaccines for protection against ASFV infection till now. The protein pNP868R was predicted to be responsible for process of mRNA 5'-end capping in ASFV, which is essential for mRNA stability and efficient translation. Here, we solved the high-resolution crystal structure of the methyltransferase (MTase) domain of pNP868R. The MTase domain structure shows a canonical class I MTase family fold and the AdoMet binds into a negative pocket. Structure-based mutagenesis studies revealed critical and conserved residues involved in AdoMet-binding and substrate RNA-binding. Notably, both the conformation and the role in MTase activities of the N-terminal extension are distinct from those of previously characterized poxvirus MTase domain. Our structure-function studies provide the basis for potential anti-ASFV inhibitor design targeting the critical enzyme.

Surface-enhanced Raman scattering by the composite structure of Ag NP-multilayer Au films separated by Al2O3.

In the present study, a nanoparticle-multilayer metal film substrate was presented with silver nanoparticles (Ag NPs) assembled on a multilayer gold (Au) film by employing alumina (Al2O3) as a spacer. The SERS performance of the proposed structures was determined. It was suggested that the SERS effect was improved with the increase in the number of layers, which was saturated at four layers. The SERS performance of the structures resulted from the mutual coupling of multiple plasmon modes [localized surface plasmons (LSPs), surface plasmon polaritons (SPPs), as well as bulk plasmon polaritons (BPPs)] generated by the Ag NP-multilayer Au film structure. Furthermore, the electric field distribution of the hybrid system was studied with COMSOL Multiphysics software, which changed in almost consistency with the experimentally achieved results. For this substrate, the limit of detection (LOD) was down to 10-13 M for the rhodamine 6G (R6G), and the proposed SERS substrate was exhibited prominently quantitatively detected capability and high reproducibility. Moreover, a highly sensitive detection was conducted on toluidine blue (TB) molecules. As revealed from the present study, the Ag NP-multilayer Au film structure can act as a dependable SERS substrate for its sensitive molecular sensing applications in the medical field.

Insights into the Neutralization and DNA Binding of Toxin-Antitoxin System ParESO-CopASO by Structure-Function Studies.

ParESO-CopASO is a new type II toxin-antitoxin (TA) system in prophage CP4So that plays an essential role in circular CP4So maintenance after the excision in Shewanella oneidensis. The toxin ParESO severely inhibits cell growth, while CopASO functions as an antitoxin to neutralize ParESO toxicity through direct interactions. However, the molecular mechanism of the neutralization and autoregulation of the TA operon transcription remains elusive. In this study, we determined the crystal structure of a ParESO-CopASO complex that adopted an open V-shaped heterotetramer with the organization of ParESO-(CopASO)2-ParESO. The structure showed that upon ParESO binding, the intrinsically disordered C-terminal domain of CopASO was induced to fold into a partially ordered conformation that bound into a positively charged and hydrophobic groove of ParESO. Thermodynamics analysis showed the DNA-binding affinity of CopASO was remarkably higher than that of the purified TA complex, accompanied by the enthalpy change reversion from an exothermic reaction to an endothermic reaction. These results suggested ParESO acts as a de-repressor of the TA operon transcription at the toxin:antitoxin level of 1:1. Site-directed mutagenesis of ParESO identified His91 as the essential residue for its toxicity by cell toxicity assays. Our structure-function studies therefore elucidated the transcriptional regulation mechanism of the ParESO-CopASO pair, and may help to understand the regulation of CP4So maintenance in S. oneidensis.

Electric Field-Modulated Surface Enhanced Raman Spectroscopy by PVDF/Ag Hybrid.

Electrically modulated surface enhanced Raman scattering (E-SERS) can be able to regulate the plasmon resonance peak of metal nanostructures, further improve the detection sensitivity of the SERS substrate. However, the E-SERS substrates require auxiliary equipment to provide the electrical potential, and most of them are non-flexible structure, which limits the application of E-SERS in the portable, in-situ and fast detection area. Here, we developed an electric field-modulated SERS substrate based on the piezoelectric effect by combining the PVDF (piezoelectric-modulated layer) and Ag nanowires (AgNWs) (SERS active layer) and investigated the SERS activity in experiment and theory. The enhanced electric field and the tunable plasmon resonance induced by the piezoelectric effect provide the additional enhancement for the SERS signal. Furthermore, we fabricated a SERS active ring with a piezoelectric field-modulated substrate and achieved the in-situ detection of glucose with a non-invasive method. This work provided innovation for the E-SERS and could greatly promote the development of the in-situ, wearable and intelligent sensors.