PaCS-Toolkit: Optimized Software Utilities for Parallel Cascade Selection Molecular Dynamics (PaCS-MD) Simulations and Subsequent Analyses.

Parallel cascade selection molecular dynamics (PaCS-MD) is an enhanced conformational sampling method conducted as a "repetition of time leaps in parallel worlds", comprising cycles of multiple molecular dynamics (MD) simulations performed in parallel and selection of the initial structures of MDs for the next cycle. We developed PaCS-Toolkit, an optimized software utility enabling the use of different MD software and trajectory analysis tools to facilitate the execution of the PaCS-MD simulation and analyze the obtained trajectories, including the preparation for the subsequent construction of the Markov state model. PaCS-Toolkit is coded with Python, is compatible with various computing environments, and allows for easy customization by editing the configuration file and specifying the MD software and analysis tools to be used. We present the software design of PaCS-Toolkit and demonstrate applications of PaCS-MD variations: original targeted PaCS-MD to peptide folding; rmsdPaCS-MD to protein domain motion; and dissociation PaCS-MD to ligand dissociation from adenosine A2A receptor.

A SERS Biosensor Based on Functionalized Au-SiNCA Integrated with a Dual Signal Amplification Strategy for Sensitive Detection of Telomerase Activity During EMT in Laryngeal Carcinoma.

Purpose: This paper aims to construct a surface-enhanced Raman spectroscopy (SERS) biosensor based on functionalized Au-Si nanocone arrays (Au-SiNCA) using a dual signal amplification strategy (SDA-CHA) to evaluate telomerase activity during epithelial-mesenchymal transition (EMT) in laryngeal carcinoma (LC). Methods: A SERS biosensor based on functionalized Au-SiNCA was designed with an integrated dual-signal amplification strategy to achieve ultrasensitive detection of telomerase activity during EMT in LC patients. Results: Labeled probes (Au-AgNRs@4-MBA@H1) and capture substrates (Au-SiNCA@H2) were prepared by modifying hairpin DNA and Raman signal molecules. Using this scheme, telomerase activity in peripheral mononuclear cells (PMNC) could be successfully detected with a limit of detection (LOD) as low as 10-6 IU/mL. In addition, biological experiments using BLM treatment of TU686 effectively mimicked the EMT process. The results of this scheme were highly consistent with the ELISA scheme, confirming its accuracy. Conclusion: This scheme provides a reproducible, selective, and ultrasensitive assay for telomerase activity, which is expected to be a potential tool for the early screening of LC in future clinical applications.

Highly sensitive and simultaneous detection of ctDNAs related to non-small cell lung cancer in serum using a catalytic hairpin assembly strategy in a SERS microfluidic chip.

Circulating tumor DNA (ctDNA) is an ideal biomarker for cancer diagnosis based on liquid biopsy, so there is an urgent need for developing an efficient, rapid, and ultrasensitive detection method to meet clinical needs. In this paper, a novel surface-enhanced Raman scattering (SERS) microfluidic chip combined with a catalytic hairpin assembly (CHA) was proposed to detect two non-small cell lung cancer (NSCLC)-related ctDNA (TP53 and PIK3CA-Q546K) simultaneously. The chip consists of six channels for parallel detection. In the reaction region, the CHA reaction between HP1 of the SERS probe and HP2 of the capture substrate was triggered by ctDNAs to form HP1-HP2 duplexes. As the reaction proceeds, more and more SERS probes are captured on the substrate. The gathered reaction products continuously form a lot of hot spots, which greatly enhance the SERS signal. This reaction was completed within 5 minutes. Through this method, the detection limits of TP53 and PIK3CA-Q546K in human serum were as low as 2.26 aM and 2.34 aM, respectively. The microfluidic chip also exhibited high specificity, reproducibility and stability. The clinical feasibility of the SERS microfluidic chip was verified by analyzing the serum samples of healthy subjects and NSCLC patients. The reliability of the experimental results was verified by the qRT-PCR test. The constructed SERS-based analytical micro-platform has great potential in dynamic monitoring of cancer staging and could be used as a clinical tool for early cancer screening.

A pump-free and high-throughput microfluidic chip for highly sensitive SERS assay of gastric cancer-related circulating tumor DNA via a cascade signal amplification strategy.

Circulating tumour DNA (ctDNA) has emerged as an ideal biomarker for the early diagnosis and prognosis of gastric cancer (GC). In this work, a pump-free, high-throughput microfluidic chip coupled with catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) as the signal cascade amplification strategy (CHA-HCR) was developed for surface-enhanced Raman scattering (SERS) assays of PIK3CA E542K and TP53 (two GC-related ctDNAs). The chip consisted of six parallel functional units, enabling the simultaneous analysis of multiple samples. The pump-free design and hydrophilic treatment with polyethylene glycol (PEG) realized the automatic flow of reaction solutions in microchannels, eliminating the dependence on external heavy-duty pumps and significantly improving portability. In the reaction region of the chip, products generated by target-triggered CHA initiated the HCR, forming long nicked double-stranded DNA (dsDNA) on the Au nanobowl (AuNB) array surface, to which numerous SERS probes (Raman reporters and hairpin DNA-modified Cu2O octahedra) were attached. This CHA-HCR strategy generated numerous active "hot spots" around the Cu2O octahedra and AuNB surface, significantly enhancing the SERS signal intensity. Using this chip, an ultralow limit of detection (LOD) for PIK3CA E542K (1.26 aM) and TP53 (2.04 aM) was achieved, and the whole process was completed within 13 min. Finally, a tumour-bearing mouse model was established, and ctDNA levels in mouse serum at different stages were determined. To verify the experimental accuracy, the gold-standard qRT-PCR assay was utilized, and the results showed a high degree of consistency. Thus, this rapid, sensitive and cost-effective SERS microfluidic chip has potential as an ideal detection platform for ctDNA monitoring.

A dual-signal amplification strategy based on pump-free SERS microfluidic chip for rapid and ultrasensitive detection of non-small cell lung cancer-related circulating tumour DNA in mice serum.

Circulating tumour DNAs (ctDNAs) have been reported to be associated with real-time information of progression; however, an accurate and sensitive method has not been established. Herein, a novel dual-signal amplification strategy based on a pump-free surface-enhanced Raman scattering (SERS) microfluidic chip and a catalytic hairpin assembly (CHA) technique was developed for the dynamic monitoring of BRAF V600E and KRAS G12V, which are two non-small cell lung cancer (NSCLC)-related ctDNAs. In the presence of targets, CHA reactions can be triggered between two hairpin DNAs, fixing Pd-Au core-shell nanorods (Pd-AuNRs) onto the magnetic beads (MBs) surface. Thereafter, the composite structures can assemble under the action of magnet, enabling dual-amplification of SERS signal. Using this strategy, the limit of detection (LOD) was low (i.e. at the aM level) in serum. Furthermore, the entire chip-based analysis process could be completed within 5 min, eliminating manual incubation and heavy-duty injection pumps. The selectivity, reproducibility and uniformity of the proposed pump-free SERS microfluidic chip were satisfactory. This superior analysis strategy was finally applied to quantify BRAF V600E and KRAS G12V in tumour-bearing nude mice serum, the results of which corresponded with those of real-time polymerase chain reaction. Overall, this study provides a promising alternative tool for the dynamic monitoring of ctDNA expression level which can benefit the clinical diagnosis of NSCLC.

Ultra-sensitive and high efficiency detection of multiple non-small cell lung cancer-related miRNAs on a single test line in catalytic hairpin assembly-based SERS-LFA strip.

Accurate quantification of multiple miRNAs biomarkers in body fluid is still a challenge for early screening of cancer. Herein, by catalytic hairpin assembly as a signal amplification strategy, we designed a novel surface-enhanced Raman scattering (SERS)-lateral flow assay (LFA) strip for ultrasensitive detection of miR-21 and miR-196a-5p in non-small cell lung cancer (NSCLC) urine on a single test (T) line. 4-mercaptobenzoic acid or 5,5'-dithiobis-2-nitrobenzoic acid as Raman molecules was labeled and two hairpin DNA sequence was modified gold nanocages (GNCs) were designed as two SERS tags. Through target miRNA-triggered catalytic hairpin assembly (CHA), the double-stranded DNAs (H1-H2 complex) formed by SERS tags and the related hairpin-structured DNA sequence 2 (H2) were immobilized on a single T line of SERS-LFA strip. This generated abundant "hot spots" because of the formation of numerous H1-H2 complex thus facilitated the SERS measurement. Through this method, two kinds of miRNAs were analyzed, resulting in limits of detection of 2.08 pM and 3.31 pM for miR-21 in PBS buffer and human urine, 1.77 pM and 2.18 pM for miR-196a-5p in PBS buffer and human urine. Significantly, the SERS-LFA strip exhibited high specificity and good repeatability toward miRNAs. The whole detection time was only 30 min, which means that the high detection efficiency of the strip. The clinical feasibility of the proposed method was also evaluated by detecting the levels of miR-21 and miR-196a-5p in urine samples from NSCLC patients and healthy subjects. The developed SERS-LFA strip has wide application prospect in biomedical research, drug development and early clinical diagnosis.

A SERS-LFA biosensor combined with aptamer recognition for simultaneous detection of thrombin and PDGF-BB in prostate cancer plasma.

An innovative surface-enhanced Raman spectroscopy and lateral flow assay (SERS-LFA) biosensor combined with aptamer recognition had been developed for the convenient, rapid, sensitive and accurate detection of thrombin and platelet-derived growth factor-BB (PDGF-BB) associated with prostate cancer simultaneously. During the biosensor operation, thrombin and PDGF-BB in the sample were recognized and combined by thiol-modified aptamers immobilized on Au-Ag hollow nanoparticles (Au-Ag HNPs) surface and biotinylated aptamers immobilized on the test lines of the biosensor. Thus, thrombin and PDGF-BB were simultaneously captured between detection aptamers and capture aptamers in a sandwich structure. Finite difference time domain simulation confirmed that 'hot spots' appeared at the gaps of Au-Ag HNPs dimer in the enhanced electromagnetic field compared to that of a single Au-Ag HNP, indicating that the aggregated Au-Ag HNPs owned a good SERS signal amplification effect. The detection limits of thrombin and PDGF-BB in human plasma were as low as 4.837 pg ml-1and 3.802 pg ml-1, respectively. Moreover, the accuracy of the biosensor which was applied to detect thrombin and PDGF-BB in prostate cancer plasma had been verified. This designed biosensor had broad application prospects in the clinical diagnosis of prostate cancer.

A label-free electrochemical aptasensor based on the core-shell Cu-MOF@TpBD hybrid nanoarchitecture for the sensitive detection of PDGF-BB.

As one of the significant serum cytokines, platelet-derived growth factor-BB (PDGF-BB) is a crucial protein biomarker overexpressed in human life-threatening tumors, the sensitive identification and quantification of which are urgently desired but challenging. Herein we report a novel core-shell nanoarchitecture consisting of Cu-based metal-organic frameworks (Cu-MOFs) and covalent organic frameworks (denoted as TpBD-COFs), which was used to prepare an aptasensor for the detection of platelet-derived growth factor-BB (PDGF-BB). The central Cu-MOFs function as signal labels with no need for extra redox media, whereas the porous TpBD serves as the shell to immobilize the PDGF-BB-targeted aptamer strands in abundance via strong interactions involving π-π stacking, electrostatic, and hydrogen bonding interactions. The proposed aptasensor based on Cu-MOF@TpBD can achieve a detection limit as low as 0.034 pg mL-1 within the dynamic detection range from 0.0001 to 60 ng mL-1. The hybridization of MOFs and COFs, together with the immobilization with the specific analyte targeted aptamer, provides a promising and propagable approach to prepare an aptasensor for the simple, sensitive, and selective detection of a specific biomarker in clinical diagnosis.

A novel surface-enhanced Raman scattering probe based on Au nanoboxes for dynamic monitoring of caspase-3 during cervical cancer cell apoptosis.

The highly sensitive and reliable detection, imaging, and monitoring of changes of intracellular caspase-3 are critical for understanding the cell apoptosis and studying the progression of caspase-3-related cervical cancer. Herein, we present a novel surface-enhanced Raman scattering (SERS) probe for the detection of caspase-3 during cervical cancer cell apoptosis, composed of Au nanoboxes modified with Nile blue A as a Raman reporter and a caspase-3-specified peptide as a molecular cross-linker. In the presence of caspase-3, the substrate peptides can be cleaved and the changed surface charge of the Au nanoboxes results in the Au nanoboxes-NBA-peptide assembling to form aggregates and a great enhancement of SERS signal. The finite-difference time-domain simulation showed that hot spots mainly located in the nanogaps of the aggregated Au nanoboxes, which in theory proved the rationality of this signal amplification method. The SERS probes exhibited excellent reproducibility and selectivity toward caspase-3. A detection limit of 0.127 fM was obtained for caspase-3, with a dynamic range from 1 fM to 1 nM. MTT assay demonstrated that the probes had no obvious cytotoxicity within a certain concentration range. HeLa cells were treated with doxorubicin to induce long-term apoptosis. Upon cellular uptake of these probes, the spatiotemporal dynamics of caspase-3 in apoptotic cells could be real-time monitored using SERS. The activity of caspase-3 increased with the prolongation of apoptosis time. The SERS results were in accordance with that of western blotting assay. This kind of probe can offer great potential for the determination of enzymatic activities in the physiological processes of cells.

Non-typhoidal Salmonella infections among children in a tertiary hospital in Ningbo, Zhejiang, China, 2012-2019.

BACKGROUND: Non-typhoidal Salmonella (NTS), a common cause of diarrheal enterocolitis, may also cause severe invasive diseases. Limited information on NTS infections in children is available in China. METHODS: We performed a retrospective study of children admitted to the Ningbo Women and Children's Hospital with culture-confirmed NTS infections between January 2012 and December 2019. Clinical and microbiological information were collected. We compared demographic, clinical and antibiotic resistance variables of invasive NTS (iNTS) infections and non-invasive NTS (non-iNTS) infections, and explored associations between hospitalizations for pediatric NTS infections and temperature and rainfall. RESULTS: A total of 166 pediatric hospitalizations due to NTS infection were identified during the 8-year study period. Most of the 166 children were <5 years old (93.4%). The primary serotype was Salmonella Typhimurium (62.6%). Of 166 children with NTS infections, 11 had invasive infection. Compared to 155 children with non-iNTS infections, we found that iNTS infections were more likely to occur in infants ≤6 months or children with an underlying medical condition of leukemia at admission, but iNTS infections less often presented with a symptom of diarrhea (P <0.05 in all cases). The resistance rates of non-iNTS isolates to ceftazidime, ceftriaxone, cefepime, and aztreonam were significantly higher than those of iNTS isolates (P <0.05 in all cases). In addition, compared with iNTS isolates, non-iNTS isolates were significantly associated with resistance to ≥4 CLSI (Clinical and Laboratory Standard Institute) classes (P = 0.041, OR: 0.089, 95% CI: 0.009-0.901) and ≥2 first-line treatment agents (P = 0.040, OR: 0.159, 95% CI: 0.028-0.916). On the other hand, we found that seasonal NTS hospitalizations were positively associated with average seasonal temperature (r = 0.961, P = 0.039) and average monthly rainfall (r = 0.921, P <0.001). CONCLUSION: Non-iNTS accounts for the majority of infections in this study; infants ≤6 months and children with underlying medical conditions of leukemia are more likely to have invasive infection. The rates of antibiotic resistance in the iNTS isolates are generally lower than those in the non-iNTS isolates. On the other hand, high temperatures and heavy rainfall are positively associated with NTS hospitalizations among children in Ningbo.

Highly sensitive detection of cytochrome c in the NSCLC serum using a hydrophobic paper based-gold nanourchin substrate.

Cytochrome c (Cyt c) is a biomarker of early apoptosis that plays a critical role in the diagnosis and therapy of non-small cell lung cancer (NSCLC). In this work, we proposed a novel surface-enhanced Raman scattering (SERS)-based biosensor to implement the ultrasensitive detection of Cyt c in the serum of NSCLC patients. The SERS-supporting substrates based on hydrophobic filter paper were composed of gold nanourchins (GNUs) surface-functionalized with the Cyt c aptamer and the cyanine 5-labeled complementary DNA. In the existence of Cyt c, it could specifically bind to its aptamer, which leads to the detachment of complementary strands modified with Cy5 and the great weakness of SERS signal. The finite-difference time domain (FDTD) simulation showed that the excellent SERS performance of GNUs aggregation was strongly dependent on a large number of "hot spots" at the tips and between the nanogaps of aggregated GNUs. Alkyl ketene dimer (AKD) was used to make the filter paper modify its property from hydrophilic to hydrophobic, which consequently increased the density of GNUs and extended the retention time of the analyte. SERS biosensors based on hydrophobic paper exhibited prominent reproducibility and selectivity. The detection limit of Cyt c in PBS was 1.148 pg/mL, while the detection limit in human serum was 1.79 pg/mL. Moreover, the analysis of the serum samples of healthy subjects and NSCLC patients confirmed the feasibility of its clinical application. The results were consistent with enzyme-linked immunosorbent assay results. This method can be a powerful strategy for quantitative detection of extracellular Cyt c, and it is expected that the SERS-based biosensors could be applied in the practical clinical diagnoses of NSCLC.

The simultaneous detection of the squamous cell carcinoma antigen and cancer antigen 125 in the cervical cancer serum using nano-Ag polydopamine nanospheres in an SERS-based lateral flow immunoassay.

The accurate analysis of tumor related biomarkers is extremely critical in the diagnosis of the early stage cervical cancer. Herein, we designed a novel and inexpensive surface-enhanced Raman scattering-based lateral flow assay (SERS-based LFA) strip with a single test line, which was applied for the rapid and sensitive quantitative simultaneous analysis of SCCA and CA125 in serum samples from patients with cervical cancer. In the presence of target antigens, the monoclonal antibody-coupled and Raman reporter-labeled nano-Ag polydopamine nanospheres (PDA@Ag-NPs) aggregated on the test line modified by the polyclonal antibody to form a double-antibody sandwich structure. The finite difference time domain simulation demonstrated that large number of "hot spots" was generated among the nanogaps of aggregated PDA@AgNPs, which resulted in a huge enhancement of the signal of the Raman reporters. Accordingly, the limit of detection was determined to be 7.156 pg mL-1 for SCCA and 7.182 pg mL-1 for CA125 in phosphate buffer and 8.093 pg mL-1 for SCCA and 7.370 pg mL-1 for CA125 in human serum, revealing high sensitivity of this SERS-based LFA strip. Significantly, the detection of SCCA and CA125 using the SERS-based LFA was observed to have high specificity and reproducibility, and the whole detection was completed within 20 min. Furthermore, the SERS-based LFA and enzyme-linked immunosorbent assay were also employed in serum samples obtained from patients with cervical cancer, cervical intraepithelial neoplasia and healthy subjects, and perfect agreement existed between both the methods. Thus, clinically, the developed SERS-based LFA strip has strong potential for the simultaneous detection of multiple cancer biomarkers in serum.

Copper(II) 1,4-naphthalenedicarboxylate on copper foam nanowire arrays for electrochemical immunosensing of the prostate specific antigen.

Nanowires of copper(II)-based metal-organic frameworks (Cu-MOFs) of type Cu(II)(1,4-naphthalenedicarboxylic acid) (1,4-NDC) were deposited on the surface of a copper foam by immersion of Cu(OH)2 nanowires in a solution of 1,4-NDC. An electrochemical immunosensor for the prostate specific antigen (PSA) is obtained by using the nanowire arrays as a redox signal probe. The signal is generated by the conversion of Cu(I) and Cu(II) of Cu-MOFs nanowires. Cu(1,4-NDC) nanowires contain many uncoordinated carboxyl groups which can bind to the amino groups of the PSA antibody. When PSA antibody binds to PSA antigen during an immune response, the current signal will decrease due to the electrical insulation of PSA antigen. The decrease of current is directly proportional to the increase of PSA concentration. The immunosensor, best operated at a voltage of typically -0.08 V (vs. Ag/AgCl), has a low limit of detection (4.4 fg·mL-1) and a wide linear range (0.1 pg·mL-1 to 20 ng·mL-1). This meets the demands of clinical diagnosis (with values <4 ng·mL-1) in serum. The method was applied to the determination of PSA in spiked serum. Graphical abstractSchematic representation of the in-situ growth of ordered Cu-MOFs wrapped with Cu(OH)2 nanowires, building the core-shell structure as the 3D electrode. A novel electrochemical immunosensor for PSA detection has been exploited, using the Cu-MOFs nanowire arrays on Cu foam as a redox signal probe for the first time.

Ultrathin nickel-metal-organic framework nanobelt based electrochemical sensor for the determination of urea in human body fluids.

Ultrathin nickel-metal-organic framework (Ni-MOF) nanobelts, [Ni20(C5H6O4)20(H2O)8]·40H2O (Ni-MIL-77), have been exploited successfully for the fabrication of a non-enzymatic urea sensor. Ni-MOF ultrathin nanobelts in alkaline media can be used as an efficient catalyst for urea electrooxidation. As a non-enzymatic urea sensor, Ni-MOF ultrathin nanobelts exhibit a high sensitivity of 118.77 µA mM-1 cm-2, wide linear range of 0.01-7.0 mM, and low detection limit of 2.23 µM (S/N = 3). The selectivity, stability and reliability of ultrathin Ni-MOF nanobelts towards urea oxidation are also investigated. Moreover, Ni-MOF ultrathin nanobelts were further used to detect urea in human body fluids. All these findings confirm that the urea sensor based on Ni-MOF ultrathin nanobelts is successfully prepared and promising for applications in medical diagnostics and environmental monitoring.

Multiplexing determination of cancer-associated biomarkers by surface-enhanced Raman scattering using ordered gold nanohoneycomb arrays.

AIM: Here, a multiplex surface-enhanced Raman scattering (SERS) based assay for simultaneous quantitation of carcinoembryonic antigen (CEA) and α-fetoprotein (AFP) was developed. METHODS: SERS tags of nanostars and SERS substrates of nanobowl arrays were functionalized with labeling and capturing antibodies, respectively. In presence of antigens, SERS tags, antigens and SERS substrates formed sandwich structure. RESULTS: The SERS-based technique showed a wide linear range from 0.5 to 100 ng/ml and detection limits were 0.41 and 0.35 ng/ml for CEA and AFP in phosphate-buffered saline buffer, respectively. Analysis results of clinical serum samples using this technique were similar to that shown in phosphate-buffered saline buffer. The LODs were 0.44 and 0.40 ng/ml for CEA and AFP, respectively. Conclusion: The precision and stability of this analysis technique were satisfactory, meanwhile, no obvious cross-reactivity could be found. What's more, it also suggested that this novel multiplex SERS-based technique could be a simple, specific, reliable, sensitive and multiplexed tool for important diagnostic and prognostic applications.

A SERS protocol as a potential tool to access 6-mercaptopurine release accelerated by glutathione-S-transferase.

The developed method for monitoring GST, an important drug metabolic enzyme, could greatly facilitate researches on relative biological fields. In this work, we have developed a SERS technique to monitor the absorbance behaviour of 6-mercaptopurine (6-MP) and its glutathione-S-transferase (GST)-accelerated glutathione (GSH)-triggered release behaviour on the surface of gold nanoflowers (GNFs), using the GNFs as excellent SERS substrates. The SERS signal was used as an indicator of absorbance or release of 6-MP on the gold surface. We found that GST can accelerate GSH-triggered release behaviour of 6-MP from the gold surface. We speculated that GST catalyzes nucleophilic GSH to competitively bind with the electrophilic substance 6-MP. Experimental results have proved that the presented SERS protocol can be utilized as an effective tool for accessing the release of anticancer drugs.

Synthesis of Au Nanostars and Their Application as Surface Enhanced Raman Scattering-Activity Tags Inside Living Cells.

This work presents the synthesis and characterization of Au nanostars (AuNSs) and demonstrates their application as surface enhanced Raman scattering (SERS)-activity tags for cellular imaging and sensing. Nile blue A (NBA) and bovine serum albumin (BSA) were used as Raman reporter molecules and capping materials, respectively. The SERS-activity tags were tested on human lung adenocarcinoma cell (A549) and alveolar type II cell (AT II) and found to present a low level of cytotoxicity and high chemical stability. These SERS-activity tags not only can be applied in multiplexed cellular imaging, including dark field imaging, transmission electron microscopy (TEM) and SERS imaging, but also can be used for cellular sensing. The SERS spectra clearly identified cellular important components such as proteins, nucleic acids, lipids, and carbohydrates. This study also shows that endocytosis is the main channel of tags internalized in cells. The AuNSs exhibiting strong surface enhanced Raman effects are utilized in the design of an efficient, stable SERS-activity tag for intracellular applications.

Novel benzothiazinones (BTOs) as allosteric modulator or substrate competitive inhibitor of glycogen synthase kinase 3ß (GSK-3ß) with cellular activity of promoting glucose uptake.

Glycogen synthase kinase 3ß (GSK-3ß) plays a key role in insulin metabolizing pathway and therefore inhibition of the enzyme might provide an important therapeutic approach for treatment of insulin resistance and type 2 diabetes. Recently, discovery of ATP noncompetitive inhibitors is gaining importance not only due to their generally increased selectivity but also for the potentially subtle modulation of the target. These kinds of compounds include allosteric modulators and substrate competitive inhibitors. Here we reported two benzothiazinone compounds (BTO), named BTO-5h (IC50=8 µM) and BTO-5s (IC50=10 µM) as novel allosteric modulator and substrate competitive inhibitor of GSK-3ß, respectively. Their different action modes were proved by kinetic experiments. Furthermore, BTO-5s was selected to check the kinases profile and showed little or even no activity to a panel of ten protein kinases at 100 µM, indicating it has good selectivity. Docking studies were performed to give suggesting binding modes which can well explain their impacts on the enzyme. Moreover, cell experiments displayed both compounds reduced the phosphorylation level of glycogen synthase in an intact cell, and greatly enhanced the glucose uptake in both HpG2 and 3T3-L1 cells. All of these results suggested BTO-5s and BTO-5h maybe have potentially therapeutic value for anti-diabetes. The results also offer a new scaffold for designing and developing selective inhibitors with novel mechanisms of action.

A method to compute the radiated sound power based on mapped acoustic radiation modes.

Application of modal expansion approach for the exterior acoustic field has drawn wide research interests in recent years. This is primarily due to the acoustic radiation modes (ARM) that can diagonalize the impedance matrix, hence significantly simplifying the computation of radiated sound power. The orthogonal ARM are typically calculated via a standard eigenvalue analysis of the impedance matrix, which normally leads to numerical difficulties especially for wideband frequency and large scale problems. In this paper, a theory of mapped ARM is proposed to avoid the cumbersome computation of ARM for convex structures. A mapping relationship between the ARM on the surface of an equivalent spherical source and the mapped ARM on the surface of a convex structure is obtained based on the equivalent source method, multipole expansion method, and boundary integral method. Furthermore, analytical expressions for the radiated sound power of structures vibrating in its mapped ARM as well as that of spheres are derived. Finally, a simple method is proposed to approximate the radiated sound power based on the modal decomposition method and the mapping relationship. Numerical simulations are conducted to validate the accuracy and efficiency of the proposed approach, and different vibrating structures with various geometries are considered. Results demonstrate that the proposed methodology for calculating the radiated sound power of convex structures is very efficient and accurate as compared with the traditional approach.

A novel label-free amperometric immunosensor for carcinoembryonic antigen based on Ag nanoparticle decorated infinite coordination polymer fibres.

In this article, for the first time, a novel, high-yield and template-free method for the synthesis of Ag nanoparticle decorated thionine/infinite coordination polymer (AgNP/THI/ICP) fibres is proposed. The thionine can be adsorbed to the AgNP/THI/ICP fibres by π-conjugation and act as the redox probe. The AgNP/THI/ICP fibres not only favor the immobilization of antibody but also facilitate the electron transfer. It is found that the AgNP/THI/ICP fibres can be designed to act as a sensitive label-free electrochemical immunosensor for carcinoembryonic antigen (CEA) determination. Under the optimized conditions, the linear range of the proposed immunosensor is estimated to be from 50 fg/mL to 100 ng/mL and the detection limit is estimated to be 0.5 fg/mL at a signal-to-noise ratio of 3, respectively. The prepared immunosensor for detection of CEA shows high sensitivity, reproducibility and stability. Our study demonstrates that the proposed immunosensor has also been used to determine CEA successfully in diluted blood samples.