Augmenting Neutrophil Extracellular Traps with Carbonized Polymer Dots: A Potential Treatment for Bacterial Sepsis.

Sepsis is a life-threatening condition that can progress to septic shock as the body's extreme response to pathogenesis damages its own vital organs. Staphylococcus aureus (S. aureus) accounts for 50% of nosocomial infections, which are clinically treated with antibiotics. However, methicillin-resistant strains (MRSA) have emerged and can withstand harsh antibiotic treatment. To address this problem, curcumin (CCM) is employed to prepare carbonized polymer dots (CPDs) through mild pyrolysis. Contrary to curcumin, the as-formed CCM-CPDs are highly biocompatible and soluble in aqueous solution. Most importantly, the CCM-CPDs induce the release of neutrophil extracellular traps (NETs) from the neutrophils, which entrap and eliminate microbes. In an MRSA-induced septic mouse model, it is observed that CCM-CPDs efficiently suppress bacterial colonization. Moreover, the intrinsic antioxidative, anti-inflammatory, and anticoagulation activities resulting from the preserved functional groups of the precursor molecule on the CCM-CPDs prevent progression to severe sepsis. As a result, infected mice treated with CCM-CPDs show a significant decrease in mortality even through oral administration. Histological staining indicates negligible organ damage in the MRSA-infected mice treated with CCM-CPDs. It is believed that the in vivo studies presented herein demonstrate that multifunctional therapeutic CPDs hold great potential against life-threatening infectious diseases.

In Situ Hybridization of Polymeric Curcumin to Arginine-Derived Carbon Quantum Dots for Synergistic Treatment of Bacterial Infections.

Effective infectious keratitis treatment must eliminate the pathogen, reduce the inflammatory response, and prevent persistent damage to the cornea. Infectious keratitis is generally treated with broad-spectrum antibiotics; however, they have the risk of causing corneal epithelial cell damage and drug resistance. In this study, we prepared a nanocomposite (Arg-CQDs/pCur) from arginine (Arg)-derived carbon quantum dots (Arg-CQDs) and polymeric curcumin (pCur). Partial carbonization of arginine hydrochloride in the solid state by mild pyrolysis resulted in the formation of CQDs, which exhibited enhanced antibacterial activity. pCur was formed by the polymerization of curcumin, and further crosslinking reduced its cytotoxicity and improved antioxidative, anti-inflammatory, and pro-proliferative activities. The pCur in situ conjugated with Arg-CQDs to form the Arg-CQDs/pCur nanocomposite, which showed a minimum inhibitory concentration of ca. 10 µg mL-1, which was >100-fold and >15-fold lower than that of the precursor arginine and curcumin, respectively, against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. The Arg-CQDs/pCur nanocomposite with combined antibacterial, antioxidative, anti-inflammatory, pro-proliferative properties, and long-term retention on cornea enabled synergistic treatment of bacterial keratitis. In a rat model, it can effectively treat P. aeruginosa-induced bacterial keratitis at a concentration 4000-fold lower than the commercially used drug, Sulmezole eye drops. Arg-CQDs/pCur nanocomposites have great potential for application in antibacterial and anti-inflammatory nanoformulations for clinical use to treat infectious diseases.

Pulsed laser irradiation induces the generation of alloy cluster ions for the screening of protease activity.

Pulsed laser irradiation can cause the fragmentation of nanoparticles, which generates cluster ions. This allows nanoparticles to be adopted as mass tag/signal amplifiers in laser desorption/ionization mass spectrometry (LDI-MS) bioassays. Herein, we demonstrate the potential of using the signal from alloy cluster ions in bioassays through a fibrin clot model to determine the activity of thrombin. A mixed solution of silver and gold nanoparticles functionalized with fibrinogen (Fg‒Ag NPs/Fg‒Au NPs) treated with thrombin can form clots composed of aggregated fibrin-Au NPs/Ag NPs. These clots analyzed with LDI-MS are noted to form intense Ag-Au alloy cluster ions, especially [Ag2Au]+, which were used to detect thrombin concentration with a dynamic range of 2.5-50 pM in human plasma. This sensing platform was further employed for the screening of direct thrombin inhibitors. This work developed a novel bioassay utilizing metallic gas-phase reactions generated from pulsed laser irradiation of aggregated nanoparticles to monitor enzymatic activity and to screen inhibitors. We believe that LDS-MS can serve as a new platform for gas-phase reaction-based bioassays.

Aminoglycoside-mimicking carbonized polymer dots for bacteremia treatment.

Bacteremia and associated bacterial sepsis are potentially fatal and occur when the host response to microbial invasion is impaired or compromised. This motivated us to develop carbonized polymer dots (CPDsMan/AA) from a mixture of mannose (Man) and positively charged amino acids [AAs; lysine, arginine (Arg), or histidine] through a one-step mild pyrolysis procedure, which effectively inhibited drug-resistant bacterial strains isolated from septic patients. The as-prepared CPDsMan/AA showed broad-spectrum antibacterial activity, including multidrug-resistant bacteria, even in human plasma. The minimal inhibitory concentration of CPDsMan/Arg is ca. 1.0 µg mL-1, which is comparable to or lower than those of other tested antibiotics (e.g., ampicillin, gentamicin, and vancomycin). In addition to directly disrupting bacterial membranes, the CPDsMan/Arg feature a structure similar to aminoglycoside antibiotics that could bind to 16S rRNA, thereby blocking bacterial protein synthesis. In vitro cytotoxic and hemolytic assays demonstrated the high biocompatibility of the CPDsMan/AA. In addition, in vivo studies on methicillin-resistant Staphylococcus aureus-infected mice treated with the CPDsMan/Arg showed a significant decrease in mortality-even better than that of antibiotics. Overall, the synthesis of the CPDsMan/AA is cost-efficient, straightforward, and effective for treating bacteremia. The polymeric features of the CPDsMan/Arg, including cationic charges and specific groups, can be recognized as a safe and broad-spectrum biocide to lessen our reliance on antibiotics to treat systemic bacterial infections in the future.

Partial carbonization of quercetin boosts the antiviral activity against H1N1 influenza A virus.

Inflenza A viruses (IAVs) are highly transmissible and pathogenic Orthomyxoviruses, which have led to worldwide outbreaks and seasonal pandemics of acute respiratory diseases, causing serious threats to public health. Currently used anti-influenza drugs may cause neurological side effects, and they are increasingly less effective against mutant strains. To help prevent the spread of IAVs, in this work, we have developed quercetin-derived carbonized nanogels (CNGsQur) that display potent viral inhibitory, antioxidative, and anti-inflammatory activities. The antiviral CNGsQur were synthesized by mild carbonization of quercetin (Qur), which successfully preserved their antioxidative and anti-inflammatory properties while also contributed enhanced properties, such as water solubility, viral binding, and biocompatibility. Antiviral assays of co-treatment, pre-treatment, and post-treatment indicate that CNGsQur interacts with the virion, revealing that the major antiviral mechanism resulting in the inhibition of the virus is by their attachment on the cell surface. Among them, the selectivity index (SI) of CNGsQur270 (>857.1) clearly indicated its great potential for clinical application in IAVs inhibition, which was much higher than that of pristine quercetin (63.7) and other clinical drugs (4-81). Compared with quercetin at the same dose, the combined effects of viral inhibition, antioxidative and anti-inflammatory activities impart the superior therapeutic effects of CNGsQur270 aerosol inhalation in the treatment of IAVs infection, as evidenced by a mouse model. These CNGsQur effectively prevent the spread of IAVs and suppress virus-induced inflammation while also exhibiting good in vivo biocompatibility. CNGsQur shows much promise as a clinical therapeutic agent against infection by IVAs.

Monitoring protein conformational changes using fluorescent nanoantennas.

Understanding the relationship between protein structural dynamics and function is crucial for both basic research and biotechnology. However, methods for studying the fast dynamics of structural changes are limited. Here, we introduce fluorescent nanoantennas as a spectroscopic technique to sense and report protein conformational changes through noncovalent dye-protein interactions. Using experiments and molecular simulations, we detect and characterize five distinct conformational states of intestinal alkaline phosphatase, including the transient enzyme-substrate complex. We also explored the universality of the nanoantenna strategy with another model protein, Protein G and its interaction with antibodies, and demonstrated a rapid screening strategy to identify efficient nanoantennas. These versatile nanoantennas can be used with diverse dyes to monitor small and large conformational changes, suggesting that they could be used to characterize diverse protein movements or in high-throughput screening applications.

Multifunctional carbonized nanogels to treat lethal acute hepatopancreatic necrosis disease.

BACKGROUND: Shrimp aquaculture has suffered huge economic losses over the past decade due to the outbreak of acute hepatopancreatic necrosis disease (AHPND), which is mainly caused by the bacteria Vibrio parahaemolyticus (V. parahaemolyticus) with the virulence pVA1 plasmid, which encodes a secretory photorhabdus insect-related (Pir) toxin composed of PirA and PirB proteins. The Pir toxin mainly attacks the hepatopancreas, a major metabolic organ in shrimp, thereby causing necrosis and loss of function. The pandemic of antibiotic-resistant strains makes the impact worse. METHODS: Mild pyrolysis of a mixture of polysaccharide dextran 70 and the crosslinker 1,8-diaminooctane at 180 â„ƒ for 3 h to form carbonized nanogels (DAO/DEX-CNGs) through controlled cross-linking and carbonization. The multifunctional therapeutic CNGs inherit nanogel-like structures and functional groups from their precursor molecules. RESULTS: DAO/DEX-CNGs manifest broad-spectrum antibacterial activity against Vibrio parahaemolyticus responsible for AHPND and even multiple drug-resistant strains. The polymer-like structures and functional groups on graphitic-carbon within the CNGs exhibit multiple treatment effects, including disruption of bacterial membranes, elevating bacterial oxidative stress, and neutralization of PirAB toxins. The inhibition of Vibrio in the midgut of infected shrimp, protection of hepatopancreas tissue from Pir toxin, and suppressing overstimulation of the immune system in severe V. parahaemolyticus infection, revealing that CNGs can effectively guard shrimp from Vibrio invasion. Moreover, shrimps fed with DAO/DEX-CNGs were carefully examined, such as the expression of the immune-related genes, hepatopancreas biopsy, and intestinal microbiota. Few adverse effects on shrimps were observed. CONCLUSION: Our work proposes brand-new applications of multifunctional carbon-based nanomaterials as efficient anti-Vibrio agents in the aquatic industry that hold great potential as feed additives to reduce antibiotic overuse in aquaculture.

Silver oxide model surface improves computational simulation of surface-enhanced Raman spectroscopy on silver nanoparticles.

Surface-enhanced Raman spectroscopy (SERS) coupled with density functional theory (DFT) computations can characterise the adsorption orientation of a molecule on a nanoparticle surface. When using DFT to simulate SERS on a silver surface, one typically employs an atom (Ag), ion (Ag+), or cluster (Agx or Agx+) as the model surface. Here, by examining the nucleobase 2,6-diaminopurine (2,6-DAP) and then generalising our strategy to three other molecules, we show that employing silver oxide (Ag2O) as the model surface can quantitatively improve the accuracy of simulated SERS.

Thermally driven formation of polyphenolic carbonized nanogels with high anticoagulant activity from polysaccharides.

We have demonstrated that alginate with negligible anticoagulant activity can be converted into carbonized nanogels with potent anticoagulant activity through a solid-state heating process. The conversion of alginate into graphene-like nanosheet (GNS)-embedded polyphenolic-alginate nanogels (GNS/Alg-NGs) has been carried out through condensation and carbonization processes. The GNS/Alg-NGs exhibit much stronger anticoagulant activity (>520-fold) compared to untreated alginate, mainly because their polyphenolic structures have a high binding affinity [dissociation constant (Kd) = 2.1 × 10-10 M] toward thrombin. In addition, the thrombin clotting time delay caused by the GNS/Alg-NGs is 10-fold longer than that of natural polyphenolic compounds, such as quercetin, catechin, naringenin, caffeic acid, and ferulic acid. The thrombin- or kaolin-activated thromboelastography of whole-blood coagulation reveals that the GNS/Alg-NGs display a much stronger anticoagulant ability than that of untreated alginate and naturally sulfated polysaccharides (fucoidan). The GNS/Alg-NGs exhibit superior biocompatibility and anticoagulant activity, as observed with an in vivo rat model, revealing their potential as a blood thinner for the treatment of thrombotic disorders.

In situ synthesis of core-shell carbon nanowires as a potent targeted anticoagulant.

We have developed a one-pot synthesis of bio-carbon nanowires from the natural product sodium alginate at low temperature, without using any catalyst, for anticoagulation applications. Sodium alginate is carbonized and sulfated/sulfonated in situ by solid state heating of a mixture of sodium alginate and ammonium sulfite. By regulating the heating temperature and the ratio of ammonium sulfite to sodium alginate, we modulated the degree of sulfation/sulfonation and carbonization, as well as the morphology of the carbon nanomaterials. The core-shell sulfated/sulfonated bio-carbon nanowires (CNWsAlg@SOx) made by the reaction of a mixture of ammonium sulfite and sodium alginate with a mass ratio of 5 (ammonium sulfite to sodium alginate) at 165 °C for 3 h, exhibit strong inhibition of thrombin activity due to their ultrahigh binding affinity towards it (dissociation constant (Kd) = 8.7 × 10-11 M). The possible formation mechanism of the carbon nanowires has been proposed. The thrombin-clotting time delay caused by CNWsAlg@SOx is ∼ 170 times longer than that caused by sodium alginate. Hemolysis and cytotoxicity assays demonstrated the high biocompatibility of CNWsAlg@SOx. Furthermore, the thromboelastography of whole-blood coagulation and rat-tail bleeding assays further reveal that CNWsAlg@SOx have a much stronger anticoagulation activity than sodium alginate and naturally sulfated polysaccharides (e.g., fucoidan). Our results suggest that the low-temperature prepared, cost-effective, and highly biocompatible CNWsAlg@SOx show great potential as an efficient anticoagulant for the prevention and treatment of diseases associated with thrombosis.

Nanoparticle-Based LDI-MS Immunoassay for the Multiple Diagnosis of Viral Infections.

Many serious public health emergencies around the globe are caused by viral epidemics. Thus, developing a reliable method for viral screening is in high demand. Multiplex assays for simultaneous detection and fast screening of high-risk pathogens are especially needed. This study employs metal nanoparticles to generate specific mass spectral signals for different RNA viruses, which enables simultaneous detection of whole viruses by laser desorption/ionization mass spectrometry (LDI-MS). We developed a nanoparticle-based sandwich immunosorbent assay as a sensing platform for the detection of viruses and viral nonstructural protein by LDI-MS. Cellulose acetate membrane (CAM) serves as the substrate for the fabrication of the sandwich immunosorbent assay with the advantages of clean mass spectra and high enrichment of analytes. Antibody-modified metal nanoparticles (Ab-MNPs; M = Au or Ag) act as metallic biocodes for the LDI-MS detection. The signal amplification readout for the virus is through the pulsed laser-induced formation of metal cluster ions ([M n]+; n = 1-3) from the Ab-MNPs which specifically bind on the CAM. Our sensing system is effective for the detection of intact viruses [Enterovirus 71 (EV71) and Japanese encephalitis virus (JEV)], nonstructural protein 1 (NS1) of Zika virus (ZIKV), EV71-spiked human serum samples, and the simultaneous detection of EV71 and ZIKV. Our probe efficiently detects EV71 in real clinical serum samples with >95% agreement with RT-qPCR results. This high-throughput LDI-MS viral detection system is simple, reliable, and high-throughput. We believe this platform has the potential to be employed for the routine screening of patients with viral infections.

Synergistically dual-functional nano eye-drops for simultaneous anti-inflammatory and anti-oxidative treatment of dry eye disease.

We have developed a rapid and straightforward topical treatment method for dry eye disease (DED) using poly(catechin) capped-gold nanoparticles (Au@Poly-CH NPs) carrying amfenac [AF; a nonsteroidal anti-inflammatory drug (NSAID)] through effective attenuation of ocular surface tissue damage in dry eyes. A dual-targeted strategy based on ocular therapeutics was adopted to simultaneously block the cyclooxygenase enzymes-induced inflammation and reactive oxygen species (ROS)-induced oxidative stress, the primary two causes of DED. The self-assembled core-shell Au@Poly-CH NPs synthesized via a simple reaction between tetrachloroaurate(iii) and catechin possess a poly(catechin) shell (∼20 nm) on the surface of each Au NP (∼60 nm). The anti-oxidant and anti-inflammatory properties of AF/Au@Poly-CH NPs were evaluated by DCFH-DA and prostaglandin E2/VEGF assays, respectively. Our results demonstrate that Au@Poly-CH NPs not only act as an anti-oxidant to suppress ROS-mediated processes, but also serve as a drug carrier of AF for a synergistic effect on anti-inflammation. In vivo biocompatibility studies show good tolerability of AF/Au@Poly-CH NPs for potential use in the treatment of ocular surface pathologies. The dual-targeted therapeutic effects of AF/Au@Poly-CH NPs lead to rapid recovery from DED in a rabbit model. Au@Poly-CH NPs loaded with NSAIDs is a promising multifunctional nanocomposite for treating various inflammation- and oxidative stress-related diseases.

Dual-functional gelatin-capped silver nanoparticles for antibacterial and antiangiogenic treatment of bacterial keratitis.

Staphylococcus aureus (S. aureus) is a leading cause of keratitis worldwide and a significant threat to healthy vision. Pathological manifestations of bacterial keratitis (BK) caused by S. aureus involve stromal opacity, edema and neovascularization of an inflamed cornea, requiring immediate medical attention. Thus, S. aureus-induced keratitis is a devastating ocular infection that can lead to blindness if effective and timely treatment is not initiated. In this study, we demonstrate gelatin-capped silver nanoparticles (G-Ag NPs) as anti-infective therapeutics for the treatment of S. aureus-induced keratitis. G-Ag NPs were prepared by simple mixing of silver nitrate, maltose and gelatin. The gelatin molecules are capped in situ on the Ag NPs (∼14 nm). Compared to uncapped Ag NPs, the G-Ag NPs possess superior stability and antibacterial activity against S. aureus. We further demonstrate that G-Ag NPs possess effective inhibition of the proliferation, migration and tube formation of human umbilical vein endothelial cells, as well as strong disturbance of the angiogenesis in chick chorioallantoic membrane and rabbit corneal neovascularization. Furthermore, intrastromal administration of highly biocompatible G-Ag NPs alleviates S. aureus-induced bacterial keratitis in rabbit eyes and bacterial infection-induced corneal neovascularization. Our results demonstrate G-Ag NPs as a promising dual functional (antimicrobial and antiangiogenic) nanotherapeutic for preclinical treatment of eye-related microbial infections.

Green synthesis of catalytic gold/bismuth oxyiodide nanocomposites with oxygen vacancies for treatment of bacterial infections.

We have developed a simple and green solution for the synthesis of catalytic gold-doped bismuth oxyiodide (Au/BiOI) nanocomposites at room temperature from an aqueous mixture of gold ions, bismuth ions, and iodide ions. Au nanoparticles (NPs) were formed in situ and doped into BiOI nanosheets. The oxygen vacancies generated in BiOI give rise to its oxidase-like activity, and Au doping facilitated the reaction leading to a 4-fold higher oxidase-like activity of the Au/BiOI nanocomposite. The Au/BiOI nanocomposites showed wide spectrum antimicrobial activity not only against non-multidrug-resistant E. coli, K. pneumoniae, S. enteritidis, S. aureus, and B. subtilis bacteria, but also against multidrug-resistant bacteria, methicillin-resistant S. aureus (MRSA). The gold doping reduced the minimal inhibitory concentration value by ∼2000-fold for the Au/BiOI nanocomposite, in comparison with only BiOI nanoparticles. The bactericidal property of the Au/BiOI nanocomposite arose from the combined effect of the disruption of the bacterial membrane through a strong interaction of the nanocomposite with the bacteria and the generation of reactive oxygen species. Also, the Au/BiOI nanocomposite is highly biocompatible, which has been demonstrated in vitro by analysis of cytotoxicity and hemolysis, and in vivo by evaluating ocular tissue responses. Furthermore, intrastromal administration of Au/BiOI nanocomposites can effectively alleviate S. aureus-induced bacterial keratitis in rabbits, suggesting a significant disinfectant benefit in preclinical studies. The Au/BiOI nanocomposites show great potential for the inactivation of bacterial pathogens in an aqueous environment and treatment of bacterial infection-induced diseases.

Self-Assembled Chiral Gold Supramolecules with Efficient Laser Absorption for Enantiospecific Recognition of Carnitine.

Stereospecific recognition of chiral molecules is ubiquitous in chemical and biological systems, thus leading to strong demand for the development of enantiomeric drugs, enantioselective sensors, and asymmetric catalysts. In this study, we demonstrate the ratio of d-Cys and l-Cys playing an important role in determining the optical properties and the structures of self-assembled Cys-Au(I) supramolecules prepared through a simple reaction of tetrachloroaurate(III) with chiral cysteine (Cys). The irregularly shaped -[d-Cys-Au(I)] n- or - [l-Cys-Au(I)] n- supramolecules with a size larger than 500 nm possessing strong absorption in the near-UV region and chiroptical characteristics were only obtained from the reaction of Au(III) with d-Cys or l-Cys. On the other hand, spindle-shaped -[d/l-Cys-Au(I)] n- supramolecules were formed when using Au(III) with mixtures of d/l-Cys. Our results have suggested that Au(I)···Au(I) aurophilic interactions, and stacked hydrogen bonding and zwitterionic interactions between d/l-Cys ligands are important in determining their structures. The NaBH4-mediated reduction induces the formation of photoluminescent gold nanoclusters (Au NCs) embedded in the chiral -[d-Cys-Au(I)] n- or -[l-Cys-Au(I)] n- supramolecules with a quantum yield of ca. 10%. The as-formed Au NCs/-[d-Cys-Au(I)] n- and Au NCs/-[l-Cys-Au(I)] n- are an enantiospecific substrate that can trap l-carnitine and d-carnitine, respectively, and function as a nanomatrix for surface-assisted laser desorption/ionization mass spectrometry (LDI-MS). The high absorption efficiency of laser energy, analyte-binding capacity, and homogeneity of the Au NCs/-[Cys-Au(I)] n- allow for quantitation of enantiomeric carnitine down to the micromolar regime with high reproducibility. The superior efficiency of the Au NCs/-[d-Cys-Au(I)] n- substrate has been further validated by quantification of l-carnitine in dietary supplements with accuracy and precision. Our study has opened a new avenue for chiral quantitation of various analytes through LDI-MS using metal nanocomposites consisting of NCs and metal-ligand complexes.

Programmable DNA switches and their applications.

DNA switches are ideally suited for numerous nanotechnological applications, and increasing efforts are being directed toward their engineering. In this review, we discuss how to engineer these switches starting from the selection of a specific DNA-based recognition element, to its adaptation and optimisation into a switch, with applications ranging from sensing to drug delivery, smart materials, molecular transporters, logic gates and others. We provide many examples showcasing their high programmability and recent advances towards their real life applications. We conclude with a short perspective on this exciting emerging field.

The Controversial Orientation of Adenine on Gold and Silver.

In which orientation does adenine adsorb on gold and silver surfaces? This question has been a matter of debate for over 30 years. Since the dawn of surface-enhanced Raman spectroscopy (SERS); it and other techniques such as tip-enhanced Raman spectroscopy (TERS), surface-enhanced infrared absorption spectroscopy (SEIRAS), scanning tunneling microscopy (STM), density functional theory (DFT) simulations, and more, have been used in many attempts to answer this seemingly straightforward, yet controversial, question. Herein, the timeline and recent advances on this topic are explored, and the frequently contradictory findings are put into context and discussed.

Visual detection of cyanide ions by membrane-based nanozyme assay.

In this paper, we report a simple one-step synthesis of well-dispersed amorphous cobalt hydroxide/oxide-modified graphene oxide (CoOxH-GO) possessing peroxidase-like catalytic activity, and its application for the detection of H2O2, glucose, and CN- ions. CoOxH is formed and deposited in situ on the GO surface through the reaction between GO (size ~ 240nm) and Co2+ in basic solution at room temperature. We investigated the enzyme-mimicking activity of the CoOxH-GO nanohybrid in detail via the H2O2-mediated oxidation of Amplex Red (AR) to form fluorescent resorufin. The peroxidase-like activity of CoOxH-GO is utilized herein for the quantitation of H2O2 in a wide concentration range, from 100nM to 100µM. When coupled with glucose oxidase (GOD), the AR/CoOxH-GO system can determine glucose level in blood samples. Interestingly, cyanide ions (CN-) significantly inhibit the catalytic activity of the CoOxH-GO nanohybrid, which allows for the construction of a probe for the detection of CN- in water samples and laboratory wastes. We fabricated a membrane-based CoOxH-GO probe for the visual detection of CN- by preparing a thin film of CoOxH-GO on a positively charged and porous nylon membrane (N+M). The CoOxH-GO/N+M operates on the principle that CN- inhibits the catalytic activity of CoOxH-GO towards the H2O2-mediated oxidation of AR to form reddish resorufin on the membrane. The intensity of the red color of the membrane decreases with increasing CN- concentration, which can be easily observed with the naked eye at the nanomolar level. This cost-effective sensing system allows for the rapid and simple determination of the concentrations of CN- in complicated wastewater samples.

DNA Modulates the Interaction of Genetically Engineered DNA-Binding Proteins and Gold Nanoparticles: Diagnosis of High-Risk HPV Infection.

Gene detection has an important role in diagnosing several serious diseases and genetic defects in modern clinical medicine. Herein, we report a fast and convenient gene detection method based on the modulation of the interaction between a heat-resistant double-stranded DNA (dsDNA)-binding protein (Sso7d) and gold nanoparticles (Au NPs). We prepared a recombinant Cys-Sso7d, which is Sso7d with an extra cysteine (Cys) residue in the N-terminus, through protein engineering to control the interaction between Sso7d and Au NPs. Cys-Sso7d exhibited a stronger affinity for Au NPs and more easily induced the aggregation of Au NPs than Sso7d. In addition, Cys-Sso7d retained its ability to bind with dsDNA. The aggregation of Au NPs induced by Cys-Sso7d was diminished in the presence of dsDNA, which could be utilized as a transduction mechanism for the detection of the polymerase chain reaction (PCR) products of human papillomavirus (HPV) gene fragments (HPV types 16 and 18). The Cys-Sso7d/Au NP probe could detect as few as 1 copy of the HPV gene. The sensitivity and specificity of the Cys-Sso7d/Au NP probe for Pap smear clinical specimens (n = 52) for HPV 16 and HPV 18 detection were 85.7%/100.0% and 85.7%/91.7%, respectively. Our results demonstrate that the Cys-Sso7d/Au NP probe can be used to diagnose high-risk HPV types in Pap smear samples with high sensitivity, specificity, and accuracy.

Satellite-like Gold Nanocomposites for Targeted Mass Spectrometry Imaging of Tumor Tissues.

We have developed a simple, rapid, high-throughput cancer diagnosis system using functional nanoparticles (NPs) consisting of poly(catechin) capped-gold NPs (Au@PC NPs) and smaller nucleolin-binding aptamer (AS1411) conjugated gold NPs (AS1411-Au NPs). The AS1411-Au NPs/Au@PC NP is used as a targeting agent in laser desorption/ionization mass spectrometry (LDI-MS)-based tumor tissue imaging. Self-assembled core-shell Au@PC NPs are synthesized by a simple reaction of tetrachloroaurate(III) with catechin. Au@PC NPs with a well-defined and dense poly(catechin) shell (~40-60 nm) on the surface of each Au core (~60-80 nm) are obtained through careful control of the ratio of catechin to gold ions, as well as the pH of the reaction solution. Furthermore, we have shown that AS1411-conjugated Au NPs (13-nm) self-assembled on Au@PC NP can from a satellite-like gold nanocomposite. The high density of AS1411-Au NPs on the surface of Au@PC NP enhances multivalent binding with nucleolin molecules on tumor cell membranes. We have employed LDI-MS to detect AS1411-Au NPs/Au@PC NPs labeled nucleolin-overexpressing MCF-7 breast cancer cells through the monitoring of Au cluster ions ([Au n ]+; 1 ≤ n ≤ 3). The ultrahigh signal amplification from Au NPs through the formation of a huge number of [Au n ]+ ions results in a sensing platform with a limit of detection of 100 MCF-7 cells mL-1. Further, we have applied the satellite-like AS1411-Au NPs/Au@PC NP nanocomposite as a labeling agent for tumor tissue imaging by LDI-MS. Our nanocomposite-assisted LDI-MS imaging platform can be extended for simultaneous analysis of different tumor markers on cell membranes when using different ligand-modified metal nanoparticles.