Effect of Penetration Enhancer on the Structure of Stratum Corneum: On-Site Study by Confocal Polarized Raman Imaging.

Keratin and lipid structures in the stratum corneum (SC) are closely related to the SC barrier function. The application of penetration enhancers (PEs) disrupts the structure of SC, thereby promoting infiltration. To quantify these PE-induced structural changes in SC, we used confocal Raman imaging (CRI) and polarized Raman imaging (PRI) to explore the integrity and continuity of keratin and lipid structures in SC. The results showed that water is the safest PE and that oleic acid (OA), sodium dodecyl sulfate (SDS), and low molecular weight protamine (LMWP) disrupted the ordered structure of keratin, while azone and liposomes had less of an effect on keratin. Azone, OA, and SDS also led to significant changes in lipid structure, while LMWP and liposomes had less of an effect. Establishing this non-invasive and efficient strategy will provide new insights into transdermal drug delivery and skin health management.

Surface-Enhanced Raman Scattering Enantioselective Detection of Gastric Cancer-Related d-Amino Acids in Saliva Based on Enzyme-Mediated Cascade Reaction.

The unusual d-amino acids (d-AAs), as the counter enantiomer of usual l-amino acids (l-AAs), have evoked increasing attention because of their potential relevance with diseases. Accordingly, it is essential to establish sensitive and selective detection methods for d-AAs without the interferences from l-AAs. The surface-enhanced Raman scattering (SERS) technique is efficacious for the detection of molecules but routinely ineffective in enantiomeric differentiation. d-Proline (d-Pro) and d-alanine (d-Ala) are regarded as biomarkers of gastric cancer. Herein, Raman-active boronate modified SERS chips are constructed to develop a d-amino acid oxidase (DAAO)-mediated cascade reaction-based SERS enantioselective assay for d-Pro and d-Ala. The principle is that DAAO selectively catalyzes the deamination of d-Pro and d-Ala, and the produced H2O2 oxidizes boronate to present a new SERS peak at 883 cm-1 for quantitative analysis in a ratiometric way. A linear range from 20 to 400 µmol/L and a limit of detection down to 14.8 µmol/L are reached. In addition, interferences from l-AAs and many other possible species coexisting in biofluids with the detection of d-Pro and d-Ala are ignorable. Enzyme-mediated cascade reaction-based SERS chips are further utilized for saliva sample analysis, and the total levels of d-Pro and d-Ala in salivary samples from gastric cancer patients are much higher than those of healthy persons. This work provides a solution for SERS enantioselective analysis and noninvasive screening chiral biomolecules for disease diagnosis.

Chirality Detection by Raman Spectroscopy: The Case of Enantioselective Interactions between Amino Acids and Polymer-Modified Chiral Silica.

In chirality research area, it is of interest to reveal the chiral feature of inorganic nanomaterials and their enantioselective interactions with biomolecules. Although common Raman spectroscopy is not regarded as a direct chirality analysis tool, it is in fact effective and sensitive to study the enantioselectivity phenomena, which is demonstrated by the enantio-discrimination of amino acid enantiomers using the polydopamine-modified intrinsically chiral SiO2 nanofibers in this work. The Raman scattering intensities of an enantiomer of cysteine are more than twice as high as those of the other enantiomer with opposite handedness. Similar results were also found in the cases of cystine, phenylalanine, and tryptophan enantiomers. In turn, these organic molecules could be used as chirality indicators for SiO2, which was clarified by the unique Raman spectra-derived mirror-image relationships. Thus, an indirect chirality detection method for inorganic nanomaterials was developed.

A flexible carbon nanotube-modified poly(styrene-butadiene)-based dopamine sensor.

In this work, a multi-walled carbon nanotube-modified flexible poly(styrene-butadiene) fiber membrane material was prepared for the sensitive and selective electrochemical detection of dopamine (DA) in human serum and DA injection. The flexible fiber membrane prepared by electrospinning technology is expected to realize its application in wearable devices. The obtained conductive film-based electrochemical sensor can effectively minimize interference caused by ascorbic acid and uric acid. Under the optimized experimental conditions of differential pulse voltammetry, DA gives a linear response in the range of 1-650 µM (R2 = 0.996). The detection limit of DA (signal-to noise ratio = 3) was determined to be 0.062 µM.

Photoelectrochemical determination of the activity of M.SssI methyltransferase, and a method for inhibitor screening.

A photoelectrochemical (PEC) method is described for the determination of the activity of M.SssI methyltransferase (MTase). The assay relies on enzyme-linkage reactions and a DNA intercalator Ru(bpy)2(dppz)2+ (where bpy is 2,2'-bipyridine, and dppz is dipyrido[3,2-a:2',3'-c]phenazine) which both serves as a PEC signal. The PEC electrode was obtained by immobilizing 5'-amino modified DNA strands (containing the methylation recognition site 5'-CCGG-3') on a polyethylenimine (PEI) coated ITO/SnO2 electrode with glutaraldehyde as crosslinking agent. In the presence of MTase and S-adenosyl-L-methionine, the 5'-CCGG-3' sequence in the DNA on the electrode is methylated. This protects the DNA strands from the shear of the methylation-sensitive restriction endonuclease HpaII. Consequently, more intact DNA strands remain on the surface of the electrode, providing more sites for Ru(bpy)2(dppz)2+ binding which in turn results in a high PEC response. The result demonstrates that the photocurrent increases linearly with the activity of MTase from 5 to 80 U·mL-1, and the limit of detection is 0.45 U·mL-1. The other MTases does not enhance the photocurrent, suggesting good selectivity of the assay. The method was also applied to rapid evaluate and screen the inhibitors of MTase. This strategy can be utilized to determinate the activity of other DNA MTases with specific DNA sequence. Graphical abstract Schematic presentation of a photoelectrochemical assay based on enzyme-linkage reactions and a photo electrochemical probe combined with the oxalic acid involved cyclic amplification system for the determination of methyltransferase activity.

Reproducible preparation of a stable polypyrrole-coated-silver nanoparticles decorated polypyrrole-coated-polycaprolactone-nanofiber-based cloth electrode for electrochemical sensor application.

A piece of conductive cloth has been successfully constructed from polypyrrole-coated silver nanoparticle (Ag@PPy) composites decorated on electrospun polycaprolactone (PCL) nanofibers that formed the core-shell structure of Ag@PPy/PCL@PPy via a photo-induced one-step redox reaction. The photochemical reaction method both accelerated the rate of formation of silver nanoparticles (Ag NPs) and enhanced the dispersion of Ag NPs at the surface of PCL@PPy film. The resulting Ag@PPy/PCL@PPy-based cloth was flexible enough to be cut and pasted onto a glass carbon electrode for the preparation of a biosensor. The resulting biosensor showed good electrochemical activity toward the reduction of H2O2 with low detection limit down to 1 µM (S/N = 3) and wide linear detection ranging from 0.01 mM to 3.5 mM (R(2) = 0.990). This sensor has been applied to detect the trace H2O2 residual in milk. The cloth electrode has been proved to exhibit long-term stability, high selectivity, and excellent reproducibility.

Dummy molecularly imprinted solid-phase extraction-SERS determination of AFB1 in peanut.

As a class I carcinogen, aflatoxin B1 (AFB1) contamination in foods and feeds accounts for 75 % of the total mycotoxin contamination. In this work, a simple and reliable surface-enhanced Raman spectroscopy (SERS) method for sensitive and selective detection of AFB1 in peanut samples integrated with dummy molecularly imprinted polymers (DMIPs) is developed. N-isopropylacrylamide (NIPAM) and 7-ethoxycoumarin (7-EOC) are chosen as monomer and dummy template, respectively and their ratio was screened through molecular design in both of kinetic and static adsorption views to form the optimal DMIPs. As-prepared dummy molecularly imprinted solid-phase extraction (DMISPE) could selectively enrich AFB1 from peanut samples. Finally, a liquid-liquid interface self-assembly constructed thioctic acid-decorated AgNPs monolayer film (TA-AgNPs MF) as a SERS-active substrate is employed to determine the amount of AFB1 eluted from DMISPE. SERS assay shows high detection sensitivity for AFB1 in peanut samples with limit of detection of 0.1 µg L-1 and a linear concentration relationship range from 0.1 to 10 µg L-1.

The IP6 micelle-stabilized small Ag cluster for synthesizing Ag-Au alloy nanoparticles and the tunable surface plasmon resonance effect.

The stable small Ag seeds (size in diameter < 10 nm) were obtained in the presence of inositol hexakisphosphoric (IP6) micelles. Then Ag-Au bimetallic nanoparticles were synthesized through a replacement reaction with the rapid interdiffusion process between such small Ag seeds in nanoclusters and HAuCl4. Adjusting the dosage of HAuCl4 resulted in different products, which possessed unique surface plasmon resonances (SPR). The morphologies of the as-made nanoparticles were observed using transmission electron microscopy and field emission scanning electron microscopy and their compositions were determined by energy-dispersive x-ray spectroscopy. Among them, the Ag-Au alloy nanoparticles with the cauliflower-like structure had a suitable SPR for highly sensitive Raman detection application as a surface-enhanced Raman scattering (SERS) substrate with a long-term stability of six months.

Molecularly imprinted Monolithic column-based SERS sensor for selective detection of cortisol in dog saliva.

Molecularly imprinted monolithic column embedded with silver nanoparticles (MIMC@Ag) was synthesized by in-situ polymerization with template and porogen inside capillary tube followed by silver precursor reduction and template/porogen removal for realizing Raman detection of cortisol. Dense silver nanoparticles generated within the monolith makes this kind of column suitable for surface enhanced Raman scattering (SERS) detection, designated as SERS-MIMC. Scanning electron microscopy and BET profiler confirmed larger pore structure in the column after template removal. The corresponding increased mass transfer/binding rate, selective adsorption and adsorptive mechanism of the MIMC were well studied with a series of adsorption experiments. The minimum Raman detectable concentration of cortisol is 1 × 10-7 mol L-1 by using MIMC@Ag with a good linear relationship in the concentration range from 1 × 10-3 to 1 × 10-7 mol L-1. SERS sigmal of cortisol can be clearly distinguished from its analogs (estradiol, cholesterol and dexamethasone), proving the selective recognition of cortisol for SERS detection by MIMC@Ag. This ease-to-prepare SERS-MIMC sensor also shows good stability and reusability. The SERS-MIMC has been successfully applied for the easy, sensitive and selective detection of cortisol in dog saliva.

Preparation of QSS@AuNPs and Solvent Inducing Enhancement Strategy for Raman Determination of Salivary Thiocyanate.

Making the substrates form highly dense, homogeneous, and stable hotspots regions is important for the sensitive detection of surface-enhanced Raman spectroscopy (SERS). A new strategy based on solvent-induced (SI) SERS substrate to form a stable interval of the hotspot for detection was explored and the enhancement factor (EF) of our SERS substrates could reach about 1.4 × 109. By preferential adsorption of alcohol solutions by Q-Sepharose microsphere (QSS) in mixed water and alcohol solutions, the size of QSS@AuNPs was dynamically adjusted and the spacing between gold nanoparticles (AuNPs) was adjusted to keep the substrate in the optimal hotspot mode for the sensitive detection of SERS in the liquid state. As a real application case, such a SI-SERS strategy was employed to determine SCN- in saliva and a limit of detection (LOD) of about 10-10 M could be achieved.

ZnO Tips Dotted with Au Nanoparticles-Advanced SERS Determination of Trace Nicotine.

Long-term exposure to nicotine causes a variety of human diseases, such as lung damage/adenocarcinoma, nausea and vomiting, headache, incontinence and heart failure. In this work, as a surface-enhanced Raman scattering (SERS) substrate, zinc oxide (ZnO) tips decorated with gold nanoparticles (AuNPs) are fabricated and designated as ZnO/Au. Taking advantage of the synergistic effect of a ZnO semiconductor with morphology of tips and AuNPs, the ZnO/Au-based SERS assay for nicotine demonstrates high sensitivity and the limit of detection 8.9 × 10-12 mol/L is reached, as well as the corresponding linear dynamic detection range of 10-10-10-6 mol/L. Additionally, the signal reproducibility offered by the SERS substrate could realize the reliable determination of trace nicotine in saliva.

Molecule-specific vibration-based chiral differentiation of Raman spectra using cysteine modified gold nanoparticles: the cases of tyrosine and phenylalanine.

The chirality of amino acids plays a key role in many biochemical processes, with the development of spectroscopic analysis methods for the chiral differentiation of amino acids being significant. Normal Raman spectroscopy is blind to chirality; however, chiral discrimination of tyrosine (Tyr) (or phenylalanine, Phe) enantiomers using Raman spectra can be achieved assisted by the construction of a simple chiral selector (i.e., cysteine (Cys)-modified Au nanoparticles (NPs)). Due to the synergetic effect between Cys and the Au NPs, the characteristic Raman scattering intensities of the Tyr (or Phe) enantiomer with the same chirality of Cys are enantioselectively boosted by over four-fold compared with those of the counter enantiomer of Tyr (or Phe). The large differences in the Raman signals allow for the determination of enantiomeric excess. Interestingly, such enantiomeric discrimination is not revealed by the common chiral analysis method of circular dichroism spectroscopy. Consequently, it is anticipated that Raman spectroscopy based on molecular vibrations will find broad applications in chirality-related detection with high sensitivity and species specificity.

Enzyme-Free Tandem Reaction Strategy for Surface-Enhanced Raman Scattering Detection of Glucose by Using the Composite of Au Nanoparticles and Porphyrin-Based Metal-Organic Framework.

In this work, an S hybrid nanosheet with multiple functions is synthesized by in situ modification of gold nanoparticles (AuNPs) onto two-dimensional (2D) metalloporphyrinic metal-organic framework (MOF) (Cu-tetra(4-carboxyphenyl)porphyrin chloride(Fe(III)), designated as AuNPs/Cu-TCPP(Fe). Cu-TCPP(Fe) nanosheets contribute peroxidase-like activity, and AuNPs have glucose oxidase (GOx) mimicking performance, which induce the cascade catalysis reactions to convert glucose into hydrogen peroxide (H2O2), and then, by using AuNP catalysis, H2O2 oxidizes the no Raman-active leucomalachite green (LMG) into the Raman-active malachite green (MG). Simultaneously, in the presence of AuNPs, sensitive and selective surface-enhanced Raman scattering (SERS) determination of glucose can be achieved. The bioenzyme-free SERS assay based on such AuNPs/Cu-TCPP(Fe) nanosheets is used for detection of glucose in saliva, showing good recovery from 96.9 to 100.8%. The work paves a new way to design a nanozyme-based SERS protocol for biomolecule analysis.

Raman tracking the activity of urease in saliva for healthcare.

The detection of urease activity in the oral cavity is considered to be an efficient way to prevent dental caries and also to screen for helicobacter pylori infection. Herein, a rapid surface enhanced Raman scattering (SERS) method is proposed to determine the activity of urease by using inositol hexaphosphate (IP6) stabilized silver nanoparticles (AgNPs@IP6) as an efficient SERS-active substrate. The determination was achieved by monitoring the SERS peak intensity of urea at 1003 cm-1. With urease increase, the response of urea at 1003 cm-1 decreases gradually, indicating the two has good correlation. A linear relationship between the absolute value of signal drop and urease concentration is observed in a range from 2.35 to 37.5 µg/mL. In addition, the rapid SERS method was used to evaluate the activity of urease in real sample of saliva without any pretreatment, exhibiting a promising potential for biomedical application.

Comprehensively evaluating the digestive performance of sludge with different lignocellulosic components in mesophilic anaerobic digester.

In excess sludge digestion, organic matters cannot be digested adequately due to its high lignocellulose content. This study attempted to comprehensively evaluate the digestive performances of sludge with different lignocellulosic components (hemicellulose, cellulose and lignin). Results show that hemicellulose/dealkaline lignin addition (S6) presents the highest methane yield of 203.6 mL/gVS. Compared to hemicellulose, dealkaline lignin is hardly degraded (lower than 10%), while its participation can promote the degradation of other organics in the system. Additionally, solo cellulose feedstock is difficult to be hydrolyzed (only 40.1%) without hemicellulose and dealkaline lignin addition. VFAs composition analysis indicates that VFA inhibition occurs in the digester with hemicellulose, cellulose and dealkaline lignin addition (S8). Microbial diversities of different digestive systems show that the relative abundance of Euryarchaeota in the digester S6 (7.2%) is much higher than others, and some specific microbes (Bacteroidetas and Firmicutes) are enriched in the S5 (74.1%) and S8 (54.7%) digesters.

Polydopamine-based functional composite particles for tumor cell targeting and dual-mode cellular imaging.

Particles which bear tumor cell targeting and multimode imaging capabilities are promising in tumor diagnosis and cancer therapy. A simple and versatile method to fabricate gold/polydopamine-Methylene Blue@Bovine Serum Albumin-glutaraldehyde-Transferrin composite particles (Au/PDA-MB@BSA-GA-Tf NPs) for tumor cell targeting and fluorescence (FL) / surface-enhanced Raman scattering (SERS) dual-modal imaging were reported in this work. Polydopamine (PDA) spheres played an important role in gold ion reduction, gold nanoparticle (Au NPs) binding and methylene blue (MB) adsorption, MB were employed as both fluorescence label and Raman reporter. In addition, glutaraldehyde (GA) crosslinked bovine serum albumin (BSA) in the outer layer of Au/PDA-MB nanoparticles can prevent MB from dissociation and leakage. The composite nanoparticles were further conjugated with transferrin (Tf) to target transferrin receptor (TfR)-overexpressed cancer cells. The targeting ability as well as the intracellular location of the probe was investigated through SERS mapping and fluorescence imaging. Their excellent biocompatibility was demonstrated by low cytotoxicity against breast cancer cell (4T1 cell).

Quartz crystal microbalance bioaffinity sensor for biotin based on mixed self-assembled monolayers and metastable molecular complex receptor.

A quartz crystal microbalance (QCM) sensor was proposed for the detection of small molecule biotin based on the mixed self-assembled monolayer (SAM) of thiols on gold substrate and the bioaffinity difference between an analyte (biotin) and an analogue compound (HABA) in binding avidin. Avidin formed a metastable complex with 2-[(4-hydroxyphenyl)azo]benzoic acid (HABA) immobilized on the crystal surface. When the sensor contacts a sample solution containing biotin, the avidin was released from the sensor surface to form a more stable complex with biotin in solution. The frequency change recorded is proportional to the desorbed mass of avidin, and there is a clear mathematic relationship between the frequency change and the biotin concentration. The use of mixed SAMs allows the stable attachment of bioreceptor molecules on the QCM, and enhances the amount of the immobilized molecules on the QCM, as a longer "space arm" in the mixed SAMs makes this monolayer membrane more accessible to capture the immobilized molecules. The proposed bioaffinity sensor has nice response to biotin in the range of 0.017-1.67 microg/mL. The sensor could be regenerated under very mild conditions simply by reimmersion of the sensor into a biotin solution to desorb the surplus avidin.

Magnetically optimized SERS assay for rapid detection of trace drug-related biomarkers in saliva and fingerprints.

New developments in the fields of human healthcare and social security call for the exploration of an easy and on-field method to detect drug-related biomarkers. In this paper, Au nanoparticles dotted magnetic nanocomposites (AMN) modified with inositol hexakisphosphate (IP6) were used as surface-enhanced Raman scattering (SERS) substrate to quickly monitor trace drug-related biomarkers in saliva and to on-site screen a trace drug biomarker in fingerprints. Due to inducing with an external magnet, such substrate presented a huge SERS activity, which has met the sensitivity requirement for assay to detect the drug biomarkers in saliva from the U.S. Substance Abuse and Mental Health Services Administration, and also the limit of detection for drug biomarker in fingerprint reached 100 nM. In addition, this AMN-based SERS assay was successfully conducted using a portable Raman spectrometer, which could be used to on-site and accurately differentiate between the smokers and drug addicts in near future.

Botanical micelle and its application for direct electrochemical biosensor.

This paper is concerned about the entrapment of horseradish peroxidase (HRP) within botanical inositol hexakisphosphoric (IP(6)) micelles for the preparation of enzyme biosensor. The good affinity of IP(6) micelles with the enzyme provides naturally biocompatible microenvironment for the enzyme immobilization, achieving the direct electron transfer between HRP and electrode surface. The resulting biosensor to H(2)O(2) detection exhibits a low detection limit of 0.1 µmol L(-1) (S/N = 3), a quick response time (3s), and a long-term stability. The apparent Michaelis-Menten constant is quite tiny about 0.0016 mmol L(-1).