A Metal Coordination-Based Supramolecular Elastomer with Shape Memory-Assisted Self-Healing Effect.

Rubber materials are widely used in aerospace, automotive, smart devices and artificial skin. It is significant to address the aging susceptibility of conventional vulcanized rubber and to impart it rapid self-healing performance for destructive crack damage. Herein, a novel supramolecular rubber elastomer is prepared by introducing metal coordination between carboxyl-terminated polybutadiene and polystyrene-vinylpyridine copolymer. Based on the metal coordination interaction, the elastomer exhibits shape memory and self-healing properties. Moreover, a rapid closure-repair process of destructive cracks is achieved by presetting temporary shapes. This shape memory-assisted self-repair model is shown to be an effective means for rapid repair of severe cracks. An approach to enhance the mechanical and self-healing properties of elastomer was demonstrated by adding appropriate amounts of oxidized carbon nano-onions (O-CNO) into the system. The tensile strength of the elastomer with an O-CNOs content of 0.5 wt% was restored to 83 ± 10% of the original sample after being repaired at 85 °C for 6 h. This study confirms that metal coordination interaction is an effective method for designing shape memory self-healing rubber elastomer. The shape memory-assisted self-healing effect provides a reference for the rapid self-repairing of severe cracks.

Detection of Protein Toxin Simulants from Contaminated Surfaces by Paper Spray Mass Spectrometry.

Proteinaceous toxins are harmful proteins derived from plants, bacteria, and other natural sources. They pose a risk to human health due to infection and also as possible biological warfare agents. Paper spray mass spectrometry (PS-MS) with wipe sampling was used to detect proteins from surfaces as a potential tool for identifying the presence of these toxins. Proteins ranging in mass between 12.4 and 66.5 kDa were tested, including a biological toxin simulant/vaccine for Staphylococcal enterotoxin B (SEBv). Various substrates were tested for these representative proteins, including a laboratory bench, a notebook cover, steel, glass, plant leaf and vinyl flooring. Carbon sputtered porous polyethylene (CSPP) was found to outperform typical chromatography paper used for paper spray, as well as carbon nanotube (CNT)-coated paper and polyethylene (PE), which have been previously shown to be well-suited for protein analysis. Low microgram quantities of the protein toxin simulant and other test proteins were successfully detected with good signal-to-noise from surfaces using a porous wipe. These applications demonstrate that PS-MS can potentially be used for rapid, sample preparation-free detection of proteins and biological warfare agents, which would be beneficial to first responders and warfighters.

A meta-analysis on correlations of OX40L variants with atherosclerotic disorders.

BACKGROUND: This meta-analysis was conducted to analyze the correlations of OX40 ligand (OX40L) variants with atherosclerotic cardio-cerebral vascular diseases (ASCVD). METHODS: Systematic literature research was conducted in PubMed, Medline, and Embase. All statistical analyses were conducted with Review Manager. RESULTS: Totally eighteen studies were enrolled for analyses. Although no any significant correlations between OX40L variants and ASCVD were detected in overall analyses. Further subgroup analyses by ethnicity revealed that rs1234314 variant was significantly associated with ASCVD in East Asians (dominant model: P = 0.03, odds ratios [OR] = 1.15, 95% confidence interval [CI], 1.01-1.31; allele model: P = 0.04, OR = 1.10, 95%CI, 1.00-1.20). When we stratified eligible studies by type of disease, positive results were observed for rs17568 variant in subjects with acute coronary syndrome (ACS) (allele model: P = 0.04, OR = 0.81, 95%CI, 0.65-0.99), for rs1234314 variant in subjects with coronary artery disease (CAD) (dominant model: P = 0.04, OR = 1.16, 95%CI, 1.00-1.35), for rs3850641 variant in subjects with CAD (recessive model: P = 0.02, OR = 1.42, 95%CI, 1.05-1.90) and myocardial infarction (MI) (recessive model: P = 0.03, OR = 1.49, 95%CI, 1.05-2.11). CONCLUSIONS: Our findings suggested that rs17568, rs1234314, and rs3850641 variants might serve as genetic biomarkers of certain types of CAD.

Targeted Protein Detection Using an All-in-One Mass Spectrometry Cartridge.

We developed a simple 3D printed cartridge for mass spectrometry (MS) targeted detection of plasma proteins, including post-translational modifications (PTMs). The cartridge uses an integrated antibody enrichment column to preconcentrate the protein target as well as a novel built-in substrate to ionize the protein targets for MS detection. We show several examples of using this cartridge to perform rapid detection of clinically significant proteoforms from plasma samples.

Ambient Ionization Mass Spectrometry Measurement of Aminotransferase Activity.

A change in enzyme activity has been used as a clinical biomarker for diagnosis and is useful in evaluating patient prognosis. Current laboratory measurements of enzyme activity involve multi-step derivatization of the reaction products followed by quantitative analysis of these derivatives. This study simplified the reaction systems by using only the target enzymatic reaction and directly detecting its product. A protocol using paper spray mass spectrometry for identifying and quantifying the reaction product has been developed. Evaluation of the activity of aspartate aminotransferase (AST) was chosen as a proof-of-principle. The volume of sample needed is greatly reduced compared with the traditional method. Paper spray has a desalting effect that avoids sprayer clogging problems seen when examining serum samples by nanoESI. This very simple method does not require sample pretreatment and additional derivatization reactions, yet it gives high quality kinetic data, excellent limits of detection (60 ppb from serum), and coefficients of variation <10% in quantitation. Graphical Abstract ᅟ.

Intestinal bacterium-derived cyp27a1 prevents colon cancer cell apoptosis.

The pathogenesis of metastasis of colon cancer (Cca) is to be further investigated. The dysfunction of apoptotic mechanism plays a role in the cancer cell over growth. This study tests a hypothesis by which intestinal bacterium-derived cyp27a1 prevents apoptosis in colon cancer cells. In this study, the levels of cyp27a1 in human stool samples were assessed by enzyme-linked immunosorbent assay. The apoptosis of Cca cells was observed by flow cytometry. The expression of cyp27a1 was assessed by real time RT-PCR and Western blotting. We observed higher levels of cyp27a1 in the stool samples of Cca patients than that from healthy subjects. Cca colon epithelial biopsy contained high levels of cyp27a1 protein, but not the cyp27a1 mRNA. Cyp27a1 prevented Cca cell apoptosis induced by vitamin D3. In conclusion, intestinal bacterium-derived cyp27a1 facilitates Cca survival by inhibiting Cca cell apoptosis.

Analysis of biofluids by paper spray MS: advances and challenges.

Paper spray MS is part of a cohort of ambient ionization or direct analysis methods that seek to analyze complex samples without prior sample preparation. Extraction and electrospray ionization occur directly from the paper substrate upon which a dried matrix spot is stored. Paper spray MS is capable of detecting drugs directly from dried blood, plasma and urine spots at the low ng/ml to pg/ml levels without sample preparation. No front end separation is performed, so MS/MS or high-resolution MS is required. Here, we discuss paper spray methodology, give a comprehensive literature review of the use of paper spray MS for bioanalysis, discuss technological advancements and variations on this technique and discuss some of its limitations.

Ionization Suppression and Recovery in Direct Biofluid Analysis Using Paper Spray Mass Spectrometry.

Paper spray mass spectrometry is a method for the direct analysis of biofluid samples in which extraction of analytes from dried biofluid spots and electrospray ionization occur from the paper on which the dried sample is stored. We examined matrix effects in the analysis of small molecule drugs from urine, plasma, and whole blood. The general method was to spike stable isotope labeled analogs of each analyte into the spray solvent, while the analyte itself was in the dried biofluid. Intensity of the labeled analog is proportional to ionization efficiency, whereas the ratio of the analyte intensity to the labeled analog in the spray solvent is proportional to recovery. Ion suppression and recovery were found to be compound- and matrix-dependent. Highest levels of ion suppression were obtained for poor ionizers (e.g., analytes lacking basic aliphatic amine groups) in urine and approached -90%. Ion suppression was much lower or even absent for good ionizers (analytes with aliphatic amines) in dried blood spots. Recovery was generally highest in urine and lowest in blood. We also examined the effect of two experimental parameters on ion suppression and recovery: the spray solvent and the sample position (how far away from the paper tip the dried sample was spotted). Finally, the change in ion suppression and analyte elution as a function of time was examined by carrying out a paper spray analysis of dried plasma spots for 5 min by continually replenishing the spray solvent. Graphical Abstract ᅟ.

Development of a Paper Spray Mass Spectrometry Cartridge with Integrated Solid Phase Extraction for Bioanalysis.

A novel paper spray cartridge with an integrated solid phase extraction (SPE) column is described. The cartridge performs extraction and pre-concentration, as well as sample ionization by paper spray, from complex samples such as plasma. The cartridge allows for selective enrichment of target molecules from larger sample volumes and removal of the matrix, which significantly improved the signal intensity of target compounds in plasma samples by paper spray ionization. Detection limits, quantitative performance, recovery, ionization suppression, and the effects of sample volume were evaluated for five drugs: carbamazepine, atenolol, sulfamethazine, diazepam, and alprazolam. Compared with direct paper spray analysis of dried plasma spots, paper spray analysis using the integrated solid phase extraction improved the detection limits significantly by a factor of 14-70, depending on the drug. The improvement in detection limits was, in large part, due to the capability of analyzing larger sample volumes. In addition, ionization suppression was found to be lower and recovery was higher for paper spray with integrated SPE, as compared to direct paper spray analysis. By spiking an isotopically labeled internal standard into the plasma sample, a linear calibration curve for the drugs was obtained from the limit of detection (LOD) to 1 µg/mL, indicating that this method can be used for quantitative analysis. The paper spray cartridge with integrated SPE could prove valuable for analytes that ionize poorly, in applications where lower detection limits are required, or on portable mass spectrometers. The improved performance comes at the cost of requiring a more complex paper spray cartridge and requiring larger sample volumes than those used in typical direct paper spray ionization.

A label-free double-amplification system for sensitive detection of single-stranded DNA and thrombin by liquid chromatography-mass spectrometry.

A label-free double amplification system has been developed by using a ternary DNA probe containing the poly(adenine-thymine) sequence assisted by exonuclease III degradation. The method achieved more than 600-fold signal amplification and allowed sensitive detection of single-stranded DNA and thrombin at the pM level by using liquid chromatography/mass spectrometry.

Highly sensitive and specific detection of histamine via the formation of a self-assembled magic number cluster with thymine by mass spectrometry.

A novel method for the detection of histamine (HIM) via the formation of a self-assembled magic number cluster with thymine (T) by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is described. The formation of the magic number cluster [T17 + HIM + 2H](2+) shifts the MS signal of histamine to the interference-free higher mass range and the signal intensity is increased by four orders of magnitude. In addition, the formation of [T17 + HIM + 2H](2+) is highly specific to histamine compared with its metabolite and other similar biogenic amines, which may be attributed to both of its amino and imidazole groups. The linear dynamic range of the method is in the range of 1 nM-20 µM, and the limit of detection can be as low as 0.1 nM. The feasibility of this method is further demonstrated by the quantitative analysis of histamine in a red wine sample. Since little sample preparation or separation is required before the analysis, this method provides a rapid new way for the sensitive and specific detection of histamine by MS.

Signal enhancement of carboxylic acids by inclusion with ß-cyclodextrin in negative high-voltage-assisted laser desorption ionization mass spectrometry.

RATIONALE: It is difficult to directly analyze carboxylic acids in complex mixtures by ambient high-voltage-assisted laser desorption ionization mass spectrometry (HALDI-MS) in negative ion mode due to the low ionization efficiency of carboxylic acids. METHODS: A method for the rapid detection of carboxylic acids in negative HALDI-MS has been developed based on their inclusion with ß-cyclodextrin (ß-CD). RESULTS: The negative HALDI-MS signal-to-noise ratios (S/Ns) of aliphatic, aromatic and hetero atom-containing carboxylic acids can all be significantly improved by forming 1:1 complexes with ß-CD. These complexes are mainly formed by specific inclusion interactions which are verified by their collision-induced dissociation behaviors in comparison with that of their corresponding maltoheptaose complexes. A HALDI-MS/MS method has been successfully developed for the detection of α-lipoic acid in complex cosmetics and ibuprofen in a viscous drug suspension. CONCLUSIONS: The negative HALDI-MS S/Ns of carboxylic acids can be improved up to 30 times via forming non-covalent complexes with ß-CD. The developed method shows the advantages of being rapid and simple, and is promising for rapid detection of active ingredients in complex samples or fast screening of drugs and cosmetics.

Laser desorption dual spray post-ionization mass spectrometry for direct analysis of samples via two informative channels.

A laser desorption dual spray post-ionization mass spectrometry method is described, and its usefulness is demonstrated with the examples of selective detection of food components, manipulation of protein charge state distribution and investigation on the formation of magic number clusters. The method is carried out by adopting two spray emitters for post-ionization of analytes desorbed by a pulsed infrared laser. Various components in a complex sample or distinct behavior of an analyte in two different spray reagents can be rapidly probed by the method quasi-simultaneously, highlighting the potential applications of this method for protein characterization, reaction study and food analysis.

Direct analysis of samples under ambient condition by high-voltage-assisted laser desorption ionization mass spectrometry in both positive and negative ion mode.

RATIONALE: With the rapid development of ambient mass spectrometry, the hybrid laser-based ambient ionization methods which can generate multiply charged ions of large biomolecules and also characterize small molecules with good signal-to-noise in both positive and negative ion modes are of particular interest. METHODS: An ambient ionization method termed high-voltage-assisted laser desorption ionization (HALDI) is developed, in which a 1064 nm laser is used to desorb various liquid samples from the sample target biased at a high potential without the need for an organic matrix. The pre-charged liquid samples are desorbed by the laser to form small charged droplets which may undergo an electrospray-like ionization process to produce multiply charged ions of large biomolecules. RESULTS: Various samples including proteins, oligonucleotides (ODNs), drugs, whole milk and chicken eggs have been analyzed by HALDI-MS in both positive and negative ion mode with little or no sample preparation. In addition, HALDI can generate intense signals with better signal-to-noise in negative ion mode than laser desorption spay post-ionization (LDSPI) from the same samples, such as ODNs and some carboxylic-group-containing small drug molecules. CONCLUSIONS: HALDI-MS can directly analyze a variety of liquid samples including proteins, ODNs, pharmaceuticals and biological fluids in both positive and negative ion mode without the use of an organic matrix. This technique may be further developed into a useful tool for rapid analysis in many different fields such as pharmaceutical, food, and biological sciences.

Online coupling of capillary electrophoresis with direct analysis in real time mass spectrometry.

The online coupling of capillary electrophoresis with ambient direct analysis in real time mass spectrometry (DART-MS) was realized by a coaxial tip interface. The analytes eluted from capillary electrophoresis (CE) were directly ionized by the metastable helium flux produced by DART and transferred into MS for the detection, with which the online separation and simultaneous detection were achieved. The CE-DART-MS can tolerate higher concentrations of detergents and salts than traditional CE-electrospray ionization (ESI)-MS and avoided the difficulties of collecting CE effluent and cleaning the interface, which simplified the experimental procedures and shortened the analysis time. The performance of the technique was successfully verified by capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) using a mixture of 4-aminoantipyrine, zolmitriptan, and quinine. This online technique showed good repeatability with the relative standard deviations (RSDs; n = 5) of 0.56-1.23% for the retention times and 2.01-7.41% for the peak areas. The quantitative analysis of 4-aminoantipyrine was accomplished in the range of 0.01-0.50 mg/mL with the linear correlation coefficient of 0.9995 and limit of detection of 14.7 fmol. Compared with CE-ESI-MS, the ion suppression effects of nonvolatile salts and detergents were efficiently minimized. The signal intensity remained constant when the concentrations reached 100 mM for sodium borate and 30 mM for SDS (in 30 mM sodium borate buffer). In addition, the proposed method was successfully applied to the detection of the endogenous caffeine in Chinese white tea.

Graphite-coated paper as substrate for high sensitivity analysis in ambient surface-assisted laser desorption/ionization mass spectrometry.

In this work, an extremely simple and quite sensitive mass spectrometric method termed ambient surface-assisted laser desorption/ionization mass spectrometry (ambient SALDI-MS) was developed to analyze different kinds of compounds, just using a piece of graphite-coated paper for the sample introduction. This provides great advantage in simplifying the analysis process. The method is quite easy to use, and there is no need to worry about the source of graphite, that is, the brands or the types of pencil. And the whole process was carried out under atmospheric pressure, offering all the merits that could occur in ambient MS. The improved sensitivity of this method is mainly because of the graphite, which serves as energy-transfer medium to absorb the energy of the photons and release it to the analytes that are adsorbed on the graphite surface. Also, three different laser wavelengths (1064, 532, and 355 nm) was tested to investigate the desorption mechanism. Fifty-one compounds, with varied chemical structures, were tried to prove that this new method possessed universal applicability to detect different kinds of small organic molecules.

Unexpected complexation reaction during analysis of proteins using laser desorption spray post-ionization mass spectrometry.

We describe complexation reactions of insulin and other proteins with metal ions generated from the substrate surface by laser irradiation in laser desorption spray post-ionization mass spectrometry (LDSPI-MS). This particular type of complexation reaction in LDSPI-MS was investigated for the first time, which indicated that the mechanistic process of LDSPI-MS might be much more complicated than that proposed before for similar methods.

[A linkage between beta-fibrinogen gene -148C/T polymorphism and cerebral infarction].

OBJECTIVE: To study the linkage between -148C/T polymorphism of beta-fibrinogen gene and plasma fibrinogen levels in patients with acute cerebral infarction. METHODS: One hundred and fifty-one patients with cerebral infarction and 101 healthy individuals were enrolled in this trial. The beta-fibrinogen gene -148C/T polymorphism was analyzed by PCR-restriction fragment length polymorphism, and plasma fibrinogen levels were obtained from prothrombin time assay. RESULTS: Plasma fibrinogen levels of patients were significantly higher than those of controls (P<0.01). In both groups, T allele carriers had higher plasma fibrinogen levels than other those did (P<0.01); and the fibrinogen level difference was still significant if both groups was based on their sex (P<0.05). Divided by age, each group of the study cases has significant difference between two genotypes (P<0.05). T -148 allele frequency of the middle age case in study group was higher than that in control group (P<0.05). CONCLUSION: High plasma fibrinogen level is a risk factor to cerebral infarction. Plasma fibrinogen level is affected by -148C/T polymorphism of beta-fibrinogen gene. With or without other risk factors and environmental factors affecting, T allele increases plasma fibrinogen level and may be a heritable risk factor to cerebral infarction.