Non-hemolytic α-AApeptides as antimicrobial peptidomimetics

We report a new class of peptide mimetics, α-AApeptides, that display broad-spectrum activity against both Gram-negative and Gram-positive bacteria and fungi. With non-hemolytic activity, resistance to protease hydrolysis, and easy sequence programmability, α-AApeptides may emerge as a novel class of antibiotics.     

Separation of exenatide analogue mono-PEGylated with 40 kDA polyethylene glycol by cation exchange chromatography

Random PEGylation usually resulted in product mixtures composed of mono-PEGylated isomers and multi-PEGylated attachments. Generally in PEGylation research, separation of the mono-PEGylated isomers was the prerequisite for finding the optimal PEGylation site. However, when peptides or proteins were PEGylated with polyethylene glycol as large as 40 kDA, the physicochemical properties like hydrophobicity and molecular size of the isomers would become too similar to make the routine separation methods, like RP-HPLC, size-exclusion chromatography, SDS-PAGE and capillary isoelectric focusing invalid. This article presented a useful method of successfully separating exenatide analogue (an incretin for diabetic therapy) isomers mono-PEGylated with 40 kDA polyethylene glycol by cation exchange chromatography, which would be a powerful tool for the PEGylation research.     

Protective effect of total aralosides of Aralia elata (Miq) Seem (TASAES) against diabetic cardiomyopathy in rats during the early stage, and possible mechanisms

Total aralosides of Aralia elata (Miq) Seem (TASAES) from Chinese traditional herb Longya Aralia chinensis L was found to improve cardiac function. The present study was to determine the protective effects of TASAES on diabetic cardiomyopathy, and the possible mechanisms. Therefore, a single dose of streptozotocin was used to induce diabetes in Wister rats. Diabetic rats were immediately treated with low, medium and high doses of TASAES at 4.9, 9.8 mg/kg and 19.6 mg/kg body weight by gavage, respectively, for eight weeks. Cardiac function was evaluated by in situ hemodynamic measurements, and patch clamp for the L-type Ca²⁺ channel current (I_Ca²⁺-L) and transient outward K⁺ channel current (I_to). Histopathological changes were observed under light and electron microscope. The expression of pro-fibrotic factor, connective tissue growth factor (CTGF) was monitored using immunohistochemistry staining. Compared with diabetic group, medium and high doses, but not low dose, of TASAES showed a significant protection against diabetes-induced cardiac dysfunction, shown by increased absolute value of left ventricular systolic pressure (LVSP) and maximum rates of pressure development (±dp/dt_max), and enhanced amplitude of I_Ca²⁺-L (P < 0.05). Histological staining indicated a significant inhibition of diabetes-caused pathological changes and up-regulation of CTGF expression (P < 0.05). The results suggest that TASAES prevents diabetes-induced cardiac dysfunction and pathological damage through up-regulating I_Ca²⁺-L in cardiac cells and decreasing CTGF expression.     

Effect of the Reaction Temperature on the Removal of Diesel Particulate Matter by Ozone Injection

This study seeks to investigate the removal efficiency of particulate matter (PM) from the actual diesel exhaust at various reaction temperatures by using non-thermal plasma (NTP). The effect of the reaction temperature on removal efficiency was reflected by the change in the concentration of particles in different modes and the weight fraction of volatile organics in PM. The Arrhenius equation was used to determine the apparent activation energies E_a of the soot in PM. In addition, the difference in the oxidation reaction at various reaction temperatures and the effect of NTP on the properties of PM were discussed. After considering the decreasing ranges of the total concentration and the weight of the PM, it was determined that 120 °C is the optimal temperature choice for PM removal. The decreasing range of the total concentration reached 57.13% and 66.79% of PM was removed when the PM is measured by weight. NTP has better effect on the removal of smaller particles. The weight fraction of the volatile fraction markedly decreases after the reaction and the apparent activation energy of soot noticeably decreased. The oxidizability of the excited species in NTP was enhanced with the increase of the reaction temperature. However, the excited species concentration declined concurrently, resulting in the occurrence of the optimized range of reaction temperature. The particles were removed by the oxidation that occurred on the surface of the primary particle and the disintegration of the structure of the particles.

     

Determination of cocaine on banknotes through an aptamer-based electrochemiluminescence biosensor

A novel electrochemiluminescence (ECL) “sandwich” biosensor has been developed to detect cocaine. The sandwich biosensor was fabricated on the basis of the fact that a single aptamer could be split into two fragments and the two dissociated parts could form a folded, associated complex in the presence of targets. One of these (capture probe), which had hexane–thiol at its 5′-terminus, was immobilized on a gold electrode via thiol–gold binding. The other one (detection probe) was labeled with the ECL reagent tris(2,2′-bipyridyl)ruthenium(II)-doped silica nanoparticles (RuSiNPs) at its 3′-terminus. Owing to the weak interaction between the two fragments, the sensor exhibited a low ECL signal in the absence of cocaine. After the target cocaine had been added to the solution, it induced association of the two fragments and stabilized the associated complexes, leading to immobilization of RuSiNPs on the electrode surface, and the ECL detected on the electrode surface was enhanced. The enhanced ECL intensity was directly proportional to the logarithm of the cocaine concentration in the range from 1.0 × 10⁻⁹ to1.0 × 10⁻¹¹ mol/L, with a detection limit of 3.7 × 10⁻¹² mol/L. The biosensor was applied to detect trace amounts of cocaine on banknotes with satisfactory results.     

Anisotropic wetting on checkerboard-patterned surfaces

A series of surfaces with microscale checkerboard patterns consisting of continuous central lines and discontinuous lateral lines were fabricated. The surface wetting properties of these checkerboard patterns were found to be anisotropic. The central continuous lines were found to have a strong influence on the dynamic wetting properties and moving trajectories of the water droplets. The droplets move more easily in the direction parallel to the central continuous lines and less easily in the direction perpendicular to the central continuous lines. Meanwhile, the droplets' moving path tends to incline toward the central continuous lines from a tilting direction. When the microsurface was modified with a layer of nanowire, the surface wettability was found to be isotropic and superhydrophobic.     

Synthesis and characterization of a 'fluorous' (fluorinated alkyl) affinity reagent that labels primary amine groups in proteins/peptides

Strong non-covalent interactions such as biotin-avidin affinity play critical roles in protein/peptide purification. A new type of 'fluorous' (fluorinated alkyl) affinity approach has gained popularity due especially to its low level of non-specific binding to proteins/peptides. We have developed a novel water-soluble fluorous labeling reagent that is reactive (via an active sulfo-N-hydroxylsuccinimidyl ester group) to primary amine groups in proteins/peptides. After fluorous affinity purification, the bulky fluorous tag moiety and the long oligoethylene glycol (OEG) spacer of this labeling reagent can be trimmed via the cleavage of an acid labile linker. Upon collision-induced dissociation, the labeled peptide ion yields a characteristic fragment that can be retrieved from the residual portion of the fluorous affinity tag, and this fragment ion can serve as a marker to indicate that the relevant peptide has been successfully labeled. As a proof of principle, the newly synthesized fluorous labeling reagent was evaluated for peptide/protein labeling ability in phosphate-buffered saline (PBS). Results show that both the aqueous environment protein/peptide labeling and the affinity enrichment/separation process were highly efficient.     

Towards a fully synthetic MUC1-based anticancer vaccine: efficient conjugation of glycopeptides with mono-, di-, and tetravalent lipopeptides using click chemistry

The membrane‐bound tumor‐associated glycoprotein MUC1 is aberrantly glycosylated in cancer cells compared with normal cells, and is therefore considered an attractive target for cancer immunotherapy. However, tumor‐associated glycopeptides from MUC1 do not elicit a sufficiently robust immune response. Therefore, antitumor vaccines were developed, which consist of MUC1 glycopeptides as the B epitopes and immune‐stimulating toll‐like receptor 2 (TLR 2) lipopeptide ligands. These fully synthetic vaccine candidates were prepared by solid‐phase synthesis of the MUC1 glycopeptides. The Pam₃Cys lipopeptide, also synthesized on solid‐phase, was C‐terminally coupled to oligovalent lysine cores, which N‐terminally incorporate O‐propargyl oligoethylene glycol acyl side chains. The MUC1 glycopeptides and lipopeptide lysine constructs were then conjugated by click chemistry to give oligovalent synthetic vaccines. Oligovalent glycopeptide–lipopeptide conjugates are considered more immunogenic than their monovalent analogues.     

Ultra-high capacity liquid chromatography chip/quadrupole time-of-flight mass spectrometry for pharmaceutical analysis

Nanoflow liquid chromatography/mass spectrometry (nano-LC/MS) has attracted increasing interest in virtue of high sensitivity, low sample consumption, and minimal matrix effect. In this work a HPLC-Chip/quadrupole time-of-flight (Q-TOF) MS device with a new ultra-high capacity small molecule chip (UHC-Chip) which features a 500 nL enrichment column and a 150 mm × 75 μm analytical column, was evaluated with a drug mixture covering a wide range of polarities. Excellent chromatographic precision with 0.1-0.5% RSD for retention time and 1.7-9.0% RSD for peak area, low limit of detection, good chip-to-chip reproducibility and linearity were obtained by using this UHC-Chip. Compared with the standard HPLC-Chip with 40 nL trapping column, the UHC-Chip showed higher enrichment capability and hence gave a higher response in signal detection. Additionally, 4-30 times increase in sensitivity was obtained compared with conventional LC/MS, which indicated that UHC-Chip/MS was a valuable tool for the quantitative analysis of low level impurities and degradation products in pharmaceuticals. Moreover, satisfactory results obtained from trace drug analysis of serum samples further proved its practicality and potential for use in drug testing and development.     

Biomimetic anchors for antifouling and antibacterial polymer brushes on stainless steel

Barnacle cement (BC) was beneficially applied on stainless steel (SS) to serve as the initiator anchor for surface-initiated polymerization. The amine and hydroxyl moieties of barnacle cement reacted with 2-bromoisobutyryl bromide to provide the alkyl halide initiator for the surface-initiated atom transfer radical polymerization (ATRP) of 2-hydroxyethyl methacrylate (HEMA). The hydroxyl groups of HEMA polymer (PHEMA) were then converted to carboxyl groups for coupling of chitosan (CS) to impart the SS surface with both antifouling and antibacterial properties. The surface-functionalized SS reduced bovine serum albumin adsorption, bacterial adhesion, and exhibited antibacterial efficacy against Escherichia coli (E. coli). The effectiveness of barnacle cement as an initiator anchor was compared to that of dopamine, a marine mussel inspired biomimetic anchor previously used in surface-initiated polymerization. The results indicate that the barnacle cement is a stable and effective anchor for functional surface coatings and polymer brushes.