Attachment of Single-Stranded DNA to Certain SERS-Active Gold and Silver Substrates: Selected Practical Tips.

Layers formed from single-stranded DNA on nanostructured plasmonic metals can be applied as "working elements" in surface-enhanced Raman scattering (SERS) sensors used to sensitively and accurately identify specific DNA fragments in various biological samples (for example, in samples of blood). Therefore, the proper formation of the desired DNA layers on SERS substrates is of great practical importance, and many research groups are working to improve the process in forming such structures. In this work, we propose two modifications of a standard method used for depositing DNA with an attached linking thiol moiety on certain SERS-active structures; the modifications yield DNA layers that generate a stronger SERS signal. We propose: (i) freezing the sample when forming DNA layers on the nanoparticles, and (ii) when forming DNA layers on SERS-active macroscopic silver substrates, using ω-substituted alkanethiols with very short alkane chains (such as cysteamine or mercaptopropionic acid) to backfill the empty spaces on the metal surface unoccupied by DNA. When 6-mercapto-1-hexanol is used to fill the unoccupied places on a silver surface (as in experiments on standard gold substrates), a quick detachment of chemisorbed DNA from the silver surface is observed. Whereas, using ω-substituted alkanethiols with a shorter alkane chain makes it possible to easily form mixed DNA/backfilling thiol monolayers. Probably, the significantly lower desorption rate of the thiolated DNA induced by alkanethiols with shorter chains is due to the lower stabilization energy in monolayers formed from such compounds.

Host-Seeking Behavior of Aphidius gifuensis (Hymenoptera: Braconidae) Modulated by Chemical Cues Within a Tritrophic Context.

Aphidius gifuensis Ashmaed is a generalist endoparasitoid that parasitizes a variety of aphid species. In China, it is widely used as a biological control agent to protect vegetables and tobaccos in open fields; control efficiency is largely dependent on its host-seeking ability. In this study, a six-choice olfactometer was used to investigate the olfactory responses of A. gifuensis to tobacco plants that had suffered damage (either varying degrees of mechanical damage or from aphid-feeding at different time intervals) and tobacco volatiles with different dosages. Furthermore, the regularity of A. gifuensis females' response toward an aphid/tobacco complex was monitored using a Y-tube olfactometer. Our findings suggest that tobacco plants are significantly attractive to A. gifuensis after they have been punctured with 50 holes, or housed with Myzus persicae (Sulzer) at a density of 400 aphids, except at an infestation time of 12 h. Moreover, aphid density had a more significant effect on the response than the time interval since aphid application. Aphidius gifuensis was found to be active during the daytime and preferred to search for their aphid hosts at 14:00 h. Five EAG-active tobacco volatiles (trans-2-hexenal, methyl salicylate, benzaldehyde, cis-3-hexen-1-ol, and 1-hexanal) were found to significantly attract A. gifuensis females at different concentration ranges. The practical implications of these results are discussed in the framework of the sustainable biological control of pest aphids in agricultural production systems.

Sensory, olfactometric and chemical characterization of the aroma potential of Garnacha and Tempranillo winemaking grapes.

Reconstituted polyphenolic and aromatic fractions (PAFs) from 33 different Garnacha and Tempranillo grapes were incubated in strict anoxia (75 °C × 24 h). Obtained hydrolyzates were characterized by sensory analysis, gas chromatography-olfactometry (GC-O) and gas chromatography-mass spectrometry (GC-MS). Five different aroma categories emerged. Garnacha may develop specific tropical/citrus fruit, kerosene and floral and Tempranillo toasty-woody and red-fruit characteristics. Those notes seem to mask alcoholic and fruit-in-syrup descriptors and the common vegetal background. Twenty-seven odorants were detected by GC-O. GC-MS data showed a clustering closely matching the one found by sensory analysis, suggesting the existence of five specific metabolomic profiles behind the five specific sensory profiles. Overall results suggest that 3-mercaptohexanol is responsible for tropical/citrus fruit, TDN for kerosene, volatile phenols for woody/toasty, ß-damascenone and massoia lactone, likely with Z-1,5-octadien-3-one for fruit-in-syrup and alcoholic notes. Nine lipid-derived unsaturated aldehydes and ketones may be responsible for the vegetal background.

Label-Free Detection of Zeptomol miRNA via Peptide Nucleic Acid Hybridization Using Novel Cyclic Voltammetry Method.

To harness the applicability of microribonucleic acid (miRNA) as a cancer biomarker, the detection sensitivity of serum miRNA needs to be improved. This study evaluated the detection sensitivity of miRNA hybridization using cyclic voltammograms (CVs) and microelectrode array chips modified with peptide nucleic acid (PNA) probes and 6-hydroxy-1-hexanethiol. We investigated the PNA probe modification pattern on array chips using fluorescently labeled cDNA. The pattern was not uniformly spread over the working electrode (WE) and had a one-dimensional swirl-like pattern. Accordingly, we established a new ion-channel sensor model wherein the WE is negatively biased through the conductive π-π stacks of the PNA/DNA duplexes. This paper discusses the mechanism underlying the voltage shift in the CV curves based on the electric double-layer capacitance. Additionally, the novel hybridization evaluation parameter ΔE is introduced. Compared to conventional evaluation using oxidation current changes, ΔE was more sensitive. Using ΔE and a new hybridization system for ultrasmall amounts of aqueous solutions (as low as 35 pL), 140 zeptomol label-free miRNA were detected without polymerase chain reaction (PCR) amplification at an adequate sensitivity. Herein, the differences in the target molar amount and molar concentration are elucidated from the viewpoint of hybridization sensitivity.

Aptamer based ratiometric electrochemical sensing of 17ß-estradiol using an electrode modified with gold nanoparticles, thionine, and multiwalled carbon nanotubes.

Gold nanoparticles (AuNPs) with a size of ~3 nm were placed on a thionine-multiwalled carbon nanotube (Thi-CNTs) conjugate to form a novel AuNP-Thi-CNTs nanocomposite. Its morphology and composition were characterized by transmission electron microscopy, atomic force microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The nanocomposite was placed on an electrode and used as a redox-active signaling interface to fabricate a ratiometric electrochemical aptasensor for 17ß-estradiol (E2). The potentiostatic insertion method was applied to insert an aptamer against E2 into a thin alkane monolayer to warrant an adequate distance between aptamers. The aptamer against E2 acts as both a collector and separator to specifically bind E2. The electrode displays two peak signals (at +0.50 V vs. SCE for E2; and at -0.32 V for Thi) which increase and decrease, respectively, in the 12 pM to 60 nM E2 concentration range. Therefore, the current ratio can be used to reliably, reproducibly, and sensitively quantify the concentration of E2. Graphical abstract Schematic presentation of a novel AuNP-Thi-CNTs nanocomposite. AuNP-Thi-CNTs showed good electrocatalytic oxidation to E2. AuNP-Thi-CNTs was used as self-redox signal interface to fabricate aptasensor. Dual signals of extrinsic E2 and inner Thi was applied to monitor the concentration of E2.

Characterisation of the Convective Hot-Air Drying and Vacuum Microwave Drying of Cassia alata: Antioxidant Activity, Essential Oil Volatile Composition and Quality Studies.

The preservation of active constituents in Cassia alata through the removal of moisture is crucial in producing a final product with high antioxidant activity. This study aims to determine the influences of various drying methods and drying conditions on the antioxidant activity, volatiles and phytosterols content of C. alata. The drying methods used were convective drying (CD) at 40 °C, 50 °C and 60 °C; freeze drying; vacuum microwave drying (VMD) at 6, 9 and 12 W/g; and two-stage convective pre-drying followed by vacuum microwave finish drying (CPD-VMFD) at 50 °C and 9 W/g. The drying kinetics of C. alata are best described by the thin-layer model (modified Page model). The highest antioxidant activity, TPC and volatile concentration were achieved with CD at 40 °C. GC-MS analysis identified the presence of 51 volatiles, which were mostly present in all samples but with quantitative variation. The dominant volatiles in fresh C. alata are 2-hexenal (60.28 mg 100 g-1 db), 1-hexanol (18.70 mg 100 g-1 db) and salicylic acid (15.05 mg 100 g-1 db). The concentration of phytosterols in fresh sample was 3647.48 mg 100 g-1 db, and the major phytosterols present in fresh and dried samples were ß-sitosterol (1162.24 mg 100 g-1 db). CPD-VMFD was effective in ensuring the preservation of higher phytosterol content in comparison with CD at 50 °C. The final recommendation of a suitable drying method to dehydrate C. alata leaves is CD at 40 °C.

High-Throughput/High-Precision Sampling of Single Cells into ICP-MS for Elucidating Cellular Nanoparticles.

In single-cell analysis with ICP-MS it is highly important to ensure precise single-cell sampling into ICP. For this purpose, a simple configured pressure-resistant MicroCross interface is developed for high-throughput/high-precision microdroplet generation and single-cell encapsulation. Aqueous cell suspension is ejected and sheared into droplets by tangent-flowing hexanol-continuous phases in the flow-focusing geometry of MicroCross, wherein to precisely trap a single cell into a droplet, with an extremely low probability of <0.005% for a single droplet encapsulating two cells. MicroCross interface is coupled with time-resolved ICP-MS (TRA-ICP-MS) for quantifying nanoparticles in single MCF-7 cells. At the optimal conditions, sufficient temporal-spatial resolution of the microdroplets is achieved facilitating high-throughput sampling of single cells into ICP. For solving the serious carbon deposition on the sampling cone and the unstable plasma torch caused by incomplete oxidation of hexanol phase in ICP, dimethyl carbonate (DMC) is for the first time used as a superb oxygen compensation reagent, which ensures adequate oxidation of hexanol, effectively eliminates the carbon deposition, and maintains a stable plasma. The single-cell analysis results indicated a remarkable discrepancy of the number of nanoparticles among the individual cells, falling into a range of 130-584 per MCF-7 cell in the case of AuNPs.

Structural basis of malodour precursor transport in the human axilla.

Mammals produce volatile odours that convey different types of societal information. In Homo sapiens, this is now recognised as body odour, a key chemical component of which is the sulphurous thioalcohol, 3-methyl-3-sulfanylhexan-1-ol (3M3SH). Volatile 3M3SH is produced in the underarm as a result of specific microbial activity, which act on the odourless dipeptide-containing malodour precursor molecule, S-Cys-Gly-3M3SH, secreted in the axilla (underarm) during colonisation. The mechanism by which these bacteria recognise S-Cys-Gly-3M3SH and produce body odour is still poorly understood. Here we report the structural and biochemical basis of bacterial transport of S-Cys-Gly-3M3SH by Staphylococcus hominis, which is converted to the sulphurous thioalcohol component 3M3SH in the bacterial cytoplasm, before being released into the environment. Knowledge of the molecular basis of precursor transport, essential for body odour formation, provides a novel opportunity to design specific inhibitors of malodour production in humans.

Fluorescence method for quickly detecting ochratoxin A in flour and beer using nitrogen doped carbon dots and silver nanoparticles.

In this paper, a FRET (Forster resonance energy transfer) based fluorescence method was developed for the quickly detection of ochratoxin A (OTA) in agricultural products (e.g., flour and beer). A highly fluorescent nitrogen doped carbon dots (CD) were served as energy donor, the DNA and MCH (6-mercapto-1-hexanol) modified Ag nanoparticles were served as energy acceptor in the FRET system. OTA can be detected in a concentration range between 10 and 5000 nM, the limit of detection is 8.7 nM. This method has three advantages: (1) an enhanced fluorescent intensity can be acquired by utilizing the nitrogen doped CD synthesized by one-step approach without sophisticated modification of nanoparticles; (2) OTA detection was accomplished quickly (less than 30 min) by using MCH as assistant molecule; (3) an extended OTA detection linear range was acquired, which may facilitate the OTA detection in real agricultural samples, and is helpful for solving food safety problems.

Molecular basis for potentiation of Cx36 gap junction channel conductance by n-alcohols and general anesthetics.

In our recent study, we have demonstrated that short carbon chain n-alcohols (up to octanol) stimulated while long carbon chain n-alcohols inhibited the conductance of connexin (Cx) 36 (Cx36) gap junction (GJ) channels. In contrast, GJ channels composed of other types of Cxs all were inhibited by n-alcohols independent of their carbon chain length. To identify the putative structural domains of Cx36, responsible for the dual effect of n-alcohols, we performed structural modeling of Cx36 protein docking with hexanol and isoflurane that stimulated as well as nonanol and carbenoxolone that inhibited the conductance of Cx36 GJs and revealed their multiple common docking sites and a single pocket accessible only to hexanol and isoflurane. The pocket is located in the vicinity of three unique cysteine residues, namely C264 in the fourth, and C92 and C87 in the second transmembrane domain of the neighboring Cx36 subunits. To examine the hypothesis that disulphide bonding might be involved in the stimulatory effect of hexanol and isoflurane, we generated cysteine substitutions in Cx36 and demonstrated by a dual whole-cell patch-clamp technique that in HeLa (human cervix carcinoma cell line) and N2A (mouse neuroblastoma cell line) cells these mutations reversed the stimulatory effect of hexanol and isoflurane to inhibitory one, typical of other Cxs that lack respective cysteines and a specific docking pocket for these compounds. Our findings suggest that the stimulatory effect of hexanol and isoflurane on Cx36 GJ conductance could be achieved by re-shuffling of the inter-subunit disulphide bond between C264 and C92 to the intra-subunit one between C264 and C87.

6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio) hexanol: a promising new anticancer compound.

The 7-nitro-2,1,3-nitrobenzoxadiazole (NBD) derivatives are a series of compounds containing the NBD scaffold that are not glutathione (GSH) peptidomimetics, and result in a strong inhibition of glutathione S-transferases (GSTs). Growing evidences highlight their pivotal roles and outstanding anticancer activity in different tumor models. In particular, 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio) hexanol (NBDHEX) is extensively studied, which is a very efficient inhibitor of GSTP1-1. It triggers apoptosis in several tumor cell lines and this cytotoxic activity is observed at micro and submicromolar concentrations. Importantly, studies have shown that NBDHEX acts as an anticancer drug by inhibiting GSTs catalytic activity, avoiding inconvenience of the inhibitor extrusion from the cell by specific pumps and disrupting the interaction between the GSTP1-1 and key signaling effectors. Additionally, some researchers also have discovered that NBDHEX can act as late-phase autophagy inhibitor, which opens new opportunities to fully exploit its therapeutic potential. In this review, we summarize the advantages, anticancer mechanisms, and analogs of this compound, which will establish the basis on the usage of NBDHEX in clinical applications in future.

Naturally Produced Defensive Alkenal Compounds Activate TRPA1.

(E)-2-alkenals are aldehydes containing an unsaturated bond between the alpha and beta carbons. 2-alkenals are produced by many organisms for defense against predators and secretions containing (E)-2-alkenals cause predators to stop attacking and allow the prey to escape. Chemical ecologists have described many alkenal compounds with 3-20 carbons common, having varied positions of double bonds and substitutions. How do these defensive alkenals act to deter predators? We have tested the effects of (E)-2-alkenals with 6-12 carbons on transient receptor potential channels (TRP) commonly found in sensory neurons. We find that (E)-2-alkenals activate transient receptor potential ankyrin subtype 1 (TRPA1) at low concentrations-EC50s 10-100 µM (in 0 added Ca(2+) external solutions). Other TRP channels were either weakly activated (TRPV1, TRPV3) or insensitive (TRPV2, TRPV4, TRPM8). (E)-2-alkenals may activate TRPA1 by modifying cysteine side chains. However, target cysteines include others beyond the 3 in the amino-terminus implicated in activation, as a channel with cysteines at 621, 641, 665 mutated to serine responded robustly. Related chemicals, including the aldehydes hexanal and decanal, and (E)-2-hexen-1-ol also activated TRPA1, but with weaker potency. Rat trigeminal nerve recordings and behavioral experiments showed (E)-2-hexenal was aversive. Our results suggest that TRPA1 is likely a major target of these commonly used defensive chemicals.

Chemical composition and biological evaluation of the volatile constituents from the aerial parts of Nephrolepis exaltata (L.) and Nephrolepis cordifolia (L.) C. Presl grown in Egypt.

The essential oil from the aerial parts of Nephrolepis exaltata and Nephrolepis cordifolia obtained by hydro-distillation were analyzed by gas chromatography/ mass spectrometry. The essential oils exhibited potential antibacterial and antifungal activities against a majority of the selected microorganisms. NEA oil showed promising cytotoxicity in breast, colon and lung carcinoma cells. The results presented indicate that NEA oil could be useful alternative for the treatment of dermatophytosis. Comparative investigation of hydro-distilled volatile constituents from aerial parts (A) of Nephrolepis exaltata (NE) and Nephrolepis cordifolia (NC) (Family Nephrolepidaceae) was carried out. Gas chromatography/mass spectrometry revealed that oils differ in composition and percentages of components. Oxygenated compounds were dominant in NEA and NCA. 2,4-Hexadien-1-ol (16.1%), nonanal (14.4%), ß-Ionone (6.7%) and thymol (2.7%) were predominant in NEA. ß-Ionone (8.0%), eugenol (7.2%) and anethol (4.6%) were the main constituents in NCA. Volatile samples were screened for their antibacterial and antifungal activities using agar diffusion method and minimum inhibitory concentrations. The cytotoxic activity was evaluated using viability assay in breast (MCF-7), colon (HCT-116) and lung carcinoma (A-549) cells by the MTT assay. The results revealed that NEA oil exhibited potential antimicrobial activity against most of the tested organisms and showed promising cytotoxicity.

Chemical composition and antibacterial activity of the essential oil from Pyrrosia tonkinensis (Giesenhagen) Ching.

The present study aimed to analyse the chemical components of the essential oil from Pyrrosia tonkinensis by GC-MS and evaluate the in vitro antibacterial activity. Twenty-eight compounds, representing 88.1% of the total essential oil, were identified and the major volatile components were trans-2-hexenal (22.1%), followed by nonanal (12.8%), limonene (9.6%), phytol (8.4%), 1-hexanol (3.8%), 2-furancarboxaldehyde (3.5%) and heptanal (3.1%). The antibacterial assays showed that the essential oil of P. tonkinensis had good antibacterial activities against all the tested microorganisms. This paper first reported the chemical composition and antimicrobial activity of the essential oil from P. tonkinensis.

Protein Synthesis in Sub-Micrometer Water-in-Oil Droplets.

Water-in-oil (w/o) emulsions are used as a cellular model because of their unique cell-like architecture. Previous works showed the capability of eukaryotic-cell-sized w/o droplets (5-50 µm) to support protein synthesis efficiently; however data about smaller w/o compartments (<1 µm) are lacking. This work focuses on the biosynthesis of the enhanced green fluorescent protein (EGFP) inside sub-micrometric lecithin-based w/o droplets (0.8-1 µm) and on its dependence on the compartments' dynamic properties in terms of solute exchange mechanisms. We demonstrated that protein synthesis is strongly affected by the nature of the lipid interface. These findings could be of value and interest for both basic and applied research.

Ratiometric electrochemical sensor for selective monitoring of cadmium ions using biomolecular recognition.

A selective, accurate, and sensitive method for monitoring of cadmium ions (Cd(2+)) based on a ratiometric electrochemical sensor was developed, by simultaneously modifying with protoporphyrin IX and 6-(ferroceney) hexanethiol (FcHT) on Au particle-deposited glassy carbon electrode. On the basis of high affinity of biomolecular recognition between protoporphyrin IX and Cd(2+), the functionalized electrode showed high selectivity toward Cd(2+) over other metal ions such as Cu(2+), Fe(3+), Ca(2+), and so on. Electroactive FcHT played the role as the inner reference element to provide a built-in correction, thus improving the accuracy for determination of Cd(2+) in the complicated environments. The sensitivity of the electrochemical sensor for Cd(2+) was enhanced by ∼3-fold through the signal amplification of electrodeposited gold nanoparticles. Accordingly, the present ratiometric method demonstrated high sensitivity, broad linear range from 100 nM to 10 µM, and low detection limit down to 10 nM (2.2 ppb), lower than EPA and WHO guidelines. Finally, the ratiometric electrochemical sensor was successfully applied in the determination of Cd(2+) in water samples, and the obtained results agreed well with those obtained by the conventional ICP-MS method.

High specific detection of osteopontin using a three-dimensional copolymer layer support based on electrochemical impedance spectroscopy.

Tumor marker detection is essential for the therapy efficiency of early stage tumors and the evaluation of disease progression. Osteopontin (OPN) is supposed to be closely related to several kinds of tumors. In the present study, we describe a label-free electrochemical detection of OPN based on a specific reaction between OPN and its relevant antibody. An artificial three-dimensional (3D) scaffold structure consisting of 11-mercaptoundecanoic acid/6-mercapto-1-hexanol, dextran amino and synthetic peptides was designed as a substrate for the immobilization of the antibody. This substrate was characterized using cyclic voltammetry, atomic force microscopy and Fourier transform infrared reflection spectroscopy. Antibody immobilization and OPN detection were conducted using electrochemical impedance spectroscopy (EIS). The low limit of detection was 0.17 nM. The concentration of cancer risk (5.77 nM) can be selectively detected with a high EIS signal. The fabricated 3D OPN sensor is proposed for application in clinical analysis.

Transesterification reaction for synthesis of palm-based ethylhexyl ester and formulation as base oil for synthetic drilling fluid.

The use of vegetable oil-based ester as a base fluid in synthetic drilling fluid has become a trend in drilling operations due to its environmental advantages. The transesterification reaction of palm oil methyl ester (POME) with 2-ethylhexanol (2EH) produced 98% of palm oil-based ethylhexyl ester in less than 30 minutes. Since the transesterification reaction of POME with 2EH is a reversible reaction, its kinetics was studied in the presence of excess EH and under vacuum. The POME-to-EH molar ratio and vacuum pressure were held constant at 1:2 and 1.5 mbar respectively and the effects of temperature (70 to 110°C) were investigated. Using excess of EH and continual withdrawal of methanol via vacuum promoted the reaction to complete in less than 10 minutes. The rate constant of the reaction (k) obtained from the kinetics study was in the range of 0.44 to 0.66 s⁻¹ and the activation energy was 15.6 kJ.mol⁻¹. The preliminary investigations on the lubrication properties of drilling mud formulated with palm oil-based 2EH ester indicated that the base oil has a great potential to substitute the synthetic ester-based oil for drilling fluid. Its high kinematic viscosity provides better lubrication to the drilling fluid compared to other ester-based oils. The pour point (-15°C) and flash point (204°C) values are superior for the drilling fluid formulation. The plastic viscosity, HPHT filtrate loss and emulsion stability of the drilling fluid had given acceptable values, while gel strength and yield point could be improved by blending it with proper additives.

What determines human body odour?

Human body odour and earwax type are genetically dependent on a single-nucleotide polymorphism (SNP) located in the ABCC11 gene. So far, it still remains to be clear how SNP in the ABCC11 gene is associated with human malodour. In a recent issue of Experimental Dermatology, Baumann et al. propose one of the underlying molecular pathways. Although one of the amino acid conjugated of the odorants, Cys-Gly-3-methyl-3-sulfanylhexanol (3M3SH), was not taken up by the transporter ABCC11, glutathione conjugate of 3MSH (SG-3MSH) was transported by ABCC11. Moreover, SG-3MSH was processed to 3M3SH by γ-glutamyl-transferase 1 (GGT1), which was abundantly expressed in apocrine sweat glands. These findings may pave a way for the pharmacogenetics of human body odour and the development of innovative deodorant products.

Derivatization of pinacolyl alcohol with phenyldimethylchlorosilane for enhanced detection by gas chromatography-mass spectrometry.

A derivatization procedure for the qualitative gas chromatography-mass spectrometry (GC-MS) analysis of pinacolyl alcohol (PA) that employs phenyldimethylchlorosilane (PhDMClS) and the promoter N-methylimidazole is described. While PA, underivatized, can be detected using conventional gas chromatographic methods, its polarity and low boiling point make its detection in complex matrices challenging. The silylation procedure described herein generates a PA-derivative exhibiting an increased on-column retention time, thus shifting its GC-MS signal away from commonly encountered, volatile, interfering analytes. Derivatized PA could be distinguished from other PhDMClS-derivatized isomeric alcohols by its unique retention time and mass spectrum. The derivatization was demonstrated to perform well in the GC-MS analysis and identification of PA in samples from Proficiency Tests administered by the Organisation for the Prohibition of Chemical Weapons (OPCW).