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Hydrogen sulfide (H 2 S) is a gaseous signaling molecule that regulates various physiological and pathological processes in the central nervous system. It is vital to develop an effective method to detect H 2 S in vivo to elucidate its critical role. However, current fluorescent probes for accurate quantification of H 2 S still face big challenges due to complicated fabrication, small Stokes shift, unsatisfactory selectivity, and especially delayed response time. Herein, based on simple postsynthetic modification, we present an innovative strategy by confining H 2 S-triggered thiolysis of dinitrophenyl (DNP) ether within a luminescent metal-organic framework (MOF) to address those issues. Due to the cleavage of the DNP moiety by H 2 S, the nanoprobe gives rise to a remarkable fluorescence turn-on signal with a large Stokes shift of 190 nm and also provides high selectivity to H 2 S against various interferents including competing biothiols. In particular, by virtue of the unique structural property of the MOF, it exhibits an ultrafast sensing ability for H 2 S (only 5 s). Moreover, the fluorescence enhancement efficiency displays a good linear correlation with H 2 S concentration in the range of 0-160 μM with a detection limit of 0.29 μM. Importantly, these superior sensing performances enable the nanoprobe to measure the basal value and monitor the change of H 2 S level in the rat brain.

Military activities and the production or disposal of ammunition often lead to soil contamination with energetic compounds (ECs) such as dinitrotoluene, trinitrotoluene, and hexogen, posing significant threats to human health and the ecosystem. Biochar has emerged as a cost-effective and widely available solution for remediating contaminated sites characterized by its capacity for pollutant removal through adsorption and conversion process, along with minimal secondary pollution. This paper provides a comprehensive review of relevant literature on biochar's efficacy in eliminating ECs, including an analysis of the underlying mechanisms. The discussion addresses challenges and opportunities associated with biochar application in ECs remediation, offering insights for future research directions. In summary, the use of biochar for ECs removal presents a promising and eco-friendly approach, facilitating the remediation of contaminated sites while promoting soil function and ecosystem recovery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Innovations in computer hardware and software capabilities have paved the way for advances in molecular modelling techniques and methods, leading to an unprecedented expansion of their potential applications. In contrast to the docking technique, which usually identifies the most stable selector-selectand (SO-SA) complex for each enantiomer, the molecular dynamics (MD) technique enables the consideration of a distribution of the SO-SA complexes based on their energy profile. This approach provides a more truthful representation of the processes occurring within the column. However, benchmark procedures and focused guidelines for computational treatment of enantioselectivity at the molecular level are still missing., Results: Twenty-eight molecular dynamics simulations were performed to study the enantiorecognition mechanisms of seven N-3,5-dinitrobenzoylated α- and β-amino acids (DNB-AAs), occurring with the two quinine- and quinidine-based (QN-AX and QD-AX) chiral stationary phases (CSPs), under polar-ionic conditions. The MD protocol was optimized in terms of box size, simulation run time, and frame recording frequency. Subsequently, all the trajectories were analyzed by calculating both the type and amount of the interactions engaged by the selectands (SAs) with the two chiral selectors (SOs), as well as the conformational and interaction energy profiles of the formed SA-SO associates. All the MDs were in strict agreement with the experimental enantiomeric elution order and allowed to establish (i) that salt-bridge and H-bond interactions play a pivotal role in the enantiorecognition mechanisms, and (ii) that the π-cation and π-π interactions are the discriminant chemical features between the two SOs in ruling the chiral recognition mechanism., Significance: The results of this work clearly demonstrate the high contribution given by MD simulations in the comprehension of the enantiorecognition mechanism with Cinchona alkaloid-based CSPs. However, from this research endeavor it clearly emerged that the MD protocol optimization is crucial for the quality of the produced results., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The microbial transcription factor YhaJ responds to 2,4-dinitrotoluene (DNT) derivatives. Here, we describe steps for overexpression and purification of the protein, characterization for the binding of a DNT derivative methylhydroquinone, and crystallization by using a random seeding technique. We then detail procedures for structure determination by employing the crystal-twin resolving processes. This protocol can also be performed using other DNT derivatives. For complete details on the use and execution of this protocol, please refer to Kim et al. 1 ., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Oryzalin (ORY) is a dinitroaniline derivative that inhibits the microtubule polymerization in plants and parasitic protozoa by selectively binding to the α-tubulin subunit. This herbicidal agent exhibits good antiprotozoal activity against major human parasites, such as Toxoplasma gondii (toxoplasmosis), Leishmania mexicana (leishmaniasis), and Plasmodium falciparum (malaria). Previous chemical mutagenesis assays on T. gondii α-tubulin (TgAT) have identified key mutations that lead to ORY resistance. Herein, we employed alchemical free energy methods and molecular dynamics simulations to determine if the ORY resistance mutations either decrease the TgAT's affinity of the compound or increase the protein stability. Our results here suggest that L136F and V202F mutations significantly decrease the affinity of ORY to TgAT, while T239I and V252L mutations diminish TgAT's flexibility. On the other hand, protein stability predictors determined that R243S mutation reduces TgAT stability due to the loss of its salt bridge interaction with E27. Interestingly, molecular dynamics simulations confirm that the loss of this key interaction leads to ORY binding site closure. Our study provides a better insight into the TgAT-ORY interaction, further supporting our recently proposed ORY-binding site., (Copyright © 2022 Elsevier Inc. All rights reserved.)

We have previously developed a universal chimeric antigen receptor (CAR), which recognizes dinitrophenyl (DNP) and can redirect T and NK cells to target cancer and HIV antigens using DNP-conjugated antibodies as adaptor molecules. However, the DNP-antibody conjugates are generated by random modification, which may not be optimal for this modular system. Here, we report the development of enhanced adaptor molecules by site-specific DNP modification. We use the genetic code expansion technology to generate single-chain fragment variable (scFv) antibodies with site-specific DNP. We compare four anti-CD19 scFv mutants and find that the one with DNP at the flexible peptide linker between V L and V H is the most effective in redirecting anti-DNP CAR-T cells against CD19 + cells. The other three mutants are ineffective in doing so due to reduced DNP exposure or abrogated CD19 binding. We also use the anti-CD22 scFv as another model adaptor molecule and again find that the peptide linker is ideal for DNP derivatization. Our approach can potentially be used to design enhanced adaptor molecules to redirect the DNP-mediated universal CAR against other tumor antigens., Competing Interests: Declaration of interest The authors declare no competing financial interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Nitroreductases are emerging as attractive bioremediation enzymes, with substrate promiscuity toward both natural and synthetic compounds. Recently, the nitroreductase NfnB from Sphingopyxis sp. strain HMH exhibited metabolic activity for dinitroaniline herbicides including butralin and pendimethalin, triggering the initial steps of their degradation and detoxification. However, the determinants of the specificity of NfnB for these herbicides are unknown. In this study, we performed structural and biochemical analyses of NfnB to decipher its substrate specificity. The homodimer NfnB is a member of the PnbA subgroup of the nitroreductase family. Each monomer displays a central α + β fold for the core domain, with a protruding middle region and an extended C-terminal region. The protruding middle region of Val75-Tyr129 represents a structural extension that is a common feature to members of the PnbA subgroup and functions as an opening wall connecting the coenzyme FMN-binding site to the surface, therefore serving as a substrate binding site. We performed mutational, kinetic, and structural analyses of mutant enzymes and found that Tyr88 in the middle region plays a pivotal role in substrate specificity by determining the dimensions of the wall opening. The mutation of Tyr88 to phenylalanine or alanine caused significant changes in substrate selectivity toward bulkier dinitroaniline herbicides such as oryzalin and isopropalin without compromising its activity. These results provide a framework to modify the substrate specificity of nitroreductase in the PnbA subgroup, which has been a challenging issue for its biotechnological and bioremediation applications., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The dinitrotoluene isomers 2,4 and 2,6-dinitrotoluene (DNT) represent highly toxic, mutagenic, and carcinogenic compounds used in explosive manufacturing and in commercial production of polyurethane foam. Bioremediation, the use of microbes to degrade residual DNT in industry wastewaters, represents a promising, low cost and environmentally friendly alternative technology to landfilling. In the present study, the effect of different bioremediation strategies on the degradation of DNT in a microcosm-based study was evaluated. Biostimulation of the indigenous microbial community with sulphur phosphate (2.3 g/kg sludge) enhanced DNT transformation (82% transformation, from 300 g/L at Day 0 to 55 g/L in week 6) compared to natural attenuation over the same period at 25 °C. The indigenous microbial activity was found to be capable of transforming the contaminant, with around 70% transformation of DNT occurring over the microcosm study. 16S rDNA sequence analysis revealed that while the original bacterial community was dominated by Gammaproteobacteria (30%), the addition of sulphur phosphate significantly increased the abundance of Betaproteobacteria by the end of the biostimulation treatment, with the bacterial community dominated by Burkholderia (46%) followed by Rhodanobacter , Acidovorax and Pseudomonas . In summary, the results suggest biostimulation as a treatment choice for the remediation of dinitrotoluenes and explosives waste.

Building on clinical case reports of the abscopal effect, there has been considerable interest in the synergistic effects of radiation and immunotherapies for the treatment of cancer. Here, the first radiolabeled antibody-recruiting small molecule that can chelate a variety of cytotoxic radionuclides is described. The platform consists of a tunable antibody-binding domain against a serum antibody of interest (e.g., dinitrophenyl hapten) to recruit endogenous antibodies that activate effector cell function, a chelate capable of binding diagnostic and therapeutic radiometals, and a tetrazine for bioorthogonal coupling with trans -cyclooctene-modified targeting vectors. The dinitrophenyl-tetrazine ligand was shown to both affect dose-dependent antibody recruitment and immune cell function (phagocytosis) in vitro , and the bisphosphonate 177 Lu-complex was shown to accumulate at sites of calcium accretion in vivo , which was achieved using both active and pretargeting strategies.

Hapten-specific endogenous antibodies are naturally occurring antibodies present in human blood. Herein, we investigated a new strategy in which small-molecule haptens were utilized as naturally occurring antibody binders for peptide half-life extension. The glucagon-like peptide 1 receptor agonist exendin 4 was site-specifically functionalized with the dinitrophenyl (DNP) hapten at the C-terminus via sortase A-mediated ligation. The resulting Ex4-DNP conjugates retained GLP-1 receptor activation potency in vitro and had a similar in vivo acute glucose-lowering effect comparable to that of native Ex4. Pharmacokinetic studies and hypoglycemic duration tests demonstrated that the Ex4-DNP conjugates displayed significantly elongated half-lives and improved long-acting antidiabetic activity in the presence of endogenous anti-DNP antibodies. In chronic treatment studies, once-daily administration of optimal conjugate 7 demonstrated more beneficial effects without prominent toxicity compared with Ex4. This strategy provides a new approach and represents an alternative to the well-established peptide-Fc fusion strategy to improve the peptide half-life and the therapeutic efficacy.

Background: Atopic dermatitis is a chronic inflammatory skin disease in humans. Although Olea europaea leaf extract (OLE) and Spirodela polyrhiza extract (SPE) have been used to protect against skin damage, the effects of their combined administration on atopic dermatitis have yet to studied., Purpose: In this study, we evaluated the potential therapeutic effects of an OLE and SPE combination on the progression of atopic dermatitis and the possible mechanisms underlying these effects in 1-chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice., Methods: Atopic dermatitis was induced by topical application of 0.2% w/v DNCB prepared in an olive oil:acetone solution (1:3), and thereafter OLE, SPE and OLE + SPE were administered orally for 5 weeks. We determined atopic dermatitis symptoms, serum IgE levels, and levels of cytokine- and gene expression in the dorsal skin and splenocytes, and performed histological and immune cell subtype analyses. The expression of skin barrier-related proteins (filaggrin, sirtuin 1, and claudin 1) was also evaluated., Results: The OLE + SPE combination significantly ameliorated atopic dermatitis symptoms, including dermatitis scores, and reduced epidermal thickness and infiltration of different inflammatory cells in mice with DNCB-induced atopic dermatitis. It also significantly reduced the number of CD4 + , CD8 + , and CD4 + /CD69 + T cells; immunoglobulin E-producing B cells (CD23 + /B220 + ) in the axillary lymph nodes; CD3 + T-cell eosinophils (chemokine-chemokine receptor 3 + /CD11b + ) in the skin; and CD3 + T cells, immunoglobulin E-producing B cells (CD23 + /B220 + ), and eosinophils in peripheral blood mononuclear cells. Additionally, the experimental combination lowered levels of serum immunoglobulin E and histamine, as well as Th2-mediated cytokines, and interleukin-4, -5, and -13, whereas it increased the levels of Th1-mediated cytokine interferon-γ in splenocytes. Furthermore, the preparation significantly restored expression of the skin barrier-related proteins filaggrin, sirtuin 1, and claudin 1, and also reduced the expression of the inflammatory cytokine interleukin-6 and chemokine-chemokine receptor 3, as well as the pruritus-related cytokine interleukin-31 and interleukin-31 receptor, in atopic dermatitis skin lesions., Conclusion: Taken together, our findings indicate that administration of a combination of OLE and SPE can alleviate atopic dermatitis symptoms by regulating immune balance and skin barrier function and may be an effective therapeutic option for the treatment of atopic dermatitis., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Herein we describe a method for inducing cancer cell death, which relies on the use of a H2O2-responsive glycan metabolic precursor in conjunction with antibody-dependent cellular cytotoxicity (ADCC) or photodynamic therapy (PDT).

Biological thiols including homocysteine (Hcy), cysteine (Cys), hydrogen sulfide (H 2 S) and glutathione (GSH) play crucial roles in various pathological and physiological processes. The development of optical probes for biothiols has been an active research area in recent years. Herein, a new turn-on fluorescence probe (HD-NBD) was designed and synthesized by fusing tetrahydro[5]helicene and 7-nitro-2,1,3-benzoxadiazole (NBD) for simultaneous discrimination of Hcy/Cys, H 2 S and GSH in aqueous solution. This probe is able to show unique absorbance enhancement at 548 nm for H 2 S and additional fluorescence enhancement at 536 nm only for Cys/Hcy, which can be used to discriminate H 2 S, Cys/Hcy and GSH simultaneously. In addition, HD-NBD also shows low background without any self-fluorescence, as well as high selectivity toward common biothiols. The low detection limits of this probe are about 0.15 μM for Hcy with a wide linear range (1-80 μM), 0.36 μM for Cys (linear range: 1-45 μM), 0.79 μM for H 2 S (linear range: 1-80 μM) and 4.44 μM for GSH (linear range: 1-60 μM). Moreover, HD-NBD can identify Hcy/Cys, H 2 S from GSH and other amino acids with high sensitivity and selectivity, therefore it could be used for detecting endogenous and exogenous Hcy/Cys under biological condition., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The formation and radical-directed dissociation of multiple hydrogen-abstracted peptide cations [M + H - mH]· + has been reported using MALDI-ISD with dinitro-substituted matrices. The MALDI-ISD of synthetic peptides using 3,5-dinitrosalicylic acid (3,5-DNSA) and 3,4-dinitrobenzoic acid (3,4-DNBA) as matrices resulted in multiple hydrogen abstraction from the analyte [M + H] + and fragment [ a ] + ions, i.e., [M + H - m H] + and [ a - m H] + ( m = 1-8). All of the ISD spectra showed unusually intense [ a ] + ions originating from cleavage at the Cα-C bond of the Leu-Xxx residues when peptides without Phe/Tyr/His/Cys residues were used. The intensity of the [ a n ] + series ions generated using 3,5-DNSA and 3,4-DNBA rapidly decreased with increasing residue number n , suggesting cleavage at multiradical sites of [M + H - m H] •+ . It was suggested that multiple hydrogen abstraction from protonated peptides [M + H] + mainly takes place from the backbone amide nitrogen.

Background: Several diseases and insects may cause damage to the normal growth of cucumber. Azoxystrobin and meptyldinocap, because of their novel mode of action, are effective against pathogens that have developed reduced sensitivity to other fungicides. Azoxystrobin is persistent in various crops and environments. However, there is a lack of research on the dissipation of these two pesticides, especially meptyldinocap., Results: Analytes could be quantified with decent recoveries of 90-101%, with relative standard deviations (RSDs) of 3.0-10.1%. The terminal residues of meptyldinocap and azoxystrobin in cucumber were all < limit of quantification (LOQ) (0.02 and 0.05 mg kg -1 ). The half-lives of meptyldinocap and azoxystrobin were 0.8-1.1 and 1.2-2.8 days, respectively. The processing factors (PFs) for washing were all < 1, but the removal rate for washing was < 29.0%. Peeling had a significant effect on the removal of pesticide. The largest residue reductions were noticed through the pickling process, but special care should be taken regarding residues in the pickling solution as pesticides could transfer to them from cucumber. A more interesting finding was that the degradation of two pesticides was accelerated by the addition of calcium oxide., Conclusion: Pesticide residues on cucumber decreased after these processes. These results enable the health-risks from dietary exposures to pesticide residues to be characterized. They enable maximum residue limits (MRLs) to be established for pesticide residues in food products. They also assist the optimization of food processing with regard to pesticide residue dissipation. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

We report herein the discovery of 3,5-dinitrophenyl 1,2,4-triazoles with excellent and selective antimycobacterial activities against Mycobacterium tuberculosis strains, including clinically isolated multidrug-resistant strains. Thorough structure-activity relationship studies of 3,5-dinitrophenyl-containing 1,2,4-triazoles and their trifluoromethyl analogues revealed the key role of the position of the 3,5-dinitrophenyl fragment in the antitubercular efficiency. Among the prepared compounds, the highest in vitro antimycobacterial activities against M. tuberculosis H 37 Rv and against seven clinically isolated multidrug-resistant strains of M. tuberculosis were found with S-substituted 4-alkyl-5-(3,5-dinitrophenyl)-4 H -1,2,4-triazole-3-thiols and their 3-nitro-5-(trifluoromethyl)phenyl analogues. The minimum inhibitory concentrations of these compounds reached 0.03 μM, which is superior to all the current first-line anti-tuberculosis drugs. Furthermore, almost all compounds with excellent antimycobacterial activities exhibited very low in vitro cytotoxicities against two proliferating mammalian cell lines. The docking study indicated that these compounds acted as the inhibitors of decaprenylphosphoryl-β-d-ribofuranose 2'-oxidase enzyme, which was experimentally confirmed by two independent radiolabeling experiments.

Triggering antibody-mediated innate immune mechanisms to kill cancer cells is an attractive therapeutic avenue. In this context, recruitment of endogenous antibodies to the cancer cell surface could be a viable alternative to the use of monoclonal antibodies. We report on antibody-recruiting polymers containing multiple antibody-binding hapten motifs and cyclooctynes that can covalently conjugate to azides introduced onto the glycocalyx of cancer cells by metabolic labeling with azido sugars.

Elucidation of the interaction between NACs and smectites is important to the understanding of the potential for transport of nitroaromatic compounds (NACs) in soils and to implementation of NAC-contaminated soil remediation. The adsorption of dinitrotoluene isomers (DNTs) and substituted dinitrobenzenes (SDNBs) by smectite was determined by batch equilibration and characterized by FTIR and XPS, along with molecular dynamics simulations. The adsorption of DNTs differed substantially among the isomers, attributed to the overall degree of nitro deflection relative to the aromatic ring plane. The substituents in SDNBs strengthened the electrostatic interaction between smectite K + and nitro groups, facilitating SDNB adsorption to smectite. The competition between 2,4-DNT and 1,3-DNB, as well as the inclusion complexation of K + by crown ether 18c6e, both reduced 2,4-DNT adsorption to smectite by weakening the K + -nitro interaction. All the results demonstrated that the electrostatic interaction between smectite K + and nitro of NACs was the predominant force in mediating their adsorption. This was supported by FTIR spectra that the N-O bands shifted due to the weakening of N-O bonds and strengthening of C-N bonds via the electron transfer to cations. The XPS of smectite further manifested the cation-nitro interactions that the binding energies of K 2p 1/2, K 2p 3/2, and Si 2p shifted higher with 1,3-DNB adsorbed. Molecular dynamics simulations indicated the aromatic planes of 2,4-DNP and 2,4-DNAs were parallel to the basal plane of smectite and the oxygens of nitro groups in the molecules were directly coordinated with smectite K + .

The development of more efficient photosensitizers with minimal damage to surrounding normal tissues has been a valuable and challenging subject during photodynamic therapy (PDT). Herein, a stimuli-activated porphyrinic photosensitizer (PEG-TPP-DNB; PEG = poly(ethylene glycol); TPP = 5,10,15,20-tetraphenylporphyrin; DNB = 2,4-dinitrobenzene) with capabilities of fluorescence and, remarkably, singlet oxygen quenching was prepared successfully for photodynamic therapy with high efficiency and biosecurity. The amphiphilic PEG-TPP-DNB could be self-assembled into nanomicelles in aqueous media and dissociated in response to reductive thiol such as glutathione. Meanwhile, the fluorescence and singlet oxygen generation of porphyrinic photosensitizer would be activated to regenerate. Moreover, the intracellular uptake and localization effectively confirmed the redox-responsive and activated behavior of PEG-TPP-DNB micelles. The cytotoxicity in vitro revealed that the micelles had low dark toxicity and great phototoxicity, and in vivo bioimaging and antitumor evaluation further indicated that the micelles possessed selective tumor imaging and targeted PDT antitumor effect as well as low systemic toxicity. Overall, this tumor microenvironment-activated photosensitizer system may provide a useful strategy for precise photodynamic therapy.

Seven cobalt(II) complexes of pyrazole derivatives and dinitrobenzoate ligands were synthesized and characterized. The single-crystal X-ray diffraction structure was determined for one of the ligands and one of the complexes. The analysis and spectral data showed that all the cobalt complexes had octahedral geometries, which was supported by DFT calculations. The complexes and their free ligands were evaluated against fungal strains of Candida albicans and emerging non- albicans species and epimastigotes of Trypanosoma cruzi . We obtained antifungal activity with a minimum inhibitory concentration (MIC) ranging from 31.3 to 250 µg mL -1 . The complexes were more active against C. krusei, showing MIC values between 31.25 and 62.5 µg mL -1 . In addition, some ligands ( L1 - L6 ) and complexes ( 5 and Co(OAc) 2 · 4H 2 O) significantly reduced the yeast to hypha transition of C. albicans at 500 µg mL -1 (inhibition ranging from 30 to 54%). Finally, the complexes and ligands did not present trypanocidal activity and were not toxic to Vero cells. Our results suggest that complexes of cobalt(II) with ligands derived from pyrazoles and dinitrobenzoate may be an attractive alternative for the treatment of diseases caused by fungi, especially because they target one of the most important virulence factors of C. albicans.

Considering the excellent photochemical properties of ruthenium(II) complexes, two new ruthenium(II) complexes, RuL 1 -DNBS and RuL 2 -DNBS, have been developed as phosphorescence probes for detection of biothiols in 100:1 (v/v) Hepes buffer (20 mM, pH = 7.2)/CH 3 CN solution. The response rate was highly improved of these two probes toward biothiols because the steric interactions between 1H-imidazo [4, 5-f] [1,10] phenanthroline group and ortho-2, 4-dinitrobenzensulfonate resulted in a relatively rapid thiol-induced S N Ar substitution reaction. RuL 1 -DNBS and RuL 2 -DNBS were weakly phosphorescent owing to the effectual photoinduced electron transfer from ruthenium(II) luminophore to the sensing group, 2,4-dinitrobenzenesulfonyl. After reacting with biothiols, the 2,4-dinitrobenzenesulfonyl group of RuL 1 -DNBS and RuL 2 -DNBS were cleavaged and the RuL 1 and RuL 2 were obtained. Meanwhile, the phosphorescence were "turn-on". Both of these two probes can detect biothiols sensitively and selectively under physiological conditions with submicromolar detection limits. Furthermore, application of RuL 2 -DNBS for detecting of intracellular biothiols has been successfully performed in living Glioma cells., (Copyright © 2019. Published by Elsevier B.V.)

Feeding experiments with stable isotopes are helpful tools for investigation of metabolic fluxes and biochemical pathways. For assessing nitrogen metabolism, the heavier nitrogen isotope, [ 15 N], has been frequently used. In plants, it is usually applied in form of [ 15 N]-nitrate, which is assimilated mainly in leaves. Thus, methods for quantification of the [ 15 N]-nitrate/[ 14 N]-nitrate ratio in leaves are useful for the planning and evaluation of feeding and pulse-chase experiments. Here we describe a simple and sensitive method for determining the [ 15 N]-nitrate to [ 14 N]-nitrate ratio in leaves. Leaf discs (8 mm diameter, approximately 10 mg fresh weight) were sufficient for analysis, allowing a single leaf to be sampled multiple times. Nitrate was extracted with hot water and derivatized with mesitylene in the presence of sulfuric acid to nitromesitylene. The derivatization product was analyzed by gas chromatography-mass spectrometry with electron ionization. Separation of the derivatized samples required only 6 min. The method shows excellent repeatability with intraday and interday standard deviations of less than 0.9 mol%. Using the method, we show that [ 15 N]-nitrate declines in leaves of hydroponically grown Crassocephalum crepidioides , an African orphan crop, with a biological half-life of 4.5 days after transfer to medium containing [ 14 N]-nitrate as the sole nitrogen source.

Nitric oxide (NO) induces a multitude of antitumor activities, encompassing the induction of apoptosis, sensitization to chemo-, radio-, or immune-therapy, and inhibition of metastasis, drug resistance, angiogenesis, and hypoxia, thus attracting much attention in the area of cancer intervention. To improve the precise targeting and treatment efficacy of NO, a glutathione (GSH)-sensitive NO donor (1,5-bis[(l-proline-1-yl)diazen-1-ium-1,2-diol- O 2 -yl]-2,4-dinitrobenzene, BPDB) coordinates with iron ions to form the nanoscale coordination polymer (NCP) via a simple precipitation and then partial ion exchange process. The obtained Fe(II)-BNCP shows desirable solubility, biocompatibility, and circulation stability. Quick NO release triggered by high concentrations of GSH in tumor cells improves the specificity of NO release in situ, thus avoiding side effects in other tissues. Meanwhile, under high concentrations of H 2 O 2 in tumors, Fe 2+ ions in BPDB-based NCP, named Fe(II)-BNCP, exert Fenton activity to generate hydroxyl radicals (·OH), which is the main contribution for chemodynamic therapy (CDT). In addition, ·O 2 - generated by the Haber-Weiss reaction of Fe 2+ ions with H 2 O 2 can quickly react with NO to produce peroxynitrite anion (ONOO - ) that is more cytotoxic than ·O 2 - or NO only. This synergistic NO-CDT effect has been proved to retard the tumor growth in Heps xenograft ICR mouse models. This work not only implements a synergistic effect of NO-CDT therapy but also offers a simple and efficient strategy to construct a coordination polymer nanomedicine via rationally designed prodrug molecules such as NO donors.

D-Amino acids are important biological molecules. Improved analytical methods for their resolution and quantification remain of keen interest. In this study, we investigated the use of Marfey's reagent (chiral) derivatization coupled with LC-MS/MS-based separation and detection of the resulting diastereomers for quantification of the 19 common L- and D-amino acids and glycine. Standard formic acid (pH 2)-based separations on reverse phase media were unable to separate all 19 amino acid DL pairs. In contrast, a water/acetonitrile/ammonium acetate (pH 6.5) solvent system allowed all 19 amino acid DL pairs to be chromatographically resolved on a 30 min gradient, with negative mode detection at pH 6.5 giving good sensitivity. Derivatization reaction rates between amino acids varied substantially, with overnight derivatization required for some amino acids. Chromatography at pH 6.5 combined with MS/MS quantification in negative mode demonstrated good linearity over a wide concentration range for all 20 amino acids. Matrix effects, assessed with an MRSA extract, were negligible. Marfey's derivatized analytes were stable for 24 h at room temperature. This method was demonstrated by determining the levels of these analytes in mid-log phase MRSA extracts. This approach provides for the chromatographic resolution and MS/MS-based quantification of all 20 common L- and D-amino acids in complex matrices. Graphical Abstract.

The determination of the stereochemistry of common and unusual amino acids is important in food chemistry, archeology, medicine, and life sciences including such diverse areas as marine biology and extraterrestrial chemistry and has greatly contributed to our current knowledge in these fields.To determine the stereochemistry of amino acids, many chromatographic methods have been developed and refined over the last decades. Here we describe a state-of-the-art indirect chromatography-based LC-MS method. Diastereomers were formed from amino acids that were reacted with chiral derivatizing agents such as Marfey's reagent (FDAA), GITC, S-NIFE, and OPA-IBLC and separated on a reversed phase column using mass spectrometry compatible buffers.

Endogenous hydrogen sulfide plays an important role in various physiological and pathological processes and the convenient and selective recognition of hydrogen sulfide has become a research hotspot. We designed and synthesized a tetrahydro[5]helicene and 2,4-dinitrobenzene conjugate (HD-DNP) as an effective fluorescent probe for selective detection of H 2 S. The selective deprotection of 2,4-dinitrophenyl ether group of HD-DNP by H 2 S led to a dramatic fluorescent enhancement (101-fold) at 500 nm and colorimetric change in DMSO-PBS solution. HD-DNP displays many advantages including low background without any self-fluorescence, as well as high selectivity towards common bio-thiols such as Cysteine, Homocysteine and Glutathione. The detection limit of this probe for H 2 S was found to be about 2.4 μM with a wide linear range (10-70 μM). The response mechanism of the probe with HS - is confirmed to be thiolysis of the dinitrophenyl ether induced by HS - through 1 H NMR comparison investigations., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Thiols, such as cysteine (Cys), homocysteine (Hcy), glutathione (GSH), hydrogen sulfide (H 2 S), and thiophenol are metabolically correlated with each other via redox reactions. As a result of the similarity of chemical properties between Cys, Hcy, GSH, H 2 S, and thiophenol, it is very challenging to develop an effective methodology to differentiate them. In this work, a triple-emission fluorescent probe, NCQ, was reported for the simultaneous detection of Cys/Hcy, GSH/H 2 S, and thiophenol with high sensitivity and selectivity. The solution of NCQ displayed distinct fluorescent signals toward Cys/Hcy, GSH/H 2 S, and thiophenol: blue and green for Cys/Hcy, blue for GSH/H 2 S, blue and red for thiophenol. Through the blue-green-red emission color combination, Cys/Hcy, GSH/H 2 S, and thiophenol could be discriminatively detected in solution and in living cells.

2,6-Dicyano-4-nitroaniline and 2-cyano-4-nitroaniline (CNNA; 2-amino-5-nitrobenzonitrile) are potent mutagens in the Ames test, even though unsubstituted nitroanilines (NAs) are no more than weak mutagens. These compounds are putative reduction products of many commercial azo dyes, including Disperse Blue 165, Disperse Blue 337, Disperse Red 73, Disperse Red 82, Disperse Violet 33, and Disperse Violet 63. We have examined the mutagenicity in strains TA98 and YG1024 of a series of commercially-available isomers of CNNA, and some related compounds, to probe the relationship between structure and genotoxic activity in this class of compounds. The potentiating effect of the cyano substituent is seen in many cases; e.g. 2-amino-4-nitrobenzonitrile is a much more potent mutagen than 3-NA. 2,4-Dinitrobenzonitrile is also highly mutagenic. Possible mechanisms for the "cyano effect" are considered, with respect to the likely structures of cyanonitroaniline-DNA adducts and the roles of the enzymes (nitroreductase and acetyl CoA:arylamine N-acetyltransferase) believed to be involved in the activation of nitroaromatic compounds. Environ. Mol. Mutagen. 59:114-122, 2018. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

Here we report a new chiral stationary phase (CSP) immobilized on silica gel based on cone calix[4]arene functionalized at the upper rim with two l-alanine units as new chiral selector that has been used in high-performance liquid chromatography. The CSP was prepared by covalently bonding the allyl groups at the lower rim of calix[4]arene to silica gel by thiol-ene click chemistry reaction. Elemental analysis of the CSP showed that 120 μmol of chiral selector bonded per gram of silica gel. 1-Hexene was used for end-capping of unreacted mercapto groups on silica gel. Since the CSP is chemically bonded to the silica, it can be used in the normal-phase and reversed-phase mode and with halogenated solvents as mobile phases, if desired. The chromatographic performance of the CSP was evaluated in the enantioseparation of the 3,5-dinitrobenzoyl derivatives of some amino acids, diclofop-methyl and dl-mandelic acid., (Copyright © 2017 John Wiley & Sons, Ltd.)

Energetic materials (EMs) bound to propellant residues can contribute to environmental risk and public health concerns. This work investigated how nitrocellulose, a common binding material in propellants, may control the release dynamics of nitroglycerin (NG) and 2,4-dinitrotoluene (2,4-DNT) from propellant residues. Batch adsorption/desorption experiments on nitrocellulose and re-interpretation on results from past leaching studies involving propellant-bound EMs were conducted. Mechanistic modeling of adsorption/desorption kinetics based on intra-particle diffusion (IPD) predicted aqueous intrinsic diffusivities (D iw ) to within a factor of 2 of expected values. Furthermore, the IPD model was able to predict effective diffusivities (D eff ) during the early leaching of NG from propellant residues to within a factor of 2 over a 3-log unit range. Prediction of leaching D eff 's associated with fired residues was less successful probably due to the neglect of compositional and morphological heterogeneity within the residues. Close correlations were found between the early and late D eff 's of residue-bound NG and between the fast- and slow-domain rate constants for both EMs, suggesting that the late leaching kinetics of bound-EMs may be empirically assessed from the early kinetics. This work illustrates that, in addition to dissolution, retarded diffusion through nitrocellulose matrix may also limit the overall release and transformation of residue-bound EMs in the field. Implications and limitations of the current study, and the steps forward are also presented., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Methamphetamine (MAMP) is a popular illicit drug abused for its central nervous system stimulating effects. MAMP is also used therapeutically in the treatment of overeating disorders, narcolepsy, attention deficit disorder, in over-the-counter (OTC) products to ease nasal congestion. MAMP exists in two enantiomeric forms, dextrorotary (d-MAMP) or levorotary (l-MAMP). The compounds are similar in chemical structure, simply differing in the orientation of functional groups around the asymmetric carbon. In part because of the availability of l-MAMP in OTC nasal inhalers, forensic guidelines require a sample to contain greater than 20% d-MAMP to consider illicit drug use when interpreting results. Standard analytical methods readily detect MAMP in biological specimens but cannot resolve the enantiomeric composition of the sample. Specialized analytical techniques based on chiral separation of the enantiomers are required to differentiate d-MAMP from l-MAMP. Our laboratory sought to develop and validate a method for the analysis of oral fluid specimens for d/l-MAMP using a chiral derivatizing agent and traditional reverse phase liquid chromatography tandem mass spectrometry (LC-MS-MS). MAMP was extracted from dilute oral fluid samples using Strata-XC solid phase extraction (SPE) cartridges and derivatized with Marfey's reagent. Chromatographic separation was achieved using Zorbax Eclipse Plus C18 columns. Linearity, accuracy and precision, recovery, matrix effects and specificity of the method were all within acceptable criteria. Intraday accuracy ranged from 93.3 to 103.4% and precision 0.1 to 1.6%. Interday accuracy ranged from 90.0 to 103.4% and precision 3.8 to 11.6%. Finally, having previously tested positive for MAMP using non-chiral analysis, 256 de-identified authentic oral fluid samples were analyzed using this validated method. 98% of all samples tested positive for d-MAMP at greater than 20%.

Conjugated microporous polymers (CMPs) with strong fluorescence are great candidates for optoelectronic applications such as photocatalysis and chemical sensing. A series of novel fluorene-based conjugated microporous polymers (FCMPs) with different electronic structures are prepared by Yamamoto coupling reactions using rationally designed monomers. The FCMPs show a high degree of microporosity, decent specific surface areas, and variable fluorescence. FCMP3, which possesses a triazine knot in the network, exhibits the highest specific surface area of 489 m 2 g -1 , the largest pore volume of 0.30 cm 3 g -1 , and the highest solid-state photoluminescence quantum yield of 11.46%. Chemical sensing performance of FCMPs is studied using a range of nitroaromatic compounds as the analytes. Among the FCMPs, FCMP3 exhibits the highest Stern-Volmer constants of 2541, 4708, and 5241 m -1 for the detection of nitrobenzene, 4-nitrotoluene, 2,4-dinitrotoluene, respectively, which are comparable to the detecting efficiency of the state-of-the-art CMP-based sensing agents., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Sorption of organic compounds on fresh black carbons (BCs) can be greatly attenuated in soil over time. We examined herein the changes in surface properties of maize straw-derived BCs (biochars) after aged in a black soil and their effects on the sorptive behaviors of naphthalene, phenanthrene and 1,3-dinitrobenzene. Dissolved fulvic and humic acids extracted from the soil were used to explore the role of dissolved organic carbon (DOC) in the aging of biochars. Chromatography analysis indicated that DOC molecules with relatively large molecular weight were preferentially adsorbed on the biochars during the aging processes. DOC sorption led to blockage of the biochar's micropores according to N 2 and CO 2 adsorption analyses. Surface chemistry of the biochars was also substantially modified, with more O-rich functional groups on the aged biochars compared to the original biochars, as evidenced by Near-edge X-ray absorption fine structure (NEXAFS) spectroscopy and X-ray photoelectron spectroscopy (XPS) analyses. The changes in both the physical and chemical surface properties of biochars by DOC led to significant attenuation of the sorption capacity and nonlinearity of the nonionic organic compounds on the aged biochars. Among the tested organic compounds, phenanthrene was the most attenuated in its sorption by the aging treatments, possibly because of its relatively large molecular size and hydrophobicity. The information can help gain a mechanistic understanding of interactions between BCs and organic compounds in soil environment., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A multicolour protein labelling technique using a protein tag and fluorogenic probes is a powerful approach for spatio-temporal analyses of proteins in living cells. Since cyanine fluorophores have attractive properties for multicolour imaging of proteins, there is a huge demand to develop fluorogenic cyanine probes for specific protein labelling in living cells. Herein, we develop fluorogenic cyanine probes for labelling a protein tag by using a dinitrobenzene fluorescence quencher. The probes enhanced fluorescence intensity upon labelling reactions and emitted orange or far-red fluorescence. Intramolecular interactions between the cyanine fluorophores and the dinitrobenzene quencher led not only to fluorescence quenching of the probes in the free state but also to promotion of labelling reactions. Furthermore, the probes successfully imaged cell-surface proteins without a washing process. These findings offer valuable information on the design of fluorogenic cyanine probes and indicate that the probes are useful as novel live-cell imaging tools.This article is part of the themed issue 'Challenges for chemistry in molecular imaging'., (© 2017 The Author(s).)

Systemic fungal infections represent an important public health concern, and new antifungal agents are highly desirable. Herein, we describe the design, synthesis, and biological evaluation of a novel class of antifungal compounds called antibody-recruiting molecules targeting fungi (ARM-Fs). Our approach relies on the use of non-peptidic small molecules, which selectively bind fungal cells and recruit endogenous antibodies to their surfaces, resulting in immune-mediated clearance. Using the opportunistic fungal pathogen Candida albicans as a model, we identified a highly specific bifunctional molecule able to mediate the engulfment and phagocytosis of C. albicans cells by human immune cells in biologically relevant functional assays. This work represents a novel therapeutic approach to treating fungal illness with significant potential to complement and/or combine with existing treatment strategies., (© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In general, the reactivity of cysteine residues of proteins is measured by 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) kinetics using spectrophotometry. Proteins with several cysteine residues may exhibit varying DTNB kinetics but residue level information can only be obtained with the prior knowledge of their three-dimensional structure. However, this method is limited in its application to the proteins containing chromophores having overlapping absorption profile with 2-nitro-5-thiobenzoic acid, such as hemoglobin (Hb). Additionally, this method is incapable of assigning cysteine reactivity at the residue levels of proteins with unknown crystal structures. However, a mass spectrometry (MS)-based platform might provide a solution to these problems. In the present study, alkylation kinetics of cysteine residues of adult human Hb (Hb A; α 2 β 2 ) and sickle cell Hb (Hb S; HBB: c.20A>T) were investigated using matrix-assisted laser desorption/ionization (MALDI) MS. Differential site-specific reactivities of cysteine residues of Hb were investigated using alkylation kinetics with iodoacetamide (IAM). The observed reactivities corroborated well with the differential surface accessibilities of cysteine residues in the crystal structures of human Hb. The proposed method might be used to investigate cysteine reactivities of all the genetic and post-translational variants of Hb discovered to date. In addition, this method can be extended to explore cysteine reactivities of proteins, irrespective of the presence of chromophores and availability of crystal structures.

Herbicides of the dinitroaniline chemical class, widely used oryzalin and trifluralin, and also nitralin were tested as inhibitors of recombinant human carbonic anhydrases (CAs). Oryzalin bound and inhibited 11 out of 12 catalytically active CA isoforms present in the human body with the affinities in the same range as clinically used CA drugs, while no effect was detected for the other two compounds. Binding of all three herbicides was examined by fluorescence-based thermal shift assay, isothermal titration calorimetry, and the inhibition of carbon dioxide hydratase activity. During the last decade, dinitroaniline compound-based therapies against protozoan diseases are being developed. Therefore, it is important to investigate their potential off-target effects, including human CAs., (© 2017 Wiley Periodicals, Inc.)

Dinitrotoluene (DNT) is a widely present pollutant in aquatic environments, and its biodegradation is an economically attractive way to effectively removal. In aquatic environments, the presence of electrochemically active bacteria (EAB) could contribute to the anaerobic bioreduction of DNT. However, the mechanism behind such a biodegradation process at gene level remains to be further elucidated. In this work, the anaerobic reduction of 2,6-dinitrotoluene (2,6-DNT) by Shewanella oneidensis MR-1, a typical EAB in aquatic environments, was investigated. S. oneidensis MR-1 was found to be able to obtain energy for growth through the anaerobic respiration on 2,6-DNT. Experimental results show that the Mtr respiratory pathway, a transmembrane electron transport chain, was involved in the 2,6-DNT bioreduction. Knockout of cymA or nfnB resulted in a substantial loss of its 2,6-DNT-reducing ability, indicating that both CymA and NfnB were the key proteins in the microbial electron transfer chain. The genetic analysis further confirms that the Mtr respiratory pathway and NfnB are mainly responsible for the anaerobic reduction of 2,6-DNT by S. oneidensis MR-1. This work is useful to better understand the anaerobic bioreduction of nitroaromatic compounds in aquatic environments and remediate the environments contaminated by nitroaromatic compounds. Biotechnol. Bioeng. 2017;114: 761-768. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Luminescent lanthanide complexes are key reagents used in the time-gated luminescence bioassay technique, but functional lanthanide complexes that can act as luminescent probes for specifically responding to analytes are very limited. In this work, we designed and synthesized a novel Eu 3+ complex-based luminescence probe for hypochlorous acid (HOCl), NPPTTA-Eu 3+ , by using terpyridine polyacid-Eu 3+ , dinitrophenyl, and hydrazine as luminophore, quencher and HOCl-recognizer moieties, respectively. In the absence of HOCl, the probe is non-luminescent due to the strong luminescence quenching of the dinitrophenyl group in the complex. However, upon reaction with HOCl, the dinitrophenyl moiety is rapidly cleaved from the probe, which affords a strongly luminescent Eu 3+ complex CPTTA-Eu 3+ , accompanied by a ∼900-fold luminescence enhancement with a long luminescence lifetime of 1.41 ms. This unique luminescence response of NPPTTA-Eu 3+ to HOCl allowed NPPTTA-Eu 3+ to be conveniently used as a probe for highly selective and sensitive detection of HOCl under the time-gated luminescence mode. In addition, by loading NPPTTA-Eu 3+ into RAW 264.7 macrophage cells and Daphnia magna, the generation of endogenous HOCl in RAW 264.7 cells and the uptake of exogenous HOCl by Daphnia magna were successfully imaged on a true-color time-gated luminescence microscope. The results demonstrated the practical applicability of NPPTTA-Eu 3+ as an efficient probe for time-gated luminescence imaging of HOCl in living cells and organisms.

Selenocysteine (Sec) carries out the majority of the functions of the various Se-containing species in vivo. Thus, it is of great importance to develop sensitive and selective assays to detect Sec. Herein, a carbon-dot-based fluorescent turn-on probe for highly selective detection of selenol in living cells is presented. The highly photoluminescent carbon dots that emit yellow-green fluorescence (Y-G-CDs; λ max = 520 nm in water) were prepared by using m-aminophenol as carbon precursor through a facile solvothermal method. The surface of Y-G-CDs was then covalently functionalized with 2,4-dinitrobenzenesulfonyl chloride (DNS-Cl) to afford the 2,4-dinitrobenzene-functionalized CDs (CD-DNS) as a nanoprobe for selenol. CD-DNS is almost nonfluorescent. However, upon treating with Sec, the DNS moiety of CD-DNS can be readily cleaved by selenolate through a nucleophilic substitution process, resulting in the formation of highly fluorescent Y-G-CDs and hence leads to a dramatic increase in fluorescence intensity. The proposed nanoprobe exhibits high sensitivity and selectivity toward Sec over biothiols and other biological species. A preliminary study shows that CD-DNS can function as a useful tool for fluorescence imaging of exogenous and endogenous selenol in living cells.

Nitro-aromatic compounds (NACs) such as 1,3-dinitrobenzene (1,3-DNB) contain the nitrogroup (-NO 2 ), in which the N with a +III oxidation state accepts electrons. Water soluble ascorbic acid (AsA) at elevated pH produces electron transfer and governs the electron-donating pathway. The influence of the NaOH/AsA molar ratio on the degradation of 1,3-DNB was investigated. Using 0.21-2 M NaOH and 20-100 mM AsA, nearly complete 1,3-DNB removals (90-100%) were achieved within 0.5 h. On the basis of intermediates identified using GC/MS, the reduction pathways of 1,3-DNB can be categorized into step-by-step electron transfer, and condensation routes. A higher NaOH/AsA molar ratio would result in relatively higher AsA decomposition, promote the condensation route into the formation of azo- and azoxy-compounds, and ultimately reduce 1,3-DNB to 1,3-phenylenediamine. Contaminated soil flushing using 500 mM NaOH/100 mM AsA revealed that 1,3-DNB was completely degraded within 2 h. Based on these test results, the alkaline AsA treatment method is a potential remediation process for NACs contaminated soils., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Co-pyrolysis of polymer and biomass wastes was investigated as a novel method for waste treatment and synthesis of enhanced biochar. Co-pyrolysis of rice straw (RS) with polypropylene (PP), polyethylene (PE) or polystyrene (PS) increased the carbon content, cation exchange capacity (CEC), surface area and pH of the biochar. As a result, the sorption of 2,4-dinitrotoluene (DNT) and Pb to polymer/RS-derived biochar was markedly enhanced. The increased aromaticity and hydrophobicity may be responsible for enhancing the DNT sorption to the polymer/RS-derived biochar. In contrast, increasing CEC, higher pH, and the newly developed surface area may account for the enhancement in Pb sorption. The addition of polymer to RS did not significantly change the catalytic role of biochar during the reduction of DNT by dithiothreitol. Our results suggest that co-pyrolysis of RS and polymer can improve the biochar properties to enhance the sorption of DNT and Pb., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In order to remediate organic contaminants in natural waters and soils, a novel zero-valent iron [Fe(0)]-included biochar was synthesized via slow pyrolysis. 2,4-Dinitrotoluene (DNT) and 2,4-dichlorophenol (DCP) were removed in water via sorption to the Fe(0)-included biochar. Compared to sorption control without Fe(0), the sorbed DNT and DCP were further transformed to reduction products by Fe(0)-included biochar. Compared to the reduction control with Fe(0), the presence of biochar promoted the reductive transformation of DNT and DCP. Increasing the pyrolysis temperature resulted in enhancing the removal of DNT and DCP, suggesting that the aromaticity of biochar may be responsible for the removal. The yields of the reduction products also indicated that unlike the direct reduction by Fe(0), different reduction pathways existed in the reduction of DNT and DCP with Fe(0)-included biochar. The results suggest that Fe(0)-included biochar is a viable option to immobilize and transform redox-sensitive organic contaminants in natural environments., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Chiral separation is crucial for investigating methamphetamine positive cases. While (S)-(+)-enantiomer of methamphetamine (S-MAMP) is a schedule II controlled substance, (R)-(-)-enantiomer (R-MAMP) is an active ingredient of a few over-the-counter drugs in the United States. Among biological specimen types, hair provides greater detection window than blood, urine or oral fluid, and are therefore regarded with particular interest. Herein we describe a novel non-chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to directly determine methamphetamine enantiomeric composition (percentage) in hair specimens. Hair samples were washed once with acetone, powdered, incubated overnight at 53°C in 0.1M hydrochloric acid (HCl), and subjected to a solid phase extraction (SPE). The extracts were derivatized using Marfey's reagent at 53°C for 60min. The final mixture was analyzed by LC-MS/MS. Chromatographic separation was achieved using a C18 Kinetex analytical column and 60% (v/v) aqueous methanol as mobile phase (isocratic). Triple quadrupole mass spectrometer was equipped with an electro-spray ionization (ESI) source operating in negative mode and the chromatograms were acquired using a multiple-reaction monitoring (MRM) approach. The results were expressed as ratio of R- to S-MAMP and then derived to composition percentages without requiring quantitating each enantiomer. The method was precise and accurate across 0-100% S-composition at a range of 80-18,000pg/mg. The performance of the new method was compared with an (S)-(-)-N-trifluoroacetylprolyl chloride (S-TPC) derivatization and gas chromatography-mass spectrometry (GC-MS) method on authentic methamphetamine-positive hair samples. Not only the new Marfey's reagent approach presented satisfactory correlation with the S-TPC approach, but it also exhibited significantly improved quality (e.g., S/N) of the chromatograms. In summary, our protocol employs cost effective and minimally hazardous Marfey's reagent to derivatize trace amounts of methamphetamine extracted from hair samples and a non-chiral LC-MS/MS approach to separate and identify the two enantiomers. The method allows determination of the methamphetamine enantiomeric composition without requiring quantitation of each enantiomer and is therefore well suited for further investigate previously determined methamphetamine positive cases. This method represents a viable tool for evaluation of long-term drug exposure., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Cellular imaging technologies employing metallic surface-enhanced Raman scattering (SERS) tags have gained much interest toward clinical diagnostics, but they are still suffering from poor controlled distribution of hot spots and reproducibility of SERS signals. Here, we report the fabrication and characterization of high narrow nanogap-containing Au@Au core-shell SERS nanoparticles (GCNPs) for the identification and imaging of proteins overexpressed on the surface of cancer cells. First, plasmonic nanostructures are made of gold nanoparticles (∼15 nm) coated with gold shells, between which a highly narrow and uniform nanogap (∼1.1 nm) is formed owing to polyA anchored on the Au cores. The well controlled distribution of Raman reporter molecules, such as 4,4'-dipyridyl (44DP) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), are readily encoded in the nanogap and can generate strong, reproducible SERS signals. In addition, we have investigated the size-dependent SERS activity of GCNPs and found that with the same laser wavelength, the Raman enhancement discriminated between particle sizes. The maximum Raman enhancement was achieved at a certain threshold of particle size (∼76 nm). High narrow nanogap-containing Au@Au core-shell SERS tags (GCTs) were prepared via the functionalization of hyaluronic acid (HA) on GCNPs, which recognized the CD44 receptor, a tumor-associated surface biomarker. And it was shown that GCTs have a good targeting ability to tumour cells and promising prospects for multiplex biomarker detection.

Hydrogen sulphide (H2S), the third endogenous gaseous signalling molecule, has attracted attention in biochemical research. The selective detection of H2S in living systems is essential for studying its functions. Fluorescence detection methods have become useful tools to explore the physiological roles of H2S because of their real-time and non-destructive characteristics. Herein we report a near-infrared fluorescent probe, NIR-HS, capable of tracking H2S in living organisms. With high sensitivity, good selectivity and low cytotoxicity, NIR-HS was able to recognize both the exogenous and endogenous H2S in living cells. More importantly, it realized the visualization of endogenous H2S generated in cells overexpressing cystathionine β-synthase (CBS), one of the enzymes responsible for producing endogenous H2S. The probe was also successfully applied to detect both the exogenous and endogenous H2S in living mice. The superior sensing properties of the probe render it a valuable research tool in the H2S-related medical research.

The phototransformation of Oryzalin was studied under UV light (λmax ≥ 290 nm) and sunlight (λmax ≥ 250 nm) in aqueous isopropanol and acetonitrile solution in absence and presence of TiO2 as sensitizer. The rate of photodegradation of Oryzalin in different solvent system followed first-order kinetics, and calculated half-lives were found to be in the range of 23.52-53.75 h for UV light and 41.23-61.43 h for sunlight. From this study, total 12 photoproducts were identified and characterized on the basis of column chromatography and Q-Tof micromass spectral data. The plausible mechanism of phototransformation involved was hydrolysis, breaking of sulfonic bond, and loss of amino and sulfonic acid group.

Partition behavior of eight small organic compounds and six proteins was examined in poly(ethylene glycol)-8000-sodium sulfate aqueous two-phase systems containing 0.215M NaCl and 0.5M osmolyte (sorbitol, sucrose, TMAO) and poly(ethylene glycol)-10000-sodium sulfate-0.215M NaCl system, all in 0.01M sodium phosphate buffer, pH 6.8. The differences between the solvent properties of the coexisting phases (solvent dipolarity/polarizability, hydrogen bond donor acidity, and hydrogen bond acceptor basicity) were characterized with solvatochromic dyes using the solvatochromic comparison method. Differences between the electrostatic properties of the phases were determined by analysis of partitioning of sodium salts of dinitrophenylated (DNP-) amino acids with aliphatic alkyl side-chain. The partition coefficients of all compounds examined (including proteins) were described in terms of solute-solvent interactions. The results obtained in the study show that solute-solvent interactions of nonionic organic compounds and proteins in polyethylene glycol-sodium sulfate aqueous two-phase system change in the presence of NaCl additive., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Two pyridoxine derivatives containing a dinitrophenyl moiety were investigated by (1)H NMR spectroscopy. Conformational dynamics in solution were studied for each compound using dynamic NMR experiments. It was shown that both compounds studied are involved into two conformational exchange processes. The first process is a transformation of the seven-membered cycle conformation between the enantiomeric P-twist and M-twist forms, and the second is a rotation of the dinitrophenyl fragment of the molecules around the C-O bond. Energy barriers of both conformational transitions were determined., (Copyright © 2015 John Wiley & Sons, Ltd.)

The erythropoietin-producing hepatocellular carcinoma receptor B4 is a receptor tyrosine kinase whose expression is preserved in various malignancies, including colon, gastric, and breast carcinoma. Hepatocellular carcinoma receptor B4 presence in tumor cells and involvement in cancer suppression makes it a potential therapeutic target for activating compounds. Moreover, modulators of its activity also have a strong potential to be used in diagnosis and therapy monitoring. We used virtual ligand screening to identify novel hepatocellular carcinoma receptor B4 kinase modulators for experimental testing. Three independent assay platforms confirmed that dinitrophenyl-L-arginine is likely to affect the kinase activity of hepatocellular carcinoma receptor B4. An enzyme-coupled spectrophotometric assay has been used to examine this possibility and may prove to be useful for assessing other novel kinase modulator candidates. Overall, our observations suggest that dinitrophenyl-L-arginine has an activating effect on hepatocellular carcinoma receptor B4 and, therefore, more efficient derivatives may have therapeutic effects in tumors where hepatocellular carcinoma receptor B4 exhibits antimalignant properties. The hepatocellular carcinoma receptor B4-activating effect is discussed with respect to previously described mechanisms, using predicted and experimental structures for docked ligands. As a novel kinase activity modulator, dinitrophenyl-L-arginine may provide new insights into molecular mechanisms by which kinases are activated or regulated, and may serve as a lead compound for the generation of novel hepatocellular carcinoma receptor B4-activating therapeutic compounds., (© 2014 John Wiley & Sons A/S.)