Search

Flavonoid-nitroxide hybrids were prepared and displayed limited antioxidant protection, performing antagonistically compared to the individual components in a peroxyl radical scavenging assay.

The versatile nature of organic conjugated materials renders their flawless integration into a diverse family of optoelectronic devices with light-harvesting, photodetection, or light-emitting capabilities. Classes of materials that offer the possibilities of two or more distinct optoelectronic functions are particularly attractive as they enable smart applications while providing the benefits of the ease of fabrication using low-cost processes. Here, we develop a novel, multi-purpose conjugated small molecule by combining boron-azadipyrromethene (aza-BODIPY) as electron acceptor with triphenylamine (TPA) as end-capping donor units. The implemented donor–acceptor–donor (D–A–D) configuration, in the form of TPA-azaBODIPY-TPA, preserves ideal charge transfer characteristics with appropriate excitation energy levels, with the additional ability to be used as either a charge transporting interlayer or light-sensing semiconducting layer in optoelectronic devices. To demonstrate its versatility, we first show that TPA-azaBODIPY-TPA can act as an excellent hole transport layer in methylammonium lead triiodide (MAPbI3)-based perovskite solar cells with measured power conversion efficiencies exceeding 17%, outperforming control solar cells with PEDOT:PSS by nearly 60%. Furthermore, the optical bandgap of 1.49 eV is shown to provide significant photodetection in the wavelength range of up to 800 nm where TPA-azaBODIPY-TPA functions as donor in near-infrared organic photodetectors (OPDs) composed of fullerene derivatives. Overall, the established versatility of TPA-azaBODIPY-TPA, combined with its robust thermal stability as well as excellent solubility and processability, provides a new guide for developing highly efficient multi-purpose electronic materials for the next-generation of smart optoelectronic devices. Ministry of Education (MOE) National Research Foundation (NRF) Published version Lam Yeng Ming would like to thank the support of the Singapore Ministry of Education Academic Research Fund Tier 2 (Grant No. MOE2019-T2-1-085). Annalisa Bruno and Enkhtur thank the National Research Foundation for the financial support (Grant No. S18-1176-SCRP). Prashant Sonar is thankful to QUT for the financial support from the Australian Research Council (ARC) for the Future Fellowship (Grant No. FT130101337) and QUT core funding (Grant No. QUT/322120-0301/07). Ajay K. Pandey acknowledges support from Australia India Strategic Research Fund (Project AISRF53820).

Biofilms are part of the natural lifecycle of bacteria and are known to cause chronic infections that are difficult to treat. Most antibiotics are developed and tested against bacteria in the planktonic state and are ineffective against bacterial biofilms. The oxazolidinones, including the last resort drug linezolid, are one of the main classes of synthetic antibiotics progressed to clinical use in the last 50 years. They have a unique mechanism of action and only develop low levels of resistance in the clinical setting. With the aim of providing insight into strategies to design more potent antibiotic compounds with activity against bacterial biofilms, we review the biofilm activity of clinically approved oxazolidinones and report on structural modifications to oxazolidinones and their delivery systems which lead to enhanced anti-biofilm activity.

An alternative synthetic route towards the widely employed electroactive poly(TEMPO methacrylate) (PTMA) via a thermally robust methoxyamine-protecting group is demonstrated herein. Protection of the radical moiety of hydroxy-TEMPO with a methyl functionality and subsequent esterification with methacrylic anhydride allows the high-yielding formation of the novel monomer methyl-TEMPO methacrylate (MTMA). The polymerization of MTMA to poly(MTMA) (PMTMA) is investigated via free radical polymerization and reversible addition-fragmentation chain-transfer polymerization (RAFT), a reversible-deactivation radical polymerization technique. Cleavage of the temperature-stable methoxyamine functionality by oxidative treatment of PMTMA with meta-chloroperbenzoic acid (mCPBA) releases the electroactive PTMA. The redox activity of PTMA was confirmed by cyclic voltammetry in lithium-ion coin cells.

Isothiazolones are widely used as biocides in industrial processing systems and personal care products, but their use to treat infections in humans has been hampered by their inherent cytotoxicity. Herein, we report a strategy to alleviate isothiazolone toxicity and improve antibacterial and antibiofilm potency by functionalization with a nitroxide moiety. Isothiazolone-nitroxide hybrids

Urinary tract infections (UTIs) are amongst the most common and prevalent infectious diseases worldwide, with uropathogenic Escherichia coli (UPEC) reported as the main causative pathogen. Fluoroquinolone antibiotics are commonly used to treat UTIs but for infections involving UPEC biofilms, which are commonly associated with catheter use and recurrent episodes, ciprofloxacin is often ineffective. Here we report the development of a ciprofloxacin–dinitroxide (CDN) conjugate with potent UPEC biofilm-eradication activity. CDN 11 exhibited a 2-fold increase in potency over the parent antibiotic ciprofloxacin against UPEC biofilms. Moreover, CDN 11 resulted in almost complete UPEC biofilm cell eradication (99.7%) at concentrations as low as 12.5 μM, and significantly potentiated ciprofloxacin's biofilm-eradication activity against UPEC upon co-administration. The biofilm-eradication activity of CDN 11 highlights the potential of nitroxide functionalized antibiotics as a promising strategy for the treatment of biofilm-related UTIs.

This work demonstrates a new pathway to the direct on-surface fabrication of surface coatings by showing that application of a plasma can lead to dehalogenative coupling of small aromatic molecules at a catalytic surface. Specifically, we show that a room temperature, atmospheric pressure plasma can be used to fabricate a coating through a surface-confined dehalogenation reaction. Plasma treatments were performed using a dielectric barrier discharge (DBD) technique under pure nitrogen with a variety of power levels and durations. Samples were analysed by optical and helium ion microscopy (HIM), X-ray photoelectron spectroscopy (XPS), optical profilometry, and contact angle measurement. By varying the plasma parameters we could control the chemistry, morphology and roughness of the film. Surface wettability also varied with the plasma parameters, with high-dose plasmas leading to a hydrophobic surface with water contact angles up to 130°.

Bacterial biofilm formation on medical devices is a threat to healthcare systems worldwide as bacteria within a biofilm are more resistant to standard antimicrobial therapies. Herein, we introduce a nitroxide-based antibiofilm coating strategy, which is specifically designed to prevent biofilm formation of Gram-negative pathogens such as Pseudomonas aeruginosa. Nitroxide-decorated hydroxyapatite surfaces were readily prepared in an aqueous dip-coating procedure using a nitroxide-functionalized catecholamine as a polymerizable coating agent. Additional spatial control over the polymer surface deposition on titanium was gained by applying a photolithographic coating setup. All nitroxide-coated surfaces exhibited excellent antibiofilm properties toward PA14 surface colonization as biofilm formation was completely suppressed. Importantly, the surrounding adhesive polymer matrix did not interfere with the nitroxide-characteristic antibiofilm properties. The herein introduced platform technology represents a bioinspired and versatile coating approach, offering a safe and prophylactic avenue to combat biofilm contamination on a variety of surfaces.

Background: The adhesion of planktonic bacteria to a surface (biotic or abiotic), and their subsequent ability to aggregate into multicellular communities called biofilms, is a major driving force of failing antibiotic therapy and persistence in chronic infections caused by a variety of pathogens (e.g., Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus) and plaguing healthcare systems worldwide. Biofilms are estimated to be involved in over 80% of all microbial infections in humans, and commonly exhibit extreme resistance to conventional antimicrobial treatments. Consequently, there is an urgent need for novel antimicrobial agents, which target biofilm residing cells. Here, we present the development and evaluation of a new generation of dual-acting nitroxide functionalised antibiotics with potent biofilm eradication activity. Methodology: Synthetic organic chemistry was utilised to produce a new generation of nitroxide functionalised antibiotics with targeted biofilm eradication capabilities. These compounds were tested for biofilm eradication and/or dispersal of several bacterial species using the MBECTM device, a reproducible high-throughput static biofilm formation system. Mature biofilms were treated with serial dilutions of the specific test agent(s) and recovered bacterial numbers were quantified by absorbance spectroscopy at 600 nm or plating for viable cell counts. Treated biofilms were also stained with Live/Dead (SYTO-9/PI) bacterial viability kit and analysed by fluorescence and confocal laser scanning microscopy. Results: Nitroxide functionalised antibiotics exhibit potent biofilm-eradication activity against a variety of medically important pathogens, including P. aeruginosa, uropathogenic E. coli, and S. aureus. In Minimal Biofilm Eradication Concentration (MBEC) assays nitroxide functionalised antibiotics were 64-fold more potent against S. aureus biofilms, and at least 2-fold more potent against uropathogenic E. coli biofilms than the parent antibiotic ciprofloxacin. Conclusions: Currently, antibiotics are often entirely ineffective against biofilm infections. Nitroxide functionalised antibiotics represent a promising new strategy, which could circumvent the resistance of Gram-positive and Gram-negative biofilms to conventional treatments.

We pioneer a versatile surface modification strategy based on mussel-inspired oxidative catecholamine polymerization for the design of nitroxide-containing thin polymer films. A 3,4-dihydroxy-l-phenylalanine (l-DOPA) monomer equipped with a 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-derived oxidation-labile hydroxylamine functional group is employed as a universal coating agent to generate polymer scaffolds with persistent radical character. Various types of materials including silicon, titanium, ceramic alumina, and inert poly(tetrafluoroethylene) (PTFE) were successfully coated with poly(DOPA-TEMPO) thin films in a one-step dip-coating procedure under aerobic, slightly alkaline (pH 8.5) conditions. Steadily growing polymer films (∼1.1 nm h–1) were monitored by ellipsometry, and their thicknesses were critically compared with those obtained from atomic force microscopic cross-sectional profiles. The heterogeneous composition of surface-adherent nitroxide scaffolds examined by X-ray photoelectron spectr...

We pioneer an approach for the visualization of pH-controlled molecular release of a fluorescent drug from a nitroxide polymer which combined yields a profluorescent system. A methacrylate-based terpolymer, bearing water-solubilizing triethylene-glycol chains, unpaired spins in the form of nitroxide and hydrazone-bound doxorubicin was synthesized to explore the possibilities of release quantification from profluorescent assemblies. Critically, we establish the fluorescence-silencing effect of polymer-bound nitroxides on covalently attached fluorophores and the overall accessibility of such constructs. The facile approach towards a functional assembly shown here provides an easy-to-assemble construct for complex delivery architectures towards applications in theranostics.

We introduce the preparation and ring-opening polymerization of a nitroxide containing carbonate monomer. Using 1,8-diazabicyclo undec-7-ene (DBU) and a thiourea as the catalyst system, we demonstrate that the free radical moiety of the monomer has no influence on the ring-opening copolymerization with D/L-lactide in a wide range of compositions between 10% and 80% of the nitroxide monomer, which can be readily controlled by the comonomer composition in the polymerization mixture. The copolymers exhibit narrow and monomodal mass distributions with number average molar masses between 6000 g·mol −1 to 7000 g·mol −1 and low dispersities below 1.1. The free radical functionality is retained during the polymerization as demonstrated by electron spin resonance (EPR) spectroscopy. High-resolution mass spectrometry coupled to size exclusion chromatography allowed an in-depth analysis of the copolymers showing well-defined linear polymer chains with excellent endgroup fidelity. Furthermore, the molar mass of the polymers was controlled by the monomer to initiator ratio which led to copolymers of up to 19 500 g·mol −1 .

Biofilms are highly organised colonies of microorganisms, at a surface or an interface, which are extremely resistant to treatment with biocides or antimicrobials. Consequently, biofilms are the primary cause of fouling and persistent infections in both industrial and clinical settings. Thus, there is a clear need to develop coatings that are able to prevent the growth of biofilms on surfaces. Herein, we report the development of polynitroxides for use as anti-biofilm coatings. We demonstrate that we can tune the composition of the nitroxide 2,2,6,6-tetramethyl-4-piperidyl methacrylate in a copolymer with methyl methacrylate in order to control the surface concentration of nitroxides in the resulting thin films. The prepared films are able to reduce biofilm fouling by Pseudomonas aeruginosa (PAO1) at nitroxide monomer concentrations as low as 30 wt%. The nitroxide containing materials show no difference in toxicity to mammalian cells, compared to poly(methyl methacrylate), in a proliferation assay with 3T3 fibroblast cells. This is the first demonstration of surface tethered nitroxides with activity against biofilms and provides new opportunities for developing antifouling surface coatings.

Organic electrochemical transistors (OECTs) are presently a focus of intense research and hold great potential in expanding the horizons of the bioelectronics industry. The notable characteristics of OECTs, including their electrolyte-gating, which offers intimate interfacing with biological environments, and aqueous stability, make them particularly suitable to be operated within a living organism (in vivo). Unlike the existing in vivo bioelectronic devices, mostly based on rigid metal electrodes, OECTs form a soft mechanical contact with the biological milieu and ensure a high signal-to-noise ratio because of their powerful amplification capability. Such features make OECTs particularly desirable for a wide range of in vivo applications, including electrophysiological recordings, neuron stimulation, and neurotransmitter detection, and regulation of plant processes in vivo. In this review, a systematic compilation of the in vivo applications is presented that are addressed by the OECT technology. First, the operating mechanisms, and the device design and materials design principles of OECTs are examined, and then multiple examples are provided from the literature while identifying the unique device properties that enable the application progress. Finally, one critically looks at the future of the OECT technology for in vivo bioelectronic applications.

As bacterial biofilms display extreme tolerance to conventional antibiotic treatments, it has become imperative to develop new antibacterial strategies with alternative mechanisms of action. Herein, we report the synthesis of a series of ciprofloxacin-nitroxide conjugates and their corresponding methoxyamine derivatives in high yield. This was achieved by linking various nitroxides or methoxyamines to the secondary amine of the piperazine ring of ciprofloxacin using amide bond coupling. Biological evaluation of the prepared compounds on preformed P. aeruginosa biofilms in flow cells revealed substantial dispersal with ciprofloxacin-nitroxide hybrid 25, and virtually complete killing and removal (94%) of established biofilms in the presence of ciprofloxacin-nitroxide hybrid 27. Compounds 25-28 were shown to be non-toxic in both human embryonic kidney 293 (HEK 293) cells and human muscle rhabdomyosarcoma (RD) cells at concentrations up to 40 μM. Significantly, these hybrids demonstrate the potential of antimicrobial-nitroxide agents to overcome the resistance of biofilms to antimicrobials via stimulation of biofilm dispersal or through direct cell killing.

A range of profluorescent nitroxides (PFNs) were tested as probes to monitor photo-induced radical-mediated damage in polymer materials. The most stable and sensitive probe of the PFNs tested was an alkyne-linked perylenediimide PFN, 6b, with napthalimide and 9,10-bis(phenylethnyl)anthranene-based versions giving lower stability and sensitivity. Results from photo-ageing of poly(1-trimethylsilyl)-1-propyne (PTMSP) and the ethylene norbornene copolymer (TOPAS®) films doped with PFN probes demonstrated that sensors employing these support materials deliver significantly enhanced sensitivity compared to traditional techniques used to monitor photo-oxidative degradation of polymers, such as infrared spectroscopy. This enhanced sensitivity for detecting polymer damage improved methods for the determination of the serviceable application lifetime of polymers.

BODIPY is a highly versatile fluorophore for biological imaging with a tunable fluorescence emission (500 - 800 nm) that overlaps with the optically transparent window for tissue (600 - 1300 nm). Herein, we describe the synthesis of optically distinct BODIPY-based profluorescent probes bearing meso- and β-substituted isoindoline nitroxides and their corresponding methoxyamine derivatives. These profluorescent nitroxide probes possess strongly suppressed fluorescence, which can be revealed upon reduction or reaction with other radicals. Examination of the pentafluorophenylhydrazine reduction of the prepared probes using tandem EPR and fluorescence spectroscopy demonstrated that the asymmetric bis-β-substituted probe 9 (λem = 603 nm) was reduced the fastest, however, the greatest difference in fluorescence emission between the nitroxide and its reduced hydroxylamine analogue was observed for probe 7 (λem = 570 nm). The significant difference in fluorescence output between the nitroxides and their corresponding diamagnetic derivatives makes these probes ideal tools for imaging reactive oxygen species in biological systems.

We introduce the high-resolution electrospray ionization mass spectrometric (ESI MS) structural elucidation of polymerized 3,4-dihydroxy-L-phenylalanine (L-DOPA) carrying 1-methoxy-2,2,6,6-tetramethylpiperidin-4-amide groups in the side chain. Critically, our powerful ESI MS platform unambiguously revealed the structurally diverse character of this multifunctional bio-adhesive polymer system.

We pioneer a self-reporting profluorescent release system driven by the thermo-reversible dynamic covalent ligation of chromophores to a lateral polymer chain, whose fluorescence is silenced by unpaired spins of nitroxide moieties prior to release. The introduced technology platform holds significant potential for imaging substrate release events.

We introduce a mass spectrometric access route to nitroxide containing polymers via high resolution electrospray ionization mass spectrometry (HR ESI MS), a polymer class that is – due to the presence of unpaired spins – highly challenging to analyze via NMR techniques. The nitroxide content within the polymer chain structure was varied between 11.3 and 29.1 mol% in a statistical copolymer consisting of styrene-stat-4-(chloromethyl)styrene (p(S-stat-CMS), 4800 ≥ Mn/g mol−1 ≥ 11 100), where 4-carboxy-2,2,6,6-tetramethylpiperidine 1-oxyl (4-carboxy-TEMPO) units were attached by post-polymerization modification. By carefully evaluating the isotopic pattern of the nitroxide containing polymers, we demonstrate that the persistent nitroxyl radical retains its structural integrity during the soft ionization process employing spray currents up to 4.3 keV and in-source collision induced dissociation energies up to 30 eV using chloride attachment technology in negative ion mode. Interestingly, high-molecular weight gas-phase aggregates are identified with increasing amounts of nitroxide side-chain functionalization. To further exemplify the power of the introduced mass spectrometric protocol, a well-defined styrene based polymer was synthesized via atom transfer radical polymerization (ATRP, Mn = 5600 g mol−1, Đ = 1.05) containing functional groups, i.e. a terminal cyanuric acid unit and Hamilton Wedge moiety as well as a free nitroxide in the penultimate position to the Hamilton wedge. The intact persistent nitroxide radical at the chain end was unambiguously identified by its isotopic pattern in a highly defined polymer structure.

Treatment of biofilm-related Staphylococcus aureus infections represents an important medical challenge worldwide, as biofilms, even those involving drug-susceptible S. aureus strains, are highly refractory to conventional antibiotic therapy. Nitroxides were recently shown to induce the dispersal of Gram-negative biofilms in vitro, but their action against Gram-positive bacterial biofilms remains unknown., Treatment of biofilm-related Staphylococcus aureus infections represents an important medical challenge worldwide, as biofilms, even those involving drug-susceptible S. aureus strains, are highly refractory to conventional antibiotic therapy. Nitroxides were recently shown to induce the dispersal of Gram-negative biofilms in vitro, but their action against Gram-positive bacterial biofilms remains unknown. Here, we demonstrate that the biofilm dispersal activity of nitroxides extends to S. aureus, a clinically important Gram-positive pathogen. Coadministration of the nitroxide CTEMPO (4-carboxy-2,2,6,6-tetramethylpiperidin-1-yloxyl) with ciprofloxacin significantly improved the biofilm eradication activity of the antibiotic against S. aureus. Moreover, covalently linking the nitroxide to the antibiotic moiety further reduced the ciprofloxacin minimal biofilm eradication concentration. Microscopy analysis revealed that fluorescent nitroxide-antibiotic hybrids could penetrate S. aureus biofilms and enter cells localized at the surface and base of the biofilm structure. No toxicity to human cells was observed for the nitroxide CTEMPO or the nitroxide-antibiotic hybrids. Taken together, our results show that nitroxides can mediate the dispersal of Gram-positive biofilms and that dual-acting biofilm eradication antibiotics may provide broad-spectrum therapies for the treatment of biofilm-related infections.

Fluorescent probes are widely used for imaging and measuring dynamic processes in living cells. Fluorescent antibiotics are valuable tools for examining antibiotic&ndash, bacterial interactions, antimicrobial resistance and elucidating antibiotic modes of action. Profluorescent nitroxides are &lsquo, switch on&rsquo, fluorescent probes used to visualize and monitor intracellular free radical and redox processes in biological systems. Here, we have combined the inherent fluorescent and antimicrobial properties of the fluoroquinolone core structure with the fluorescence suppression capabilities of a nitroxide to produce the first example of a profluorescent fluoroquinolone-nitroxide probe. Fluoroquinolone-nitroxide (FN) 14 exhibited significant suppression of fluorescence (&gt, 36-fold), which could be restored via radical trapping (fluoroquinolone-methoxyamine 17) or reduction to the corresponding hydroxylamine 20. Importantly, FN 14 was able to enter both Gram-positive and Gram-negative bacterial cells, emitted a measurable fluorescence signal upon cell entry (switch on), and retained antibacterial activity. In conclusion, profluorescent nitroxide antibiotics offer a new powerful tool for visualizing antibiotic&ndash, bacterial interactions and researching intracellular chemical processes.

Urinary tract infections (UTIs) are amongst the most common and prevalent infectious diseases worldwide, with uropathogenic Escherichia coli (UPEC) reported as the main causative pathogen. Fluoroquinolone antibiotics are commonly used to treat UTIs but for infections involving UPEC biofilms, which are commonly associated with catheter use and recurrent episodes, ciprofloxacin is often ineffective. Here we report the development of a ciprofloxacin–dinitroxide (CDN) conjugate with potent UPEC biofilm-eradication activity. CDN 11 exhibited a 2-fold increase in potency over the parent antibiotic ciprofloxacin against UPEC biofilms. Moreover, CDN 11 resulted in almost complete UPEC biofilm cell eradication (99.7%) at concentrations as low as 12.5 μM, and significantly potentiated ciprofloxacin's biofilm-eradication activity against UPEC upon co-administration. The biofilm-eradication activity of CDN 11 highlights the potential of nitroxide functionalized antibiotics as a promising strategy for the treatment of biofilm-related UTIs.

Profluorescent nitroxides (PFNs) have emerged as an important class of imaging agents for monitoring intracellular redox status and levels of oxidative stress. However, the fast reduction of nitroxides upon incubation within cells limits the window of opportunity for detection. By increasing the concentration of nitroxides per fluorophore, their reduction to the corresponding hydroxylamines and the subsequent switch-on of fluorescence can be delayed. Herein the preparation of nitroxide-containing polymers of different chain length coupled to a fluorophore is reported and their reduction with pentafluorophenylhydrazine is examined. The fluorescence switch-on kinetics and radical concentrations are monitored by fluorescence and electron paramagnetic resonance spectroscopy and compared to a conventional PFN bearing a single nitroxide moiety. The polymeric PFNs display significant delays in reduction and fluorescence switch-on and higher turn-on ratios than their single-nitroxide counterparts. The results of this study indicate that polymeric PFNs are a promising architecture for future imaging agents.

Here we report the first example of an alkoxyamine derived from an azaphenalene nitroxide, which when exposed to UV-light, readily undergoes homolysis to efficiently and cleanly re-form a nitroxide. This process selectively cleaves the C–O bond of the alkoxyamine and occurs orders of magnitude more rapidly than any other alkoxyamine homolysis previously described in the literature. We demonstrate the use of light to generate a persistent radical effect (PRE) and apply this to undertake radical insertion, radical exchange and proof of concept polymerization reactions.

Novel profluorescent mono- and bis-isoindoline nitroxides linked to napthalimide and perylene diimide structural cores are described. These nitroxide-fluorophore probes display strongly suppressed fluorescence in comparison to their corresponding non-radical diamagnetic methoxyamine derivatives. The perylene-based probe possessing two isoindoline systems tethered through ethynyl linkages was shown to be the most photostable in solution, demonstrating significantly enhanced longevity over the 9,10-bis(phenylethynyl)anthracene fluorophore used in previous profluorescent nitroxide probes.

The utility of profluorescent nitroxides (PFNs) as sensitive probes to detect early stage photodegradation in a cyclic olefin copolymer, TOPAS®, during both natural aging and accelerated aging under laboratory conditions is reported. PFN additives in TOPAS® capture radicals to form fluorescent adducts as the material degrades. The levels of fluorescence detectable from the polymer reflect the degree of free-radical degradation in the material, with the PFN probes delivering enhanced sensitivity over traditional analytical methods for the detection of photodegradation of TOPAS®. The probes are able to highlight polymer degradation occurring within the oxidation “induction” period, where little change can be observed using infrared spectroscopy; however, their efficacy does not extend far beyond this period. The effective probe lifetime however can be significantly extended through the use of common additives such as the UV absorber (Tinuvin P) and a hindered amine stabiliser analogue (1,1,3,3-tetramethylisoindol-2-yloxyl, TMIO).

A substrate‐independent and versatile coating platform for (spatially resolved) surface functionalization, based on nitroxide radical coupling (NRC) reactions and the formation of thermo‐labile alkoxyamine functional groups, was introduced. Nitroxide‐decorated poly(glycidyl methacrylate) (PGMA) microspheres, obtained through bioinspired copolymer surface deposition using dopamine and a nitroxide functional dopamine derivative as monomers, were conjugated with small functional groups in a rewritable process. Reversible coding of the nitroxide functional microspheres by NRC and decoding through thermal alkoxyamine fission were monitored and characterized by electron paramagnetic resonance (EPR) spectroscopy and X‐ray photoelectron spectroscopy (XPS). In addition, this nitroxide coating system was exploited in “grafting‐to” polymer surface ligations of poly(methyl methacrylate) (PMMA) and poly(2,2,2‐trifluoroethyl methacrylate) (PTFEMA) in spatially confined areas. Polymer strands terminated with an Irgacure 2959 (2‐hydroxy‐4′‐(2‐hydroxyethoxy)‐2‐methylpropiophenone) photoinitiator were obtained through chain‐transfer polymerization, and subsequently coupled to nitroxide‐immobilized poly(dopamine) (PDA)‐coated silicon substrates by using rapid photoclick NRC reactions. Light‐driven polymer surface coding was visualized by time‐of‐flight secondary ion mass spectrometry (ToF‐SIMS) and XPS imaging.

Dual-acting hybrid anti-oxidant/anti-inflammatory agents were developed employing the principle of pharmacophore hybridization. Hybrid agents were synthesized by combining stable anti-oxidant nitroxides with conventional non-steroidal anti-inflammatory drugs (NSAIDs). Several of the hybrid nitroxide-NSAID conjugates displayed promising anti-oxidant and anti-inflammatory effects on two Non-Small Cell Lung Cancer (NSCLC) cells (A549 and NCI-H1299) and in ameliorating oxidative stress induced in 661 W retinal cells. One ester-linked nitroxide-aspirin analogue (27) delivered better anti-inflammatory effects (cyclooxygenase inhibition) than the parent compound (aspirin), and also showed similar reactive oxygen scavenging activity to the anti-oxidant, Tempol. In addition, a nitroxide linked to the anti-inflammatory drug indomethacin (39) significantly ameliorated the effects of oxidative stress on 661 W retinal neurons at efficacies greater or equal to the anti-oxidant Lutein. Other examples of the hybrid conjugates displayed promising anti-cancer activity, as demonstrated by their inhibitory effects on the proliferation of A549 NSCLC cells.

A simple modular strategy for the synthesis of profluorescent nitroxide containing polymers is described. The incorporation of an epoxide as a pendant functionality on a polymer backbone synthesized using ATRP and subsequent nucleophilic ring-opening with sodium azide gave hydroxyl and azide functionality within a 3-bond radius. Orthogonal coupling chemistry then allowed the independent attachment of fluorophore and nitroxide groups in close proximity, giving rise to a profluorescent polymer. Validation of the viability of these materials as fluorescent sensors is demonstrated through efficient fluorescence switch-on observed when the materials are exposed to a model reductant or carbon-centred radical source.

In aerosol research, a common approach for the collection of particulate matter (PM) is the use of filters in order to obtain sufficient material to undertake analysis. For subsequent chemical and toxicological analyses, in most of cases the PM needs to be extracted from the filters. Sonication is commonly used to most efficiently extract the PM from the filters. Extraction protocols generally involve 10 - 60 min of sonication. The energy of ultrasonic waves causes the formation and collapse of cavitation bubbles in the solution. Inside the collapsing cavities the localised temperatures and pressures can reach extraordinary values. Although fleeting, such conditions can lead to pyrolysis of the molecules present inside the cavitation bubbles (gases dissolved in the liquid and solvent vapours), which results in the production of free radicals and the generation of new compounds formed by reactions with these free radicals. For example, simple sonication of pure water will result in the formation of detectable levels of hydroxyl radicals. As hydroxyl radicals are recognised as playing key roles as oxidants in the atmosphere the extraction of PM from filters using sonication is therefore problematic. Sonication can result in significant chemical and physical changes to PM through thermal degradation and other reactions. In this article, an overview of sonication technique as used in aerosol research is provided, the capacity for radical generation under these conditions is described and an analysis is given of the impact of sonication-derived free radicals on three molecular probes commonly used by researchers in this field to detect Reactive Oxygen Species in PM.

The powerful oxidant HOCl (hypochlorous acid and its corresponding anion, (-)OCl) generated by the myeloperoxidase (MPO)-H2O2-Cl(-) system of activated leukocytes is strongly associated with multiple human inflammatory diseases; consequently there is considerable interest in inhibition of this enzyme. Nitroxides are established antioxidants of low toxicity that can attenuate oxidation in animal models, with this ascribed to superoxide dismutase or radical-scavenging activities. We have shown (M.D. Rees et al., Biochem. J. 421, 79-86, 2009) that nitroxides, including 4-amino-TEMPO (4-amino-2,2,6,6-tetramethylpiperidin-1-yloxyl radical), are potent inhibitors of HOCl formation by isolated MPO and activated neutrophils, with IC50 values of ~1 and ~6 µM respectively. The utility of tetramethyl-substituted nitroxides is, however, limited by their rapid reduction by biological reductants. The corresponding tetraethyl-substituted nitroxides have, however, been reported to be less susceptible to reduction. In this study we show that the tetraethyl species were reduced less rapidly than the tetramethyl species by both human plasma (89-99% decreased rate of reduction) and activated human neutrophils (62-75% decreased rate). The tetraethyl-substituted nitroxides retained their ability to inhibit HOCl production by MPO and activated neutrophils with IC50 values in the low-micromolar range; in some cases inhibition was enhanced compared to tetramethyl substitution. Nitroxides with rigid structures (fused oxaspiro rings) were, however, inactive. Overall, these data indicate that tetraethyl-substituted nitroxides are potent inhibitors of oxidant formation by MPO, with longer plasma and cellular half-lives compared to the tetramethyl species, potentially allowing lower doses to be employed.

Recent advances in helium ion microscopy (HIM) have enabled the use of fine-focused He+ beams to image and shape materials at the nanoscale. In addition to traditional ion milling, the beam can also be used to induce reactions, such as cross-linking, in films of organic molecules. Here, we compare the use of focused ion and electron beams to fabricate spatially-defined cross-linked features in nanometre-thick films of tetracene. Ion and electron beam treatments were performed using the focussed energetic beams in a HIM and a scanning electron microscope, respectively. The patterned samples were analysed by optical microscopy, HIM, atomic force microscopy and nanoindentation. For samples fabricated using both energetic beams, the total deposited particle dose could be used to modify the optical properties, thickness and hardness of the dosed regions. X-ray photoelectron spectroscopy revealed that the dosed regions exhibited a higher sp3 content, consistent with crosslinking; rinsing in solvent showed that the patterned regions were insoluble and could be isolated by removing the unmodified film through dissolution. These molecular nanopatterns demonstrate the promise for ultrahigh resolution chemical lithography, and for fabrication of nanocomponents with tailored physical properties.

The ability of nitric oxide (NO) to induce biofilm dispersion has been well established. Here, we investigated the effect of nitroxides (sterically hindered nitric oxide analogues) on biofilm formation and swarming motility in Pseudomonas aeruginosa . A transposon mutant unable to produce nitric oxide endogenously ( nirS ) was deficient in swarming motility relative to the wild type and the complemented strain. Moreover, expression of the nirS gene was upregulated by 9.65-fold in wild-type swarming cells compared to planktonic cells. Wild-type swarming levels were substantially restored upon the exogenous addition of nitroxide containing compounds, a finding consistent with the hypothesis that NO is necessary for swarming motility. Here, we showed that nitroxides not only mimicked the dispersal activity of NO but also prevented biofilms from forming in flow cell chambers. In addition, a nirS transposon mutant was deficient in biofilm formation relative to the wild type and the complemented strain, thus implicating NO in the formation of biofilms. Intriguingly, despite its stand-alone action in inhibiting biofilm formation and promoting dispersal, a nitroxide partially restored the ability of a nirS mutant to form biofilms.

A new method for the direct aryl iodination of isoindolines and isoindoline nitroxides which utilizes periodic acid and potassium iodide in sulfuric acid is presented. Di-iodo functionalized tetramethyl and tetraethyl isoindolines and a di-iodo tetramethyl isoindoline nitroxide were prepared in high yield (70-82%). The analogous mono-iodo species were afforded in modest yield (34-48%). Iodinated nitrones were also obtained from a tetraethyl isoindoline nitroxide.

The robust and diversely useful isoindoline nitroxide, 5-carboxy-1,1,3,3-tetramethylisoindolin-2-yloxyl (1; CTMIO), has previously been synthesised in low-to-moderate yields from phthalic anhydride (3). Recent interest in its biological potential as a potent antioxidant and in other areas has seen an increased demand for its production. Herein, three new synthetic routes to CTMIO are presented and their efficiencies assessed. Two routes, via the nitrile 9 and the formyl compound 11, derive from 5-bromo-1,1,3,3-tetramethylisoindoline (6). The third approach starts from the readily accessible starting material, 4-methylphthalic anhydride (12), and proceeds by a methylarene oxidation with potassium permanganate. The three new approaches yield CTMIO in comparable overall yields (16–18 %); however, the synthetic efficiency is most improved when employing the nitrile intermediate 9.

Particles emitted by vehicles are known to cause detrimental health effects, with their size and oxidative potential among the main factors responsible. Therefore, understanding the relationship between traffic composition and both the physical characteristics and oxidative potential of particles is critical. To contribute to the limited knowledge base in this area, we investigated this relationship in a 4.5 km road tunnel in Brisbane, Australia. On-road concentrations of ultrafine particles (

As bacterial biofilms are often refractory to conventional antimicrobials, the need for alternative and/or novel strategies for the treatment of biofilm related infections has become of paramount importance. Herein, we report the synthesis of novel hybrid molecules comprised of two different hindered nitroxides linked to the piperazinyl secondary amine of ciprofloxacin via a tertiary amine linker achieved utilising reductive amination. The corresponding methoxyamine derivatives were prepared alongside their radical-containing counterparts as controls. Subsequent biological evaluation of the hybrid compounds on preformed P. aeruginosa flow cell biofilms divulged significant dispersal and eradication abilities for ciprofloxacin-nitroxide hybrid compound 10 (up to 95% eradication of mature biofilms at 40 μM). Importantly, these hybrids represent the first dual-action antimicrobial-nitroxide agents, which harness the dispersal properties of the nitroxide moiety to circumvent the well-known resistance of biofilms to treatment with antimicrobial agents., Asian Office of Aerospace Research and Development (Grant 15IOA087-154020, Queensland University of Technology, Fundación Ramón Areces (Postdoctoral Scholarship)