Imperial College London

ProfessorAlexandraPorter

Faculty of EngineeringDepartment of Materials

Professor of Bio-imaging and Analysis
 
 
 
//

Contact

 

+44 (0)20 7594 9691a.porter

 
 
//

Location

 

B341 Royal School of MinesRoyal School of MinesSouth Kensington Campus

//

Summary

 

Publications

Publication Type
Year
to

141 results found

Gonzalez Carter D, Goode A, Kiryushko D, Masuda S, Hu S, Lopes Rodrigues R, Dexter D, Shaffer MSP, Porter AEet al., 2019, Quantification of blood-brain barrier transport and neuronal toxicity of unlabelled multiwalled carbon nanotubes as a function of surface charge, Nanoscale, ISSN: 2040-3364

Nanoparticles capable of penetrating the blood-brain barrier (BBB) will greatly advance the delivery of therapies against brain disorders. Carbon nanotubes hold great potential as delivery vehicles due to their high aspect-ratio and cell-penetrating ability. Studies have shown multiwalled carbon nanotubes (MWCNT) cross the BBB, however they have largely relied on labelling methods to track and quantify transport, or on individual electron microscopy images to qualitatively assess transcytosis. Therefore, new direct and quantitative methods, using well-defined and unlabelled MWCNT, are needed to compare BBB translocation of different MWCNT types. Using highly controlled anionic (-), cationic (+) and non-ionic (0) functionalized MWCNT (fMWCNT), we correlate UV-visible spectroscopy with quantitative transmission electron microscopy, quantified from c. 270 endothelial cells, to examine cellular uptake, BBB transport and neurotoxicity of unlabelled fMWCNT. Our results demonstrate that: i) a large fraction of cationic and non-ionic, but not anionic fMWCNT become trapped at the luminal brain endothelial cell membrane; ii) despite high cell association, fMWCNT uptake by brain endothelial cells is low (< 1.5% ID) and does not correlate with BBB translocation, iii) anionic fMWCNT have highest transport levels across an in vitro model of the human BBB compared to non-ionic or cationic nanotubes; and iv) fMWCNT are not toxic to hippocampal neurons at relevant abluminal concentrations; however, fMWCNT charge has an effect on carbon nanotube neurotoxicity at higher fMWCNT concentrations. This quantitative combination of microscopy and spectroscopy, with cellular assays, provides a crucial strategy to predict brain penetration efficiency and neurotoxicity of unlabelled MWCNT and other nanoparticle technologies relevant to human health.

Journal article

Gomez-Gonzalez MA, Koronfel MA, Goode AE, Al-Ejji M, Voulvoulis N, Parker JE, Quinn PD, Scott TB, Xie F, Yallop ML, Porter AE, Ryan MPet al., 2019, Spatially resolved dissolution and speciation changes of ZnO nanorods during short-term in situ incubation in a simulated wastewater environment, ACS Nano, Vol: 13, Pages: 11049-11061, ISSN: 1936-0851

Zinc oxide engineered nanomaterials (ZnO ENMs) are used in a variety of applications worldwide due to their optoelectronic and antibacterial properties with potential contaminant risk to the environment following their disposal. One of the main potential pathways for ZnO nanomaterials to reach the environment is via urban wastewater treatment plants. So far there is no technique that can provide spatiotemporal nanoscale information about the rates and mechanisms by which the individual nanoparticles transform. Fundamental knowledge of how the surface chemistry of individual particles change, and the heterogeneity of transformations within the system, will reveal the critical physicochemical properties determining environmental damage and deactivation. We applied a methodology based on spatially resolved in situ X-ray fluorescence microscopy (XFM), allowing observation of real-time dissolution and morphological and chemical evolution of synthetic template-grown ZnO nanorods (∼725 nm length, ∼140 nm diameter). Core-shell ZnO-ZnS nanostructures were formed rapidly within 1 h, and significant amounts of ZnS species were generated, with a corresponding depletion of ZnO after 3 h. Diffuse nanoparticles of ZnS, Zn3(PO4)2, and Zn adsorbed to Fe-oxyhydroxides were also imaged in some nonsterically impeded regions after 3 h. The formation of diffuse nanoparticles was affected by ongoing ZnO dissolution (quantified by inductively coupled plasma mass spectrometry) and the humic acid content in the simulated sludge. Complementary ex situ X-ray absorption spectroscopy and scanning electron microscopy confirmed a significant decrease in the ZnO contribution over time. Application of time-resolved XFM enables predictions about the rates at which ZnO nanomaterials transform during their first stages of the wastewater treatment process.

Journal article

O'Connell RA, Porter AE, Higgins JS, Cabral JTet al., 2019, Phase behaviour of poly(2, 6-diphenyl-p-phenylene oxide) (PPPO) in mixed solvents, Polymer, Vol: 180, Pages: 121652-121652, ISSN: 0032-3861

Journal article

Leo BF, Fearn S, Gonzalez-Carter D, Theodorou I, Ruenraroengsak P, Goode A, Mcphail D, Dexter DT, Shaffer MSP, Chung KF, Porter AE, Ryan MPet al., 2019, Label-free TOF-SIMS imaging of sulfur producing enzymes inside microglia cells following exposure to silver nanowires, Analytical Chemistry, Vol: 91, Pages: 11098-11107, ISSN: 0003-2700

There are no methods sensitive enough to detect enzymes within cells, without the use of analyte labelling. Here we show that it is possible to detect protein ion signals of three different H2S-synthesizing enzymes inside microglia after pre-treatment with silver nanowires (AgNW) using time of flight-secondary ion mass spectrometry (TOF-SIMS). Protein fragment ions, including the fragment of amino acid (C4H8N+ - 70 amu), fragments of the sulfur producing cystathionine-containing enzymes and the Ag+ ion signal could be detected without the use of any labels; the cells were mapped using the C4H8N+ amino acid fragment. Scanning electron microscopy imaging and energy dispersive x-ray chemical analysis showed that the AgNWs were inside the same cells imaged by TOF-SIMS and transformed chemically into crystalline Ag2S within cells in which the sulfur producing proteins were detected. The presence of these sulfur producing cystathionine-containing enzymes within the cells was confirmed by Western Blots and confocal microscopy images of fluorescently labelled antibodies against the sulfur producing enzymes. Label-free ToF-SIMS is very promising for the label-free identification of H2S-contributing enzymes and their cellular localization in biological systems. The technique could in future be used to identify which of these enzymes are most contributory.

Journal article

Ruenraroengsak P, Kiryushko D, Theodorou IG, Klosowski MM, Taylor ER, Niriella T, Palmieri C, Yagüe E, Ryan MP, Coombes RC, Xie F, Porter AEet al., 2019, Frizzled-7-targeted delivery of zinc oxide nanoparticles to drug-resistant breast cancer cells, Nanoscale, Vol: 11, Pages: 12858-12870, ISSN: 2040-3364

There is a need for novel strategies to treat aggressive breast cancer subtypes and overcome drug resistance. ZnO nanoparticles (NPs) have potential in cancer therapy due to their ability to potently and selectively induce cancer cell apoptosis. Here, we tested the in vitro chemotherapeutic efficacy of ZnONPs loaded via a mesoporous silica nanolayer (MSN) towards drug-sensitive breast cancer cells (MCF-7: estrogen receptor-positive, CAL51: triple-negative) and their drug-resistant counterparts (MCF-7TX, CALDOX). ZnO-MSNs were coated on to gold nanostars (AuNSs) for future imaging capabilities in the NIR-II range. Electron and confocal microscopy showed that MSN-ZnO-AuNSs accumulated close to the plasma membrane and were internalized by cells. High-resolution electron microscopy showed that MSN coating degraded outside the cells, releasing ZnONPs that interacted with cell membranes. MSN-ZnO-AuNSs efficiently reduced the viability of all cell lines, and CAL51/CALDOX cells were more susceptible than MCF7/MCF-7-TX cells. MSN-ZnO-AuNSs were then conjugated with the antibody to Frizzled-7 (FZD-7), the receptor upregulated by several breast cancer cells. We used the disulphide (S-S) linker that could be cleaved with a high concentration of glutathione normally observed within cancer cells, releasing Zn2+ into the cytoplasm. FZD-7 targeting resulted in approximately three-fold amplified toxicity of MSN-ZnO-AuNSs towards the MCF-7TX drug-resistant cell line with the highest FZD-7 expression. This study shows that ZnO-MSs are promising tools to treat triple-negative and drug-resistant breast cancers and highlights the potential clinical utility of FZD-7 for delivery of nanomedicines and imaging probes specifically to these cancer types.

Journal article

Parichart N, Tsigkou O, Li S, Porter A, Jones Jet al., 2019, Human mesenchymal stem cells (hMSCs) differentiate into an osteogenic lineage in presence of strontium containing bioactive glass nanoparticles, Acta Biomaterialia, Vol: 90, Pages: 373-392, ISSN: 1742-7061

While bioactive glass and ions released during its dissolution are known to stimulate osteoblast cells, the effect bioactive glass has on human stem cells is not clear. Here, we show that spherical monodispersed strontium containing bioactive nanoparticles (Sr-BGNPs) of composition 90.6 mol% SiO2, 5.0 mol% CaO, 4.4% mol% SrO (4.4%Sr-BGNPs) and 88.8 mol% SiO2, 1.8 mol% CaO, and 9.4 mol% SrO (9.4%Sr-BGNPs) stimulate bone marrow derived human stem cell (hMSC) differentiation down an osteogenic pathway without osteogenic supplements. The particles were synthesised using a modified Stӧber process and had diameters of 90 ± 10 nm. Previous work on similar particles that did not contain Sr (80 mol% SiO2, 20 mol% CaO) showed stem cells did not differentiate when exposed to the particles. Here, both compositions of the Sr-BGNPs (up to concentration of 250 μg/mL) stimulated the early-, mid-, and late-stage markers of osteogenic differentiation and accelerated mineralisation in the absence of osteogenic supplements. Sr ions play a key role in osteogenic stem cell differentiation. Sr-BGNP dissolution products did not adversely affect hMSC viability and no significant differences in viability were measured between each particle composition. Confocal and transmission electron microscopy (TEM) demonstrated that monodispersed Sr- BGNPs were internalised and localised within vesicles in the cytoplasm of hMSCs. Degradation of particles inside the cells was observed, whilst maintaining effective cations (Ca and Sr) in their silica network after 24 h in culture. The uptake of Sr-BGNPs by hMSCs was reduced by inhibitors of specific routes of endocytosis, indicating that the Sr-BGNPs uptake by hMSCs was probably via mixed endocytosis mechanisms. Sr-BGNPs have potential as injectable therapeutic devices for bone regeneration or treatment of conditions such as osteoporosis, because of their ability deliver a sustained release of osteogenic inorganic cations, e.g. calcium (Ca) or a

Journal article

Koronfel MA, Goode AE, Gomez-Gonzalez MA, Nelson Weker J, Simões TA, Brydson R, Quinn P, Toney MF, Hart A, Porter AE, Ryan MPet al., Chemical Evolution of CoCrMo Wear Particles: an In-Situ Characterisation Study, The Journal of Physical Chemistry C, ISSN: 1932-7447

Journal article

Singh M, Nabavi E, Zhou Y, Gallina ME, Zhao H, Ruenraroengsak P, Porter AE, Ma D, Cass AEG, Hanna GB, Elson DSet al., 2019, Laparoscopic fluorescence image-guided photothermal therapy enhances cancer diagnosis and treatment, Nanotheranostics, Vol: 3, Pages: 89-102, ISSN: 2206-7418

Endoscopy is the gold standard investigation in the diagnosis of gastrointestinal cancers and the management of early and pre-malignant lesions either by resection or ablation. Recently gold nanoparticles have shown promise in cancer diagnosis and therapeutics (theranostics). The combination of multifunctional gold nanoparticles with near infrared fluorescence endoscopy for accurate mapping of early or pre-malignant lesions can potentially enhance diagnostic efficiency while precisely directing endoscopic near infrared photothermal therapy for established cancers. The integration of endoscopy with near infrared fluorescence imaging and photothermal therapy was aided by the accumulation of our multifunctionalized PEG-GNR-Cy5.5-anti-EGFR-antibody gold nanorods within gastrointestinal tumor xenografts in BALB/c mice. Control mice (with tumors) received either gold nanorods or photothermal therapy, while study mice received both treatment modalities. Local (tumor-centric) and systemic effects were examined for 30 days. Clear endoscopic near infrared fluorescence signals were observed emanating specifically from tumor sites and these corresponded precisely to the tumor margins. Endoscopic fluorescence-guided near infrared photothermal therapy successfully induced tumor ablations in all 20 mice studied, with complete histological clearance and minimal collateral damage. Multi-source analysis from histology, electron microscopy, mass spectrometry, blood, clinical evaluation, psychosocial and weight monitoring demonstrated the inherent safety of this technology. The combination of this innovative nanotechnology with gold standard clinical practice will be of value in enhancing the early optical detection of gastrointestinal cancers and a useful adjunct for its therapy.

Journal article

Theodorou I, Ruenraroengsak P, Carter D, Jiang Q, Yague E, Aboagye E, Coombes RC, Porter AE, Ryan M, Xie Fet al., 2019, Towards multiplexed near-infrared cellular imaging using gold nanostar arrays with tunable fluorescence enhancement, Nanoscale, Vol: 11, Pages: 2079-2088, ISSN: 2040-3364

Sensitive detection of disease biomarkers expressed by human cells is critical to the development of novel diagnostic and therapeutic methods. Here we report that plasmonic arrays based on gold nanostar (AuNS) monolayers enable up to 19-fold fluorescence enhancement for cellular imaging in the near-infrared (NIR) biological window, allowing the application of low quantum yield fluorophores for sensitive cellular imaging. The high fluorescence enhancement together with low autofluorescence interference in this wavelength range enable higher signal-to-noise ratio compared to other diagnostic modalities. Using AuNSs of different geometries and therefore controllable electric field enhancement, cellular imaging with tunable enhancement factors is achieved, which may be useful for the development of multicolour and multiplexed platforms for a panel of biomarkers, allowing to distinguish different subcell populations at the single cell level. Finally, the uptake of AuNSs within HeLa cells and their high biocompatibility, pave the way for novel high-performance in vitro and in vivo diagnostic platforms.

Journal article

Gonzalez-Carter DA, Ong ZY, McGilvery CM, Dunlop IE, Dexter DT, Porter AEet al., 2019, L-DOPA functionalized, multi-branched gold nanoparticles as brain-targeted nano-vehicles, Nanomedicine: Nanotechnology, Biology and Medicine, Vol: 15, Pages: 1-11, ISSN: 1549-9634

The blood-brain barrier (BBB) is a protective endothelial barrier lining the brain microvasculature which prevents brain delivery of therapies against brain diseases. Hence, there is an urgent need to develop vehicles which efficiently penetrate the BBB to deliver therapies into the brain. The drug L-DOPA efficiently and specifically crosses the BBB via the large neutral amino acid transporter (LAT)-1 protein to enter the brain. Thus, we synthesized L-DOPA-functionalized multi-branched nanoflower-like gold nanoparticles (L-DOPA-AuNFs) using a seed-mediated method involving catechols as a direct reducing-cum-capping agent, and examined their ability to cross the BBB to act as brain-penetrating nanovehicles. We show that L-DOPA-AuNFs efficiently penetrate the BBB compared to similarly sized and shaped AuNFs functionalized with a non-targeting ligand. Furthermore, we show that L-DOPA-AuNFs are efficiently internalized by brain macrophages without inducing inflammation. These results demonstrate the application of L-DOPA-AuNFs as a non-inflammatory BBB-penetrating nanovehicle to efficiently deliver therapies into the brain.

Journal article

Theodorou I, Jiang Q, Malms L, Xie X, Coombes RC, Aboagye E, Porter AE, Ryan M, Xie Fet al., 2018, Fluorescence enhancement from single gold nanostars: towards ultra-bright emission in the first and second near-infrared biological windows, Nanoscale, Vol: 10, Pages: 15854-15864, ISSN: 2040-3364

Gold nanostars (AuNSs) are promising agents for the development of high-performance diagnostic devices, by enabling metal enhanced fluorescence (MEF) in the physiological near-infrared (NIR) and second near-infrared (NIR-II) windows. The local electric field near their sharp tips and between their branches can be enhanced by several orders of magnitude, holding great promise for large fluorescence enhancements from single AuNS particles, rather than relying on interparticle coupling in nanoparticle substrates. Here, guided by electric field simulations, two different types of AuNSs with controlled morphologies and plasmonic responses in the NIR and NIR-II regions are used to investigate the mechanism of MEF from colloidal AuNSs. Fluorophore conjugation to AuNSs allows significant fluorescence enhancement of up to 30 times in the NIR window, and up to 4-fold enhancement in the NIR-II region. Together with other inherent advantages of AuNSs, including their multispike morphology offering easy access to cell membranes and their large surface area providing flexible multifunctionality, AuNS are promising for the development of in vivo imaging applications. Using time-resolved fluorescence measurements to deconvolute semi-quantitatively excitation enhancement from emission enhancement, we show that a combination of enhanced excitation and an increased radiative decay rate, both contribute to the observed large enhancement. In accordance to our electric field modelling, however, excitation enhancement is the component that varies most with particle morphology. These findings provide important insights into the mechanism of MEF from AuNSs, and can be used to further guide particle design for high contrast enhancement, enabling the development of MEF biodetection technologies.

Journal article

Kiryushko D, Pankratova S, Klingelhofer J, Dmytriyeva O, Owczarek S, Renziehausen A, Syed N, Porter A, Dexter D, Kiryushko Det al., 2018, The S100A4 protein signals through the ErbB4 receptor to promote neuronal survival., Theranostics, Vol: 8, Pages: 3977-3990, ISSN: 1838-7640

Understanding the mechanisms of neurodegeneration is crucial for development of therapies to treat neurological disorders. S100 proteins are extensively expressed in the injured brain but S100's roleand signalling in neural cells remain elusive. We recently demonstrated that the S100A4 protein protects neurons in brain injury and designed S100A4-derived peptides mimicking its beneficial effects. Here we show that neuroprotection by S100A4 involves the growth factor family receptorErbB4 and its ligand Neuregulin 1 (NRG), key regulators of neuronal plasticity and implicated in multiple brain pathologies. The neuroprotective effect of S100A4 depends on ErbB4 expression andthe ErbB4 signalling partners ErbB2/Akt, and is reduced by functional blockade of NRG/ErbB4 in cell models of neurodegeneration. We also detect binding of S100A4 with ErbB1 (EGFR) and ErbB3. S100A4-derived peptides interact with, and signal through ErbB, are neuroprotective inprimary and immortalized dopaminergic neurons, and do not affect cell proliferation/motility - features which make them promising as potential neuroprotectants. Our data suggest that the S100- ErbB axis may be an important mechanism regulating neuronal survival and plasticity

Journal article

Ellis T, Chiappi M, García-Trenco A, Al-Ejji M, Sarkar S, Georgiou TK, Shaffer MSP, Tetley TD, Schwander S, Ryan MP, Porter AEet al., 2018, Multimetallic microparticles increase the potency of rifampicin against intracellular mycobacterium tuberculosis, ACS Nano, Vol: 12, Pages: 5228-5240, ISSN: 1936-0851

Mycobacterium tuberculosis ( M.tb) has the extraordinary ability to adapt to the administration of antibiotics through the development of resistance mechanisms. By rapidly exporting drugs from within the cytosol, these pathogenic bacteria diminish antibiotic potency and drive the presentation of drug-tolerant tuberculosis (TB). The membrane integrity of M.tb is pivotal in retaining these drug-resistant traits. Silver (Ag) and zinc oxide (ZnO) nanoparticles (NPs) are established antimicrobial agents that effectively compromise membrane stability, giving rise to increased bacterial permeability to antibiotics. In this work, biodegradable multimetallic microparticles (MMPs), containing Ag NPs and ZnO NPs, were developed for use in pulmonary delivery of antituberculous drugs to the endosomal system of M.tb-infected macrophages. Efficient uptake of MMPs by M.tb-infected THP1 cells was demonstrated using an in vitro macrophage infection model, with direct interaction between MMPs and M.tb visualized with the use of electron FIB-SEM tomography. The release of Ag NPs and ZnO NPs within the macrophage endosomal system increased the potency of the model antibiotic rifampicin by as much as 76%, realized through an increase in membrane disorder of intracellular M.tb. MMPs were effective at independently driving membrane destruction of extracellular bacilli located at the exterior face of THP1 macrophages. This MMP system presents as an effective drug delivery vehicle that could be used for the transport of antituberculous drugs such as rifampicin to infected alveolar macrophages, while increasing drug potency. By increasing M.tb membrane permeability, such a system may prove effectual in improving treatment of drug-susceptible TB in addition to M.tb strains considered drug-resistant.

Journal article

Botelho D, Leo BF, Massa C, Sarkar S, Tetley T, Chung KF, Chen S, Ryan MP, Porter A, Atochina-Vasserman EN, Zhang J, Schwander S, Gow AJet al., 2018, Exposure to silver nanospheres leads to altered respiratory mechanics and delayed immune response in an in vivo Murine model, Frontiers in Pharmacology, Vol: 9, ISSN: 1663-9812

Here we examine the organ level toxicology of both carbon black (CB) and silver nanoparticles (AgNP). We aim to determine metal-specific effects to respiratory function, inflammation and potential interactions with lung lining fluid (LLF). C57Bl6/J male mice were intratracheally instilled with saline (control), low (0.05 μg/g) or high (0.5 μg/g) doses of either AgNP or CB 15 nm nanospheres. Lung histology, cytology, surfactant composition and function, inflammatory gene expression, and pulmonary function were measured at 1, 3, and 7 days post-exposure. Acutely, high dose CB resulted in an inflammatory response, increased neutrophilia and cytokine production, without alteration in surfactant composition or respiratory mechanics. Low dose CB had no effect. Neither low nor high dose AgNPs resulted in an acute inflammatory response, but there was an increase in work of breathing. Three days post-exposure with CB, a persistent neutrophilia was noted. High dose AgNP resulted in an elevated number of macrophages and invasion of lymphocytes. Additionally, AgNP treated mice displayed increased expression of IL1B, IL6, CCL2, and IL10. However, there were no significant changes in respiratory mechanics. At day 7, inflammation had resolved in AgNP-treated mice, but tissue stiffness and resistance were significantly decreased, which was accompanied by an increase in surfactant protein D (SP-D) content. These data demonstrate that the presence of metal alters the response of the lung to nanoparticle exposure. AgNP-surfactant interactions may alter respiratory function and result in a delayed immune response, potentially due to modified airway epithelial cell function.

Journal article

Koronfel MA, Goode AE, Weker JN, Tay SER, Stitt CA, Simoes TA, Mosselmans JFW, Quinn P, Brydson R, Hart A, Toney MF, Porter AE, Ryan MPet al., 2018, Understanding the reactivity of CoCrMo-implant wear particles, npj Materials Degradation, Vol: 2, ISSN: 2397-2106

CoCrMo-based metal-on-metal hip implants experienced unexpectedly high failure rates despite the high wear and corrosion resistance of the bulk material. Although they exhibit a lower volumetric wear compared to other implant materials, CoCrMo-based implants produced a significantly larger 'number' of smaller wear particles. CoCrMo is nominally an extremely stable material with high Cr content providing passivity. However, despite the Co:Cr ratio in the original alloy being 2:1; chemical analyses of wear particles from periprosthetic tissue have found the particles to be composed predominately of Cr species, with only trace amounts of Co remaining. Here a correlative spectroscopy and microscopy approach has shown that these particles dissolve via a non-stoichiometric, and geometrically inhomogeneous, mechanism similar to de-alloying. This mechanism is previously unreported for this material and was not apparent in any of the regulatory required tests, suggesting that such tests are insufficiently discriminating.

Journal article

Kenney JPL, Ellis T, Nicol FS, Porter AE, Weiss DJet al., 2018, The effect of bacterial growth phase and culture concentration on U(VI) removal from aqueous solution, Chemical Geology, Vol: 482, Pages: 61-71, ISSN: 0009-2541

Bacteria play a key role in controlling the mobility of contaminants, such as uranium (U), in the environment. Uranium could be sourced from disposed radioactive waste, derived either from surface disposal trenches for Low Level Waste (LLW) that, because of the waste type and disposal concept, would typically present acidic conditions or from the geological disposal of LLW or Intermediate Level Waste (ILW) that, because of the waste type and the disposal concept, would typically present alkaline conditions. In disposed radioactive waste, there could be variable amounts of cellulosic material. Bacterial cells may be living in a range of different growth phases, depending on the growth conditions and nutrients available at the time any waste-derived U migrated to the cells. A key knowledge gap to date has been the lack of a mechanistic understanding of how bacterial growth phases (exponential, stationary, and death phase) affect the ability of bacteria to remove U(VI) from solution. To address this, we first characterised the cells using potentiometric titrations to detect any differences in proton binding to proton active sites on Pseudomonas putida cells at each growth phase under aerobic conditions, or under anaerobic conditions favourable to U(IV) reoxidation. We then conducted batch U(VI) removal experiments with bacteria at each phase suspended in 1 and 10 ppm U aqueous solutions with the pH adjusted from 2 to 12 as well as with culture concentrations from 0.01 to 10 g/L, to identify the minimal concentration of bacteria in solution necessary to affect U removal. We found that, in death phase, P. putida cells exhibited double the concentration of proton active sites than bacteria grown to exponential and stationary phase. However, we did not see a difference in the extent of U(VI) removal, from a 10 ppm U solution, between the different growth phases as a function of pH (2 to 12). Culture concentration affected U removal between pH 2–8, where U removal dec

Journal article

Klosowski M, Carzaniga R, Shefelbine S, Porter A, McComb Det al., 2018, Nanoanalytical electron microscopy of events predisposing to mineralisation of turkey tendon, Scientific Reports, Vol: 8, ISSN: 2045-2322

The macro- and micro-structures of mineralised tissues hierarchy are well described and understood. However, investigation of their nanostructure is limited due to the intrinsic complexity of biological systems. Preceding transmission electron microscopy studies investigating mineralising tissues have not resolved fully the initial stages of mineral nucleation and growth within the collagen fibrils. In this study, analytical scanning transmission electron microscopy and electron energy-loss spectroscopy were employed to characterise the morphology, crystallinity and chemistry of the mineral at different stages of mineralization using a turkey tendon model. In the poorly mineralised regions, calcium ions associated with the collagen fibrils and ellipsoidal granules and larger clusters composed of amorphous calcium phosphate were detected. In the fully mineralised regions, the mineral had transformed into crystalline apatite with a plate-like morphology. A change in the nitrogen K-edge was observed and related to modifications of the functional groups associated with the mineralisation process. This transformation seen in the nitrogen K-edge might be an important step in maturation and mineralisation of collagen and lend fundamental insight into how tendon mineralises.

Journal article

Nobakhti S, Depalle B, Porter A, Shefelbine Set al., 2017, The role of osteopontin in the mechanics of the bone across length-scales., Annual Meeting of the American-Society-for-Bone-and-Mineral-Research (ASBMR), Publisher: WILEY, Pages: S190-S190, ISSN: 0884-0431

Conference paper

Ong ZY, Chen S, Nabavi E, Regoutz A, Payne DJ, Elson DS, Dexter DT, Dunlop IE, Porter AEet al., 2017, Multibranched Gold Nanoparticles with Intrinsic LAT-1 Targeting Capabilities for Selective Photothermal Therapy of Breast Cancer., ACS Applied Materials and Interfaces, Vol: 9, Pages: 39259-39270, ISSN: 1944-8244

Because of the critical role of the large neutral amino acid transporter-1 (LAT-1) in promoting tumor growth and proliferation, it is fast emerging as a highly attractive biomarker for the imaging and treatment of human malignancies, including breast cancer. While multibranched gold nanoparticles (AuNPs) have emerged as a promising modality in the photothermal therapy (PTT) of cancers, some of the key challenges limiting their clinical translation lie in the need to develop reproducible and cost-effective synthetic methods as well as the selective accumulation of sufficient AuNPs at tumor sites. In this study, we report a simple and direct seed-mediated synthesis of monodispersed multibranched AuNPs using the catechol-containing LAT-1 ligands, L- and D-dopa, to confer active cancer targeting. This route obviates the need for additional conjugation with targeting moieties such as peptides or antibodies. Nanoflower-like AuNPs (AuNF) with diameters of approximately 46, 70, and 90 nm were obtained and were found to possess excellent colloidal stability and biocompatibility. A significantly higher intracellular accumulation of the L- and D-dopa functionalized AuNFs was observed in a panel of breast cancer cell lines (MCF-7, MDA-MB-231, MDA-MB-468, and MDA-MB-453) when compared to the nontargeting control AuNFs synthesized with dopamine and 4-ethylcatechol. Importantly, no significant difference in uptake between the targeting and nontargeting AuNFs was observed in a non-tumorigenic MCF-10A breast epithelial cell line, hence demonstrating tumor selectivity. For PTT of breast cancer, Ag(+) was introduced during synthesis to obtain L-dopa functionalized nanourchin-like AuNPs (AuNUs) with strong near-infrared (NIR) absorbance. The L-dopa functionalized AuNUs mediated selective photothermal ablation of the triple negative MDA-MB-231 breast cancer cell line and sensitized the cells to the anticancer drugs cisplatin and docetaxel. This work brings forward an effective strategy

Journal article

Naruphontjirakul P, Porter AE, Jones JR, 2017, In vitro osteogenesis by intracellular uptake of strontium containing bioactive glass nanoparticles., Acta Biomaterialia, Vol: 66, Pages: 67-80, ISSN: 1742-7061

Monodispersed strontium containing bioactive glass nanoparticles (Sr-BGNPs) with two compositions were synthesised, through a modified sol-gel Stöber process, wherein silica nanoparticles (SiO2-NPs) were formed prior to incorporation of calcium and strontium, with diameters of 90 ± 10 nm. The osteogenic response of a murine preosteoblast cell line, MC3T3-E1, was investigated in vitro for a nanoparticle concentration of 250 µg/mL with compositions of 87 mol% SiO2, 7 mol% CaO, 6 mol% SrO and 83 mol% SiO2, 3 mol% CaO, 14 mol% SrO. Dissolution studies in minimum essential media (α-MEM) at pH 7.4 and artificial lysosomal fluid (ALF) at pH 4.5 showed that the particles dissolved and that Sr2+ ions were released from Sr-BGNPs in both environments. Both particle compositions and their ionic dissolution products enhanced the alkaline phosphatase (ALP) activity of the cells and calcium deposition. Immunohistochemistry (IHC) staining of Col1a1, osteocalcin (OSC) and osteopontin (OSP) showed that these proteins were expressed in the MC3T3-E1 cells following three weeks of culture. In the basal condition, the late osteogenic differentiation markers, OSC and OSP, were more overtly expressed by cells cultured with Sr-BGNPs with 14 mol% SrO and their ionic release products than in the control condition. Col1a1 expression was only slightly enhanced in the basal condition, but was enhanced further by the osteogenic supplements. These data demonstrate that Sr-BGNPs accelerate mineralisation without osteogenic supplements. Sr-BGNPs were internalised into MC3T3-E1 cells by endocytosis and stimulated osteogenic differentiation of the pre-osteoblast cell line. Sr-BGNPs are likely to be beneficial for bone regeneration and the observed osteogenic effects of these particles can be attributed to their ionic release products. STATEMENT OF SIGNIFICANCE: We report, for the first time, that monodispersed bioactive glass nanoparticles (∼90 nm) are internalised into pre

Journal article

Harris-Birtill D, Singh M, Zhou Y, Shah A, Ruenraroengsak P, Gallina ME, Hanna GB, Cass AEG, Porter AE, Bamber J, Elson DSet al., 2017, Gold nanorod reshaping in vitro and in vivo using a continuous wave laser., PLoS ONE, Vol: 12, ISSN: 1932-6203

Gold nanorods (GNRs) are increasingly being investigated for cancer theranostics as they possess features which lend themselves in equal measures as contrast agents and catalysts for photothermal therapy. Their optical absorption spectral peak wavelength is determined by their size and shape. Photothermal therapy using GNRs is typically established using near infrared light as this allows sufficient penetration into the tumour matrix. Continuous wave (CW) lasers are the most commonly applied source of near infrared irradiation on GNRs for tumour photothermal therapy. It is perceived that large tumours may require fractionated or prolonged irradiation. However the true efficacy of repeated or protracted CW irradiation on tumour sites using the original sample of GNRs remains unclear. In this study spectroscopy and transmission electron microscopy are used to demonstrate that GNRs reshape both in vitro and in vivo after CW irradiation, which reduces their absorption efficiency. These changes were sustained throughout and beyond the initial period of irradiation, resulting from a spectral blue-shift and a considerable diminution in the absorption peak of GNRs. Solid subcutaneous tumours in immunodeficient BALB/c mice were subjected to GNRs and analysed with electron microscopy pre- and post-CW laser irradiation. This phenomenon of thermally induced GNR reshaping can occur at relatively low bulk temperatures, well below the bulk melting point of gold. Photoacoustic monitoring of GNR reshaping is also evaluated as a potential clinical aid to determine GNR absorption and reshaping during photothermal therapy. Aggregation of particles was coincidentally observed following CW irradiation, which would further diminish the subsequent optical absorption capacity of irradiated GNRs. It is thus established that sequential or prolonged applications of CW laser will not confer any additional photothermal effect on tumours due to significant attenuations in the peak optical absorpt

Journal article

Luongo G, Perez JE, Kosel J, Georgiou TK, Regoutz A, Payne DJ, Stevens MM, Porter AE, Dunlop IEet al., 2017, Scalable high-affinity stabilization of magnetic iron oxide nanostructures by a biocompatible antifouling homopolymer, ACS Applied Materials and Interfaces, Vol: 9, Pages: 40059-40069, ISSN: 1944-8244

Iron oxide nanostructures have been widely developed for biomedical applications, due to their magnetic properties and biocompatibility. In clinical application, the stabilization of these nanostructures against aggregation and non-specific interactions is typically achieved using weakly anchored polysaccharides, with better-defined and more strongly anchored synthetic polymers not commercially adopted due to complexity of synthesis and use. Here, we show for the first time stabilization and biocompatibilization of iron oxide nanoparticles by a synthetic homopolymer with strong surface anchoring and a history of clinical use in other applications, poly(2-methacryloyloxyethy phosphorylcholine) (poly(MPC)). For the commercially important case of spherical particles, binding of poly(MPC) to iron oxide surfaces and highly effective individualization of magnetite nanoparticles (20 nm) are demonstrated. Next-generation high-aspect ratio nanowires (both magnetite/maghemite and core-shell iron/iron oxide) are furthermore stabilized by poly(MPC)-coating, with nanowire cytotoxicity at large concentrations significantly reduced. The synthesis approach is exploited to incorporate functionality into the poly(MPC) chain is demonstrated by random copolymerization with an alkyne-containing monomer for click-chemistry. Taking these results together, poly(MPC) homopolymers and random copolymers offer a significant improvement over current iron oxide nanoformulations, combining straightforward synthesis, strong surface-anchoring and well-defined molecular weight.

Journal article

Theodorou IG, Muller KH, Chen S, Goode AE, Yufit V, Ryan MP, Porter AEet al., 2017, Silver Nanowire Particle Reactivity with Human Monocyte-Derived Macrophage Cells: Intracellular Availability of Silver Governs Their Cytotoxicity, ACS BIOMATERIALS SCIENCE & ENGINEERING, Vol: 3, Pages: 2336-2347, ISSN: 2373-9878

Silver nanowires (AgNWs) are increasingly being used in the production of optoelectronic devices, with manufacturing processes posing a risk for occupational exposures via inhalation. Although some studies have explored the environmental effects of AgNWs, few data exist on human health effects. Alveolar macrophages are central in the clearance of inhaled fibers from the lungs, with frustrated phagocytosis often stated as a key determinant for the onset of inflammatory reactions. However, the mechanisms through which fully ingested AgNWs interact with, degrade, and transform within primary macrophages over time, and whether the reactivity of the AgNWs arises due to ionic or particulate effects, or both, are poorly understood. Here, a combination of elemental quantification, 3D tomography, analytical transmission electron microscopy (TEM), and confocal microscopy were employed to monitor the uptake, intracellular Ag+ availability, and processing of AgNWs of two different lengths (1 and 10 μm) inside human monocyte-derived macrophages (HMMs). Using AgNO3 and spherical silver nanoparticles (AgNPs) as a comparison, the amount of total bioavailable/intracellular Ag highly correlated to the cytotoxicity of AgNWs. The 10 μm AgNWs were completely internalized in HMMs, with numerous lysosomal vesicles observed in close vicinity to the AgNWs. Following cellular uptake, AgNWs dissolved and transformed intracellularly, with precipitation of AgCl as well as Ag2S. These transformation processes were likely due to AgNW degradation in the acidic environment of lysosomes, leading to the release of Ag+ ions that rapidly react with Cl– and SH– species of the cell microenvironment. Our data suggest that, in HMMs, not only frustrated phagocytosis but also the extent of intracellular uptake and dissolution of AgNWs dictates their cytotoxicity.

Journal article

Theodorou I, Jawad Z, Jiang Q, Aboagye E, Porter A, Ryan M, Xie Fet al., 2017, Gold Nanostar Substrates for Metal Enhanced Fluorescence through the First and Second Near-Infrared Windows, Chemistry of Materials, Vol: 29, Pages: 6916-6926, ISSN: 1520-5002

Gold nanostars (AuNSs) are receiving increasing attention for their potential applications in bionanotechnology because of their unique optical properties related to their complex branched morphology. Their sharp features allow strong localized surface plasmon resonances, tunable in the near-infrared (NIR) region, and large enhancements of local electromagnetic fields. Here, the application of AuNSs in metal-enhanced fluorescence (MEF) in the NIR and second NIR (NIR-II) biological windows is explored for the first time. NIR/NIR-II fluorophores are incorporated onto monolayers of AuNSs with tunable plasmonic responses. Over 320-fold fluorescence enhancement is achieved in the NIR, confirming that AuNS substrates are promising NIR-MEF platforms for the development of ultrasensitive biosensing applications. Using fluorescence lifetime measurements to semiquantitatively deconvolute the excitation enhancement from emission enhancement, as well as modeling to simulate the electric field enhancement, we show that a combination of enhanced excitation and an increased radiative decay rate, accompanied by an increase in the quantum yield, contribute to the observed large enhancement. AuNSs with different morphological features exhibit significantly different excitation enhancements, indicating the important role of the particle morphology on the magnitude of electromagnetic field enhancement and the resulting enhancement factor. Importantly, enhancement factors of up to 50-fold are also achieved in the NIR-II region, suggesting that this system holds promise for the future development of bright probes for NIR/NIR-II biosensing and bioimaging.

Journal article

Foglia F, Karan S, Nania M, Jiang Z, Porter AE, Barker R, Livingston AG, Cabral JTet al., 2017, Neutron Reflectivity and Performance of Polyamide Nanofilms for Water Desalination, ADVANCED FUNCTIONAL MATERIALS, Vol: 27, ISSN: 1616-301X

The structure and hydration of polyamide (PA) membranes are investigated with a combination of neutron and X-ray reflectivity, and their performance is benchmarked in reverse osmosis water desalination. PA membranes are synthesized by the interfacial polymerization of m-phenylenediamine (MPD) and trimesoyl chloride (TMC), varying systematically reaction time, concentration, and stoichiometry, to yield large-area exceptionally planar films of ≈10 nm thickness. Reflectivity is employed to precisely determine membrane thickness and roughness, as well as the (TMC/MPD) concentration profile, and response to hydration in the vapor phase. PA film thickness is found to increase linearly with reaction time, albeit with a nonzero intercept, and the composition cross-sectional profile is found to be uniform, at the conditions investigated. Vapor hydration with H2O and D2O from 0 to 100% relative humidity results in considerable swelling (up to 20%), but also yields uniform cross-sectional profiles. The resulting film thickness is found to be predominantly set by the MPD concentration, while TMC regulates water uptake. A favorable correlation is found between higher swelling and water uptake with permeance. The data provide quantitative insight into the film formation mechanisms and correlate reaction conditions, cross-sectional nanostructure, and performance of the PA active layer in RO membranes for desalination.

Journal article

Robinson RK, Birrell MA, Adcock JJ, Wortley MA, Dubuis ED, Chen S, McGilvery CM, Hu S, Shaffer MSP, Bonvini SJ, Maher SA, Mudway IS, Porter AE, Carlsten C, Tetley TD, Belvisi MGet al., 2017, Mechanistic link between diesel exhaust particles and respiratory reflexes, Journal of Allergy and Clinical Immunology, Vol: 141, Pages: 1074-1084.e9, ISSN: 1097-6825

BackgroundDiesel exhaust particles (DEPs) are a major component of particulate matter in Europe's largest cities, and epidemiologic evidence links exposure with respiratory symptoms and asthma exacerbations. Respiratory reflexes are responsible for symptoms and are regulated by vagal afferent nerves, which innervate the airway. It is not known how DEP exposure activates airway afferents to elicit symptoms, such as cough and bronchospasm.ObjectiveWe sought to identify the mechanisms involved in activation of airway sensory afferents by DEPs.MethodsIn this study we use in vitro and in vivo electrophysiologic techniques, including a unique model that assesses depolarization (a marker of sensory nerve activation) of human vagus.ResultsWe demonstrate a direct interaction between DEP and airway C-fiber afferents. In anesthetized guinea pigs intratracheal administration of DEPs activated airway C-fibers. The organic extract (DEP-OE) and not the cleaned particles evoked depolarization of guinea pig and human vagus, and this was inhibited by a transient receptor potential ankyrin-1 antagonist and the antioxidant N-acetyl cysteine. Polycyclic aromatic hydrocarbons, major constituents of DEPs, were implicated in this process through activation of the aryl hydrocarbon receptor and subsequent mitochondrial reactive oxygen species production, which is known to activate transient receptor potential ankyrin-1 on nociceptive C-fibers.ConclusionsThis study provides the first mechanistic insights into how exposure to urban air pollution leads to activation of guinea pig and human sensory nerves, which are responsible for respiratory symptoms. Mechanistic information will enable the development of appropriate therapeutic interventions and mitigation strategies for those susceptible subjects who are most at risk.

Journal article

Li Y, Klosowski MM, McGilvery CM, Porter AE, Livingston AG, Cabral JTet al., 2017, Probing flow activity in polyamide layer of reverse osmosis membrane with nanoparticle tracers, JOURNAL OF MEMBRANE SCIENCE, Vol: 534, Pages: 9-17, ISSN: 0376-7388

We investigate the flow activity of the nanostructured polyamide layer in reverse osmosis (RO) membrane, using gold nanoparticle (NP) tracers of 1–40 nm diameter. Following a detailed structural examination of a commercial SW30RH membrane selected for this study, NP solutions were infiltrated from either the polyamide front or the polysulfone support side. The permeate was then analyzed spectroscopically while the entrapment of NPs within the membrane was mapped by high resolution electron microscopy. Results show that back-filtered NPs exhibited a fractionated distribution according to size: 1 nm nanoparticles permeate across the polyamide-polysulfone interface reaching the interior of the polyamide corrugations, while the larger ones (>10 nm) are retained within the polysulfone and gradually arrested at approximately 100 nm below the polyamide-polysulfone interface. Intermediate-sized 5 nm nanoparticles reached the undulating folds just below the polyamide layer. Permeation pathways across polyamide layer appear to exclude all tracers above 1 nm, which become selectively distributed across the polyamide layer: positively charged NPs label the outer surface of the polyamide film (expected to be carboxylate-rich), while negatively charged particles are uniformly distributed within the layer. Diafiltration measurements quantify the transient kinetics of NP retention and permeation. Overall, our results establish the flow activity of the polyamide nodular surface and provide estimates for the dimensions of permeation pathways.

Journal article

Gonzalez Carter DA, Leo BF, Ruenraroengsak P, Chen S, Goode A, Theodorou I, Chung KF, Carzaniga R, Shaffer M, Dexter D, Ryan M, Porter Aet al., 2017, Silver nanoparticles reduce brain inflammation and related neurotoxicity through induction of H2S-synthesizing enzymes, Scientific Reports, Vol: 7, ISSN: 2045-2322

Silver nanoparticles (AgNP) are known to penetrate into the brain and cause neuronal death. However, there is a paucity in studies examining the effect of AgNP on the resident immune cells of the brain, microglia. Given microglia are implicated in neurodegenerative disorders such as Parkinson’s disease (PD), it is important to examine how AgNPs affect microglial inflammation to fully assess AgNP neurotoxicity. In addition, understanding AgNP processing by microglia will allow better prediction of their long term bioreactivity. In the present study, the in vitro uptake and intracellular transformation of citrate-capped AgNPs by microglia, as well as their effects on microglial inflammation and related neurotoxicity were examined. Analytical microscopy demonstrated internalization and dissolution of AgNPs within microglia and formation of non-reactive silver sulphide (Ag2S) on the surface of AgNPs. Furthermore, AgNP-treatment up-regulated microglial expression of the hydrogen sulphide (H2S)-synthesizing enzyme cystathionine-γ-lyase (CSE). In addition, AgNPs showed significant anti-inflammatory effects, reducing lipopolysaccharide (LPS)-stimulated ROS, nitric oxide and TNFα production, which translated into reduced microglial toxicity towards dopaminergic neurons. Hence, the present results indicate that intracellular Ag2S formation, resulting from CSE-mediated H2S production in microglia, sequesters Ag+ ions released from AgNPs, significantly limiting their toxicity, concomitantly reducing microglial inflammation and related neurotoxicity.

Journal article

Chung KF, Seiffert J, Chen S, Theodorou IG, Goode AE, Leo BF, McGilvery CM, Hussain F, Wiegman C, Rossios C, Zhu J, Gong J, Tariq F, Yufit V, Monteith AJ, Hashimoto T, Skepper JN, Ryan MP, Zhang J, Tetley TD, Porter AEet al., 2017, Inactivation, clearance, and functional effects of lung-instilled short and long silver nanowires in rats, ACS Nano, Vol: 11, Pages: 2652-2664, ISSN: 1936-086X

There is a potential for silver nanowires (AgNWs) to be inhaled, but there is little information on their health effects and their chemical transformation inside the lungs in vivo. We studied the effects of short (S-AgNWs; 1.5 μm) and long (L-AgNWs; 10 μm) nanowires instilled into the lungs of Sprague–Dawley rats. S- and L-AgNWs were phagocytosed and degraded by macrophages; there was no frustrated phagocytosis. Interestingly, both AgNWs were internalized in alveolar epithelial cells, with precipitation of Ag2S on their surface as secondary Ag2S nanoparticles. Quantitative serial block face three-dimensional scanning electron microscopy showed a small, but significant, reduction of NW lengths inside alveolar epithelial cells. AgNWs were also present in the lung subpleural space where L-AgNWs exposure resulted in more Ag+ve macrophages situated within the pleura and subpleural alveoli, compared with the S-AgNWs exposure. For both AgNWs, there was lung inflammation at day 1, disappearing by day 21, but in bronchoalveolar lavage fluid (BALF), L-AgNWs caused a delayed neutrophilic and macrophagic inflammation, while S-AgNWs caused only acute transient neutrophilia. Surfactant protein D (SP-D) levels in BALF increased after S- and L-AgNWs exposure at day 7. L-AgNWs induced MIP-1α and S-AgNWs induced IL-18 at day 1. Large airway bronchial responsiveness to acetylcholine increased following L-AgNWs, but not S-AgNWs, exposure. The attenuated response to AgNW instillation may be due to silver inactivation after precipitation of Ag2S with limited dissolution. Our findings have important consequences for the safety of silver-based technologies to human health.

Journal article

Calderon L, Han TT, McGilvery CM, Yang L, Subramaniam P, Lee K-B, Schwander S, Tetley TD, Georgopoulos PG, Ryan M, Porter AE, Smith R, Chung KF, Lioy PJ, Zhang J, Mainelis Get al., 2017, Release of airborne particles and Ag and Zn compounds from nanotechnology-enabled consumer sprays: Implications for inhalation exposure, ATMOSPHERIC ENVIRONMENT, Vol: 155, Pages: 85-96, ISSN: 1352-2310

The increasing prevalence and use of nanotechnology-enabled consumer products have increased potential consumer exposures to nanoparticles; however, there is still a lack of data characterizing such consumer exposure. The research reported here investigated near-field airborne exposures due to the use of 13 silver (Ag)-based and 5 zinc (Zn)-based consumer sprays. The products were sprayed into a specially designed glove box, and all products were applied with equal spraying duration and frequency. Size distribution and concentration of the released particles were assessed using a Scanning Mobility Particle Sizer and an Aerodynamic Particle Sizer. Inductively coupled plasma mass spectrometry (ICP-MS) was used to investigate the presence of metals in all investigated products. Spray liquids and airborne particles from select products were examined using transmission electron microscopy (TEM) and Energy Dispersive X-ray Spectroscopy (EDS). We found that all sprays produced airborne particles ranging in size from nano-sized particles (<100 nm) to coarse particles (>2.5 μm); however, there was a substantial variation in the released particle concentration depending on a product. The total aerosol mass concentration was dominated by the presence of coarse particles, and it ranged from ∼30 μg/m3 to ∼30,000 μg/m3. The TEM verified the presence of nanoparticles and their agglomerates in liquid and airborne states. The products were found to contain not only Ag and Zn compounds - as advertised on the product labeling - but also a variety of other metals including lithium, strontium, barium, lead, manganese and others. The results presented here can be used as input to model population exposures as well as form a basis for human health effects studies due to the use nanotechnology-enabled products.

Journal article

This data is extracted from the Web of Science and reproduced under a licence from Thomson Reuters. You may not copy or re-distribute this data in whole or in part without the written consent of the Science business of Thomson Reuters.

Request URL: http://wlsprd.imperial.ac.uk:80/respub/WEB-INF/jsp/search-html.jsp Request URI: /respub/WEB-INF/jsp/search-html.jsp Query String: respub-action=search.html&id=00302152&limit=30&person=true