Browsing by Author "Holden, PJ"

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    Application of polyethylene glycol to promote cellular biocompatibility of polyhydroxybutyrate films
    (Hindawi Publishing Corporation, 2011-08-22) Chan, RTH; Marçal, H; Russell, RA; Holden, PJ; Foster, LJR
    Polyhydroxybutyrate (PHB) is a biomaterial with potential for applications in biomedical and tissue engineering; however, its brittle nature and high crystallinity limit its potential. Blending PHB with a variety of PEGs produced natural-synthetic composite films composed of FDA-approved polymers with significant reductions in crystallinity, from 70.1% for PHB films to 41.5% for its composite with a 30% (w/w) loading of PEG2000. Blending also enabled manipulation of the material properties, increasing film flexibility with an extension to break of 2.49±1.01% for PHB films and 8.32±1.06% for films containing 30% (w/w) PEG106. Significant changes in the film surface properties, as measured by porosity, contact angles, and water uptake, were also determined as a consequence of the blending process, and these supported greater adhesion and proliferation of neural-associated olfactory ensheathing cells (OECs). A growth rate of 7.2×105 cells per day for PHB films with 30% (w/w) PEG2000 loading compared to 2.5×105 for PHB films was observed. Furthermore, while cytotoxicity of the films as measured by lactate dehydrogenase release was unaffected, biocompatibility, as measured by mitochondrial activity, was found to increase. It is anticipated that fine control of PEG composition in PHB-based composite biomaterials can be utilised to support their applications in medicinal and tissue engineering applications. Copyright © 2011 Rodman T. H. Chan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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    The Australian National Deuteration Facility for structure function applications using neutrons
    (International Conference on Neutron Scattering, 2017-07-12) Darwish, TA; Yepuri, NR; Heuer, AK; Duff, AP; Wilde, KL; Holden, PJ
    There have been limited global initiatives in the field of molecular deuteration where the majority of these programs focus on biological deuteration of proteins and lipids, while more complex deuterated small molecules haven’t been widely available to the neutron community. This has limited the experiments that can be performed, and formed a bottle-neck for advancing the applications of neutron scattering. In this paper we will discuss the recent advancements and the impact of deuteration on the research outcomes achieved by using deuterated molecules produced by the National Deuteration Facility of the Australian Nuclear Science and Technology Organisation. Recent high-impact case studies in the fields of molecular electronics, structural biology, and biotechnology will be presented which reveal the exciting and diverse characterisation studies which are now available for the neutron community.
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    Bacterial nano-particle uptake under extreme white-beam irradiation conditions
    (Australian Institute of Physics, 2009-02-05) Wilde, KL; Asquith, NL; Graham, SM; Holden, PJ; Stampfl, APJ; Kempson, I; Yang, BW; Hwu, YK
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    Biodegradable polyhydroxyalkanoates/SWNT nanocomposite spheres and films prepared by spray-drying method
    (American Chemical Society, 2008-04-06) Yun, SI; Gadd, GE; Russell, RA; Holden, PJ
    The nanocomposite microspheres and films of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxyoctanoate) (PHO) with single wall carbon nanotubes (SWNTs), were prepared by spray-drying method. The TEM immages showed that SWNTS well dispersed in PHB abd PHO indicating that rapid evaporation by spray drying effectively prevents aggregation of SWNTs. The Optical microscopy showed that the crystalline size substantially decreased for PHB/SWNTs nanocomposites with a 1% weight fraction of SWNTs relative to PHB (PHB/(1%)SWNTs), indicating the effective nucleation of PHB crystallization by SWNTs. Mechanical properties of the nanocomposite films were measured by nanoindentation. Both polymer nanocomposite films showed an increase in hardness (H) and Young's modulus (E), with SWNTs concentration.
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    BioPEGylation of polyhydroxybutyrate promotes nerve cell health and migration
    (ACS Publications, 2013-12-03) Chan, RTH; Russell, RA; Marçal, H; Lee, TH; Holden, PJ; Foster, LJR
    This study reports on the superior suitability of Polyhydroxybutyrate-polyethylene glycol hybrid polymers biosynthesised by Cupriavidus necator over PHB as biomaterials for tissue engineering. Incorporation of PEG106 (DEG) during PHB biosynthesis reduced crystallinity, molecular weight, and hydrophobicity while improving mechanical properties. In vitro olfactory ensheathing cell (OEC) proliferation was enhanced by cultivation on PHB-b-DEG films. Cultivation on PHB and PHB-b-DEG films showed no cytotoxic responses and cell viability and membrane integrity was sustained. PHB-b-DEG films promoted OECs entering into the DNA replication (S) phase and mitotic (G2-M) phase during the cell growth cycle and apoptosis was low. This study also confirmed an association between the level of neurite-outgrowth inhibitory protein (Nogo) and receptor pair Ig-like receptor B (PirB) expression and cell proliferation, both being down-regulated in cells grown on hybrid films when compared with PHB and asynchronous growth. Thus, DEG-terminated PHB-based biomaterials have great potential as biological scaffolds supporting nerve repair. © 2013 American Chemical Society.
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    Biopolymer deuteration for neutron scattering and other isotope-sensitive techniques
    (Elsevier, 2015) Russell, RA; Garvey, CJ; Darwish, TA; Foster, LJR; Holden, PJ
    The use of microbial biosynthesis to produced deuterated recombinant proteins is a well-established practice in investigations of the relationship between molecular structure and function using neutron scattering and nuclear magnetic resonance spectroscopy. However, there have been few reports of using microbial synthetic capacity to produce labeled native biopolymers. Here, we describe methods for the production of deuterated polyhydroxyalkanoate biopolyesters in bacteria, the polysaccharide chitosan in the yeast Pichia pastoris, and cellulose in the bacterium Gluconacetobacter xylinus. The resulting molecules offer not only multiple options in creating structural contrast in polymer blends and composites in structural studies but also insight into the biosynthetic pathways themselves. © 2015 Elsevier Inc.
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    Biosynthesis and characterization of deuterated polyhydroxyoctanoate
    (American Chemical Society, 2006-04) Foster, LJR; Russell, RA; Sanguanchaipaiwong, V; Stone, DJM; Hook, JM; Holden, PJ
    The synthesis of a polyhydroxyalkanoate with medium chain length alkyl substituents by Pseudomonas oleovorans was investigated using protonated and deuterated forms of octanoic acid in a minimal salts medium. Cultivation with deuterated octanoic acid resulted in a reduced rate of polymer accumulation compared to that with its protonated counterpart (107 and 207 mg of polymer L-1 of medium h-1 of cultivation, respectively). Nuclear magnetic resonance and gas chromatography coupled mass spectrometry of the derivatized polymer was used to establish the extent and distribution of deuterium in the biopolymer. A partially deuterated heteropolymer with 3-hydroxyoctanoic acid as the main constituent was produced. Deuteration is an important tool for contrast variation studies using neutron scattering, but predicates that the deuterated polymer is otherwise comparable in its physiochemical and material properties to its protonated counterpart. In studies reported here, the deuterated biopolymer exhibited an additional diffraction maximum at 7.55 Å and slight differences in its melting point (60 and 55 °C) and glass transition temperature (−39 and −36 °C) when compared to its protonated equivalent. While significant differences between the protonated and deuterated biopolymers were determined, our results support the use of this deuterated polyhydroxyalkanoate in its application in investigations using analytical neutron scattering techniques. © 2006, American Chemical Society
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    Carbon nanotube mediated miscibility of polyhydroxyalkanoate blends and chemical imaging using deuterium-labelled poly(3-hydroxyoctanoate)
    (Elsevier, 2018-08) Russell, RA; Foster, LJR; Holden, PJ
    Biopolymers have potential as scaffolds supporting regrowth of damaged tissues, however their material properties may limit the range of applications. Blending polymers with different thermomechanical properties has been demonstrated to extend the range of possible applications for polyhydroxyalkanoate (PHA) polymers, while the addition of nanoparticles can be used to modulate miscibility which influences strength and flexibility of the blend. Here we report on the blending of Poly(3-hydroxybutyrate) and Poly(3-hydroxyoctanoate) which possess different thermomechanical properties, and the effect of single wall carbon nanotubes (SWCNT) on their miscibility, electrical conductivity and thermomechanical performance. The apparent perturbation of phase boundaries in nanocomposite films observed by Scanning Electron Microscopy (SEM) was complemented by chemical mapping of film cross sections containing a deuterium-labelled poly(3-hydroxyoctanoate) phase in the blend using Infrared Microspectroscopy (IRM), suggesting increased miscibility due to nanoparticle addition. The electrical percolation threshold in nanocomposite films was observed between 0.5 and 1 wt% SWCNT, where the surface resistivity was reduced by eight orders of magnitude compared to the insulating polymer blend. Addition of SWCNT did not impact significantly on mechanical properties of films containing up to 2.5 wt% SWCNT. A solvent cast bionanocomposite film containing optimally 1 wt% SWCNT yielded a material with improved electrical conductivity compared to the SWCNT-free blend and which supported growth of Olfactory Ensheathing Cells, providing a basis for developing biopolymer scaffolds capable of conducting electrical stimulation. Crown Copyright © 2018. Published by Elsevier Ltd.
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    Chemical deuteration and neutrons for structure function applications
    (Australian Institute of Nuclear Science and Engineering, 2016-11-29) Darwish, TA; Yepuri, NR; Heuer, AK; Cagnes, MP; Holden, PJ
    In small angle neutron scattering and neutron reflectometry studies, the use of mixtures of deuterated and hydrogenated solvents to manipulate scattering length density and achieve contrast variation is widespread. This approach, while useful is less effective for multicomponent organic systems containing molecules of similar scattering length densities. In such systems molecular deuteration is necessary to achieve contrast between the different components and it significantly increases the options in structure function investigations. There have been limited global initiatives in the field of molecular deuteration where the majority of these programs focus on biological deuteration of proteins and lipids, while more complex deuterated small molecules haven’t been widely available to the neutron community. This has limited the experiments that can be performed, and formed a bottle neck for advancing the applications of neutron scattering. In this paper we will discuss the recent advancements and the impact of deuteration on the research outcomes achieved by using deuterated molecules produced by the chemical deuteration laboratories at ANSTO’s National Deuteration Facility. Recent high-impact case studies will be presented which reveal the exciting and diverse characterisation studies which are now available for the neutron community. The chemical deuteration of surfactants, sugars, heterocyclic and aromatic compounds has made possible a wide range of investigations systems in the fields of molecular electronics, structural biology, and biotechnology.
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    Correction to structural characterization of a model gram-negative bacterial surface using lipopolysaccharides from rough strains of Escherichia coli
    (American Chemical Society, 2014-07-24) Le Brun, AP; Clifton, LA; Halbert, CE; Lin, B; Meron, M; Holden, PJ; Lakey, JH; Holt, SA
    In the original publication, the schematic of lipopolysaccharide (LPS) from Escherichia coli in Figure 1 is incorrect. A corrected version of the figure and accompanying legend is below. Figure 1. Schematic of the organization of Escherichia coli LPS. LPS was from the rough mutant J5 strain of E. coli O111:B4, which produces an Rc chemotype with a core oligosaccharide as described by Müller-Loennies et al. (1) The original R mutants, which defined the different chemotypes were from Salmonella minnesota, so in this paper we use the terms Ra/Rc to denote the chemotype of E. coli LPS used according to this convention. Kdo, 2-keto-3-deoxyoctonic acid; Hep, l-glycero-D-manno heptose; Glc, glucose; Gal, galactose; GlcN, glucosamine. The Lipid A tails consists of four (R)-3-hydroxy-mystic acids, one myristic acid, and one lauric acid. Additional phosphates and ethanolamines on Kdo and Hep have been omitted for clarity. © 2014 American Chemical Society. CC-BY - Open Access
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    Demonstration of the use of Scenedesmus and Carteria biomass to drive bacterial sulfate reduction by Desulfovibrio alcoholovorans isolated from an artificial wetland
    (Elsevier, 2003-10) Russell, RA; Holden, PJ; Wilde, KL; Neilan, BA
    A major factor limiting application of bacterial sulfate reduction to removal of sulfate and heavy metals in wetland systems is the requirement to supply carbon and energy to drive the process. Primary production by aquatic plants and algae is a cheap option for driving sustainable bacterial sulfate reduction and most operational systems have relied on plants. The use of harvested, non-growing algal biomass to support bacterial sulfate reduction was investigated. Two genera of green algae, strains N9 and A3, were isolated from treatment cells from the Artificial Wetland Filter at the Ranger uranium mine (Northern Territory, Australia) which successfully removes UO22+, Mn2+ and nitrate, but little sulfate, from mine waters. These algae were identified as Carteria sp. and Scenedesmus sp. and were used as the sole carbon and energy source to enrich a sulfate-reducing mixed bacterial culture from the constructed wetland. Bacterial sulfate reduction supported solely by degradation of algal biomass was demonstrated at laboratory scale using both algae. In excess of 300 mg/L, sulfate was reduced in 17 days following an initial period of approximately 8 days during which sulfate levels did not decrease. The amount and rate of reduction was shown to be dependent on the concentration of algal biomass added. Carteria algae at low concentration showed reduction earlier; however, yields at higher concentration were affected by unknown inhibition. Scenedesmus strain N9 produced a maximum specific yield of 94.3 g of sulfate reduced per gram biomass added compared with 43.5 for Carteria strain A3. Sequence analysis of the 16S rRNA gene of members of the bacterial consortium indicated that the sulfate-reducing bacteria (SRB) showed highest homology (98.5%) with Desulfovibrio alcoholovorans. A second bacterium, which showed homologies of 91–92% with organisms of the Clostridial assemblage, was also present in the culture and represents a new species, or possibly a new genus. Crown Copyright 2003 Published by Elsevier B.V.
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    Determination of solution properties of poly(3-hydroxybutyrate) and deuterated poly(3-hydroxybutyrate) by SANS.
    (American Chemical Society, 2008-04-06) Yun, SI; Russell, RA; Holden, PJ
    Polyhydroxyalkanoates (PHAs) are natural, linear, thermoplastic polyesters synthesized by micro-organisms as intracellular carbon reserves and ion sinks in response to limited nutrient availability and imposed stress conditions. We bio-synthesized PHB and deuterated PHB (d-PHB) and performed SANS on the dilute solutions of these molecules. In this paper, we report the conformational properties of PHB and d-PHB in d-chloroform and chloroform, respectively.
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    Deuterated polymers for probing phase separation using infrared microspectroscopy
    (ACS Publications, 2013-12-23) Russell, RA; Darwish, TA; Puskar, L; Martin, DE; Holden, PJ; Foster, LJR
    Infrared (IR) microspectroscopy has the capacity to determine the extent of phase separation in polymer blends. However, a major limitation in the use of this technique has been its reliance on overlapping peaks in the IR spectra to differentiate between polymers of similar chemical compositions in blends. The objective of this study was to evaluate the suitability of deuteration of one mixture component to separate infrared (IR) absorption bands and provide image contrast in phase separated materials. Deuteration of poly(3-hydroxyoctanoate) (PHO) was achieved via microbial biosynthesis using deuterated substrates, and the characteristic C–D stretching vibrations provided distinct signals completely separated from the C–H signals of protonated poly(3-hydroxybutyrate) (PHB). Phase separation was observed in 50:50 (% w/w) blends as domains up to 100 μm through the film cross sections, consistent with earlier reports of phase separation observed by scanning electron microscopy (SEM) of freeze-fractured protonated polymer blends. The presence of deuterated phases throughout the film suggests there is some miscibility at smaller length scales, which increased with increasing PHB content. These investigations indicate that biodeuteration combined with IR microspectroscopy represents a useful tool for mapping the phase behavior of polymer blends.
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    Effect of carbon nanotube functionalization on the structure and properties of poly(3-hydroxybutyrate)/MWCNTs biocomposites
    (Springer Link, 2014-07) Huh, M; Jung, MH; Park, YS; Kim, BJ; Kang, M; Holden, PJ; Yun, SI
    Multi walled carbon nanotubes (MWCNTs) covalently functionalized with an alkyl chain exhibited a better dispersion in poly(3-hydroxybutyrate) (PHB) solutions and cast films as compared with acid-treated MWCNTs (MWCNT-COOH) due to the much improved solubility in chloroform. The alkylated MWCNTs more effectively strengthened PHB composites than non-alkylated MWCNTs due to their uniform dispersion as well as stronger interaction of alkylated MWCNTs with the PHB matrix. Both acid-treated and alkylated MWCNTs added to PHB matrix facilitated crystallization kinetics. However the crystallization kinetics were found to be slower for the alkylated MWCNTs/PHB composites than acid-treated MWCNTs composites. The results may be ascribed to the inhibitory effect on PHB crystallization caused by the steric hindrance of the long alkyl chains attached to MWCNTs. © The Polymer Society of Korea and Springer Sciene+Business Media Dordrecht 2014.
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    Effect of sol-gel encapsulation on lipase structure and function: a small angle neutron scattering study
    (Springer, 2005-01) Rodgers, LE; Holden, PJ; Knott, RB; Finnie, KS; Bartlett, JR; Foster, LJR
    The application of small angle neutron scattering (SANS) to the characterisation of sol–gel hosts containing biomolecules offers the opportunity to explore the relationship between gel structure and catalyst. A model system involving the immobilisation of Candida antarctica lipase B (CALB) was investigated. Gels were produced by fluoride-catalysed hydrolysis of fixed ratios of tetramethylorthosilicate (TMOS) and methyltrimethoxysilane (MTMS). Phase separation between the enzyme and the evolving sol–gel matrix was minimised by incorporating glycerol into the sol–gel precursor solution. The potential stabilising effect of the NaF catalyst upon the enzyme was also investigated. Scattering studies were conducted on both immobilised lipase, and lipase in free solution. Scattering studies on free enzyme provided evidence of multiple populations of enzyme aggregates and showed that choice of solvent affected the degree of aggregation. Both NaF and glycerol affected neutron scattering, indicating changes in lipase conformation. Increasing glycerol concentration increased the degree of aggregation and produced differences in solvent packing on the surface of protein molecules. Initial evidence from SANS data indicated that the presence of the enzyme during gel formation conferred structural changes on the gel matrix. Modelling the effect of sol–gel encapsulation on lipase requires comparison of data from free enzyme to the immobilised form. Removal of the enzyme from the sol–gel structure, post gelation, is necessary to better characterise the modified matrix. This methodological problem will be the subject of future investigations. © 2005, Springer.
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    The effect of sulfate-reducing bacteria on adsorption of 137Cs by soils from arid and tropical regions
    (Elsevier, 2004-03-05) Russell, RA; Holden, PJ; Payne, TE; McOrist, GD
    Soils from different climatic regions of Australia were studied to determine their adsorption of 137Cs, and the effect of microbial sulfate reduction on this adsorption. The soils consisted of a surface and regolith samples from the site of a proposed low and intermediate level radioactive waste repository in arid South Australia, and two red earth loam soils from an experimental plot in the tropical Northern Territory. The process of bacterial sulfate reduction substantially decreased the adsorption of 137Cs to the arid and tropical soils, although extended incubation resulted in greater adsorption to the regolith sample. This could have implications for the mobility of radionuclides entering these soil ecosystems. Crown Copyright 2004 Published by Elsevier Ltd.
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    The electronic structure of S-layer proteins from Lactobacillus brevis
    (IEEE, 2008-07-28) Graham, SM; Asquith, NL; Wilde, KL; Short, KT; Holden, PJ; Stampfl, APJ; Holmes, AJ; Ruys, AJ; Stojanov, P; Riley, JD; Fang, LJ; Yang, YW; Hwu, YK
    The valence electronic structure of the S-layer of Lactobacillus brevis is determined using synchrotron-based photoelectron spectroscopy and soft X-ray absorption spectroscopy. Spectra are compared to experimental work on amino-acids and S-layers of Bacillus sphaericus. While it is indeed possible to identify energy levels with those of natural amino-acids, distinct energy shifts are indeed observed which cannot be reconciled using such simple comparisons. Furthermore a strong nitrogen signal observed in both the occupied and unoccupied energy levels suggests that the Lactobacillus brevis protein is amine-terminated. A discussion of the surface of this protein is given. © 2008 IEEE
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    Evolution of the neutron-scattering capability on the OPAL reactor at ANSTO
    (Taylor & Francis Online, 2016-04-29) Klose, F; Constantine, P; Kennedy, SJ; Schulz, JC; Robinson, RA; Holden, PJ; McIntyre, GJ
    Australia is currently in the very privileged position of having the world's newest, fully operating research reactor, OPAL. As a consequence, the suite of neutron-beam instruments is also amongst the youngest and most advanced in the world, with full advantage taken in their construction of lessons learned at reactors elsewhere to develop state-of-the-art instruments that are best suited to the local and regional user communities. There are two thermal-neutron beam ports, two cold-neutron beam ports, and two (future) hot-neutron beam ports around the OPAL reactor core (see Fig. 3 in [1] and [2]). One each of the thermal-neutron and cold-neutron ports feed into a set of three guide bundles that serve the present (first) guide hall. In the initial construction phase, completed in 2007, only the outer two (TG1 and TG3) of the thermal guides, and the outer two (CG1 and CG3) of the cold guides were installed. The other thermal-neutron (TG4) and cold-neutron (CG4) guides are relatively short, to serve single instruments within the reactor beam hall with the highest flux but without eliminating line-of-sight [2]. Table 1 lists the current suite of instruments at the OPAL reactor, with brief technical details and the principal features. Figure 1 shows the layout of the current suite, with the anticipated location of the next instrument, BioRef (vide infra), indicated. We now briefly outline the evolution of the suite. © 2021 Informa UK Limited
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    Fabrication of honeycomb-structured porous films from poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) via the breath figures method
    (Wiley, 2011-12-27) Huh, M; Jung, MH; Park, YS; Kang, TB; Nah, C; Russell, RA; Holden, PJ; Yun, SI
    Formation of porous films from poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) using the breath figures (BF) method was investigated by evaporating solutions in chloroform in humid air and examining film structure using optical and scanning electron microscopy (SEM). BF films were successfully fabricated from PHB (Mw = 486,000 g/mol) and displayed hexagonal arrays of pores, which varied in diameter (D = 7–2 μm) with solution concentrations (0.5–2.00%). SEM of fractured films also showed subsurface closed nano-pores (D = 500–700 nm). BF films cast from PHBV (5% HV) formed arrays with smaller pores and apparent surface defects. Differential scanning calorimetry showed that porous PHB and PHBV films produced using the BF method were more crystalline than nonporous solvent cast films of PHB and PHBV. POLYM. ENG. SCI., 2011. © 2011 Society of Plastics Engineers
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    Fluorine-18 radiolabelling and in vitro / in vivo metabolism of [18F]D4-PBR111
    (John Wiley & Sons, Inc, 2019-05-26) Wyatt, NA; Safavi-Naeini, M; Wotherspoon, ATL; Arthur, A; Nguyen, AP; Parmar, A; Hamze, H; Day, CM; Zahra, D; Matesic, L; Davis, E; Rahardjo, GL; Yepuri, NR; Shepherd, R; Murphy, RB; Pham, TQ; Nguyen, VH; Callaghan, PD; Holden, PJ; Grégoire, MC; Darwish, TA; Fraser, BH
    Objectives The purinergic receptor P2X ligand-gated ion channel type 7 (P2X7R) is an adenosine triphosphate (ATP)-gated ion-channel, and P2X7R is a key player in inflammation. P2X7R is an emerging therapeutic target in central nervous system (CNS) diseases including Alzheimer's disease (AD) and Parkinson's disease (PD), because P2X7R also plays a pivotal role in neuroinflammation. P2X7R represents a potential molecular imaging target for neuroinflammation via biomedical imaging technique positron emission tomography (PET), and several radioligands targeting P2X7R have been developed and evaluated in animals. In our previous work, we have developed and characterized [11C]GSK1482160 as a P2X7R radioligand for neuroinflammation,2 clinical evaluation of [11C]GSK1482160 in healthy controls and patients is currently underway, and the estimation of radiation dosimetry for [11C]GSK1482160 in normal human subjects has been reported.3 Since the half-life (t1/2) of radionuclide carbon-11 is only 20.4 min, it is attractive for us to develop derivatives of [11C]GSK1482160, which can be labeled with the radionuclide fluorine-18 (t1/2, 109.7 min), and a fluorine-18 ligand would be ideal for widespread use.4 To this end, a series of [18F]fluoroalkyl including [18F]fluoromethyl (FM), [18F]fluoroethyl (FE), and [18F]fluoropropyl (FP) derivatives of GSK1482160 have been prepared and examined as new potential P2X7R radioligands. © 2019 The Authors