Optimizing dose enhancement with Ta2O5 nanoparticles for synchrotron microbeam activated radiation therapy

Microbeam Radiation Therapy (MRT) exploits tumour selectivity and normal tissue sparing with spatially fractionated kilovoltage X-ray microbeams through the dose volume effect. Experimental measurements with Ta₂O₅ nanoparticles (NPs) in 9L gliosarcoma treated with MRT at the Australian Synchrotron, increased the treatment efficiency. Ta₂O₅ NPs were observed to form shells around cell nuclei which may be the reason for their efficiency in MRT. In this article, our experimental observation of NP shell formation is the basis of a Geant4 radiation transport study to characterise dose enhancement by Ta₂O₅ NPs in MRT. Our study showed that NP shells enhance the physical dose depending microbeam energy and their location relative to a single microbeam. For monochromatic microbeam energies below ∼70 keV, NP shells show highly localised dose enhancement due to the short range of associated secondary electrons. Low microbeam energies indicate better targeted treatment by allowing higher microbeam doses to be administered to tumours and better exploit the spatial fractionation related selectivity observed with MRT. For microbeam energies above ∼100 keV, NP shells extend the physical dose enhancement due to longer-range secondary electrons. Again, with NPs selectively internalised, the local effectiveness of MRT is expected to increase in the tumour. Dose enhancement produced by the shell aggregate varied more significantly in the cell population, depending on its location, when compared to a homogeneous NP distribution. These combined simulation and experimental data provide first evidence for optimising MRT through the incorporation of newly observed Ta₂O₅ NP distributions within 9L cancer cells.     

Local dose enhancement of proton therapy by ceramic oxide nanoparticles investigated with Geant4 simulations

Nanoparticles (NPs) have been shown to enhance X-ray radiotherapy and proton therapy of cancer. The effectiveness of radiation damage is enhanced in the presence of high atomic number (high-Z) NPs due to increased production of low energy, higher linear energy transfer (LET) secondary electrons when NPs are selectively internalized by tumour cells. This work quantifies the local dose enhancement produced by the high-Z ceramic oxide NPs Ta₂O₅ and CeO₂, in the target tumour, for the first time in proton therapy, by means of Geant4 simulations. The dose enhancement produced by the ceramic oxides is compared against gold NPs. The energy deposition on a nanoscale around a single nanoparticle of 100 nm diameter is investigated using the Geant4-DNA extension to model particle interactions in the water medium. Enhancement of energy deposition in nano-sized shells of water, local to the NP boundary, ranging between 14% and 27% was observed for proton energies of 5 MeV and 50 MeV, depending on the NP material. Enhancement of electron production and energy deposition can be correlated to the direct DNA damage mechanism if the NP is in close proximity to the nucleus.     

First proof of bismuth oxide nanoparticles as efficient radiosensitisers on highly radioresistant cancer cells

This study provides the first proof of the novel application of bismuth oxide as a radiosensitiser. It was shown that on the highly radioresistant 9L gliosarcoma cell line, bismuth oxide nanoparticles sensitise to both kilovoltage (kVp) or megavoltage (MV) X-rays radiation. 9L cells were exposed to a concentration of 50 μg.mL⁻¹ of nanoparticle before irradiation at 125 kVp and 10 MV. Sensitisation enhancement ratios of 1.48 and 1.25 for 125 kVp and 10 MV were obtained in vitro, respectively. The radiation enhancement of the nanoparticles is postulated to be a combination of the high Z nature of the bismuth (Z = 83), and the surface chemistry. Monte Carlo simulations were performed to elucidate the physical interactions between the incident radiation and the nanoparticle. The results of this work show that Bi₂O₃ nanoparticles increase the radiosensitivity of 9L gliosarcoma tumour cells for both kVp and MV energies. Monte Carlo simulations demonstrate the advantage of a platelet morphology.     

Dietary supplementation of green synthesized manganese-oxide nanoparticles and its effect on growth performance,muscle composition and digestive enzyme activities of the giant freshwater prawn Macrobrachium rosenbergii

The green synthesized Mn₃O₄ nanoparticles (manganese-oxide nanoparticles) using Ananas comosus (L.) peel extract was characterized by various techniques. HR-SEM photograph showed that manganese-oxide nanoparticles (Mn-oxide NPs) were spherical in shape, with an average size of 40–50 nm. The Zeta potential revealed the surface charge of Mn-oxide NPs to be negative. Further, the Mn-oxide NPs were dietary supplemented for freshwater prawn Macrobrachium rosenbergii. The experimental basal diets were supplemented with Mn-oxide NPs at the rates of 0 (control), 3.0, 6.0, 9.0, 12, 15 and 18 mg/kg dry feed weight. The as-supplemented Mn-oxide NPs were fed in M. rosenbergii for a period of 90 days. The experimental study demonstrated that prawns fed with diet supplemented with 3–18 mg Mn-oxide NPs/kg shows enhanced (P < 0.05) growth performance, including final weight and weight gain (WG). Significant differences (P < 0.05) in feed conversion ratio (FCR) were observed in prawn fed with different diets. Additionally, prawns fed with 3.0–18 mg/kg Mn-oxide NPs supplemented diets achieved significant (P < 0.05) improvement in growth performance, digestive enzyme activities and muscle biochemical compositions, while, the prawns fed with 16 mg/kg of Mn-oxide NPs showed enhanced performance. Prawns fed on diet supplemented with 16 mg/kg Mn-oxide NPs showed significantly (P < 0.05) higher total protein level. The antioxidants enzymatic activity (SOD and CAT) metabolic enzymes status in muscle and hepatopancreas showed no significant (P > 0.05) alterations in prawns fed with 3.0–18 mg/kg of Mn-oxide NPs supplemented diets. Consequently, the present work proposed that 16 mg/kg of Mn-oxide NPs could be supplemented for flexible enhanced survival, growth and production of M. rosenbergii. Therefore, the data of the present study recommend the addition of 16 mg/kg of Mn-oxide NPs diet to developed prawn growth and antioxidant defense system.     

Antioxidant and cytotoxic effect of biologically synthesized selenium nanoparticles in comparison to selenium dioxide

The present study was designed to evaluate antioxidant and cytotoxic effect of selenium nanoparticles (Se NPs) biosynthesized by a newly isolated marine bacterial strain Bacillus sp. MSh-1. An organic–aqueous partitioning system was applied for purification of the biogenic Se NPs and the purified Se NPs were then investigated for antioxidant activity using DPPH scavenging activity and reducing power assay. Cytotoxic effect of the biogenic Se NPs and selenium dioxide (SeO₂) on MCF-7 cell line was assesed by MTT assay. Tranmission electron micrograph (TEM) of the purified Se NPs showed individual and spherical nanostructure in size range of about 80–220 nm. The obtained results showed that, at the same concentration of 200 μg/mL, Se NPs and SeO₂ represented scavenging activity of 23.1 ± 3.4% and 13.2 ± 3.1%, respectively. However, the data obtained from reducing power assay revealed higher electron-donating activity of SeO₂ compared to Se NPs. Higher IC₅₀ of the Se NPs (41.5 ± 0.9 μg/mL) compared to SeO₂ (6.7 ± 0.8 μg/mL) confirmed lower cytotoxicity of the biogenic Se NPs on MCF-7 cell line.     

Experimental assessment of the influence of beam hardening filters on image quality and patient dose in volumetric 64-slice X-ray CT scanners

Beam hardening filters have long been employed in X-ray Computed Tomography (CT) to preferentially absorb soft and low-energy X-rays having no or little contribution to image formation, thus allowing the reduction of patient dose and beam hardening artefacts. In this work, we studied the influence of additional copper (Cu) and aluminium (Al) flat filters on patient dose and image quality and seek an optimum filter thickness for the GE LightSpeed VCT 64-slice CT scanner using experimental phantom measurements. Different thicknesses of Cu and Al filters (0.5–1.6 mm Cu, 0.5–4 mm Al) were installed on the scanner’s collimator. A planar phantom consisting of 13 slabs of Cu having different thicknesses was designed and scanned to assess the impact of beam filtration on contrast in the intensity domain (CT detector’s output). To assess image contrast and image noise, a cylindrical phantom consisting of a polyethylene cylinder having 16 holes filled with different concentrations of K₂HPO₄ solution mimicking different tissue types was used. The GE performance and the standard head CT dose index (CTDI) phantoms were also used to assess image resolution characterized by the modulation transfer function (MTF) and patient dose defined by the weighted CTDI. A 100 mm pencil ionization chamber was used for CTDI measurement. Finally, an optimum filter thickness was determined from an objective figure of merit (FOM) metric. The results show that the contrast is somewhat compromised with filter thickness in both the planar and cylindrical phantoms. The contrast of the K₂HPO₄ solutions in the cylindrical phantom was degraded by up to 10% for a 0.68 mm Cu filter and 6% for a 4.14 mm Al filter. It was shown that additional filters increase image noise which impaired the detectability of low density K₂HPO₄ solutions. It was found that with a 0.48 mm Cu filter the 50% MTF value is shifted by about 0.77 lp/cm compared to the case where the filter is not used. An added Cu filter with approximately 0.5 mm thickness accounts for 50% reduction in radiation-absorbed dose as measured by the weighted CTDI. The FOM results indicate that with an additional filter of 0.5 mm Cu or minimum 4 mm Al, a good compromise between image quality and patient dose is achieved for CT images acquired at tube voltages of 120 and 140 kVp. The results seem to indicate that an optimum filter for high kVp acquisitions, routinely used in cardiovascular imaging, should be 0.5 mm copper or 4 mm aluminium minimum.     

Impact of surface grafting density of PEG macromolecules on dually fluorescent silica nanoparticles used for the in vivo imaging of subcutaneous tumors

BackgroundIn the context of systematically administered nanomedicines, the physicochemistry of NP surfaces must be controlled as a prerequisite to improve blood circulation time, and passive and active targeting. In particular, there is a real need to develop NP stealth and labelling for both in vivo and microscopic fluorescence imaging in a mice model.MethodsWe have synthesized NIR/red dually fluorescent silica nanoparticles of 19 nm covalently covered by a PEG layer of different grafting density in the brush conformational regime by using a reductive amination reaction. These particles were characterized by TEM, DRIFT, DLS, TGA, ζ potential measurements, UV-vis and fluorescence spectroscopy. Prostate tumors were generated in mice by subcutaneous injection of RM1-CMV-Fluc cells. Tumor growth was monitored by BLI after a D-luciferin injection. Four samples of PEGylated fluorescent NPs were individually intravenously injected into 6 mice (N = 6, total 24 mice). Nanoparticle distribution was investigated using in vivo fluorescence reflectance imaging (FRI) over 48 h and microscopy imaging was employed to localize the NPs within tumors in vitro.ResultsFluorescent NP accumulation, due to the enhanced permeability and retention (EPR) effect, increases gradually as a function of increased PEG surface grafting density with a huge difference observed for the highest density grafting. For the highest grafting density, a blood circulation time of up to 24 h was observed with a strong reduction in uptake by the liver. In vivo experimental results suggest that the biodistribution of NPs is very sensitive to slight variations in surface grafting density when the NPs present a high curvature radius.ConclusionThis study underlines the need to compensate a high curvature radius with a PEG-saturated NP surface to improve blood circulation and accumulation within tumors through the EPR effect. Dually fluorescent NPs PEGylated to saturation display physical properties useful for assessing the susceptibility of tumors to the EPR effect.General significanceControl of the physicochemical features of nanoparticle surfaces to improve blood circulation times and monitoring of the EPR effect. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader.     

Potential of using cerium oxide nanoparticles for protecting healthy tissue during accelerated partial breast irradiation (APBI)

The purpose of this study is to investigate the feasibility of using cerium oxide nanoparticles (CONPs) as radical scavengers during accelerated partial breast irradiation (APBI) to protect normal tissue. We hypothesize that CONPs can be slowly released from the routinely used APBI balloon applicators—via a degradable coating—and protect the normal tissue on the border of the lumpectomy cavity over the duration of APBI. To assess the feasibility of this approach, we analytically calculated the initial concentration of CONPs required to protect normal breast tissue from reactive oxygen species (ROS) and the time required for the particles to diffuse to various distances from the lumpectomy wall. Given that cerium has a high atomic number, we took into account the possible inadvertent dose enhancement that could occur due to the photoelectric interactions with radiotherapy photons. To protect against a typical MammoSite treatment fraction of 3.4 Gy, 5 ng·g⁻¹ of CONPs is required to scavenge hydroxyl radicals and hydrogen peroxide. Using 2 nm sized NPs, with an initial concentration of 1 mg·g⁻¹, we found that 2–10 days of diffusion is required to obtain desired concentrations of CONPs in regions 1–2 cm away from the lumpectomy wall. The resultant dose enhancement factor (DEF) is less than 1.01 under such conditions. Our results predict that CONPs can be employed for radioprotection during APBI using a new design in which balloon applicators are coated with the NPs for sustained/controlled in-situ release from within the lumpectomy cavity.     

High activity and selectivity immobilized lipase on Fe3O4 nanoparticles for banana flavour synthesis

Lipase (E.C.3.1.1.3) from Thermomyces lanuginosus (TL) was directly bonded, through multiple physical interactions, on citric acid functionalized monodispersed Fe₃O₄ nanoparticles (NPs) in presence of a small amount of hydrophobic functionalities. A very promising scalable synthetic approach ensuring high control and reproducibility of the results, and an easy and green immobilization procedure was chosen for NPs synthesis and lipase anchoring. The size and structure of magnetic nanoparticles were characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). The samples at different degree of functionalization were analysed through thermogravimetric measurements. Lipase immobilization was further confirmed by enzymatic assay and Fourier transform infrared (FT-IR) spectra. Immobilized lipase showed a very high activity recovery up to 144% at pH = 7 and 323% at pH = 7.5 (activity of the immobilized enzyme compared to that of its free form). The enzyme, anchored to the Fe₃O₄ nanoparticles, to be easy recovered and reused, resulted more stable than the native counterpart and useful to produce banana flavour. The immobilized lipase results less sensitive to the temperature and pH, with the optimum temperature higher of 5 °C and optimum pH up shifted to 7.5 (free lipase optimum pH = 7.0). After 120 days, free and immobilized lipases retained 64% and 51% of their initial activity, respectively. Ester yield at 40 °C for immobilized lipase reached 88% and 100% selectivity.     

Fluorescence microscopy colocalization of lipid–nucleic acid nanoparticles with wildtype and mutant Rab5–GFP: A platform for investigating early endosomal events

Endosomal entrapment is known to be a major bottleneck to successful cytoplasmic delivery of nucleic acids (NAs) using cationic liposome–NA nanoparticles (NPs). Quantitative measurements of distributions of NPs within early endosomes (EEs) have proven difficult due to the sub-resolution size and short lifetime of wildtype EEs. In this study we used Rab5–GFP, a member of the large family of GTPases which cycles between the plasma membrane and early endosomes, to fluorescently label early endosomes. Using fluorescence microscopy and quantitative image analysis of cells expressing Rab5–GFP, we found that at early time points (t < 1 h), only a fraction (≈ 35%) of RGD-tagged NPs (which target cell surface integrins) colocalize with wildtype EEs, independent of the NP's membrane charge density. In comparison, a GTP-hydrolysis deficient mutant, Rab5–Q79L, which extends the size and lifetime of EEs yielding giant early endosomes (GEEs), enabled us to resolve and localize individual NPs found within the GEE lumen. Remarkably, nearly all intracellular NPs are found to be trapped within GEEs implying little or no escape at early time points. The observed small degree of colocalization of NPs and wildtype Rab5 is consistent with recycling of Rab5–GDP to the plasma membrane and not indicative of NP escape from EEs. Taken together, our results show that endosomal escape of PEGylated nanoparticles occurs downstream of EEs i.e., from late endosomes/lysosomes. Our studies also suggest that Rab5–Q79L could be used in a robust imaging assay which allows for direct visualization of NP interactions with the luminal membrane of early endosomes.     

Pancreatic lipase inhibitory and antioxidative constituents from the aerial parts of Paeonia lactiflora Pall. (Ranunculaceae)

To date, there have been reports mostly about research results of the peony root in comparison to the aerial parts. According to our study, the aerial parts of P.lactiflora showed superior anti-oxidative and pancreatic lipase inhibitory activities than its root. Especially, the water extract and the ethyl acetate fraction of the ethanol extract exhibited potent pancreatic lipase inhibitory activity by 53.11 ± 1.22% and 46.16 ± 1.55% at the same dose of orlistat (62.5 ± 1.27%). The ethanol extract exhibited the best anti-oxidative activity with IC₅₀ of 17.08 ± 0.9 μg/mL, and the ethyl acetate fraction 19.75 ± 0.02 μg/mL, respectively, comparing to the positive control rutin (IC₅₀, 22.66 ± 0.29 μg/mL). From the anti-oxidative and pancreatic lipase inhibitory active fractions three new compounds, monplacphloroside (1), monplachydroxyquinoside (2) and herbacetin-7-O-β-d-sophoroside (3) were isolated along with 19 (4-22) known ones.Compounds, PGG (14), 1-O-methyl-2,3,4,6-tetra-O-galloyl-β-d-glucopyranose (17) and ethylgallate (9) were found to be the strongest antioxidants and pancreatic lipase inhibitors. Monoterpenes, albiflorin R₂ (19) and albiflorin (20) were determined for the first time as strong pancreatic lipase inhibitors. The presence of the esterified galloyl moiety, with its increasing numbers or the β-lactone cycle within the molecular structure plays an essential role for the enhancement of the pancreatic lipase enzyme inhibitory activity.     

Oxygen regulates the effective diffusion distance of nitric oxide in the aortic wall

Endothelium-derived nitric oxide (NO) is critical in maintaining vascular tone. Accumulating evidence shows that NO bioavailability is regulated by oxygen concentration. However, it is unclear to what extent the oxygen concentration regulates NO bioavailability in the vascular wall. In this study, a recently developed experimental setup was used to measure the NO diffusion flux across the aortic wall at various oxygen concentrations. It was observed that for a constant NO concentration at the endothelial surface, the measured NO diffusion flux out of the adventitial surface at [O₂] = 0 μM is around fivefold greater than at [O₂] = 150 μM, indicating that NO is consumed in the aortic wall in an oxygen-dependent manner. Analysis of experimental data shows that the rate of NO consumption in the aortic wall is first order with respect to [NO] and first order with respect to [O₂], and the rate constant k₁ was determined as (4.0 ± 0.3) × 10³ M^?1 s^?1. Computer simulations demonstrate that NO concentration distribution significantly changes with oxygen concentration and the effective NO diffusion distance at low oxygen level ([O₂] ≤ 25 μM) is significantly longer than that at high oxygen level ([O₂] = 200 μM). These results suggest that oxygen-dependent NO consumption may play an important role in dilating blood vessels during hypoxia by increasing the effective NO diffusion distance.     

Hypoxia induced CA9 inhibitory targeting by two different sulfonamide derivatives including Acetazolamide in human Glioblastoma

HIF-1α regulated genes are mainly responsible for tumour resistance to radiation- and chemo-therapy. Among these genes, carbonic anhydrase isoform IX (CA9) is highly over expressed in many types of cancer especially in high grade brain cancer like Glioblastoma (GBM). Inhibition of the enzymatic activity by application of specific chemical CA9 inhibitor sulphonamides (CAI) like Acetazolamide (Aza.), the new sulfonamide derivative carbonic anhydrase inhibitor (SU.D2) or indirect inhibitors like the HIF-1α inhibitor Chetomin or molecular inhibitors like CA9-siRNA are leading to an inhibition of the functional role of CA9 during tumorigenesis. Human GBM cells were treated with in vitro hypoxia (1, 6, or 24 h at 0.1%, O₂). Aza. application was at a range between 250 and 8000 nM and the HIF-1α inhibitor Chetomin at a concentration range of 150–500 nM. Cell culture plates were incubated for 24 h under hypoxia (0.1% O₂). Further, CA9-siRNA constructs were transiently transfected into GBM cells exposed to extreme hypoxic aeration conditions. CA9 protein expression level was detectable in a cell-type specific manner under normoxic conditions. Whereas U87-MG exhibited a strong aerobic expression, U251 and U373 displayed moderate and GaMG very weak normoxic CA9 protein bands. Aza. as well as SU.D2 displayed inhibitory characteristics to hypoxia induced CA9 expression in the four GBM cell lines for 24 h of hypoxia (0.1% O₂) at concentrations between 3500 and 8000 nM, on both the protein and mRNA level. Parallel experiments using CA9-siRNA confirmed these results. Application of 150–500 nM of the glycolysis inhibitor Chetomin under similar oxygenation conditions led to a sharply reduced expression of both CA IX protein and CA9 mRNA levels, indicating a clear glucose availability involvement for the hypoxic HIF-1α and CA9 expression in GBM cells. Hypoxia significantly influences the behaviour of human tumour cells by activation of genes involved in the adaptation to hypoxic stress. The main objective in malignant GBM therapy is either to eradicate the tumour or to convert it into a controlled, quiescent chronic disease. Aza., SU.D2, Chetomin or CA9-siRNA possesses functional CA9 inhibitory characteristics when applied against human cancers with hypoxic regions like GBM. They may be used as alternative or in conjunction with other direct inhibitors possessing similar functionality, thereby rendering them as potential optimal tools for the development of an optimized therapy in human brain cancer treatment.     

Immobilization of glucose oxidase onto cobalt based on silica core/shell nanoparticles as carrier

Co–B/SiO₂/NH₂ magnetic nanoparticles (NPs) were prepared from a silica shell-coated Co–B core using the Stöber method and amine-modification on the surface. Glucose oxidase (GOD) was covalently immobilized on the surface of Co–B/SiO₂/NH₂ NPs using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC) as an activating agent. The magnetic NPs characteristics, such as the synthesis of Co–B/SiO₂/NH₂ NPs, effect of pH, temperature, and concentration of buffer for enzyme immobilization, were investigated. The optimal reaction conditions for immobilization were determined to be 0.1 M of phosphate buffer solution, pH 7.0, and 5 °C. In the case of immobilized GOD without d-glucose and with 0.1 M of d-glucose for blocking, 22.98 U/g and 24.83 U/g of their original activity were retained after 7 reuses, respectively.     

Toxicity of silver nanoparticles on the brain of Oreochromis niloticus and Tilapia zillii

BackgroundSilver nanoparticles (Ag-NPs) are widely used nowadays in a variety of commercial applications including medical, health care, textiles and household supplies.ObjectivesThe current study was designed to determine the median lethal dose (LC₅₀) of Ag-NPs on Oreochromis niloticus and Tilapia zillii.MethodsAcute and sub-acute toxicity study of the Ag-NPs on brain tissues was carried out using different concentrations of the NPs at 2 mg L and 4 mg L. These concentrations were dispersed in deionized water with the exception of the control groups in the experiments. Biochemical and molecular analysis were conducted on tissue homogenates in order to evaluate the potential effects of NPs on the antioxidant system.ResultsThe Ag-NP acute toxicity (96 h LC₅₀) values of 19.5 ± 2 and 20 ± 2.4 mg/L were reported for O. niloticus and T. zillii respectively. Fish exposed to 2 mg/L Ag-NPs did not show any significant change in the levels of reduced glutathione (GSH), total glutathione (tGSH) levels, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activity or genes expression and malondialdehyde (MDA) level. In contrary, a dose of 4 mg/L showed a significant reduction in the levels all the above-mentioned parameters except in MDA level where it was significantly induced.ConclusionResults indicate that exposure of O. niloticus and T. zillii to Ag-NPs (4 mg/L) has deleterious effects on brain antioxidant system, whereas a dose of 2 mg/L has no effects.     

C60 fullerenes disrupt cellular signalling leading to TRPC4 and TRPC6 channels opening by the activation of muscarinic receptors and G-proteins in small intestinal smooth muscles

The effect of water-soluble pristine C₆₀ fullerene nanoparticles (C₆₀NPs) on receptor-operated cation channels formed by TRPC4/C6 proteins in ileal smooth muscle cells was investigated for the first time. Activation of these channels subsequent to acetylcholine binding to the expressed in these cells M₂ and M₃ muscarinic receptors represents the key event in the parasympathetic control of gastrointestinal smooth muscle motility and cholinergic excitation-contraction coupling. Experiments were performed on single collagenase-dispersed mouse ileal myocytes using patch-clamp techniques with symmetrical 125 mM Cs⁺ solutions and [Ca^2 +]_i ‘clamped’ at 100 nM in order to isolate the muscarinic cation current (mI_CAT). The current was induced by intracellular infusion of 200 μM GTPγS, which activates G-proteins directly, i.e. bypassing the muscarinic receptors. C₆₀NPs applied at 10^− 6 M at peak response to activation of G-proteins caused mI_CAT inhibition by 47.0 ± 3.5% (n = 9). The inhibition developed rather slowly, with the time constant of 119 ± 16 s, was voltage-independent and irreversible. Thus, C₆₀NPs are unlikely to cause any direct block of TRPC4/C6 channels; rather, they may accumulate in the membrane and disrupt G-protein signalling leading to mI_CAT generation. C₆₀NPs may represent a novel class of biocompatible molecules for the treatment of disorders associated with enhanced gastrointestinal motility.     

Radiological properties of 3D printed materials in kilovoltage and megavoltage photon beams

PurposeThis study evaluates the radiological properties of different 3D printing materials for a range of photon energies, including kV and MV CT imaging and MV radiotherapy beams.MethodsThe CT values of a number of materials were measured on an Aquilion One CT scanner at 80 kVp, 120 kVp and a Tomotherapy Hi Art MVCT imaging beam. Attenuation of the materials in a 6 MV radiotherapy beam was investigated.ResultsPlastic filaments printed with various infill densities have CT values of −743 ± 4, −580 ± 1 and −113 ± 3 in 120 kVp CT images which approximate the CT values of low-density lung, high-density lung and soft tissue respectively. Metal-infused plastic filaments printed with a 90% infill density have CT values of 658 ± 1 and 739 ± 6 in MVCT images which approximate the attenuation of cortical bone. The effective relative electron density RED_eff is used to describe the attenuation of a megavoltage treatment beam, taking into account effects relating to the atomic number and mass density of the material. Plastic filaments printed with a 90% infill density have RED_eff values of 1.02 ± 0.03 and 0.94 ± 0.02 which approximate the relative electron density RED of soft tissue. Printed resins have RED_eff values of 1.11 ± 0.03 and 1.09 ± 0.03 which approximate the RED of bone mineral.Conclusions3D printers can model a variety of body tissues which can be used to create phantoms useful for both imaging and dosimetric studies.     

Characterization of a spore surface lipase from the biocontrol agent Metarhizium anisopliae

A Metarhizium anisopliae spore surface lipase (MASSL) strongly bound to the fungal spore surface has been purified by ion exchange chromatography on DEAE sepharose followed by ultrafiltration and hydrophobic interaction chromatography on phenyl sepharose. Electrophoretic analyses showed that the molecular weight of this lipase is ～66 kDa and pI is 5.6. Protein sequencing revealed that identified peptides in MASSL shared identity with several lipases or lipase-related sequences. The enzyme was able to hydrolyze triolein, the animal lipid cholesteryl stearate and all ρNP ester substrates tested with some preference for esters with a short acyl chain. The values of K_m and V_max for the substrates ρNP palmitate and ρNP laurate were respectively 0.474 mM and 1.093 mMol min^?1 mg^?1 and 0.712 mM and 5.696 mMol min^?1 mg^?1. The optimum temperature of the purified lipase was 30 °C and the enzyme was most stable within the most acid pH range (pH 3–6). Triton X-100 increased and SDS reduced enzyme lipolytic activity. MASSL activity was stimulated by Ca²⁺, Mg²⁺ and Co²⁺ and inhibited by Mn²⁺. The inhibitory effect on activity exerted by EDTA and EGTA was limited, while the lipase inhibitor Ebelactone B completely inhibited MASSL activity as well as PMSF. Methanol 0.5% apparently did not affect MASSL activity while β-mercaptoethanol activated the enzyme.     

Biodistribution of 125I-labeled polymeric vaccine carriers after subcutaneous injection

Polymeric nanoparticles (NPs) comprised of hydrophilic poly(γ-glutamic acid) in the main chain and hydrophobic phenylalanine in the side chain (γ-PGA-Phe) are a promising vaccine carrier for various kinds of diseases. However, little is known about the fate of subcutaneously administered γ-PGA-Phe NPs. Therefore, we newly synthesized γ-PGA graft phenylalanine and tyrosine conjugates (γ-PGA-Phe-Tyr), and then γ-PGA-Phe-Tyr NPs were labeled with ¹²⁵I for monitoring their biodistribution (γ-PGA-Phe-Tyr(¹²⁵I) NPs). Dynamic light scattering (DLS) measurements showed that γ-PGA-Phe-Tyr(¹²⁵I) NPs showed 200 nm in diameter and a negative ζ-potential, which was comparable to those of their precursors. γ-scintigraphic images showed that in mice, subcutaneously injected γ-PGA-Phe-Tyr(¹²⁵I) NPs were mainly observed at the site of injection (SOI), but not other organs 1 h after administration. However, γ-PGA-PheTyr(¹²⁵I) NPs were almost undetectable at the SOI and other organs at 11 days postinjection. Similar results were observed when γ-PGA-Phe-Tyr(¹²⁵I) NPs were subcutaneously injected into rats. Furthermore, at 11 days postinjection, 73 ± 3% of the injected dose of γ-PGA-Phe-Tyr(¹²⁵I) NPs was detected in the feces (14 ± 1%) and urine (59 ± 1%). These results clearly showed that subcutaneously injected γ-PGA-Phe-Tyr(¹²⁵I) NPs were cleared from the body, and γ-PGA-Phe NPs were safe and effective vaccine carriers.