Triple channel analysis of Gafchromic EBT3 irradiated with clinical carbon-ion beams

Self-developing radiochromic film is widely used in radiotherapy QA procedures. To compensate for typical film inhomogeneities, the triple channel analysis method is commonly used for photon-irradiated film. We investigated the applicability of this method for Gafchromic^TMEBT3 (Ashland) film irradiated with a clinically used carbon-ion beam. Calibration curves were taken from EBT3 film specimens irradiated with monoenergetic carbon-ion beams of different doses. Measurements of the lateral field shape and homogeneity were performed in the middle of a passively modulated spread-out Bragg peak and compared to simultaneous characterization by means of a 2D ionization chamber array. Additional measurements to investigate the applicability of EBT3 for quality assurance (QA) measurement in carbon-ion beams were performed. The triple-channel analysis reduced the relative standard deviation of the doses in a uniform carbon ion field by 30% (from 1.9% to 1.3%) and reduced the maximum deviation by almost a factor of 3 (from 28.6% to 9.8%), demonstrating the elimination of film artifacts. The corrected film signal showed considerably improved image quality and quantitative agreement with the ionization chamber data, thus providing a clear rationale for the usage of the triple channel analysis in carbon-beam QA.     

Effect of verification imaging on in vivo dosimetry results using Gafchromic EBT3 film

In this work, the apparent treatment dose that kV planar or CBCT imaging contributes to Gafchromic EBT3 film used for in vivo dosimetry, was investigated. Gafchromic EBT3 film pieces were attached to a variety of phantoms and irradiated using the linear accelerator’s built-in kV imaging system, in both kV planar mode and CBCT mode. To evaluate the sensitivity of the film in the clinical scenario where dose contributions are received from both imaging and treatment, additional pieces of film were irradiated using base doses of 50 cGy and then irradiated using selected kV planar and CBCT techniques. For kV planar imaging, apparent treatment doses of up to 3.4 cGy per image pair were seen. For CBCT, apparent treatment doses ranged from 0.22 cGy to 3.78 cGy. These apparent doses were reproducible with and without the inclusion of the 50 cGy base dose. The contribution of apparent treatment dose from both planar kV as well as CBCT imaging can be detected, even in conjunction with an actual treatment dose. The magnitude of the apparent dose was found to be dependent on patient geometry, scanning protocol, and measurement location. It was found that the apparent treatment dose from the imaging could add up to 8% of additional uncertainty to the in vivo dosimetry result, if not taken into account. It is possible for this apparent treatment dose to be accounted for by subtraction of the experimentally determined apparent doses from in vivo measurements, as demonstrated in this work.     

Assessment of clinically relevant dose distributions in pelvic IOERT using Gafchromic EBT3 films

PurposeIn IOERT a single dose of radiation is delivered to the tumour site during surgery. Manual dose calculations are used and the irradiation target volume, electron energy and applicator are decided on site by the radiation oncologist. This work assesses the effect that irregular and curved surfaces, typical of pelvic IOERT, may have on the expected dose distribution.MethodsThe feasibility of using Gafchromic EBT3 films and a slab phantom to obtain 2D dose distributions was investigated. Different set-ups were tested by comparison with water tank measurements, applying the gamma function analysis with 2% and 2 mm criteria. The validated set-up was then used to obtain reference dose distributions, which were converted to colour-coded graphical representations. Phantoms with step-like and curved surfaces were created to simulate typical pelvic IOERT irradiation surfaces, and the dose distributions were obtained and compared with the reference distributions.ResultsGood agreement with water tank measurements was obtained for all applicators below 2 mm, using the chosen setup in reference conditions. In non-reference conditions, the presence of a step-like surface creates an adjacent hotspot, followed by a quick reduction of the dose in depth. With curved surfaces, the dose distribution is shifted forward, becoming curved and deeper, but when the applicator is larger than the hole, hotspots are also observed.ConclusionsThe shape of the irradiation surfaces alters the dose distribution. Visualization of these effects is important to assess target coverage and interpret in vivo measurements in pelvic IOERT.     

Evaluation of Gafchromic EBT2 film for the measurement of anisotropy function for high-dose-rate 192Ir brachytherapy source with respect to thermoluminescent dosimetry

Aim

The aim of this work was to assess the suitability of the use of a Gafchromic EBT2 film for the measurement of anisotropy function for microSelectron HDR ¹⁹²Ir (classic) source with a comparative dosimetry method using a Gafchromic EBT2 film and thermoluminescence dosimeters (TLDs).

Background

Sealed linear radiation sources are commonly used for high dose rate (HDR) brachytherapy treatments. Due to self-absorption and oblique filtration of radiation in the source capsule material, an inherent anisotropy is present in the dose distribution around the source which can be described by a measurable two-dimensional anisotropy function, F(r, θ).

Materials and methods

Measurements were carried out in a specially designed and locally fabricated PMMA phantom with provisions to accommodate miniature LiF TLD rods and EBT2 film dosimeters at identical radial distances with respect to the ¹⁹²Ir source.

Results

The data of anisotropy function generated by the use of the Gafchromic EBT2 film method are in agreement with their TLD measured values within 4%. The produced data are also consistent with their experimental and Monte Carlo calculated results for this source available in the literature.

Conclusion

Gafchromic EBT2 film was found to be a feasible dosimeter in determining anisotropy in the dose distribution of ¹⁹²Ir source. It offers high resolution and is a viable alternative to TLD dosimetry at discrete points. The method described in this paper is useful for comparing the performances of detectors and can be applied for other brachytherapy sources as well.     

Under-response correction for EBT3 films in the presence of proton spread out Bragg peaks

We present a study of the under-response of the new Gafchromic EBT3 films and a procedure to accurately perform 2D and 3D proton dosimetry measurements for both pristine and spread out Bragg peaks (SOBP) of any energy. These new films differ from the previous EBT2 generation by a slightly different active layer composition, which we show has not effected appreciably their response. The procedure and the beam quality correction factor curve have been benchmarked using 29 MeV modulated proton beams. In order to show the correction to apply when EBT3 films are used as treatment verification tools in anthropomorphic phantoms, two simulation studies involving clinical energies are presented: a SOBP for eye treatments and a SOBP to treat 20 cm deep and 5 cm thick tumours. We find maximum under-responses of 37%, 30% and 7.7% for the modulated 29 MeV beam, eye and deep tumour treatment, respectively, which were attained close to the end of the peak tails, due to a higher proportion of very low energy protons. The maximum deviations between corrected and uncorrected doses were for the three cases, respectively, 20.7%, 8.3% and 2.1% of the average dose across flat region of the SOBP. These values were obtained close to the distal edge of the SOBPs, where the proportion of low energy protons was not as high as on the tail, but there still was a number of protons high enough to deposit a reasonable amount of dose in the films.     

Radiation dose verification of an X-ray based blood irradiator using EBT3 radiochromic films calibrated using Gamma Knife machine

AimBlood irradiators (BI) initial acceptance testing and routine annual dosimetry checks require radiation dose measurements in order to comply with regulatory requirements.BackgroundTraditionally thermo-luminescence dosimeters (TLD) have been used to measure the dose. The EBT3 film is reported to be a better dosimeter for low energy X-rays than its predecessors EBT2 and EBT. To the best of our knowledge, the use of EBT3 films to perform dosimetry on X-ray based BI has not been reported yet.Materials and methodsWe performed routine radiation dosimetry checks using EBT3 films on a new X-ray based BI and compared the results with TLD dosimetry. Calibration films were irradiated with radiation beam from a Co-60 Gamma Knife (GK) radiosurgery machine and, alternatively, using an Ir-192 high dose rate (HDR) brachytherapy device. The films were calibrated to cover a wide dose range from 1 to 40 Gy. Such a wide dose range has not been reported yet in BI film dosimetry.ResultsWe obtained a relative difference of about 6.6% between doses measured using TLD and those measured using EBT3 films. Both irradiation methods using GK or HDR were found to be adequate for the calibration of the EBT3 Gafchromic films.ConclusionsWe recommend the use of EBT3 films in routine X-ray based BI dosimetry checks. The presented method takes advantage of available radiotherapy equipment that can be efficiently used for EBT3 films calibration. The method is fast, reproducible and saves valuable medical physicist's time.     

Surface dose measurements with GafChromic EBT film for 6 and 18 MV photon beams

The aim of this study was to determine the surface doses using GafChromic EBT films and compare them with plane-parallel ionization chamber measurements for 6 and 18 MV high energy photon beams. The measurements were made in a water equivalent solid phantom in the build-up region of the 6 and 18 MV photon beams at 100 cm SSD for various field sizes. Markus type plane-parallel ion chamber with fixed-separation between collecting electrodes was used to measure the percent depth doses. GafChromic EBT film measurements were performed both on the phantom surface and maximum dose depth at the same geometry with ion chamber measurements. The surface doses found using GafChromic EBT film were 15%, 20%, 29%and 39% ± 2% (1SD) for 6 MV photons, 6%, 11%, 23% and 32% ± 2% (1SD) for 18 MV photons at 5, 10, 20 and 30 cm² field sizes, respectively. GafChromic EBT film provides precise measurements for surface dose in the high energy photons. Agreement between film and plane-parallel chamber measurements was found to be within ±3% for 18 MV photon beams. There was 5% overestimate on the surface doses when compared with the plane-parallel chamber measurements for all field sizes in the 6 MV photon beams.     

Energy and dose dependence of GafChromic EBT3-V3 film across a wide energy range

AimTo determine the energy and dose dependence of GafChromic EBT3-V3 film over an energy range 0.2 mm Al HVL to 6 MV.BackgroundThe decay scheme of a brachytherapy source may be complex and the spectrum of energy can be wide. LiF TLDs are the golden standard recommended for dosimetric measures in brachytherapy, for their energy independence, but TLDs could be not available in some centres. An alternative way to perform dose measurements is to use GafChromic films, but they show energy dependence.Methods and materialsFilms have been irradiated at increasing dose with three different beams: 6 MV beam, TPR20, 10 = (0.684 ± 0.01), HVL = (2.00 ± 0.01)mmAl and HVL = (0.20 ± 0.01)mmAl. Calibration curves were generated using the same dose range (0cGy to 850cGy) for the three energies. Using the 6 MV calibration curve as reference, the film response in terms of net optical density (OD) was evaluated.ResultsThe difference in the calibration curve obtained by irradiating the film with 6 MV and 2 mm Al HVL energy beams is less than 3 %, within the calibration uncertainty, in the dose range 500-850cGy. The OD of EBT3-V3 film is significantly lower at 0.2 mmAl HVL compared to 6 MV, showing differences up to 25 %.ConclusionWithin the range 6 MV-2 mm Al HVL and dose higher than 500cGy, GafChromic EBT3-V3 films are energy independent. In this dose range, films can be calibrated in a simple geometry, using a 6 MV Linac beam, and can be used for brachytherapy sources dose measures. The use of EBT3 films can be extended to reference dosimetry in Ir-192 clinical brachytherapy.     

Measurement and comparison of head scatter factor for 7 MV unflattened (FFF) and 6 MV flattened photon beam using indigenously designed columnar mini phantom

Aim

To measure and compare the head scatter factor for 7 MV unflattened and 6 MV flattened photon beam using a home-made designed mini phantom.

Background

The head scatter factor (Sc) is one of the important parameters for MU calculation. There are multiple factors that influence the Sc values, like accelerator head, flattening filter, primary and secondary collimators.

Materials and methods

A columnar mini phantom was designed as recommended by AAPM Task Group 74 with high and low atomic number material for measurement of head scatter factors at 10 cm and d_max dose water equivalent thickness.

Results

The Sc values measured with high-Z are higher than the low-Z mini phantoms observed for both 6MV-FB and 7MV-UFB photon energies. Sc values of 7MV-UFB photon beams were smaller than those of the 6MV-FB photon beams (0.6–2.2% (Primus), 0.2–1.4% (Artiste) and 0.6–3.7% (Clinac iX (2300CD))) for field sizes ranging from 10 cm × 10 cm to 40 cm × 40 cm. The SSD had no influence on head scatter for both flattened and unflattened beams. The presence of wedge filters influences the Sc values. The collimator exchange effects showed that the opening of the upper jaw increases Sc irrespective of FF and FFF.

Conclusions

There were significant differences in Sc values measured for 6MV-FB and unflattened 7MV-UFB photon beams over the range of field sizes from 10 cm × 10 cm to 40 cm × 04 cm. Different results were obtained for measurements performed with low-Z and high-Z mini phantoms.     

Optimized methodology for extraction of (1 → 3)(1 → 6)-β-d-glucan from Saccharomyces cerevisiae and in vitro evaluation of the cytotoxicity and genotoxicity of the corresponding carboxymethyl derivative

The cell wall of Saccharomyces cerevisiae is an important source of β-d-glucan, a glucose homopolymer with immunostimulant properties. The standard methodologies described for its extraction involve acid and alkaline washings, which degrade part of its glucose chains and reduce the final yield. In the present study, an optimized methodology for extraction of β-d-glucan from S. cerevisiae cells, involving sonication and enzyme treatment, with a yield of 11.08 ± 0.19%, was developed. The high-purity (1 → 3)(1 → 6)-β-d-glucan was derivatized to carboxymethyl-glucan (CM-G). In vitro tests with CM-G in Chinese hamster epithelial cells (CHO-k1) did not reveal any cytotoxic or genotoxic effects or influences of this molecule on cell viability. The method described here is a convenient alternative for the extraction of (1 → 3)(1 → 6)-β-d-glucan under mild conditions without the generation of wastes that could be potentially harmful to the environment.     

Interdependent Nucleocytoplasmic Trafficking and Interactions of Dis3 with Rrp6, the Core Exosome and Importin‐α3

Subcellular compartmentalization of exoribonucleases (RNAses) is an important control mechanism in the temporal and spatial regulation of RNA processing and decay. Despite much progress towards understanding RNAse substrates and functions, we know little of how RNAses are transported and assembled into functional, subcellularly restricted complexes. To gain insight into this issue, we are studying the exosome‐binding protein Dis3, a processive 3′ to 5′ exoribonuclease. Here, we examine the interactions and subcellular localization of the Drosophila melanogaster Dis3 (dDis3) protein. N‐terminal domain mutants of dDis3 abolish associations with the ‘core’ exosome, yet only reduce binding to the ‘nuclear’ exosome‐associated factor dRrp6. We show that nuclear localization of dDis3 requires a C‐terminal classic nuclear localization signal (NLS). Consistent with this, dDis3 specifically co‐precipitates the NLS‐binding protein importin‐α3. Surprisingly, dDis3 constructs that lack or mutate the C‐terminal NLS retain importin‐α3 binding, suggesting that the interaction is indirect. Finally, we find that endogenous dDis3 and dRrp6 exhibit coordinated nuclear enrichment or exclusion, suggesting that dDis3, Rrp6 and importin‐α3 interact in a complex independent of the core. We propose that the movement and deposition of this complex is important for the subcellular compartmentalization and regulation of the exosome core.     

Selection of mutants resistant to Alternaria blotch from in vitro -cultured apple shoots irradiated with X- and γ-rays

We produced mutants resistant to Alternaria blotch disease in several cultivars of apple (Malus × domestica Borkh.) by irradiation with X- or γ-rays. An efficient in vitro assay method was established using chemically-synthesized AM-toxin I of Alternaria alternata (Fr.) Keissler to screen for mutants resistant to Alternaria blotch disease. The frequency of necrotic lesions was investigated by applying various concentrations of AM-toxin I to leaf discs of the first, third, and fifth leaves from the shoot apex of several apple cultivars, including Jonathan, Fuji, Oorin, and Indo. In vitro-grown apple shoots of susceptible cultivars were then treated with various doses of X- or γ-ray irradiation. Several mutants resistant to AM-toxin I were obtained by combining the techniques for tissue culture of apple shoots with the AM-toxin I screening method. Following a repeat second screening test with AM-toxin I, mutant plants were sprayed with a spore suspension of A. alternata and found resistant to be the fungal pathogen. These mutants showed normal phenotypic appearance, and so far, no difference has been observed between the original plants and mutants except for the susceptibility to Alternaria blotch.     

Studies on the 5α-androstane-3β, 17β-diol-6α-hydroxylase and on the 5α-androstane-3β, 17β-diol-7α-hydroxylase in anterior hypophysis of male rats.

In anterior pituitaries from male rats, it appeared that 5α-androstane-3β, 17β-diol was quickly metabolized into 5α-androstane-3β,6α-17β-triol and 5α-androstane-3β,7α, 17β-triol by action of 6α- and 7α-hydroxylases. Hydroxysteroid hydroxylases were located in endoplasmic reticulum and were dependent on NADPH⁺. Their optimum pH was 8.0, optima temperature, 37°C, and their apparent Km was 2.7 μM. Hydroxylative reactions were not reversible and not modified by gonadectomy. Hydroxylation seemed an efficient control of the pituitary level of 5α-andros-tane-3β, 17β-diol.     

Persistent luminescence of CaMgSi2O6:Eu2+,Dy3+ and CaMgSi2O6:Eu2+,Ce3+ phosphors prepared using the solid‐state reaction method

CaMgSi₂O₆:Eu²⁺,Dy³⁺ and CaMgSi₂O₆:Eu²⁺,Ce³⁺ phosphors were synthesized using the solid‐state reaction method. X‐Ray diffraction (XRD) and photoluminescence (PL) analyses were used to characterize the phosphors. The XRD results revealed that the synthesized CaMgSi₂O₆:Eu²⁺,Dy³⁺ and CaMgSi₂O₆:Eu²⁺,Ce³⁺ phosphors were crystalline and are assigned to the monoclinic structure with a space group C2/c. The calculated crystal sizes of CaMgSi₂O₆:Eu²⁺,Dy³⁺ and CaMgSi₂O₆:Eu²⁺,Ce³⁺ phosphors with a main (221) diffraction peak were 44.87 and 53.51 nm, respectively. Energy‐dispersive X‐ray spectroscopy (EDX) confirmed the proper preparation of the sample. The PL emission spectra of CaMgSi₂O₆:Eu²⁺,Dy³⁺ and CaMgSi₂O₆:Eu²⁺,Ce³⁺ phosphors have a broad band peak at 444.5 and 466 nm, respectively, which is due to electronic transition from 4f⁶5d¹ to 4f⁷. The afterglow results indicate that the CaMgSi₂O₆:Eu²⁺,Dy³⁺ phosphor has better persistence luminescence than the CaMgSi₂O₆:Eu²⁺,Ce³⁺ phosphor. Copyright © 2015 John Wiley & Sons, Ltd.     

Effects of rare earth ions (Tb,Ce, Eu,Dy) on the thermoluminescence characteristics of sol–gel derived and γ‐irradiated SiO2 nanoparticles

Highly pure SiO₂ and SiO₂:RE nanoparticles were synthesized by the sol–gel method. The morphological, structural and optical properties of the nanoparticles were characterized by X‐ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). XRD results indicate that all the samples studied were free from impurities. SEM/TEM results indicate that the samples were well dispersed. Surface characterization of the nanocrystals by Fourier transform infrared spectroscopy has been carried out and the structure of surface‐bound SiO₂ based on spectral analysis is proposed. Thermoluminescence (TL) characteristics were investigated to study the influence of rare earth dopants (Tb, Ce, Eu, Dy) on SiO₂ matrix subjected to 0.5 kG (1 h) γ‐irradiation. Among these rare earth elements, Eu³⁺ was found to be the most efficient dopant for SiO₂ showing maximum thermoluminescence intensity. SiO₂:Eu_0.5 seems to be a promising candidate for use as a TL dosimeter. Copyright © 2013 John Wiley & Sons, Ltd.     

Novel high-performance liquid chromatographic assay using fluorescence detection for the quantitation of plasma γ-methylene-10-deazaaminopterin and its major metabolite, 7-hydroxy-γ-methylene-10-deazaaminopterin, in patients with solid cancers in a phase I trial

γ-Methylene-10-deazaaminopterin (MDAM), a unique dihydrofolate reductase inhibitor, has demonstrated antitumor activity against a broad spectrum of human solid tumors in preclinical studies. A novel reversed-phase, ion-pair high-performance liquid chromatography (HPLC) assay that uses fluorescence detection has been developed to quantitate levels of MDAM and its major metabolite, 7-hydroxy-γ-methylene-10-deazaaminopterin (7-OH-MDAM), in human plasma. The recovery of MDAM and 7-OH-MDAM from plasma was >97% by a simple one-step deproteinization process using tetrabutylammonium bromide (TBABr) and methanol. MDAM and 7-OH-MDAM remained stable in plasma over a 28-day test period at ambient temperatures, and neither compound was light-sensitive. The limit of quantitation was 0.005 μM for both MDAM and 7-OH-MDAM. This assay has been found to be simple, sensitive and reproducible in determining plasma concentrations of MDAM and 7-OH-MDAM in patients with solid cancers in a phase I trial.     

Models of Monoamine Oxidase A and B Active Sites Obtained by using 3D QSAR with CoMFA Analysis

Abstract

The monoamine oxidase catalyses the oxidative deamination of neuroactive amines. This enzyme exists in two forms A and B, which differ by substrates preference and inhibitors specificity. Investigation of the structures of these enzymes and design new selective inhibitors are of greatly interesting since MAO A inhibitors are used in therapeutic practice as antidepressants and MAO B inhibitors – in the treatment Parkinson's diseases. The three dimension structures of monoamine oxidases are still unknown. Therefore, one of the most perspective approach to define significant features of structure active site is method based on analysis of structure-activity relationship (3D QSAR) with comparison of molecular fields analysis (CoMFA) allowing to get the spatial distribution of important properties affecting the activity.

In present study we investigate the structures of active sites MAO A and B using 16 pyrazinocarbazole derivatives in variant conformation. Majority of pyrazinocarbazole derivatives have a rigit conformation, but three of those is sufficiently flexible. The latters can be in two conformation types: long molecules (substitution accommodate along axis of main structure) and short molecules (substitution accommodate at acute angle about of main structure). Several 3D QSAR and CoMFA models of MAO A and B active sites were design for data sets containing various types of flexible molecules conformation. All obtained models are statistical reliable and have sufficient predictive power for tested compound tetrindole. The best MAO A model that include two flexible molecules in long conformations was obtained, and the longest one of those in short conformation. In contrast, for MAO B model containing all flexible molecules in the short conformations is more preferred.

On the basis of obtained data the schematic models of MAO A and B active sites structures are proposed. According to these models MAO A active site have the narrow long cavity that accommodate long molecules, while MAO B active site is broader and shorter.