Modulation of oxidative phosphorylation (OXPHOS) by radiation- induced biophotons.

Radiation-induced biophotons are an electromagnetic form of bystander signalling. In human cells, biophoton signalling is capable of eliciting effects in non-irradiated bystander cells. However, the mechanisms by which the biophotons interact and act upon the bystander cells are not clearly understood. Mitochondrial energy production and ROS are known to be involved but the precise interactions are not known. To address this question, we have investigated the effect of biophoton emission upon the function of the complexes of oxidative phosphorylation (OXPHOS). The exposure of bystander HCT116 p53 +/+ cells to biophoton signals emitted from ß-irradiated HCT116 p53 +/+ cells induced significant modifications in the activity of Complex I (NADH dehydrogenase or NADH:ubiquinone oxidoreductase) such that the activity was severely diminished compared to non-irradiated controls. The enzymatic assay showed that the efficiency of NADH oxidation to NAD+ was severely compromised. It is suspected that this impairment may be linked to the photoabsorption of biophotons in the blue wavelength range (492-455 nm). The photobiomodulation to Complex I was suspected to contribute greatly to the inefficiency of ATP synthase function since it resulted in a lower quantity of H+ ions to be available for use in the process of chemiosmosis. Other reactions of the ETC were not significantly impacted. Overall, these results provide evidence for a link between biophoton emission and biomodulation of the mitochondrial ATP synthesis process. However, there are many aspects of biological modulation by radiation-induced biophotons which will require further elucidation.

COMP report: CPQR technical quality control guidelines for radiation treatment centers.

The Canadian Organization of Medical Physicists (COMP), in close partnership with the Canadian Partnership for Quality Radiotherapy (CPQR) has developed a series of Technical Quality Control (TQC) guidelines for radiation treatment equipment. These guidelines outline the performance objectives that equipment should meet in order to ensure an acceptable level of radiation treatment quality. The TQC guidelines have been rigorously reviewed and field tested in a variety of Canadian radiation treatment facilities. The development process enables rapid review and update to keep the guidelines current with changes in technology. This announcement provides an introduction to the guidelines, describing their scope and how they should be interpreted. Details of recommended tests can be found in separate, equipment specific TQC guidelines published in the JACMP (COMP Reports), or the website of the Canadian Partnership for Quality Radiotherapy (www.cpqr.ca).

Measurement of skin surface dose distributions in radiation therapy using poly(vinyl alcohol) cryogel dosimeters.

In external beam radiation therapy (EBRT), skin dose measurement is important to evaluate dose coverage of superficial target volumes. Treatment planning systems (TPSs) are often inaccurate in this region of the patient, so in vivo measurements are necessary for skin surface dose estimation. In this work, superficial dose distributions were measured using radiochromic translucent poly(vinyl alcohol) cryogels. The cryogels simultaneously served as bolus material, providing the necessary buildup to achieve the desired superficial dose. The relationship between dose to the skin surface and dose measured with the bolus was established using a series of oblique irradiations with gantry angles ranging from 0° to 90°. EBT-2 Gafchromic film was placed under the bolus, and the ratio of bolus-film dose was determined ranging from 0.749 ± 0.005 to 0.930 ± 0.002 for 0° and 90° gantry angles, respectively. The average ratio over 0-67.5° (0.800 ± 0.064) was used as the single correction factor to convert dose in bolus to dose to the skin surface. The correction factor was applied to bolus measurements of skin dose from head and neck intensity-modulated radiation therapy (IMRT) treatments delivered to a RANDO phantom. The resulting dose distributions were compared to film measurements using gamma analysis with a 3%/3 mm tolerance and a 10% threshold. The minimum gamma pass rate was 95.2% suggesting that the radiochromic bolus may provide an accurate estimation of skin surface dose using a simple correction factor. This study demonstrates the suitability of radiochromic cryogels for superficial dose measurements in megavoltage photon beams.

Translucent poly(vinyl alcohol) cryogel dosimeters for simultaneous dose buildup and monitoring during chest wall radiation therapy.

Chest wall radiation therapy treatment delivery was monitored using a 5 mm thick radiochromic poly(vinyl alcohol) cryogel that also provided buildup material. The cryogels were used to detect positioning errors and measure the impact of shifts for a chest wall treatment that was delivered to a RANDO phantom. The phantom was shifted by ± 2, ± 3, and ± 5 mm from the planned position in the anterior/posterior (A/P) direction; these shifts represent setup errors and the uncertainty associated with lung filling during breath-hold. The two-dimensional absolute dose distributions measured in the cryogel at the planned position were compared with the distributions at all shifts from this position using gamma analysis (3%/3 mm, 10% threshold). For shifts of ± 2, ± 3, and ± 5 mm the passing rates ranged from 94.3% to 95.6%, 74.0% to 78.8%, and 17.5% to 22.5%, respectively. These results are consistent with the same gamma analysis performed on dose planes calculated in the middle of the cryogel and on the phantom surface using our treatment plan-ning system, which ranged from 94.3% to 95.0%, 76.8% to 77.9%, and 23.5% to 24.3%, respectively. The Pinnacle dose planes were then scaled empirically and compared to the cryogel measurements. Using the same gamma metric, the pass rates ranged from 97.0% to 98.4%. The results of this study suggest that cryogels may be used as both a buildup material and to evaluate errors in chest wall treat-ment positioning during deep-inspiration breath-hold delivery. The cryogels are sensitive to A/P chest wall shifts of less than 3 mm, which potentially allows for the detection of clinically relevant errors.

Production, review, and impact of technical quality control guidelines in a national context.

A close partnership between the Canadian Partnership for Quality Radiotherapy (CPQR) and the Canadian Organization of Medical Physicist's (COMP) Quality Assurance and Radiation Safety Advisory Committee (QARSAC) has resulted in the development of a suite of Technical Quality Control (TQC) guidelines for radiation treatment equipment; they outline specific performance objectives and criteria that equipment should meet in order to assure an acceptable level of radiation treatment quality. The adopted framework for the development and maintenance of the TQCs ensures the guidelines incorporate input from the medical physics com-munity during development, measures the workload required to perform the QC tests outlined in each TQC, and remain relevant (i.e., "living documents") through subsequent planned reviews and updates. The framework includes consolidation of existing guidelines and/or literature by expert reviewers, structured stages of public review, external field-testing, and ratification by COMP. This TQC develop-ment framework is a cross-country initiative that allows for rapid development of robust, community-driven living guideline documents that are owned by the com-munity and reviewed to keep relevant in a rapidly evolving technical environment. Community engagement and uptake survey data shows 70% of Canadian centers are part of this process and that the data in the guideline documents reflect, and are influencing, the way Canadian radiation treatment centers run their technical quality control programs. For a medium-sized center comprising six linear accelerators and a comprehensive brachytherapy program, we evaluate the physics workload to 1.5 full-time equivalent physicists per year to complete all QC tests listed in this suite.

A Randomized Phase II Trial of Prostate Boost Irradiation With Stereotactic Body Radiotherapy (SBRT) or Conventional Fractionation (CF) External Beam Radiotherapy (EBRT) in Locally Advanced Prostate Cancer: The PBS Trial (NCT03380806).

Standard therapy for high-risk (HR) prostate cancer (PrCa) involves androgen deprivation therapy (ADT) and pelvic conventional fractionation (CF) external beam radiotherapy (EBRT) followed by boost CF-EBRT treatment to prostate for a total of 78 to 80 Gy in 39 to 40 fractions. This is a long and inconvenient treatment for patients. Brachytherapy boost treatment studies indicate that escalation of biological dose of radiotherapy (RT) can improve outcomes in HR-PrCa. However, brachytherapy is an invasive treatment associated with increased toxicity and requires specialized resources. Stereotactic body radiotherapy (SBRT) is a promising, non-invasive alternative to brachytherapy. However, its impact on patient quality of life (QoL) and RT-associated toxicity has not been investigated in a randomized setting. In this study, we investigate SBRT as a boost treatment, following pelvic CF-EBRT, in patients with HR-PrCa treated with ADT. One hundred patients with locally advanced PrCa will be randomized to receive daily CF-EBRT of 45 to 46 Gy in 23 to 25 fractions followed by either daily CF-EBRT of 32 to 33 Gy in 15 to 16 fractions (control arm) or SBRT boost treatment of 19.5 to 21 Gy in 3 fractions (1 fraction per week) (experimental arm). The primary objective of the PBS trial is early bowel and urinary QoL (expanded prostate index composite [EPIC], up to 6 months after RT). This phase II randomized study (PBS) provides an appropriate setting to investigate effectively the impact of SBRT boost on QoL and toxicity in patients with HR-PrCa, before this modality can be compared against the current standard of care in larger phase III protocols.

Stability of Intracavitary Applicator Placement for HDR Brachytherapy of Cervix Cancer.

INTRODUCTION/BACKGROUND: Cervical cancer is often treated with a combination of external beam radiation therapy and high-dose-rate intracavitary brachytherapy. An intrauterine ring and tandem applicator is used for intracavitary brachytherapy. The dose is prescribed to the high-risk clinical target volume. The goals of this study were to investigate the stability of intracavitary applicator placement during patient transfer and to evaluate the dosimetric impact of displacement. METHODS: Fourteen patients with cervical cancer were analyzed. Three sets of orthogonal fluoroscopic radiographs were obtained in the high-dose-rate suite after the insertion and before treatment: pre-computed tomography (CT) fluoroscopic radiograph with patient in the lithotomy position, pre-CT fluoroscopic radiograph with patient in the legs down position, and post-CT fluoroscopic radiograph with patient in the legs down position. Applicator position after CT was compared with the pre-CT radiographs to determine if the position changed during patient transfer. The displacement was measured in the anterior-posterior, medio-lateral, and superior-inferior directions, as well as the degree of pitch, roll, and yaw. To study the impact of applicator shifts on dose to organs at risk (OARs), the ring and tandem applicator was shifted virtually in the BrachyVision treatment planning system. The OARs studied included the small bowel, sigmoid colon, rectum, and bladder. Five millimeter shifts were made in the superior-inferior, medio-lateral, and anterior-posterior direction. Three degree rotations were made in the pitch, yaw, and roll directions. Applicator shifts were analyzed in only one direction at a time. The dosimetric impact on OARs was evaluated by comparing the original and shifted/rotated plans to dose-volume histogram-based criteria. RESULTS: The average displacements were 1.9 ± 0.5 mm laterally, 3.0 ± 0.6 mm longitudinally, and 9.5 ± 1.5 mm anterior-posterior. The average applicator rotation on the posterior-anterior radiograph was 1.0 ± 0.2° and 2.6 ± 0.6° on the lateral radiograph. Five millimeter anterior-posterior shifts had the greatest effect on dose to OARs. On average, 5 mm anterior shifts had the greatest effect on the small bowel dose, where there was a 13.7% (79.6 cGy) increase in D2cc. Five millimeter anterior shifts also affected bladder dose, with a 36.5% (141.1 cGy) increase in D2cc. Five millimeter POST shifts increased the rectal D2cc by 28.6% (168.7 cGy). Other directional shifts had negligible effects on dose. The largest effect on OAR dose arising from rotations was to the sigmoid colon, when the applicator rotated in the POST pitch direction. As a result, the dose increased by 4.7% (7.6 cGy). All other rotations had minimal impact on OAR doses. CONCLUSION: Patient transfer resulted in applicator shifts and rotations that had a measurable effect on dose to OARs. The displacements were the result of either a direct shift or rotation of the applicator. Additional tracking of these shifts and rotations may clarify the sources of these unwanted motions and suggest possible mitigation strategies.

Effect of liposomal confinement on photothermal and photo-oximetric fluorescence lifetimes of photosensitizers with varying hydrophilicity.

The time-resolved fluorescence of photosensitizers (PSs) of varying hydrophobicities, di-and tetrasulfonated Al phthalocyanines (Al-2 and Al-4), and Photochlor (HPPH), was investigated in liposomes used as cell-mimetic models. Using frequency-and time-domain apparatus, the fluorescence lifetime, tau(fluo), was compared for PSs free in aqueous solution and in a liposome-associated state at varied temperatures (25 to 78 degrees C) and oxygen concentrations (0-190 microM). The analysis of tau(fluo) revealed different decay behaviors for the free-solution and liposome-confined PSs, most significantly for the lipophilic HPPH. Hydrophilic PS drugs (Al-4, Al-2) were less affected by the liposomal confinement, depending on the relative hydrophilicity of the compound and the consequent localization in liposomes. Changes in the emission decay due to confinement were detected as differences in the lifetime between the bulk solution and the liposome-localized PS in response to heating and deoxygenation. Specifically, hydrophilic Al-4 produced an identical lifetime trend as a function of temperature both in solu and in a liposome-confined state. Hydrophobic HPPH exhibited a fundamental transformation in its fluorescence decay kinetics, transitioning from a multiexponential (in free solution) to single-exponential (in liposome) decay. Deoxygenation resulted in a ubiquitous tau(fluo) increase for all PSs in free solution, while the opposite, a tau(fluo) decrease, occurred in all liposomal PSs.

Effect of liposomal confinement on photochemical properties of photosensitizers with varying hydrophilicity.

Preferential tumor localization and the aggregation state of photosensitizers (PSs) can depend on the hydrophilic/hydrophobic nature of the molecule and affect their phototoxicity. In this study, three PSs of different hydrophilicity are introduced in liposomes to understand the structure-photochemistry relationship of PSs in this cellular model system. Absorbance and fluorescence spectra of amphiphilic aluminum (III) phthalocyanine disulfonate chloride adjacent isomer (Al-2), hydrophilic aluminum (III) phthalocyanine chloride tetrasulfonic acid (Al-4), and lipophilic 2-(1-hexyloxyethyl)-2-devinyl pyropheophorbide (HPPH) are compared in a liposomal confined state with free PS in bulk solution. For fluorescence measurements, a broad range of concentrations of both bulk and liposomal confined PSs are examined to track the transition from monomers to dimers or higher order aggregates. Epifluorescence microscopy, absorbance, and fluorescence measurements all confirm different localization of the PSs in liposomes, depending on their hydrophilicity. In turn, the localization affects the aggregation of molecules inside the liposome cell model. Data obtained with such cellular models could be useful in optimizing the photochemical properties of photosensitizing drugs based on their structure-dependent interactions with cellular media and subcellular organelles.

Quantification of fluorophore concentration in vivo using two simple fluorescence-based measurement techniques.

The effect of photodynamic therapy treatments depends on the concentration of photosensitizer at the treatment site; thus a simple method to quantify concentration is desirable. This study compares the concentration of a fluorophore and sensitizer, aluminum phthalocyanine tetrasulfonate (AlPcS4), measured by two simple fluorescence-based techniques in vivo to post mortem chemical extraction and fluorometric assay of those tissues: skin, muscle, fascia, liver, and kidney (cortex and medulla). Fluorescence was excited and detected by a single optical fiber, or by an instrument that measured the ratio of the fluorescence and excitation reflectance. The in vivo measurements were compared to calibration measurements made in tissue-simulating phantoms to estimate the tissue concentrations. Reasonable agreement was observed between the concentration estimates of the two instruments in the lighter colored tissues (skin, muscle, and fascia). The in vivo measurements also agreed with the chemical extractions at low (< 0.6 microg/g) tissue concentrations, but underestimated higher tissue concentrations. Measurements of fluorescence lifetime in vivo demonstrated that AlPcS4 retains its mono-exponential decay in skin, muscle, and fascia tissues with a lifetime similar to that measured in aqueous tissue-simulating phantoms. In liver and kidney an additional short lifetime component was evident.

5-aminolevulinic acid for quantitative seek-and-treat of high-grade dysplasia in Barrett's esophagus cellular models.

High-grade dysplasia (HGD) in Barrett's esophagus (BE) poses increased risk for developing esophageal adenocarcinoma. To date, early detection and treatment of HGD regions are still challenging due to the sampling error from tissue biopsy and relocation error during the treatment after histopathological analysis. In this study, CP-A (metaplasia) and CP-B (HGD) cell lines were used to investigate the "seek-and-treat" potential using 5-aminolevulinic acid-induced protoporphyrin IX (PpIX). The photodynamic therapy photosensitizer then provides both a phototoxic effect and additional image contrast for automatic detection and real-time laser treatment. Complementary to our studies on automatic classification, this work focused on characterizing subcellular irradiation and the potential phototoxicity on both metaplasia and HGD. The treatment results showed that the HGD cells are less viable than metaplastic cells due to more PpIX production at earlier times. Also, due to mitochondrial localization of PpIX, a better killing effect was achieved by involving mitochondria or whole cells compared with just nucleus irradiation in the detected region. With the additional toxicity given by PpIX and potential morphological/textural differences for pattern recognition, this cellular platform serves as a platform to further investigate real-time "seek-and-treat" strategies in three-dimensional models for improving early detection and treatment of BE.

Measurement of fluorophore concentrations and fluorescence quantum yield in tissue-simulating phantoms using three diffusion models of steady-state spatially resolved fluorescence.

Steady-state diffusion theory models of fluorescence in tissue have been investigated for recovering fluorophore concentrations and fluorescence quantum yield. Spatially resolved fluorescence, excitation and emission reflectance Carlo simulations, and measured using a multi-fibre probe on tissue-simulating phantoms containing either aluminium phthalocyanine tetrasulfonate (AlPcS4), Photofrin meso-tetra-(4-sulfonatophenyl)-porphine dihydrochloride The accuracy of the fluorophore concentration and fluorescence quantum yield recovered by three different models of spatially resolved fluorescence were compared. The models were based on: (a) weighted difference of the excitation and emission reflectance, (b) fluorescence due to a point excitation source or (c) fluorescence due to a pencil beam excitation source. When literature values for the fluorescence quantum yield were used for each of the fluorophores, the fluorophore absorption coefficient (and hence concentration) at the excitation wavelength (mu(a,x,f)) was recovered with a root-mean-square accuracy of 11.4% using the point source model of fluorescence and 8.0% using the more complicated pencil beam excitation model. The accuracy was calculated over a broad range of optical properties and fluorophore concentrations. The weighted difference of reflectance model performed poorly, with a root-mean-square error in concentration of about 50%. Monte Carlo simulations suggest that there are some situations where the weighted difference of reflectance is as accurate as the other two models, although this was not confirmed experimentally. Estimates of the fluorescence quantum yield in multiple scattering media were also made by determining mu(a,x,f) independently from the fitted absorption spectrum and applying the various diffusion theory models. The fluorescence quantum yields for AlPcS4 and TPPS4 were calculated to be 0.59 +/- 0.03 and 0.121 +/- 0.001 respectively using the point source model, and 0.63 +/- 0.03 and 0.129 +/- 0.002 using the pencil beam excitation model. These results are consistent with published values.

Evaluation of a ferrous benzoic xylenol orange transparent PVA cryogel radiochromic dosimeter.

A stable cryogel dosimeter was prepared using ferrous benzoic xylenol orange (FBX) in a transparent poly-(vinyl alcohol) (PVA) cryogel matrix. Dose response was evaluated for different numbers of freeze-thaw cycles (FTCs), different concentrations of PVA, and ratios of water/dimethyl sulfoxide. Linear relationships between dose and absorbance were obtained in the range of 0-1000 cGy for all formulations. Increasing the concentration of PVA and number of FTCs resulted in increased absorbance and sensitivity. The effects of energy and dose rate were also evaluated. No significant dose rate dependence was observed over the range 1.05 to 6.33 Gy min(-1). No energy response was observed over photon energies of 6, 10, and 18 MV.

Monitoring photosensitizer uptake using two photon fluorescence lifetime imaging microscopy.

Photodynamic Therapy (PDT) provides an opportunity for treatment of various invasive tumors by the use of a cancer targeting photosensitizing agent and light of specific wavelengths. However, real-time monitoring of drug localization is desirable because the induction of the phototoxic effect relies on interplay between the dosage of localized drug and light. Fluorescence emission in PDT may be used to monitor the uptake process but fluorescence intensity is subject to variability due to scattering and absorption; the addition of fluorescence lifetime may be beneficial to probe site-specific drug-molecular interactions and cell damage. We investigated the fluorescence lifetime changes of Photofrin(®) at various intracellular components in the Mat-LyLu (MLL) cell line. The fluorescence decays were analyzed using a bi-exponential model, followed by segmentation analysis of lifetime parameters. When Photofrin(®) was localized at the cell membrane, the slow lifetime component was found to be significantly shorter (4.3 ± 0.5 ns) compared to those at other locations (cytoplasm: 7.3 ± 0.3 ns; mitochondria: 7.0 ± 0.2 ns, p < 0.05).

Quantification of fluorophore concentration in tissue-simulating media by fluorescence measurements with a single optical fiber.

Quantifying fluorescent compounds in turbid media such as tissue is made difficult by the effects of multiple scattering and absorption of the excitation and emission light. The approach that we used was to measure fluorescence using a single 200-microm optical fiber as both the illumination source and the detector. Fluorescence of aluminum phthalocyanine tetrasulfonate (AlPcS4) was measured over a wide range of fluorophore concentrations and optical properties in tissue-simulating phantoms. A root-mean-square accuracy of 10.6% in AlPcS4 concentration was attainable when fluorescence was measured either interstitially or at the phantom surface. The individual effects of scattering, absorption, and the scattering phase function on the fluorescence signal were also studied by experiments and Monte Carlo simulations.