LETd Optimization Verification With an SOI Microdosimeter.

PURPOSE: A first of its kind experimental verification of dose-averaged linear energy transfer (LETd) optimized treatment plans for proton therapy has been carried out using a silicon-on-insulator microdosimeter at the Massachusetts General Hospital (MGH), Boston, USA. METHODS AND MATERIALS: Three clinical treatment plans of a typical ependymoma structure set were designed using the standard clinical approach, the proposed protocol approach, and a one-field approach. The plans were then reoptimized to reduce the LETd-weighted dose in the brain stem. All six plans were delivered in a solid water phantom and the experimental yD‾ measured. RESULTS: After LETd optimization, a reduction in yD‾ was found within the brain stem by an average of 12%, 19%, and 4% for the clinical, protocol, and one-field plans, respectively, while maintaining adequate coverage of the tumor structure. The experimental LETd-weighted doses were in agreement with the treatment planning system calculations and Monte Carlo simulations and reinforced the improvement of the optimization. CONCLUSIONS: This work demonstrates the first experimental verification of the clinical implementation of LETd optimization for patient treatment with proton therapy.

The impact of sensitive volume thickness for silicon on insulator microdosimeters in hadron therapy.

Compact silicon on insulator (SOI) microdosimeters have been used to characterise the radiation field of many different hadron therapy beams. SOI devices are particularly attractive in hadron therapy fields due to their spatial resolution being well suited to the sharp dose gradients at the end of the primary beam's range. Due to the small size of SOI's sensitive volumes (SVs), which are usually ∼1-10 [Formula: see text]m thick, the fabrication of these devices can present challenges which are not as common for more conventional thickness silicon devices such as silicon spectroscopy detectors. Microdosimetry is the study of the energy deposition in micrometre sized volumes representing biological sites and is a powerful approach to estimate the biological effect of radiation on the micron-scale level, in a cell. However, cell sizes vary extensively translating in different energy deposition spectra. This work studies SV thicknesses between 1 and 100 [Formula: see text]m using Geant4 and examines the impact of SV dimensions on microdosimetric quantities. The quantities studied were the frequency mean lineal energy, [Formula: see text], and the dose mean lineal energy, [Formula: see text]. Additionally the relative biological effectiveness (RBE), estimated by the microdosimetric kinetic model (MKM), is also investigated. To study the impact of the SV thickness, SOI microdosimeters were irradiated with proton, [Formula: see text] and [Formula: see text] ion beams with ranges of ∼160 mm, with the microdosimeter being set at various positions along the Bragg curve. It was found that [Formula: see text] was influenced the least in proton beams and increased for heavier ion beams. Conversely, [Formula: see text] was impacted by the SV thickness the most in proton beams and [Formula: see text] was the least. Similar to [Formula: see text], protons were impacted the most by the SV thickness when estimating the RBE using the MKM. The cause of these differences was largely due to the different densities of the delta electron track structure for the case of [Formula: see text] and the energy transferred to the medium from the primary beam for [Formula: see text].

Validation of Geant4 for silicon microdosimetry in heavy ion therapy.

Microdosimetry is a particularly powerful method to estimate the relative biological effectiveness (RBE) of any mixed radiation field. This is particularly convenient for therapeutic heavy ion therapy (HIT) beams, referring to ions larger than protons, where the RBE of the beam can vary significantly along the Bragg curve. Additionally, due to the sharp dose gradients at the end of the Bragg peak (BP), or spread out BP, to make accurate measurements and estimations of the biological properties of a beam a high spatial resolution is required, less than a millimetre. This requirement makes silicon microdosimetry particularly attractive due to the thicknesses of the sensitive volumes commonly being ∼10 [Formula: see text]m or less. Monte Carlo (MC) codes are widely used to study the complex mixed HIT radiation field as well as to model the response of novel microdosimeter detectors when irradiated with HIT beams. Therefore it is essential to validate MC codes against experimental measurements. This work compares measurements performed with a silicon microdosimeter in mono-energetic [Formula: see text], [Formula: see text] and [Formula: see text] ion beams of therapeutic energies, against simulation results calculated with the Geant4 toolkit. Experimental and simulation results were compared in terms of microdosimetric spectra (dose lineal energy, [Formula: see text]), the dose mean lineal energy, y  D and the RBE10, as estimated by the microdosimetric kinetic model (MKM). Overall Geant4 showed reasonable agreement with experimental measurements. Before the distal edge of the BP, simulation and experiment agreed within ∼10% for y  D and ∼2% for RBE10. Downstream of the BP less agreement was observed between simulation and experiment, particularly for the [Formula: see text] and [Formula: see text] beams. Simulation results downstream of the BP had lower values of y  D and RBE10 compared to the experiment due to a higher contribution from lighter fragments compared to heavier fragments.

Optimisation of the design of SOI microdosimeters for hadron therapy quality assurance.

Silicon-on-insulator (SOI) microdosimeters offer a promising method for routine quality assurance (QA) for hadron therapy due to their ease of operation and high spatial resolution. However, one complication which has been shown previously is that the traditional use of the mean chord length, [Formula: see text], calculated using Cauchy's formula, for SOI devices in clinical carbon ion fields is not appropriate due to the strong directionality of the radiation field. In a previous study, we demonstrated that the mean path length, [Formula: see text], which is the mean path of charged particles in the sensitive volume (SV), is a more appropriate method to obtain microdosimetric quantities and biological relevant values, namely the relative biological effectiveness (RBE) by means of the microdosimetric kinetic model. The previous work, which was limited to mono-energetic [Formula: see text] ion beams typical of heavy ion therapy (HIT), is extended here to investigate the [Formula: see text] in a pristine proton beam as well as for spread out Bragg peaks (SOBP) for both proton and carbon ion clinical beams. In addition, the angular dependence of the SOI device for a number of different SV designs is also investigated to quantify the effects which the alignment has on the [Formula: see text]. It is demonstrated that the [Formula: see text] can be accurately estimated along the depth of a pristine or SOBP using the energy deposition spectra for both proton and [Formula: see text] ion beams. This observation allows a quick and accurate estimation of the [Formula: see text] for experimental use.

Feasibility study on the use of 3D silicon microdosimeter detectors for microdosimetric analysis in boron neutron capture therapy.

This paper presents the feasibility study of a novel 3D mesa bridge microdosimeter and its use for BNCT dosimetry. The performance of the microdosimeter was studied using Monte Carlo simulation. The clinical BNCT field at Kyoto University Reactor (KUR) using both thermal and epithermal irradiation modes were used in this study. Results show that this microdosimeter can be utilised as an effective tool to measure microdosimetric spectrum in the BNCT field and experimental validation will follow once KUR is operational.

Recessive mutations in NDUFA2 cause mitochondrial leukoencephalopathy.

Deficiencies of mitochondrial respiratory chain complex I frequently result in leukoencephalopathy in young patients, and different mutations in the genes encoding its subunits are still being uncovered. We report 2 patients with cystic leukoencephalopathy and complex I deficiency with recessive mutations in NDUFA2, an accessory subunit of complex I. The first patient was initially diagnosed with a primary systemic carnitine deficiency associated with a homozygous variant in SLC22A5, but also exhibited developmental regression and cystic leukoencephalopathy, and an additional diagnosis of complex I deficiency was suspected. Biochemical analysis confirmed a complex I deficiency, and whole-exome sequencing revealed a homozygous mutation in NDUFA2 (c.134A>C, p.Lys45Thr). Review of a biorepository of patients with unsolved genetic leukoencephalopathies who underwent whole-exome or genome sequencing allowed us to identify a second patient with compound heterozygous mutations in NDUFA2 (c.134A>C, p.Lys45Thr; c.225del, p.Asn76Metfs*4). Only 1 other patient with mutations in NDUFA2 and a different phenotype (Leigh syndrome) has previously been reported. This is the first report of cystic leukoencephalopathy caused by mutations in NDUFA2.

MICRODOSIMETRIC APPLICATIONS IN PROTON AND HEAVY ION THERAPY USING SILICON MICRODOSIMETERS.

Using the CMRP 'bridge' µ+ probe, microdosimetric measurements were undertaken out-of-field using a therapeutic scanning proton pencil beam and in-field using a 12C ion therapy field. These measurements were undertaken at Mayo Clinic, Rochester, USA and at HIMAC, Chiba, Japan, respectively. For a typical proton field used in the treatment of deep-seated tumors, we observed dose-equivalent values ranging from 0.62 to 0.99 mSv/Gy at locations downstream of the distal edge. Lateral measurements at depths close to the entrance and along the SOBP plateau were found to reach maximum values of 3.1 mSv/Gy and 5.3 mSv/Gy at 10 mm from the field edge, respectively, and decreased to ~0.04 mSv/Gy 120 mm from the field edge. The ability to measure the dose-equivalent with high spatial resolution is particularly relevant to healthy tissue dose calculations in hadron therapy treatments. We have also shown qualitatively and quantitively the effects critical organ motion would have in treatment using microdosimetric spectra. Large differences in spectra and RBE10 were observed for treatments where miscalculations of 12C ion range would result in critical structures being irradiated, showing the importance of motion management.

P-113D, an antimicrobial peptide active against Pseudomonas aeruginosa, retains activity in the presence of sputum from cystic fibrosis patients.

Antimicrobial peptides are a source of novel agents that could be useful for treatment of the chronic lung infections that afflict cystic fibrosis (CF) patients. Efficacy depends on antimicrobial activity against the major pathogens of CF patients, Pseudomonas aeruginosa, Staphylococcus aureus, and Haemophilus influenzae, in the environment of the CF patient's airway. We describe the in vitro efficacies of derivatives of histatins, which are histidine-rich peptides produced by the salivary glands of humans and higher primates. P-113, a peptide containing 12 of the 24 amino acid residues of the parent molecule, histatin 5, retained full antibacterial activity and had a good spectrum of activity in vitro against the prominent pathogens of CF patients. However, P-113 was not active in the presence of purulent sputum from CF patients. In contrast, P-113D, the mirror-image peptide with the amino acid residues in the D configuration, was stable in sputum, was as active as P-113 against pathogens of CF patients in the absence of sputum and retained significant activity in the presence of sputum from CF patients. Recombinant human DNase, which effectively liquefies sputum, enhanced the activity of P-113D in undiluted sputum against both exogenous (added) bacteria and endogenous bacteria. Because of its properties, P-113D shows potential as an inhalant in chronic suppressive therapy for CF patients.

Epidemiological study of Candida spp. colonization in cardiovascular surgical patients.

Candida infections involve multiple risk factors. Among the independent risk factors identified, the degree of colonization of Candida spp. allows the prediction of subsequent severe candidosis in surgical patients. The aim of this study was to assess among 13 selected variables, those that would best predict the perioperative variation of the colonization index (CI) of Candida spp. in cardiovascular surgical patients. The colonization index took into account the number of sites colonized and the density of growth. The results showed that 56.8% of our patients were colonized perioperatively. A total of 116 isolates were identified and Candida albicans accounted for 76.7% of the strains. Among the patients who developed post-surgical Candida infections, 57.1% had an increase of the CI early after the operation. By univariate analysis, three factors were significantly associated with an increase of the CI in patients after surgery; sex (female), the duration of central intravascular catheterization and the length of stay in the surgical intensive care unit (SICU). Epidemiological data could help predict those patients who are at risk of developing Candida infections.

Cytoskeletal regulation of Caco-2 intestinal monolayer paracellular permeability.

An abnormal increase in intestinal paracellular permeability may be an important pathogenic factor in various intestinal diseases. The intracellular factors and processes that regulate and cause alteration of intestinal paracellular permeability are not well understood. The purpose of this study was to examine some of the intracellular processes involved in cytoskeletal regulation of intestinal epithelial paracellular permeability using the filter-grown Caco-2 intestinal epithelial monolayers. Cytochalasin-b and colchicine were used to disrupt the cytoskeletal elements, actin microfilaments, and microtubules. Cytochalasin-b (5 micrograms/ml) and colchicine (2 x 10(-5) M) at the doses used caused marked depolymerization and disruption of actin microfilaments and microtubules, respectively. Cytochalasin-b-induced disruption of actin microfilaments resulted in perturbation of tight junctions and desmosomes and an increase in Caco-2 monolayer paracellular permeability. The cytochalasin-b-induced disruption of actin microfilaments and subsequent changes in intercellular junctional complexes and paracellular permeability were not affected by inhibitors of protein synthesis (actinomycin-D or cycloheximide) or microtubule function (colchicine), but were inhibited by metabolic energy inhibitors (2,4-dinitrophenol or sodium azide). The cytochalasin-b-induced disturbance in Caco-2 actin microfilaments and intercellular junctional complexes and increase in paracellular permeability were rapidly reversed. The paracellular pathway "re-tightening" following cytochalasin-b removal was not affected by actinomycin-D, cycloheximide, or colchicine, but was inhibited by 2,4-dinitrophenol and sodium azide. The colchicine-induced disruption of microtubules did not have significant effect on actin microfilaments, intercellular junctions, or paracellular permeability. These findings suggest that cytochalasin-b-induced increase in Caco-2 monolayer paracellular permeability was due to actin microfilament mediated perturbation of intercellular junctional complexes. The re-tightening of paracellular pathways (following removal of cytochalasin-b) resulted from energy-mediated re-assembly of pre-existing actin microfilaments and intercellular junctional complexes. This re-closure process did not require protein synthesis or microtubule-mediated shuttling process.

Activity of urokinase diluted in 0.9% sodium chloride injection or 5% dextrose injection and stored in glass or plastic syringes.

The effects of the diluent, the container, the i.v. set, and the drug concentration on the adsorption of urokinase to i.v. administration systems were studied, along with the compatibility of urokinase with plastic and glass syringes. Solutions of urokinase 1500 and 5000 IU/mL in 0.9% sodium chloride injection and 5% dextrose injection in glass and polyvinyl chloride (PVC) containers were sampled at 2 and 30 minutes. Administration sets were attached to PVC containers containing the urokinase-5% dextrose injection solutions, and samples were collected at 90 and 150 minutes. Glass and polypropylene syringes containing urokinase 5000 IU/mL in 0.9% sodium chloride injection or 5% dextrose injection were sampled at 0, 4, 8, and 24 hours. Urokinase activity was measured by an in vitro clot lysis assay. No urokinase diluted in 0.9% sodium chloride injection adsorbed to glass or PVC containers. For urokinase 1500 IU/mL in 5% dextrose injection, a loss of 15% to 20% occurred almost instantaneously in PVC containers; additional losses to the infusion sets were minimal. However, for urokinase 5000 IU/mL in 5% dextrose injection, no losses were observed in the PVC systems. No drug loss to glass bottles was seen for urokinase 1500 or 5000 IU/mL in 5% dextrose injection. Urokinase potency remained constant in polypropylene and glass syringes for 24 hours. To minimize urokinase sorption to PVC containers, higher concentrations of urokinase diluted in 5% dextrose injection should be used, provided that clinical safety and efficacy are not compromised. The use of 0.9% sodium chloride injection as a diluent also prevents sorption losses.