Methods to Assess Autophagy and Chronological Aging in Yeast.

Autophagy is a catabolic process that is crucial for cellular homeostasis and adaptive response to changing environments. Importantly, autophagy has been shown to be induced in many longevity-associated scenarios and to be required to maintain lifespan extension. Notably, autophagy is a highly conserved cellular process among eukaryotes, and the yeast Saccharomyces cerevisiae has become a universal model system for unraveling the molecular machinery underlying autophagic mechanisms. Here, we discuss different protocols to monitor survival and autophagy of yeast cells upon chronological aging. These include the use of propidium iodide to assess the loss of cell membrane integrity, as well as clonogenic assays to directly determine survival rates. Additionally, we describe methods to quantify autophagic flux, including the alkaline phosphatase activity or the GFP liberation assays, which measure the delivery of autophagosomal cargo to the vacuole. In sum, we have recapped established protocols used to evaluate a link between lifespan extension and autophagy in yeast.

Simultaneous irradiation of the breast and regional lymph nodes in prone position using helical tomotherapy.

OBJECTIVE: We investigated dosimetric advantages of using helical tomotherapy to simultaneously irradiate the breast and regional lymph nodes for patients positioned prone, and compared tomotherapy plan qualities for the prone position with those previously published for the supine position. METHODS: Tomotherapy plans for 11 patients (5 left breast, 6 right) simulated with the involved breast suspended downward were generated. Each target (ipsilateral breast and supraclavicular, axillary and internal mammary chain nodes) was to receive 45 Gy. RESULTS: For targets, V(40.5)≥99.9% and V(42.8)≥99.5% for all patients, where V(40.5) and V(42.8) denote the relative target volume receiving at least 40.5 and 42.8 Gy, respectively. The targets' maximum dose was, on average, approximately 49.5 Gy. The mean doses to the contralateral lung and heart were lower for right-breast cases (2.8 Gy lung, 2.7 Gy heart) than for left-breast cases (3.8 Gy lung, 8.7 Gy heart). Mean organ doses to the ipsilateral lung (9.3 Gy) and contralateral breast (2.3 Gy) from the prone breast tomotherapy plans were similar to those reported for conventional radiotherapy techniques. For the left breast with regional nodes, tomotherapy plans for prone-positioned patients yielded lower mean doses to the contralateral breast and heart than previously reported data for tomotherapy plans for supine-positioned patients. CONCLUSION: Helical tomotherapy with prone breast positioning can simultaneously cover the breast and regional nodes with acceptable uniformity and can provide reduced mean dose to proximal organs at risk compared with tomotherapy with supine position. The similarity of plan quality to existing data for conventional breast radiotherapy indicates that this planning approach is appropriate, and that the risk of secondary tumour formation should not be significantly greater.

SU-E-T-438: Optimal Proton Beam Energy to Treat Adult CNS, Adult Head and Neck, and Pediatric Cancers.

PURPOSE: Proton therapy machines with lower beam energy requirements are expected to be easier and more cost-efficient to design, construct, and operate, particularly when using novel proton acceleration technology (e.g., dielectric wall). We determine, for adult central nervous system (CNS) and head and neck (HN) and pediatric cases, the optimal proton beam energies that can lead to clinically acceptable plan quality. METHODS: Proton treatment plans for various adult CNS and HN and pediatric cases, previously treated at our institution using helical tomotherapy, were generated using a commercial planning system (XiO, v. 4.62, Elekta) with a passively-scattered proton beam model. Proton beam orientations were chosen such that the distance from the skin to the distal target edge was minimal; however, beams could not pass through areas with significant surface irregularity (e.g. ear), nor through OARs with strict dose limitations (e.g. lens). For a given beam direction, the planning system would calculate an optimum range and modulation to cover a given target volume. Beam weights were adjusted so that 95% of the target volume received the prescribed dose. All plans were limited to two proton beams. RESULTS: An optimal proton energy exists for each case studied. Among all cases, target dose conformity (the ratio of target volume to volume encompassed by theprescribed dose) ranged from 0.74 to 0.94, and target dose uniformity (the ratio of the D5 and D95 doses) ranged from 1.01 to 1.04, with the optimal proton energies. Among the CNS and HN cases and for most pediatric cases, the maximum proton energy required was 133 MeV. For a pediatric abdomen case, 159 MeV protons were required. CONCLUSIONS: For protontreatment of most adult CNS and HN and pediatric cases, the optimal energies that are capable of generating good quality plans are only as high as 130 MeV.

The dosimetric and delivery advantages of a new 160-leaf MLC.

The purpose of this study was to conduct a measurement and treatment planning study on the dosimetric and delivery advantages of a new 160-leaf multileaf collimator (MLC). Recently, a new 160-leaf multileaf collimator (Siemens 160 MLC(TM)) was introduced. The 160-MLC is a single focused design that consists of 160-leafs (80 pairs), each 95 mm thick with a projected leaf width of 5 mm at the machine isocenter. Compared to its double focused predecessors, the 82-leaf MLC (Siemens OPTIVIEW((tm)) MLC) and 58-leaf MLC (Siemens 3-D MLC((tm))), the 160-MLC has leaf widths of half the size. The most notable difference is the new slanted leaf design that replaced the tongue and groove system and allows for complete interdigitation. A systematic study that compared the dosimetric and delivery differences among the 160-MLC, 58-MLC, and divergent Cerrobend blocks was performed. Dosimetric conformity for each collimator type was determined by conforming each to circular targets of various diameters. The effective penumbra for each collimator type was calculated by conforming each, at various collimator angles, to a square stationary target. The quality of 3D conformal radiotherapy treatment (3D-CRT) plans and the quality intensity modulated radiation treatment (IMRT) plans were respectively compared with each collimator type. The 160-MLC was found to have improved dosimetric conformity over the 58-MLC. The divergent Cerrobend block showed marginal dosimetric conformity improvement over the 160-LMC. Overall, the 160-MLC had a 45% and 29% reduction in the 20/80 and 30/90 effective penumbra over the 58-MLC, respectively, while exhibiting only a slightly larger effective penumbra over the divergent Cerrobend block. Comparing 3D-CRT plans generated for small lesions of the head and neck, the V100 for the PTV of the plans generated with the Cerrobend blocks, the 58-MLC, and the 160-MLC were 97.78%, 92.51%, and 99.18%, respectively, while with regards to the OARs, the three produced similar DVHs. IMRT plans generated with the 160-MLC were found to significantly reduce the total delivered monitor units by up to 14.7% and the number of segments by as much as 10.7% compared to the 58-MLC. The average delivery time for the direct aperture optimized (DAO) IMRT plans generated with the 160-MLC was approximately 5 minutes. Overall, compared to the 58-MLC, the new 160-MLC was found to improve dosimetric conformity and IMRT delivery efficiency.

Dual scattering foil design for poly-energetic electron beams.

The laser wakefield acceleration (LWFA) mechanism can accelerate electrons to energies within the 6-20 MeV range desired for therapy application. However, the energy spectrum of LWFA-generated electrons is broad, on the order of tens of MeV. Using existing laser technology, the therapeutic beam might require a significant energy spread to achieve clinically acceptable dose rates. The purpose of this work was to test the assumption that a scattering foil system designed for a mono-energetic beam would be suitable for a poly-energetic beam with a significant energy spread. Dual scattering foil systems were designed for mono-energetic beams using an existing analytical formalism based on Gaussian multiple-Coulomb scattering theory. The design criterion was to create a flat beam that would be suitable for fields up to 25 x 25 cm2 at 100 cm from the primary scattering foil. Radial planar fluence profiles for poly-energetic beams with energy spreads ranging from 0.5 MeV to 6.5 MeV were calculated using two methods: (a) analytically by summing beam profiles for a range of mono-energetic beams through the scattering foil system, and (b) by Monte Carlo using the EGS/BEAM code. The analytic calculations facilitated fine adjustments to the foil design, and the Monte Carlo calculations enabled us to verify the results of the analytic calculation and to determine the phase-space characteristics of the broadened beam. Results showed that the flatness of the scattered beam is fairly insensitive to the width of the input energy spectrum. Also, results showed that dose calculated by the analytical and Monte Carlo methods agreed very well in the central portion of the beam. Outside the useable field area, the differences between the analytical and Monte Carlo results were small but significant, possibly due to the small angle approximation. However, these did not affect the conclusion that a scattering foil system designed for a mono-energetic beam will be suitable for a poly-energetic beam with the same central energy. Further studies of the dosimetric properties of LWFA-generated electron beams will be done using Monte Carlo methods.

Dose properties of x-ray beams produced by laser-wakefield-accelerated electrons.

Given that laser wakefield acceleration (LWFA) has been demonstrated experimentally to accelerate electron beams to energies beyond 25 MeV, it is reasonable to assess the ability of existing LWFA technology to compete with conventional radiofrequency linear accelerators in producing electron and x-ray beams for external-beam radiotherapy. We present calculations of the dose distributions (off-axis dose profiles and central-axis depth dose) and dose rates of x-ray beams that can be produced from electron beams that are generated using state-of-the-art LWFA. Subsets of an LWFA electron energy distribution were propagated through the treatment head elements (presuming an existing design for an x-ray production target and flattening filter) implemented within the EGSnrc Monte Carlo code. Three x-ray energy configurations (6 MV, 10 MV and 18 MV) were studied, and the energy width deltaE of the electron-beam subsets varied from 0.5 MeV to 12.5 MeV. As deltaE increased from 0.5 MeV to 4.5 MeV, we found that the off-axis and central-axis dose profiles for x-rays were minimally affected (to within about 3%), a result slightly different from prior calculations of electron beams broadened by scattering foils. For deltaE of the order of 12 MeV, the effect on the off-axis profile was of the order of 10%, but the central-axis depth dose was affected by less than 2% for depths in excess of about 5 cm beyond d(max). Although increasing deltaE beyond 6.5 MeV increased the dose rate at d(max) by more than 10 times, the absolute dose rates were about 3 orders of magnitude below those observed for LWFA-based electron beams at comparable energies. For a practical LWFA-based x-ray device, the beam current must be increased by about 4-5 orders of magnitude.

Dose properties of a laser accelerated electron beam and prospects for clinical application.

Laser wakefield acceleration (LWFA) technology has evolved to where it should be evaluated for its potential as a future competitor to existing technology that produces electron and x-ray beams. The purpose of the present work is to investigate the dosimetric properties of an electron beam that should be achievable using existing LWFA technology, and to document the necessary improvements to make radiotherapy application for LWFA viable. This paper first qualitatively reviews the fundamental principles of LWFA and describes a potential design for a 30 cm accelerator chamber containing a gas target. Electron beam energy spectra, upon which our dose calculations are based, were obtained from a uniform energy distribution and from two-dimensional particle-in-cell (2D PIC) simulations. The 2D PIC simulation parameters are consistent with those reported by a previous LWFA experiment. According to the 2D PIC simulations, only approximately 0.3% of the LWFA electrons are emitted with an energy greater than 1 MeV. We studied only the high-energy electrons to determine their potential for clinical electron beams of central energy from 9 to 21 MeV. Each electron beam was broadened and flattened by designing a dual scattering foil system to produce a uniform beam (103%>off-axis ratio>95%) over a 25 x 25 cm2 field. An energy window (deltaE) ranging from 0.5 to 6.5 MeV was selected to study central-axis depth dose, beam flatness, and dose rate. Dose was calculated in water at a 100 cm source-to-surface distance using the EGS/BEAM Monte Carlo algorithm. Calculations showed that the beam flatness was fairly insensitive to deltaE. However, since the falloff of the depth-dose curve (R10-R90) and the dose rate both increase with deltaE, a tradeoff between minimizing (R10-R90) and maximizing dose rate is implied. If deltaE is constrained so that R10-R90 is within 0.5 cm of its value for a monoenergetic beam, the maximum practical dose rate based on 2D PIC is approximately 0.1 Gy min(-1) for a 9 MeV beam and 0.03 Gy min(-1) for a 15 MeV beam. It was concluded that current LWFA technology should allow a table-top terawatt (T3) laser to produce therapeutic electron beams that have acceptable flatness, penetration, and falloff of depth dose; however, the dose rate is still 1%-3% of that which would be acceptable, especially for higher-energy electron beams. Further progress in laser technology, e.g., increasing the pulse repetition rate or number of high energy electrons generated per pulse, is necessary to give dose rates acceptable for electron beams. Future measurements confirming dosimetric calculations are required to substantiate our results. In addition to achieving adequate dose rate, significant engineering developments are needed for this technology to compete with current electron acceleration technology. Also, the functional benefits of LWFA electron beams require further study and evaluation.

Avoiding patient self-blame.

There is need for shared responsibility between health care providers and patients. However, an unintended consequence of today's therapies that focus on mind/body interaction is the risk of patients' guilt and self-blame for contracting a disease (such as cancer) and/or for failing to heal themselves. This article suggests ways practitioners of complementary therapies can approach medical patients with a constructive attitude that minimizes the chance of unintentional psychological harm.

Lambda spectra in 11.6A GeV/c Au-Au collisions.

E896 has measured Lambda production in 11.6A GeV/c Au-Au collisions over virtually the whole rapidity phase space. The midrapidity p(t) distributions have been measured for the first time at this energy and appear to indicate that the Lambda hyperons have different freeze-out conditions than protons. A comparison with the relativistic quantum molecular dynamics model shows that while there is good shape agreement at high rapidity the model predicts significantly different slopes of the m(t) spectra at midrapidity. The data, where overlap occurs, are consistent with previously reported measurements.

The role of the psychologist in the evaluation and treatment of infertility.

The experience of infertility can be devastating for the couple desiring a child. For women, pregnancy and motherhood are developmental milestones that are highly emphasized by our culture. This paper will explain psychological issues in infertility, including evaluation and intervention. Specific issues such as privacy and disclosure, and deciding when to end infertility treatment, will also be discussed. A psychologist, or other mental health professional on the health care team, is essential to treatment of the biopsychosocial nature of infertility.