COMPARISON OF PROLIFERATING CELL NUCLEAR ANTIGEN (PCNA) STAINING AND BRDURD-LABELING INDEX UNDER DIFFERENT PROLIFERATIVE CONDITIONS IN-VITRO BY FLOW-CYTOMETRY

PC10 is a monoclonal antibody against proliferating cell nuclear antigen (PCNA). The staining pattern in immunochemistry depends on fixation and detergent extraction treatment. The aim of this study was to validate the flow cytometric PCNA assay against Bromodeoxyuridine-labelling index (BrdUrd-LI) under different proliferative conditions in vitro. Expression of PCNA in methanol fixed cells with, and without, prior detergent extraction with EDTA/Triton was compared to BrdUrd-labelling index in NIH-3T3 fibroblasts and human Caski tumour cells in exponential phase and under confluent conditions. Serum stimulation and serum starvation conditions were studied. The results for BrdUrd-LI and PCNA-index after extraction showed good correlation for 3T3 fibroblasts and for Caski cells, with some differences for serum withdrawn Caski cells. There was no correlation between the number of cells that were positive for PCNA without extraction and BrdUrd-LI. Spheroid cells with G(1)-DNA-content showed an almost synchronous recruitment and progression through the cell cycle after trypsination and replating. Tightly bound PCNA paralleled this synchronicity whereas total PCNA did not change significantly. The results demonstrate that immunochemical detection of non-extractable PCNA-index gives similar results as compared with BrdUrd-labelling index under different proliferative conditions in vitro for different monolayer cell lines, whereas without extraction PCNA does not correlate with BrdUrd-LI in these fast growing cell lines due to its long half-life. PCNA expression parallels the progression through the cell cycle in V79 spheroids, a primitive model of tumour growth

Robustness of sweeping-window arc therapy treatment sequences against intrafractional tumor motion

Purpose: Due to the potentially periodic collimator dynamic in volumetric modulated arc therapy (VMAT) dose deliveries with the sweeping-window arc therapy (SWAT) technique, additional manifestations of dosimetric deviations in the presence of intrafractional motion may occur. With a fast multileaf collimator (MLC), and a flattening filter free dose delivery, treatment times close to 60 s per fraction are clinical reality. For these treatment sequences, the human breathing period can be close to the collimator sweeping period. Compared to a random arrangement of the segments, this will cause a further degradation of the dose homogeneity. Methods: Fifty VMAT sequences of potentially moving target volumes were delivered on a two dimensional ionization chamber array. In order to detect interplay effects along all three coordinate axes, time resolved measurements were performed twice-with the detector aligned in vertical (V) or horizontal (H) orientation. All dose matrices were then moved within a simulation software by a time-dependent motion vector. The minimum relative equivalent uniform dose EUDr,m for all breathing starting phases was determined for each amplitude and period. Furthermore, an estimation of periods with minimum EUD was performed. Additionally, LINAC logfiles were recorded during plan delivery. The MLC, jaw, gantry angle, and monitor unit settings were continuously saved and used to calculate the correlation coefficient between the target motion and the dose weighed collimator motion component for each direction (CC, LR, AP) separately. Results: The resulting EUDr,m were EUDr,m(CCV) = (98.3 +/- 0.6)%, EUDr,m(CCH) = (98.6 +/- 0.5)%, EUDr,m(AP(V)) = (97.7 +/- 0.9)%, and EUDr,m(LRH) = (97.8 +/- 0.9)%. The overall minimum relative EUD observed for 360. arc midventilation treatments was 94.6%. The treatment plan with the shortest period and a minimum relative EUD of less than 97% was found at T = 6.1 s. For a partial 120 degrees arc, an EUDr,m = 92.0% was found. In all cases, a correlation coefficient above 0.5 corresponded to a minimum in EUD. Conclusions: With the advent of fast VMAT delivery techniques, nonrobust treatment sequences for human breathing patterns can be generated. These sequences are characterized by a large correlation coefficient between a target motion component and the corresponding collimator dynamic. By iteratively decreasing the maximum allowed dose rate, a low correlation coefficient and consequentially a robust treatment sequence are ensured. (C) 2015 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 Unported License

Coherent molecule formation in anharmonic potentials near confinement-induced resonances

We perform a theoretical and experimental study of a system of two ultracold atoms with tunable interaction in an elongated trapping potential. We show that the coupling of center-of-mass and relative motion due to an anharmonicity of the trapping potential leads to a coherent coupling of a state of an unbound atom pair and a molecule with a center of mass excitation. By performing the experiment with exactly two particles we exclude three-body losses and can therefore directly observe coherent molecule formation. We find quantitative agreement between our theory of inelastic confinement-induced resonances and the experimental results. This shows that the effects of center-of-mass to relative motion coupling can have a significant impact on the physics of quasi-1D quantum systems.Comment: 7 pages, 4 figure

Fermionization of two distinguishable fermions

In this work we study a system of two distinguishable fermions in a 1D harmonic potential. This system has the exceptional property that there is an analytic solution for arbitrary values of the interparticle interaction. We tune the interaction strength via a magnetic offset field and compare the measured properties of the system to the theoretical prediction. At the point where the interaction strength diverges, the energy and square of the wave function for two distinguishable particles are the same as for a system of two identical fermions. This is referred to as fermionization. We have observed this phenomenon by directly comparing two distinguishable fermions with diverging interaction strength with two identical fermions in the same potential. We observe good agreement between experiment and theory. By adding one or more particles our system can be used as a quantum simulator for more complex few-body systems where no theoretical solution is available

Quantification and Assessment of Interfraction Setup Errors Based on Cone Beam CT and Determination of Safety Margins for Radiotherapy

Introduction To quantify interfraction patient setup-errors for radiotherapy based on cone-beam computed tomography and suggest safety margins accordingly. Material and Methods Positioning vectors of pre-treatment cone-beam computed tomography for different treatment sites were collected (n = 9504). For each patient group the total average and standard deviation were calculated and the overall mean, systematic and random errors as well as safety margins were determined Results The systematic (and random errors) in the superior-inferior, left-right and anterior-posterior directions were: for prostate, 2.5(3.0), 2.6(3.9) and 2.9(3.9) mm; for prostate bed, 1.7(2.0), 2.2(3.6) and 2.6(3.1) mm; for cervix, 2.8(3.4), 2.3(4.6) and 3.2(3.9) mm; for rectum, 1.6(3.1), 2.1(2.9) and 2.5(3.8) mm; for anal, 1.7(3.7), 2.1(5.1) and 2.5(4.8) mm; for head and neck, 1.9(2.3), 1.4(2.0) and 1.7(2.2) mm; for brain, 1.0(1.5), 1.1(1.4) and 1.0(1.1) mm; and for mediastinum, 3.3(4.6), 2.6(3.7) and 3.5(4.0) mm. The CTV-to-PTV margins had the smallest value for brain (3.6, 3.7 and 3.3mm) and the largest for mediastinum (11.5, 9.1 and 11.6mm). For pelvic treatments the means (and standard deviations) were 7.3 (1.6), 8.5 (0.8) and 9.6 (0.8) mm. Conclusions Systematic and random setup-errors were smaller than 5mm. The largest errors were found for organs with higher motion probability. The suggested safety margins were comparable to published values in previous but often smaller studies