Differential signaling through p190 and p210 BCR-ABL fusion proteins revealed by interactome and phosphoproteome analysis.

Two major types of leukemogenic BCR-ABL fusion proteins are p190BCR-ABLand p210BCR-ABL. Although the two fusion proteins are closely related, they can lead to different clinical outcomes. A thorough understanding of the signaling programs employed by these two fusion proteins is necessary to explain these clinical differences. We took an integrated approach by coupling protein-protein interaction analysis using biotinylation identification with global phosphorylation analysis to investigate the differences in signaling between these two fusion proteins. Our findings suggest that p190BCR-ABL and p210BCR-ABL differentially activate important signaling pathways, such as JAK-STAT, and engage with molecules that indicate interaction with different subcellular compartments. In the case of p210BCR-ABL, we observed an increased engagement of molecules active proximal to the membrane and in the case of p190BCR-ABL, an engagement of molecules of the cytoskeleton. These differences in signaling could underlie the distinct leukemogenic process induced by these two protein variants.

SU-E-T-411: Dosimetric Comparison Between Two Multileaf Collimator Systems for Stereotactic Radiosurgery and Radiotherapy.

PURPOSE: This aim of this study is to compare the dosimetric parameters of two multileaf collimator (MLC) systems: (1) the beam modulator (BM), which is the MLC commercial name for Elekta 'Synergy S' linear accelerator; and (2) Radionics micro-MLC (MMLC). METHODS: Dosimetric parameters of percentage depth dose (PDD), in-plane and cross-plane beam profile, penumbra, MLC leakage and transmission for a 6 MV photon beam with different field sizes and depths were measured using ionization chamber, film, solid water phantom and water tank. At the same time, the BM and MMLC were modeled using the BEAMnrc code and the above dosimetric parameters were calculated using Monte Carlo simulations. Energy fluence spectra for the two MLC were determined using the BEAMnrc and BEAMDP. RESULTS: We found that dosimetric parameters (PDD, beam profile, energy fluence spectra, leakage and transmission) of the two MLC were similar, except for penumbra. The leaf-side and leaf-end 20%-80% penumbras at 10 cm depth for a 10×10 cm2 field were 4.8 and 5.1 mm for the MMLC and 5.3 mm and 6.3 mm for the BM, respectively. The maximum percentage of the leakage for the BM and MMLC are 1.3% and 1.2%, while the average percentage of leakage for the BM and MMLC are 0.9% and 1%. CONCLUSIONS: Based on their dosimetric characteristics for stereotactic radiosurgery and radiotherapy, it can be concluded that both the BM and MMLC can be used effectively, though the latter showed slightly sharper dose penumbra especially in the leaf-end direction. However, the BM has the advantages of producing considerably larger field at isocenter and having a greater isocenter clearance compared to the MMLC. The dosimetric data in this study should help radiotherapy staff to appreciate dependence of dosimetry on the MLC design and configuration for stereotactic radiosurgery and radiotherapy. Actual or potential conflicts of interest do not exist.

Stereotactic radiotherapy for treatment of juxtapapillary choroidal melanoma: 3-year follow-up.

AIMS: To report the treatment outcomes and complication rates of stereotactic radiotherapy in the management of patients with juxtapapillary choroidal melanoma. METHODS: A retrospective review of 64 consecutive patients with juxtapapillary choroidal melanoma, located within 2 mm of the optic disc, treated with stereotactic radiotherapy at Princess Margaret Hospital between October 1998 and January 2006. RESULTS: The median age was 63 years. The median tumour height was 4.2 mm, and median largest basal diameter was 9.8 mm. The prescribed radiation dose was 70 Gy in five fractions over 10 days, and the median follow-up was 37 months. Post-treatment, the actuarial rates of local tumour control, metastases and survival at 37 months were 94%, 15% and 90%, respectively. Actuarial rates of radiation-induced complications at 37 months were neovascular glaucoma 42%, cataract 53%, retinopathy 81% and optic neuropathy 64%. Secondary enucleation was necessary for 10 patients (16%), in four patients for tumour recurrence and in six for painful neovascular glaucoma. CONCLUSIONS: Stereotactic radiotherapy offers a non-invasive alternative to enucleation and brachytherapy in the management of juxtapapillary choroidal melanoma with a high tumour control rate, however, at the expense of a significant rate of long-term ocular complications.

Novel splice variants of sFlt1 are upregulated in preeclampsia.

Soluble fms-like tyrosine kinase-1 (sFlt1) is a truncated splice variant of Flt1, which is upregulated in preeclampsia. In this study we sought to characterize the unique C-terminus of sFlt. Through bioinformatic analyses, we identified two novel sFlt1 splice variants and two previously described sFlt1 splice variants. The novel variants are identical to the previously described sFlt1_v1 through exon 13, but then diverge to unique 3' termini consisting of a novel exon 15 (sFlt1_v2 and sFlt1_v3) or an extension of exon 14 (sFlt1_v4). Quantitative PCR showed that three out of four sFlt variants were upregulated in placenta of women with preeclampsia. Mass spectrometry analysis of sFlt1 purified from placental serum confirmed the presence of sFlt1_v1 protein, and an additional variant which includes sequence derived from exon 14. siRNA experiments targeting each variant confirmed that three of the four variants contribute significantly to total sFlt1 expression by cytotrophoblasts in vitro. These findings provide evidence that human placenta expresses a family of sFlt1 splice variants, at least three of which are expressed as proteins, and which appear to be globally upregulated in preeclampsia.

Dosimetric evaluation of a dedicated stereotactic linear accelerator using measurement and Monte Carlo simulation.

Dosimetric parameters of a dedicated stereotactic linear accelerator have been investigated using measurements and Monte Carlo simulations. This linac has a unique built in multileaf collimation (MLC) system with the maximum opening of 16 x 21 cm2 and 4 mm leaf width at the isocenter and has successfully been modeled for the first time using the Monte Carlo simulation. The high resolution MLC, combined with its relatively large maximum field size, opens up a new opportunity for expanding stereotactic radiation treatment techniques from traditionally treating smaller targets to larger ones for both cranial and extracranial lesions. Dosimetric parameters of this linac such as accuracy of leaf positioning and field shaping, leakage and transmission, percentage depth doses, off-axes dose profiles, and dose penumbras were measured and calculated for different field sizes, depths, and source to surface distances. In addition, the ability of the linac in accurate dose delivery of several treatment plans, including intensity modulated radiation therapy (IMRT), performed on phantom and patients was determined. Ionization chamber, photon diode detector, films, several solid water phantoms, and a water tank were used for the measurements. The MLC leaf positioning to any particular point in the maximum aperture was accurate with a standard deviation of 0.29 mm. Maximum and average leakages were 1.7% and 1.1% for the reference field of 10.4 x 9.6 cm2. Measured penumbra widths (80%-20%) for this field at source axis distance (SAD) of 100 cm at a depth of 1.5 cm (dmax) were 3.2 and 4 mm for the leaf-sides and leaf-ends, respectively. The corresponding results at 10 cm depth and SAD =100 cm were 5.4 and 6.3 mm. Monte Carlo results generally agreed with the measurements to within 1% and or 1 mm, with respective uncertainties of 0.5% and 0.2 mm. The linac accuracy in delivering non-IMRT treatment plans was better than 1%. Ionization chamber dosimetry results for a phantom IMRT plan in the high dose and low dose regions were -0.5% and +3.6%, respectively. Dosimetry results at isocenter for three patients' IMRT plans were measured to be within 3% of their corresponding treatment plans. Film dosimetry was also used to compare dose distributions of IMRT treatment plans and delivered cumulative doses at different cross sectional planes.

Comprehensive quality assurance for stereotactic radiosurgery treatments.

We have used a commercially available high precision Lucy phantom to perform comprehensive quality assurance for stereotactic radiosurgery treatments. The quantitative evaluation of system uncertainties included imaging, planning and treatment delivery systems. The quality assurance tests showed that the well-defined targets were identified to within +/-1 mm in all the imaging modalities. The pre-known target volumes were reproduced within 2 cm3 in both MR and CT. The planned target was delivered within 2% of the prescribed dose and to within 2 mm accuracy. The inaccuracy in the isocentre position at the Linac was less than 1.2 mm. The maximum error observed in the depth helmet was 0.5 mm and the overall uncertainty was within 0.23 mm. We have also established a quality assurance program based on the study and proposed the tolerance and the frequency of the tests required to be carried out. The tests were carried out using a Radionics planning system and delivered on a Varian Clinac 2100 linear accelerator machine. These tests also established a base line for future comparisons.

Persistent right sinus venosus valve producing right ventricular outflow tract obstruction.

A 12-year-old girl presented with exercise intolerance. Spinnaker formation of a persistent right sinus venosus valve produced right ventricular outflow tract obstruction. The heart was otherwise normal, and surgical correction was successful.

Head-upright tilt table testing for children with vasovagal syncope.

Vasovagal (neurocardiogenic) syncope is the most common type of syncope in the pediatric population. An extensive and expensive workup is not indicated in patients with a consistent history. By reproducing the effects of gravitational stress, tilt table testing confirms the diagnosis and avoids unnecessary and often unrewarding investigations.

A comparison of dosimetry techniques in stereotactic radiosurgery.

Accurate dosimetry of small-field photon beams used in stereotactic radiosurgery (SRS) can be made difficult because of the presence of lateral electronic disequilibrium and steep dose gradients. In the published literature, data acquisition for radiosurgery is mainly based on diode and film dosimetry, and sometimes on small ionization chamber or thermolominescence dosimetry. These techniques generally do not provide the required precision because of their energy dependence and/or poor resolution. In this work PTW diamond detectors and Monte Carlo (EGS4) techniques have been added to the above tools to measure and calculate SRS treatment planning requirements. The validity of the EGS4 generated data has been confirmed by comparing results to those obtained with an ionization chamber, where the field size is large enough for electronic equilibrium to be established at the central axis. Using EGS4 calculations, the beam characteristics under the experimental conditions have also been quantified. It was shown that diamond detectors are potentially ideal for SRS and yield more accurate results than the above traditional modes of dosimetry.

Dose rate dependence of a PTW diamond detector in the dosimetry of a 6 MV photon beam.

The dose rate dependence and current/voltage characteristics of a PTW Riga diamond detector in the dosimetry of a 6 MV photon beam have been investigated. Diamond detectors are radiosensitive resistors whose conductivity (i) varies almost in proportion to dose rate and (ii) is almost independent of bias voltage for a constant dose rate. At the recommended bias of +100 V, and also at +200 V, the detector is operating with incomplete charge collection due to the electron-hole recombination time being shorter that the maximum time for an electron to be collected by the anode. As dose rate is varied by changing FSD or depth (changing dose per pulse), detector current and dose rate are related by the expression i alpha Ddelta where delta is approximately 0.98. This manifests itself in an overestimate in percentage depth-dose at a depth of 30 cm of approximately 1% when compared to ionization chamber results. A similar sublinearity is seen when pulse repetition frequency is varied, indicating that the dependence is an on average rather than an instantaneous dose rate. The dose rate dependence is attributed to the reduction in recombination time as dose rate increases.

Pulmonary valve stenosis associated with hypertrophic cardiomyopathy.

The association of pulmonic stenosis with hypertrophic cardiomyopathy is rare in infancy. Presented here is an infant with atypical picture of pulmonic stenosis and echocardiographic evidence of hypertrophic cardiomyopathy. At eight months of age, she had a successful percutaneous balloon valvuloplasty and has subsequently been managed with propranolol.