Downregulation of toll-like receptor 4 and IL-6 following irradiation of the rat urinary bladder.

The pathophysiology behind radiation cystitis is poorly understood. Here we investigated whether bladder irradiation affects the immune system of the rat urinary bladder. Female rats were sedated and exposed to one single radiation dose of 20 Gy or only sedated (controls) and killed 16 h to 14 days later. Rats were placed in a metabolic cage at 16 h, 3 days, 7 days and 14 days following bladder irradiation. The urinary bladders were harvested and analysed with qPCR, immunohistochemistry and/or Western blot for the expression of interferon (IFN)-γ, interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, nitric oxide synthases (eNOS, iNOS and nNOS), tumour necrosis factor (TNF)-α and toll-like receptor 4 (TLR4). Urine was collected and analysed for IL-6 and nitrite (reflecting nitric oxide activity) with ELISA and the Griess reaction, respectively. Irradiation increased bladder frequency and decreased voiding volumes 14 days following bladder irradiation. Bladder irradiation increased the expression of IL-10 and collagen in the bladder, while TLR4 and IL-6 expressions were decreased in the urothelium concomitantly with a decrease in mast cells in the submucosa and urine levels of IL-6 and nitrite. The present findings show that bladder irradiation leads to urodynamic changes in the bladder and may suppress important immunoregulatory pathways in the urinary bladder.

Cylindrical ionization chamber response in static and dynamic 6 and 15 MV photon beams.

Purpose.To investigate the response of the CC13 ionization chamber under non-reference photon beam conditions, focusing on penumbra and build-up regions of static fields and on dynamic intensity-modulated beams.Methods. Measurements were performed in 6 MV 100 × 100, 20 × 100, and 20 × 20 mm2static fields. Monte Carlo calculations were performed for the static fields and for 6 and 15 MV dynamic beam sequences using a Varian multi-leaf collimator. The chamber was modelled using EGSnrc egs_chamber software. Conversion factors were calculated by relating the absorbed dose to air in the chamber air cavity to the absorbed dose to water. Correction and point-dose correction factors were calculated to quantify the conversion factor variations.Results. The correction factors for positions on the beam central axis and at the penumbra centre were 0.98-1.02 for all static fields and depths investigated. The largest corrections were obtained for chamber positions beyond penumbra centre in the off-axis direction. Point-dose correction factors were 0.54-0.71 at 100 mm depth and their magnitude increased with decreasing field size and measurement depth. Factors of 0.99-1.03 were obtained inside and near the integrated penumbra of the dynamic field at 100 mm depth, and of 0.92-0.94 beyond the integrated penumbra centre. The variations in the ionization chamber response across the integrated dynamic penumbra qualitatively followed the behaviour across penumbra of static fields.Conclusions. Without corrections, the CC13 chamber was of limited usefulness for profile measurements in 20-mm-wide fields. However, measurements in dynamic small irregular beam openings resembling the conditions of pre-treatment patient quality assurance were feasible. Uncorrected ionization chamber response could be applied for dose verification at 100 mm depth inside and close to large gradients of dynamically accumulating high- and low-dose regions assuming 3% tolerance between measured and calculated doses.

Effects of lung tissue characterization in radiotherapy of breast cancer under deep inspiration breath hold when using Monte Carlo dosimetry.

PURPOSE: To investigate the sensitivity of Monte Carlo (MC) calculated lung dose distributions to lung tissue characterization in external beam radiotherapy of breast cancer under Deep Inspiration Breath Hold (DIBH). METHODS: EGSnrc based MC software was employed. Mean lung densities for one hundred patients were analysed. CT number frequency and clinical dose distributions were calculated for 15 patients with mean lung density below 0.14 g/cm3. Lung volume with a pre-defined CT numbers was also considered. Lung tissue was characterized by applying different CT calibrations in the low-density region and air-lung tissue thresholds. Dose impact was estimated by Dose Volume Histogram (DVH) parameters. RESULTS: Mean lung densities below 0.14 g/cm3 were found in 10% of the patients. CT numbers below -960 HU dominated the CT frequency distributions with a high rate of CT numbers at -990 HU. Mass density conversion approach influenced the DVH shape. V4Gy and V8Gy varied by 7% and 5% for the selected patients and by 9% and 3.5% for the pre-defined lung volume. V16Gy and V20Gy, were within 2.5%. Regions above 20 Gy were affected. Variations in air- lung tissue differentiation resulted in DVH parameters within 1%. Threshold at -990 HU was confirmed by the CT number frequency distributions. CONCLUSIONS: Lung dose distributions were more sensitive to variations in the CT calibration curve below lung (inhale) density than to air-lung tissue differentiation. Low dose regions were mostly affected. The dosimetry effects were found to be potentially important to 10% of the patients treated under DIBH.

Irradiation-induced progenitor cell death in the developing brain is resistant to erythropoietin treatment and caspase inhibition.

One hemisphere of postnatal day 8 (P8) rats or P10 mice was irradiated with a single dose of 4-12 Gy, and animals were killed from 2 h to 8 weeks after irradiation (IR). In the subventricular zone (SVZ) and the granular cell layer (GCL) of the dentate gyrus, harboring neural and other progenitor cells, nitrosylation and p53 peaked 2-12 h after IR, followed by markers for active caspase-3, apoptosis-inducing factor and TUNEL (6-24 h). Ki67-positive (proliferating) cells had disappeared by 12 h and partly reappeared by 7 days post-IR. The SVZ and GCL areas decreased approximately 50% 7 days after IR. The development of white matter was hampered, resulting in 50-70% less myelin basic protein staining. Pretreatment with erythropoietin did not confer protection against IR. Caspase inhibition by overexpression of XIAP prevented caspase-9 and caspase-3 activation but not cell death, presumably because of increased caspase-independent cell death.

Measurement of bone mineral using multiple-energy x-ray absorptiometry.

Our laboratory has previously reported a method of determining the amount of bone mineral using triple-energy absorptiometry with a continuous x-ray spectrum. In the present study, the experimental properties of the technique were examined. The accuracy, the influence of fat content and body thickness and the in vitro and in vivo precision were analysed. The results found in this investigation showed that despite the complexity of the technique, the amount of bone mineral can be accurately determined. The in vivo precision was determined to be 3.4%, expressed as the coefficient of variation (CV), for different skeletal parts. The in vitro precision was found to be 2.1% (CV). Neither the fat content nor the body thickness had any effect on the measured bone mineral values. Excellent linearity and a close correlation were found between the true and the measured bone mineral values.

The feasibility of triple-energy absorptiometry for the determination of bone mineral, Ca and P in vivo.

The theoretical feasibility of triple-energy absorptiometry in general and the experimental conditions when using triple-energy absorptiometry for the determination of bone mineral, elemental calcium and phosphorus content in vivo have been investigated. A theoretical analysis of the decomposition of the mass attenuation coefficients is presented and discussed. The main obstacle to the effective use of triple-energy absorptiometry in vivo is the large number of pulses which must be detected to reduce the statistical fluctuations.

Determination of bone mineral density in the third lumbar vertebral body using photon absorptiometry techniques.

Dual-photon absorptiometry and triple-energy X-ray absorptiometry were used to investigate the total bone mineral content and density as well as the trabecular bone mineral density in the third lumbar vertebral body. Both anteroposterior (AP) and lateral (LAT) measurements were performed. By combining the two projections it was found that the mean trabecular bone mineral density for all 202 subjects included in the study was 52% (SD +/- 20%) of the total bone mineral density in the third lumbar vertebral body. The mean trabecular bone mineral density as a fraction of the total vertebral body bone mineral density decreased as a function of age. The relative annual change in this fraction differed between males and females. It was also found that neither trabecular nor total bone mineral density differed significantly between male and female subjects aged 25-35 years, and bone mineral density (BMD), expressed in g/cm3, showed no correlation to subject height, body weight or body mass index (BMI). Male and female individuals showed different rates of change of trabecular bone mineral density with age.

Apoptosis-inducing factor deficiency decreases the proliferation rate and protects the subventricular zone against ionizing radiation.

Cranial radiotherapy in children often leads to progressive cognitive decline. We have established a rodent model of irradiation-induced injury to the young brain. A single dose of 8 Gy was administered to the left hemisphere of postnatal day 10 (P10) mice. Harlequin (Hq) mice, carrying the hypomorphic apoptosis-inducing factor AIF(Hq) mutation, express 60% less AIF at P10 and displayed significantly fewer dying cells in the subventricular zone (SVZ) 6 h after IR, compared with wild type (Wt) littermates. Irradiated cyclophilin A-deficient (CypA(-/-)) mice confirmed that CypA has an essential role in AIF-induced apoptosis after IR. Hq mice displayed no reduction in SVZ size 7 days after IR, whereas 48% of the SVZ was lost in Wt mice. The proliferation rate was lower in the SVZ of Hq mice. Cultured neural precursor cells from the SVZ of Hq mice displayed a slower proliferation rate and were more resistant to IR. IR preferentially kills proliferating cells, and the slower proliferation rate in the SVZ of Hq mice may, at least partly, explain the protective effect of the Hq mutation. Together, these results indicate that targeting AIF may provide a fruitful strategy for protection of normal brain tissue against the detrimental side effects of IR.

Bone mineral and migratory patterns in uncemented total knee arthroplasties: a randomized 5-year follow-up study of 38 knees.

We measured the amount of bone mineral in the medial tibial condyle 1 week postoperatively, after 1 year and after 4-5 years in 38 arthrotic knees randomized to a Freeman-Samuelson hydroxyapatite-coated (FS HA) or a Miller-Galante II (MG II) total knee arthroplasty. Clinically excellent results were recorded in both groups after 5 years. At the last follow-up, the overall decrease in bone mineral was 26%, as measured by triple-energy X-ray absorptiometry. The decrease was larger in FS HA knees than in MG II knees after 4-5 years, indicating stress-shielding of the proximal tibia. Radiostereometry at 1 and 5 years showed smaller maximum total point motion, maximum subsidence and varus or valgus tilt in the FS HA group. There was a tendency towards a reversed relationship between subsidence and change in bone mineral after 1 year, but not after 4-5 years. Distal fixation of the stem in the Freeman-Samuelson hydroxyapatite-coated (FS HA) components might explain the more pronounced loss of bone mineral in the medial tibial condyle.