[en] The cellular stress response, whereby very low doses of cytotoxic agents induce resistance to much higher doses, is an evolutionary defence mechanism and is stimulated following challenges by numerous chemical, biological and physical agents including particularly radiation, drugs, heat and hypoxia. There is much homology in the effects of these agents which are manifest through the up-regulation of various genetic pathways. Low-dose radiation stress influences processes involved in cell-cycle control, signal transduction pathways, radiation sensitivity, changes in cell adhesion and cell growth. There is also homology between radiation and other cellular stress agents, particularly hypoxia. Whereas traditionally, hypoxia was regarded mainly as an agent conferring resistance to radiation, there is now much evidence illustrating the cytokine-like properties of hypoxia as well as radiation. Stress phenomena are likely to be important in risks arising from low doses of radiation. Conversely, exploitation of the stress response in settings appropriate to therapy can be particularly beneficial not only in regard to radiation alone but in combinations of radiation and drugs. Similarly, tissue hypoxia can be exploited in novel ways of enhancing therapeutic efficacy. Bioreductive drugs, which are cytotoxically activated in hypoxic regions of tissue, can be rendered even more effective by hypoxia-induced increased expression of enzyme reductases. Nitric oxide pathways are influenced by hypoxia thereby offering possibilities for novel vascular based therapies. Other approaches are discussed

[en] Purpose: To determine the oxygen enhancement ratio (OER) and shape of the oxygen sensitization curve of mouse foot skin, the extent to which glutathione (GSH) depletion radiosensitized skin, and the dependence of such sensitization on the ambient oxygen tension. Methods and Materials: The feet of WHT mice were irradiated with single doses of 240 kVp x-rays while mice were exposed to carbogen or gases with oxygen/nitrogen mixtures containing 8-100% O₂. The anoxic response was obtained by occluding the blood supply to the leg of anesthetized mice with a tourniquet, surrounding the foot with nitrogen, and allowing the mice to breathe 10% O₂. Further experiments were performed to assess the efficacy of this method to obtain an anoxic response. Radiosensitivity of skin was assessed using the acute skin-reaction assay. Glutathione levels were modified using two schedules of dl-buthionine sulphoximine (BSO) and diethylmaleate (DEM), which were considered to produce extensive and intermediate levels of GSH depletion in the skin of the foot during irradiation. Results: Carbogen caused the greatest radiosensitization of skin, with a reproducible enhancement of 2.2 relative to the anoxic response. The OER of 2.2 is lower than other reports for mouse skin. This may indicate that the extremes of oxygenation were not produced, although there was no direct evidence for this. When skin radiosensitivity was plotted against the logarithm of the oxygen tension in the ambient gas, a sigmoid curve with a K value of 17-21% O₂ in the ambient gas was obtained. Depletion of GSH caused minimal radiosensitization when skin was irradiated under anoxic or well-oxygenated conditions. Radiosensitization by GSH depletion was maximal at intermediate oxygen tensions of 10-21% O₂ in the ambient gas. Increasing the extent of GSH depletion led to increasing radiosensitization, with sensitization enhancement ratios of 1.2 and 1.1, respectively, for extensive and intermediate levels of GSH depletion. In mice exposed to 100% O₂, a significant component of skin radiosensitivity was due to diffusion of oxygen directly through the skin. Pentobarbitone anesthesia radiosensitized skin in mice exposed to 100% O₂ by a factor of 1.2, but did not further sensitize skin in mice exposed to carbogen. Conclusions: Glutathione levels and the local oxygen tension at the time of irradiation were important determinants of mouse foot skin radiosensitivity. The extent to which GSH levels altered the radiosensitivity of skin was critically dependent on the local oxygen tension. These results have significant implications for potential clinical application of GSH depletion

[en] The rate and early pain pattern of development of radiation-induced renal damage has been determined in the mouse by measuring reductions in both haematocrit and excretion of ⁵¹Cr-EDTA, and increases in both urination frequency and in urine volume. Kidneys of CBA mice were irradiated bilaterally with 2 fractions of X-rays, one week apart. Renal function was determined immediately prior to irradiation and at 3-4 weekly intervals to 22 weeks post-irradiation. Onset of damage was detected as early as 3-6 weeks using the urination frequency assay. This was confirmed by estimating the volume of urine excreted. A significant fall in haematocrit was not detected until 6-9 weeks post-treatment and a fall in isotope clearance was not detected significantly until 12 weeks. This early detection of damage was consistent with reports using both mouse and other species. The time at which damage was detected first was independent of radiation dose for the frequency and haematocrit assays. For ⁵¹Cr-EDTA clearance, there was the suggestion of earlier functional loss for the higher doses. Following the onset of damage, a steady, dose-dependent decline in renal function was measured by all assays. The latency period is defined as the time required to reach a given level of functional damage. This time decreased with increasing radiation dose, to a minimum value set by the time of onset of damage, which varied from 3-12 weeks, depending on the assay used. The differences in response measured prior to 12 weeks post-irradiation represent the first occasion on which a dissociation between these 3 assays has been detected. The radiation-induced anaemia was characterised as normocytic with no evidence of haemolysis. (author). 21 refs.; 5 figs.; 1 tab

No abstract available

[en] Purpose: A model is introduced to integrate biological factors such as cell migration and bystander effects into physical dose distributions, and to incorporate spatial dose information in plan analysis and optimization. Methods: The model consists of a dose convolution filter (DCF) with single parameter σ. Tissue response is calculated by an existing NTCP model with DCF-applied dose distribution as input. The authors determined σ of rat spinal cord from published data. The authors also simulated the GRID technique, in which an open field is collimated into many pencil beams. Results: After applying the DCF, the NTCP model successfully fits the rat spinal cord data with a predicted value of σ=2.6±0.5 mm, consistent with 2 mm migration distances of remyelinating cells. Moreover, it enables the appropriate prediction of a high relative seriality for spinal cord. The model also predicts the sparing of normal tissues by the GRID technique when the size of each pencil beam becomes comparable to σ. Conclusions: The DCF model incorporates spatial dose information and offers an improved way to estimate tissue response from complex radiotherapy dose distributions. It does not alter the prediction of tissue response in large homogenous fields, but successfully predicts increased tissue tolerance in small or highly nonuniform fields.

[en] Purpose: Intensity-modulated radiotherapy (IMRT) is delivered using a variety of techniques with differing temporal dose characteristics. Spatial dose metrics are generally used to evaluate treatment plan quality. However, the use of this information alone neglects the effects of the significant differences in dose delivery duration and dose accumulation patterns, both of which can impact cell survival. This study uses the linear-quadratic model with dose protraction corrections to evaluate the biological effectiveness of different IMRT delivery techniques, including fixed gantry IMRT in SMLC (step-and-shoot) and DMLC (sliding window) modes and a rotational IMRT technique (helical tomotherapy) for the treatment of prostate and head/neck sites. Methods: The temporal dose pattern was measured using a small volume ion chamber (A1SL--0.057 cm³) to calculate the protraction factor, and biological equivalent dose (BED) was calculated for a range of repair half-times and α/β ratios. The treatment BED is compared to an ideal delivery of the target prescription dose, in which dose is delivered instantaneously (G(t₀)=1), to evaluate loss in biological effectiveness due to protraction in delivery. In the case of a conventional prescription, the loss in biological effectiveness was further evaluated using published tumor control probability (TCP) data. Results: With SMLC and DMLC IMRT delivery, for both prostate and head/neck, the expected additional loss in BED is about 1% compared to 3D CRT, which corresponds to a predicted 2%-3% reduction in TCP. For tomotherapy, the prostate BED loss is smaller in comparison to 3D CRT; hence, the authors expect a TCP increase of the order of 2%-3%. The aforementioned differences are due to the dose accumulation time. Conclusions: While it is theoretically possible to compensate for changes in biologically effective dose, this would be hindered by large uncertainties in parameters used for such calculations; therefore, it is advantageous to irradiate target volume elements as rapidly as possible. The results of this study indicate that temporal dose delivery pattern is an important component in determining the biological effects of IMRT treatment.

[en] Background and purpose: Initial promising results of 3D conformal neutron radiotherapy (3D-CNRT) were subsequently limited by high normal tissue toxicities. It is now possible to deliver intensity modulated neutron radiotherapy (IMNRT). The present work compares photon IMRT, 3D-CNRT and IMNRT for three prostate patients to quantify the benefits of IMNRT. Materials and methods: We compare updated 3D-CNRT plans, IMNRT plans, and conventional IMRT plans by translating neutron DVHs into effective photon DVHs using the dose dependent radiobiological effectiveness (RBE) for each structure. RBE curves are parameterized for a range of normal tissue and prostate tumor values. Generalized equivalent uniform dose (gEUD) and gEUD in 2 Gy fractions (gEUD₂) is calculated for each structure, plan, and parameterization. Rectal sparing and dose to prostate-GTV are compared for 3D-CNRT, IMNRT, and IMRT as a function of normal tissue and prostate RBE. Results: The closer the RBE values of prostate tumor and normal tissue, the greater the advantage of IMNRT over 3D-CNRT. The rectal sparing achieved using IMNRT ranged from ∼5% to 13% depending upon the choice of RBE for rectum and the α/β value of prostate tumor. IMNRT may provide a theoretical dose advantage over photon IMRT if the α/β value of prostate is 1.5 and the RBEs of prostate and rectum differ by more than 5%. For higher values of prostate α/β any advantages of IMNRT over IMRT could require that the RBEs of prostate and rectum differ by as much as 20%. Conclusions: IMNRT provides a clear normal tissue sparing advantage over 3D-CNRT. The advantage increases when the RBEs of the target structure and the normal tissue are similar. This RBE translation method could help identify clinical sites where the dose sparing advantages of IMNRT would allow for the exploitation of the radiobiological advantages of high-LET neutron radiotherapy.

[en] Purpose: The shape of the ionizing radiation response curve at very low doses has been the subject of considerable debate. Linear-no-threshold (LNT) models are widely used to estimate risks associated with low-dose exposures. However, the low-dose hyperradiosensitivity (HRS) phenomenon, in which cells are especially sensitive at low doses but then show increased radioresistance at higher doses, provides evidence of nonlinearity in the low-dose region. HRS is more prominent in the G2 phase of the cell cycle than in the G0/G1 or S phases. Here we provide the first cytogenetic mechanistic evidence of low-dose HRS in human peripheral blood lymphocytes using structural chromosomal aberrations. Methods and Materials: Human peripheral blood lymphocytes from 2 normal healthy female donors were acutely exposed to cobalt 60 γ rays in either G0 or G2 using closely spaced doses ranging from 0 to 1.5 Gy. Structural chromosomal aberrations were enumerated, and the slopes of the regression lines at low doses (0-0.4 Gy) were compared with doses of 0.5 Gy and above. Results: HRS was clearly evident in both donors for cells irradiated in G2. No HRS was observed in cells irradiated in G0. The radiation effect per unit dose was 2.5- to 3.5-fold higher for doses ≤0.4 Gy than for doses >0.5 Gy. Conclusions: These data provide the first cytogenetic evidence for the existence of HRS in human cells irradiated in G2 and suggest that LNT models may not always be optimal for making radiation risk assessments at low doses

[en] Highlights: • Human cells were irradiated in G1 or G2 and evaluated for micronuclei and bridges. • Cells irradiated in G2 but not in G1 exhibit low dose hyper-radiosensitivity. • Response curves of cells irradiated in G2 do not fit a linear-no-threshold model. • Response curves of cells irradiated in G1 fit a linear-no-threshold model. - Abstract: The dose-effect relationships of cells exposed to ionizing radiation are frequently described by linear quadratic (LQ) models over an extended dose range. However, many mammalian cell lines, when acutely irradiated in G2 at doses ≤0.3 Gy, show hyper-radiosensitivity (HRS) as measured by reduced clonogenic cell survival, thereby indicating greater cell lethality than is predicted by extrapolation from high-dose responses. We therefore hypothesized that the cytogenetic response in G2 cells to low doses would also be steeper than predicted by LQ extrapolation from high doses. We tested our hypothesis by exposing four normal human lymphoblastoid cell lines to 0–400 cGy of Cobalt-60 gamma radiation. The cytokinesis block micronucleus assay was used to determine the frequencies of micronuclei and nucleoplasmic bridges. To characterize the dependence of the cytogenetic damage on dose, univariate and multivariate regression analyses were used to compare the responses in the low- (HRS) and high-dose response regions. Our data indicate that the slope of the response for all four cell lines at ≤20 cGy during G2 is greater than predicted by an LQ extrapolation from the high-dose responses for both micronuclei and bridges. These results suggest that the biological consequences of low-dose exposures could be underestimated and may not provide accurate risk assessments following such exposures

[en] Purpose: An association between low-dose hyper-radiosensitivity (HRS) and the 'early' G2/M checkpoint has been established. An improved molecular understanding of the temporal dynamics of this relationship is needed before clinical translation can be considered. This study was conducted to characterize the dose response of the early G2/M checkpoint and then determine whether low-dose radiation sensitivity could be increased by synchronization or chemical inhibition of the cell cycle. Methods and Materials: Two related cell lines with disparate HRS status were used (MR4 and 3.7 cells). A double-thymidine block technique was developed to enrich the G2-phase population. Clonogenic cell survival, radiation-induced G2-phase cell cycle arrest, and deoxyribonucleic acid double-strand break repair were measured in the presence and absence of inhibitors to G2-phase checkpoint proteins. Results: For MR4 cells, the dose required to overcome the HRS response (approximately 0.2 Gy) corresponded with that needed for the activation of the early G2/M checkpoint. As hypothesized, enriching the number of G2-phase cells in the population resulted in an enhanced HRS response, because a greater proportion of radiation-damaged cells evaded the early G2/M checkpoint and entered mitosis with unrepaired deoxyribonucleic acid double-strand breaks. Likewise, abrogation of the checkpoint by inhibition of Chk1 and Chk2 also increased low-dose radiosensitivity. These effects were not evident in 3.7 cells. Conclusions: The data confirm that HRS is linked to the early G2/M checkpoint through the damage response of G2-phase cells. Low-dose radiosensitivity could be increased by manipulating the transition of radiation-damaged G2-phase cells into mitosis. This provides a rationale for combining low-dose radiation therapy with chemical synchronization techniques to improve increased radiosensitivity.