Nonuniform irradiation of the canine intestine. I. Effects

To investigate the effects of nonuniform irradiation on the small intestine, we prepared 24 dogs for continent isoperistaltic ileostomies under aseptic surgical conditions and general anesthesia. After a 3-week recovery period, the ileum was catheterized with a fiberoptic endoscope to observe the intestinal mucosa and to harvest mucosal biopsies. The baseline macroscopic and microscopic appearance of the intestinal mucosa was determined. Two weeks later, the ileum was catheterized with a 100-cm soft tube containing 40 groups of three thermoluminescent dosimeters placed at equally spaced intervals, and a dose of either 4.5, 8, 10, 11, or 15 Gy 60Co gamma rays was delivered to the right abdomen (nonuniform exposure). This method allowed a direct and precise assessment of the dose received at 40 sites located in the 100-cm intestinal segment. The intestinal mucosa was again evaluated 1, 4, and 6 days after irradiation. All animals exposed to 4.5 and 8 Gy survived, whereas none survived after 11 and 15 Gy. After exposure to 10 Gy, 60% of the animals died within 4-6 days and 40% survived with symptoms associated with both the intestinal and the hematopoietic syndromes. Crypt cell necrosis, blunting of villi, and reduction of the mucosal lining increased between 1 and 4 days after irradiation, and mucosal damage was correlated with intraintestinal dosimetry at Day 6. The granulocyte counts at Day 4 were significantly lower than baseline level in animals that died within 4-6 days but not in survivors. The present model appears to be realistic and clinically relevant, allowing the concurrent study of the intestinal and hematopoietic effects of high-dose nonuniform irradiation similar to that received by patients during radiation therapy as well as by radiation accident victims.     

Survival after total-body irradiation. I. Effects of partial small bowel shielding

The small intestine of the rat was shielded during total-body irradiation (TBI) to evaluate the effects of radiation dose and length of intestine shielded on survival. Sprague-Dawley rats were anesthetized in groups of 10. Using aseptic surgical procedures 80, 40, 20, or 10 cm, or none of the proximal or distal small intestine were temporarily exteriorized and shielded during irradiation with photons from an 18 MeV linear accelerator. Less than 17% of the dose was delivered to the shielded intestines. In unshielded animals deaths occurred from Days 4 to 6 with 13, 15, or 17 Gy and from Days 8 to 30 with 9, 11, and 12 Gy. However, in all animals exposed to 15 Gy with all or part of the small intestine shielded, survival was increased to between 5 and 9 days. Shielding of the distal small intestine was more effective in prolonging survival than shielding of the proximal small intestine. The previously identified target of radiation damage in the small intestine is the crypt stem cell. In this study, the analysis of histological specimens of shielded and irradiated small intestine suggested that humoral factors also influence intestinal histology and survival after irradiation. These humoral factors are thought to originate from the irradiated body tissues, the shielded proximal intestine, and the shielded distal intestine. Further studies are required to identify these factors and to determine their mode of action and their therapeutic potential after radiation damage to the small intestine.     

Quantification of gut injury with diamine oxidase activity: development of a fission neutron RBE and measurements with combined injury in mouse models

Plasma and small intestine diamine oxidase (DAO) activities were measured on Days 2, 4, and 6 following irradiation of mice with a range of doses of fission neutrons and 60Co. With increasing doses of radiation, plasma DAO activity increased on Day 2 and intestinal DAO activity decreased on Day 4; moreover, the approximate relative biological effectiveness values for these changes in activity were 5.81 for plasma DAO activity on Day 2 and 3.88 for intestinal DAO activity on Day 4. On Day 6 relatively high levels of radiation caused DAO activity in the small intestine to remain depressed whereas low levels resulted in recovery with activities at or near controls. In animals with combined injury (radiation plus 30% surface burn or wound), changes in DAO activity in the intestine were similar to those with radiation alone; plasma DAO activity, in contrast to radiation alone, did not show an increase at the 2-day mark. These dose-dependent relationships should provide a basis for using DAO as a potential indicator of biological damage from radiation exposure within the lethal range.     

Neutron and X-ray effects on small intestine summarized by using a mathematical model or paradigm.

The responses of intestinal tissues to ionizing radiation can be described by comparing irradiated cell populations qualitatively or quantitatively with corresponding controls. This paper describes quantitative data obtained from resin-embedded sections of neutron-irradiated mouse small intestine at different times after treatment. Information is collected by counting cells or structures present per complete circumference. The data are assessed by using standard statistical tests, which show that early mitotic arrest precedes changes in goblet, absorptive, endocrine and stromal cells and a decrease in crypt numbers. The data can also produce ratios of irradiated: control figures for cells or structural elements. These ratios, along with tissue area measurements, can be used to summarize the structural damage as a composite graph and table, including a total figure, known as the Morphological Index. This is used to quantify the temporal response of the wall as a whole and to compare the effects of different qualities of radiation, here X-ray and cyclotron-produced neutron radiations. It is possible that such analysis can be used predictively along with other reference data to identify the treatment, dose and time required to produce observed tissue damage.     

Simulation modelling of the postradiation restoration of the crypt-villus system

Basic principles have been developed for a discrete stochastic simulation model of an elementary proliferative unit of the intestinal epithelium, a "crypt-villus" system. The analysis of the results obtained after a single exposure of the animal's abdomen to 3 and 6 Gy radiation has demonstrated that the dynamics of the number of cells that synthesize DNA in a small intestine crypt of exposed mice depends on the rate of radiation damage repair (50 to 100 h following irradiation). The rate of repair after 6 Gy irradiation is 1.5 times lower that after 3 Gy. The changes in the shape of the labeled mitoses curve, followed up during the postirradiation recovery of the intestinal epithelium, may occur with the time parameters of the cell mitotic cycle being invariable.     

Functional and structural alterations of epithelial barrier properties of rat ileum following X-irradiation

Irradiation of the digestive system leads to alterations of the small intestine. We have characterized the disruption of the barrier integrity in rat ileum from 1 to 14 days following irradiation ranging from 6 to 12 Gy. The intestinal permeability to 14C-mannitol and 3H-dextran 70 000 was measured in vitro in Ussing chambers. In parallel to these functional studies, immunohistochemical analyses of junctional proteins (ZO-1 and beta-catenin) of ileal epithelium were performed by confocal microscopy. Irradiation with 10 Gy induced a marked decrease in epithelial tissue resistance at three days and a fivefold increase in mannitol permeability, without modifications of dextran permeability. A disorganization of the localization for ZO-1 and beta-catenin was also observed. At 7 days after irradiation, we observed a recovery of the organization of junctional proteins in parallel to a return of intestinal permeability to control value. In addition to these time-dependent effects, a gradual effect on epithelial integrity of the radiation doses was observed 3 days after irradiation. This study shows a disruption of the integrity of the intestinal barrier in rat ileum following abdominal X-irradiation, depending on the time postirradiation and on the delivered dose. The loss of barrier integrity was characterized by a disorganization of proteins of tight and adherent junctions, leading to increased intestinal permeability to mannitol.     

In situ biological dose mapping estimates the radiation burden delivered to 'spared' tissue between synchrotron X-ray microbeam radiotherapy tracks

Microbeam radiation therapy (MRT) using high doses of synchrotron X-rays can destroy tumours in animal models whilst causing little damage to normal tissues. Determining the spatial distribution of radiation doses delivered during MRT at a microscopic scale is a major challenge. Film and semiconductor dosimetry as well as Monte Carlo methods struggle to provide accurate estimates of dose profiles and peak-to-valley dose ratios at the position of the targeted and traversed tissues whose biological responses determine treatment outcome. The purpose of this study was to utilise γ-H2AX immunostaining as a biodosimetric tool that enables in situ biological dose mapping within an irradiated tissue to provide direct biological evidence for the scale of the radiation burden to 'spared' tissue regions between MRT tracks. Γ-H2AX analysis allowed microbeams to be traced and DNA damage foci to be quantified in valleys between beams following MRT treatment of fibroblast cultures and murine skin where foci yields per unit dose were approximately five-fold lower than in fibroblast cultures. Foci levels in cells located in valleys were compared with calibration curves using known broadbeam synchrotron X-ray doses to generate spatial dose profiles and calculate peak-to-valley dose ratios of 30-40 for cell cultures and approximately 60 for murine skin, consistent with the range obtained with conventional dosimetry methods. This biological dose mapping approach could find several applications both in optimising MRT or other radiotherapeutic treatments and in estimating localised doses following accidental radiation exposure using skin punch biopsies.     

The effect of chronic gamma irradiation in an exponentially decreasing dose rate on the nucleic acid content in the hematopoietic organs and blood of rats]

The effect of continuous gamma irradiation at exponentially decreased dose rates (from 562 mGy/h to 13 mGy/h with a total cumulative dose of 14.355 Gy delivered over a period of 10 days) on the nucleic acid content of rat hemopoietic tissues and blood was followed up. The radiation model used simulated a decrease in the radioactivity of a fission mixture in the contaminated environment resulting from a nuclear device accident. We have found that the dynamics of the changes seems to be similar to that observed after acute exposure, and the hemopoiesis recovery starts just at the time of irradiation. In evaluating the damage and recovery extent after accidental irradiation, we consider it expedient to complement the biological dosimetry with the indices studied work including the determination of DNA and RNA concentrations in blood of irradiated human beings.     

The effect of heme oxygenase-1 induction by octreotide on radiation enteritis

Radiation enteritis occurs as a response to abdominal radiation, which can cause mucosal damage in the gastrointestinal mucosal epithelium. The small intestine is one of the most radiosensitive organs in the abdomen. The present study was undertaken to investigate the effect of octreotide (OCT) administration on heme oxygenase-1 (HO-1) expression of the radiation enteritis model. Rats received 50 mg/kg/day OCT for 4 days before irradiation and continued for 3 days after irradiation. Intestinal myeloperoxidase (MPO) activities, malondialdehyde (MDA) levels are indicators of oxidative damage while caspase-3 activities reveal apoptosis degree of the small intestine. At histological examination, the terminal ileum tissue was analyzed for morphological changes. Irradiation significantly increased the intestinal MPO and caspase-3 activities, MDA levels and HO-1 expression in comparison to sham control group. OCT treatment was associated with increased HO-1 expression and caspase-3 activity, decreased MPO activity and MDA levels. Histological examination revealed that the intestinal mucosal structure was preserved in the OCT treated group. OCT appears to have protective effects against radiation-induced intestinal damage. This protective effect is, in part, mediated by modification of the inflammatory response and the induction of HO-1 expression.     

Photodimerization of uracil in films and their use for dosimetry of genetically active ultraviolet irradiation

On the basis of the known photodimerization of uracil it is proposed to use its films obtained by sublimation in vacuum for the dosimetry of genetically active UV radiation. Experimental estimations are presented of radiation dose rate resulting in a change of uracil optical density delta D/D0 under irradiation due to photodimerization for erythemal lamp lo-30. The data obtained are used for dosimetry of lethal damage of Chlamydomonas reinhardii. Estimations are also presented of the sun radiation dose rate which induce uracil photodimerization on the earth surface and in the extraterrestrial atmosphere.     

The protective effects of XH‐105 against radiation‐induced intestinal injury

Radiation‐induced intestinal injury is one of the major side effects in patients receiving radiation therapy. There is no specific treatment for radiation enteritis in the clinic. We designed and synthesized a new compound named XH‐105, which is expected to cleave into polyphenol and aminothiol in vivo to mitigate radiation injury. In the following study, we describe the beneficial effects of XH‐105 against radiation‐induced intestinal injury. C57BL/6J mice were treated by gavage with XH‐105 1 hour before total body irradiation (TBI), and the survival rate was monitored. Histological changes were examined, and survival of Lgr5⁺ intestinal stem cells Ki67⁺ cells, villi⁺ enterocytes and lysozymes was determined by immunohistochemistry. DNA damage and cellular apoptosis in intestinal tissue were also evaluated. Compared to vehicle‐treated mice after TBI, XH‐105 treatment significantly enhanced the survival rate, attenuated structural damage of the small intestine, decreased the apoptotic rate, reduced DNA damage, maintained cell regeneration and promoted crypt proliferation and differentiation. XH‐105 also reduced the expression of Bax and p53 in the small intestine. These data suggest that XH‐105 is beneficial for the protection of radiation‐induced intestinal injury by inhibiting the p53‐dependent apoptosis signalling pathway.     

Radiation injury: some aspects of the oncogenic effects.

The late effects or irradiation stem from cell killing, mutation, and malignant transformation. Cancer is the major somatic late effect of exposure to low dose levels of radiation, and estimates of risk of cancer in man after irradiation are based entirely on human experience. The data for dose-response relationships for the induction of tumors by external irradiation in man have been obtained from a single exposure or a small number of exposures delivered at high dose rates. In contrast, exposure to environmental irradiation is mainly protracted over a long period of time and is delivered at a low dose rate. As yet no allowance has been made for the effect of protraction of the exposure time in estimating the risk of cancer, although an adjustment has been made in the case of estimates of genetic risk. Incidence of tumors has been the only parameter used for risk estimates, but latent period and degree of malignancy, which are probably both dose and dose-rate dependent, influence the nature of the risk from radiation. As the knowledge about the effects of low-level radiation has been accumulated and assimilated over the last 70 years, so has the concern for reasonable standards of safety. There are still problems in the estimation of radiation risks, but at least many of the relevant questions can now be framed. The problems of estimating risks for chemical carcinogens are clearly greater, but the experience gained from radiation studies should help in the design of the necessary experiments.     

Modelling slow wave activity in the small intestine

We have developed an anatomically based model to simulate slow wave activity in the small intestine. Geometric data for the human small intestine were obtained from the Visible Human project. These data were used to create a one-dimensional finite element mesh of the entire small intestine using an iterative fitting procedure. The electrically active components of the intestinal walls were modelled using a modified Fitzhugh-Nagumo cell model embedded within a longitudinal smooth muscle layer and a layer containing Interstitial Cells of Cajal. Within these layers, the monodomain equation was used to describe slow wave propagation. To solve the monodomain equation, a high-resolution finite difference grid, with an average spatial resolution of 0.95 mm, was embedded within each finite element. The resulting simulations of intestinal activity agree with the experimental observation that slow wave frequency gradually declines from 12 cycles per minute (cpm) in the duodenum to 8 cpm at the terminal ileum. Furthermore, the simulations demonstrated a decrease in conduction velocity with distance along the small intestine (10.7 cm/s in the duodenum, 5.1cm/s in the jejunum and 1.4 cm/s in the ileum), matching experimental recordings from the canine small intestine. We conclude that the framework presented here is capable of qualitatively simulating normal slow wave activity in an anatomical model of the small intestine.     

A dosimetric method of total body irradiation

The determination of the dose delivered to the body during Total Body Irradiation (TBI) is not easy, as direct measurements are impossible. This article presents the intention to develop a systematic and simple method for absorbed dose determination in(60)Co TBI using mid plane dose factors (MDF) and external measurements, i.e. entrance and exit dose readings by semiconductor detectors. In order to perform the radiation field analysis prior to TBI treatment, a special silicon n-type detector with increased sensitivity (1200 nC/Gy) was used. The dependence of the calibration factor of this detector exposed to different dose rates in standard and TBI conditions was investigated. A theoretical model of anthropomorphic phantoms of cylindrical ellipsoid shape was established. The software made it possible to generalize the applicability of TBI dosimetry to any individual case of real patient data. A quality assurance analysis of dosimetric results of 350 patients, who underwent TBI during a 17-year period, was performed.     

In vivo monitoring of total skin electron dose using optically stimulated luminescence dosimeters

AimThis study retrospectively analysed the results of using optically stimulated radiation dosimeters (OSLDs) for in vivo dose measurements during total skin electron therapy (TSET, also known as TSEI, TSEB, TSEBT, TSI or TBE) treatments of patients with mycosis fungoides.BackgroundTSET treatments are generally delivered to standing patients, using treatment plans that are devised using manual dose calculations that require verification via in vivo dosimetry. Despite the increasing use of OSLDs for radiation dosimetry, there is minimal published guidance on the use of OSLDs for TSET verification.Materials and methodsThis study retrospectively reviewed in vivo dose measurements made during treatments of nine consecutive TSET patients, treated between 2013 and 2018. Landauer nanoDot OSLDs were used to measure the skin dose at reference locations on each patient, as well as at locations of clinical interest such as the head, hands, feet, axilla and groin.Results1301 OSLD measurements were aggregated and analysed, producing results that were in broad agreement with previous TLD studies, while providing additional information about the variation of dose across concave surfaces and potentially guiding future refinement of treatment setup. In many cases these in vivo measurements were used to identify deviations from the planned dose in reference locations and to identify anatomical regions where additional shielding or boost treatments were required.ConclusionsOSLDs can be used to obtain measurements of TSET dose that can inform monitor unit adjustments and identify regions of under and over dosage, while potentially informing continuous quality improvement in TSET treatment delivery.     

Radioprotective properties of ATP and modification of acid phosphatase response after a lethal dose of whole body p(66MeV)/Be neutron radiation to BALB/c mice.

The intraperitoneal administration of exogenous ATP prior to a lethal dose (7 Gy) of whole body neutron irradiation increased the radioresistance of BALB/c mice. This radiation used the beam from a neutron therapy facility produced by the reaction p(66 MeV)/Be. Survival of the mice, determined 7 days post-irradiation as the endpoint, was increased from 26% to 86% by the action of the exogenous ATP. Furthermore, the response of acid phosphatase activity as an indicator of the acute radiation effects showed a marked augmentation in both tissues studied, testes and small intestine. The activity of the enzyme after neutron irradiation with prior administration of ATP showed smaller increases when compared with the increases observed after neutron irradiation alone. This implies that exogenous ATP reduces the effect of the lytic enzyme and, hence, damage. Finally, changes were observed in the activity of acid phosphatase in the testes and intestine with different concentrations of exogenous ATP. In both tissues there was a monotonic decrease in the activity of the enzyme with increase of the concentration of exogenous ATP administrated before radiation. These results reflect the protective ability of exogenous ATP as an adaptive defence mechanism to reduce radiation damage in normal tissues after a lethal dose of neutron radiation.     

Methylene blue protects intestinal mucosa from free radical-mediated sublethal radiation damage

Susceptibility of the intestine to radiation damage is a primary reason for the failure of external beam radiation to cure intra-abdominal cancer. High-dose irradiation causes intestinal denudation, fluid loss, and resultant shock. Simple and effective methods for protecting the intestine from irradiation damage are not available. Many biological effects of ionizing irradiation are caused by free radical intermediates. We have previously reported that many of the toxicities of doxorubicin, a classic free radical generating anticancer agent, are blocked by methylene blue pretreatment. We have now found that pretreatment with methylene blue protects rats from intestinal damage, as measured histologically and by quantitative stool blood determinations. Whereas the exact mechanisms of this protection remain elusive, we believe this method of modulating the therapeutic index of ionizing radiation deserves additional preclinical and clinical study.     

Hypophysectomy-induced inhibition of augmented acetylcholine responses of the rat bowel following adrenalectomy and/or whole body irradiation

Summary Functional changes in the intestinal responsiveness to a fixed. dose of acetylcholine were studied in muscle strips removed from young adult male rats previously exposed to whole body gamma radiation. In the irradiated rat the responsiveness to a fixed dose of acetylcholine was found to be augmented in the small intestine but not in the colon. Similar motor patterns for the small intestine were found when muscle strips from adrenalectomized rats were studied. Preradiation adrenalectomy further exaggerated the post-radiation sensitivity of the rat small intestine to acetylcholine. Hypophysectomy prior to either adrenalectomy and/or whole body radiation was associated with an absence of augmented small intestinal motor activity following administration of acetylcholine. The response of the large bowel to acetylcholine, however, was not modified by adrenalectomy and/or hypophysectomy. These observations suggest an endocrine component to the acute 3–5 day intestinal radiation syndrome.This work was presented at the 25th Annual Meeting of the Radiation Research Society, San Juan, Puerto Rico, May 1977     

Preliminary investigation on the use of the MOSFET dosimeter in proton beams

Metal Oxide Semiconductor (MOS) device structures can be used to measure ionizing radiation through the mechanism of hole trapping in the oxide layer leading to changing of electrical characteristic of the device. They are a new type of direct reading semiconductor dosimeters. Due to their extremely small physical size, ability to permanently store the accumulated dose, dose-rate independence and their ease of use make them very promising for in vivo dosimetry. They are attractive for dosimetry in small radiation fields used in modern radiation oncology modalities, as conformal radiotherapy, IMRT, stereotactic radiotherapy/radiosurgery and proton therapy. Preliminary results on the use of commercial MOSFET dosimeters (TN-502RD, Thomson & Nielsen Electronics Ltd, Canada) irradiated on therapeutic 62 MeV proton beams are presented. Linearity with absorbed dose, sensibility and energy dependence were investigated. Moreover, the possibility to use of MOSFET dosimeters in order to measure the Output Factors (OF) for very small irradiation fields was verified. The comparison of OF obtained using MOSFETs and other dosimetry systems is reported.