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A single x-ray dose of 1,200 to 1,600 rads to the mouse kidney is associated with definite morphologic alteration but minimal functional impairment at six months; this progresses to profound structural and functional impairment by one year after irradiation. Subjective morphologic assessment of renal damage at six months correlates well with total radiation dose, fractionation schedule and energy characteristics of the radiation beam but does not provide adequate quantitative numerical data for sophisticated statistical tests of significance or for comparisons of effect variability at given dose levels. This investigation assessed the applicability of computer-assisted morphometric analysis (CAMA) for quantitation of effects and in making statistical comparisons of significance between kidneys subjectively classified as to degree of histologic alterations. Images of renal cortex tubular nuclei from the various histologic grades were digitized, recorded and analyzed with the CAMA system. Results indicate that the reliability of specific grade assignment by CAMA for individual nuclei was inadequate but that separation of irradiated and unirradiated renal tissue (bivariate group means) was quite distinct and of high reliability. Differences were present among the four irradiated histologic grades, but they were not marked, especially among the three highest grades. More accurate quantitation of nuclear size variations was achieved, and chromatin textural differences were detected that were not apparent to the eye. Computer-assisted morphometric analysis appears to have a valuable application in the quantification and analysis of chronic radiation effects.

A system is described for comparing various modalities and fractionation schedules of radiation by means of their long-term morphologic effects upon the mouse kidney. The comparison system utilizes a grading scale for histopathologic changes in which a given histologic grade depends upon meeting defined threshold criteria, rather than quantitation of a particular measurement. Renal tubular alterations served as the basis for comparison, since they appeared more reliably defined than glomerular changes. The radiation dose that induced a specific threshold effect in kidneys from 50% of the animals at 6 months was defined as the effective dose-50%, or ED50.ED50 was found for x-rays and negative pimesons administered in 1, 2, or 5 fractions. From these data, the relative biologic effectiveness (RBE) of negative pi-mesons with respect to x-rays was determined for each fractionation schedule.

The evaluation of radiation injury in complex organs has tended to lag behind comparable areas of investigative interest. This observation is somewhat surprising in view of the increased use of radiotherapy as a primary or adjuvant mode of therapy for malignant disease. In part, this problem appears to relate to the difficulty of identifying and quantifying the morphologic consequences of radiation injury in complex organs, in which the various component tissues exhibit a broad spectrum of radiosensitivities. Two approaches have been employed to address this problem: (1) utilization of sophisticated probes to evaluate the functional and morphologic sequelae of radiation injury and (2) segmentation of complex tissues into their component parts, which are then evaluated individually. Both approaches are illustrated in the papers presented in this issue. The purpose of this overview is to call attention to some of the attendant difficulties of the former approach, as seen in an ongoing investigative program concerned with radiation injury of the kidney.

Late radiation injury manifests itself in all morphologic compartments of the kidney, but loss of cell mass is most significant in the proximal convoluted tubules. Development of an end stage or nonfunctional kidney requires 12 or more months after single fraction X ray exposures of about 12 Gy (2, 5, 13) and is associated with marked morphologic alterations of renal tubules. Radiation induced changes were studied at 6 months after irradiation, a time interval when histological alterations appear minor, but in previous studies were shown to correlate with the later end stage alterations (5, 8, 9). Renal alterations were graded objectively based on renal weight, variation in size of tubule cell nuclei, and glomerular nuclear volume fraction. Irradiation was associated with loss of renal weight, increased variability of tubule nuclear size, and previously unappreciated changes in glomerular nuclear volume fraction. A classification index derived from a weighted combination of renal weight ratio and tubule cell nuclear variability correlates with radiation dose and with previously established subjective histologic grading of renal damage, and allows objective comparisons of various fractionation schedules.

Histologic quantitation of renal radiation injury based on an intact mouse model correlates well with loss of renal mass, as measured by the ratio of right to left renal weight, radiation dose, and time after irradiation. Use of the radiation dose at which 50% of animals show renal tubular changes of a specified grade (ED50), allows comparison of 1, 5 and 15 fraction exposures. The X-ray dose necessary to effect significant injury (Grade 3 change) by six months after irradiation (ED50) was 11.6, 25.7, and 44.0 Gy for single, 5, and 15 fractions, respectively. Isoeffect plots of total radiation dose as a function of fraction number are linear on logarithmic coordinates, indicating that this relationship followed a power function. When plotted as inverse total dose versus dose per fraction, the six month post-irradiation data were linear, but the 12 month data were not. Lack of linearity is not in agreement with the multi-fraction isoeffect linear quadratic model.