Dandamudi V. Rao

Macroscopic dosimetry for radioimmunotherapy: nonuniform activity distributions in solid tumors

In the context of radioimmunotherapy of cancer, there is a need for continued improvement of dosimetry of radionuclides localized in tumors. Current methods assume uniform distribution of radionuclides in the tumor despite experimental evidence indicating nonuniormity. We have developed a model in which nonuniform distribution of radioactivity in the tumor is taken into account. Spherically symmetric radionuclide distributions, depending linearly and exponentially on the radial position, are considered. Dose rate profiles in the tumor are calculated for potentially useful beta-emitting radionuclides, including 32P, 67Cu, 90Y, 111Ag, 131I, and 188Re, and for 193mPt, an emitter of conversion electrons and low-energy Auger electrons. For the radionuclide distributions investigated, high-energy beta emitters, such as 90Y, are most effective in treating large tumors (diameter, d greater than approximately 1 cm), whereas for small tumors (d approximately 1 mm), medium energy beta emitters such as 67Cu are better suited. Very small tumors (d less than 1 mm), and micrometastases are best handled with low-energy electron emitters such as 193mPt.

Dosimetry of Auger-electron-emitting radionuclides: report no. 3 of AAPM Nuclear Medicine Task Group No. 6.

The biological effects of Auger-electron-emitting radionuclides can be as severe as those of alpha particles of high linear energy transfer. A great deal of effort has been expended in exploring the biological effects of Auger electron emitters. Much of this effort has been devoted to improving theoretical and experimental techniques required to calculate absorbed doses and correlate them with the observed biological effects. Given that the main purpose of dosimetry is to obtain a physical descriptor with which to correlate radiation toxicity, then nowhere is this challenge greater than when biological specimens are subject to Auger electron cascades. The dense shower of short-range Auger electrons released by radionuclides, which decay by electron capture or internal conversion, results in biological damage that is highly dependent on the location of the decay site within the cell. In this report, different approaches to Auger electron dosimetry are described and compared. Methods to calculate the absorbed dose from Auger electron emitters at the DNA, cellular, multicellular, and organ levels are described as they relate to the biological effects. The concept of a radiation weighting factor for Auger electrons to be used in the calculation of equivalent dose is reviewed. The importance of subcellular distribution of Auger emitters in determining the biological effects of these radionuclides is emphasized and incorporated into the equivalent dose formalism. The Task Group recommends that a preliminary radiation weighting factor of 10 be used for deterministic effects of Auger electrons, and a value of 20 for stochastic effects.

Vitamins as radioprotectors in vivo II. Protection by vitamin A and soybean oil against radiation damage caused by internal radionuclides

Tissue-incorporated radionuclides impart radiation energy over extended periods of time depending on their effective half-lives. The capacity of vitamin A dissolved in soybean oil to protect against the biological effects caused by internal radionuclides is investigated. The radiochemicals examined are DNA-binding 125IdU, cytoplasmically localized H125IPDM and the alpha-particle emitter 210Po citrate. As in our previous studies, spermatogenesis in mice is used as the experimental model and spermatogonial cell survival is the biological end point. Surprisingly, soybean oil itself provides substantial and equal protection against the Auger effect of 125IdU, which is comparable to a high-LET radiation effect, as well as the low-LET effects of H125IPDM, the dose modification factors (DMFs) being 3.6 +/- 0.9 (SEM) and 3.4 +/- 0.9, respectively. The protection afforded by the oil against the effects of 5.3 MeV alpha particles emitted by 210Po is also significant (DMF = 2.2 +/- 0.4). The presence of vitamin A in the oil further enhanced the radioprotection against the effect of 125IdU (DMF = 4.8 +/- 1.3) and H125IPDM (DMF = 5.1 +/- 0.6); however, no enhancement is provided against the effects of alpha particles. These interesting results with soybean oil and vitamin A, together with data on the subcellular distribution of the protectors, provide clues regarding the mechanistic aspects of the protection. In addition, the data for vitamin A reaffirm our earlier conclusion that the mechanism by which DNA-bound Auger emitters impart biological damage is primarily indirect in nature.

Radiotoxicity of Platinum-195m-Labeled trans-Platinum (II) in Mammalian Cells

The chemotoxicity and radiotoxicity of trans-dichlorodiammineplatinum (II) labeled with 195mPt (trans-195mPt) are investigated to ascertain the potential of radioplatinum coordination complexes as antineoplastic agents. Platinum-195m, with a half-life of about 4 days, is a prolific emitter of low-energy Auger electrons because of the high probability of internal conversion in its isomeric transitions. The kinetics of cellular uptake and retention after incubation and the radiotoxicity of this Auger electron emitter in the form of trans-195mPt is investigated using cells of the Chinese hamster V79 cell line. The cellular uptake of 195mPt reaches a plateau in about 3 to 5 h of incubation and varies nonlinearly with the extracellular concentration of radioactivity. The radioactivity is eliminated from the cells after incubation with an effective half-life of 24 h. Cell survival data, when corrected for the chemical toxicity of nonradiolabeled trans-platinum, give a cell survival curve typical for radiations with high linear energy transfer. At 37% survival, the mean lethal cellular uptake is about 1.0 mBq/cell. Dosimetric considerations, based on subcellular distribution of the radionuclide, yield a value of 4.8 for the relative biological effectiveness when compared with 250 kVp X rays. Theoretical Monte Carlo track-structure calculations indicate that the density of radical species produced in liquid water in the immediate vicinity of a 195mPt decay site is substantially greater than the density of species along the track of a 5.3 MeV alpha particle. This explains qualitatively the efficacy of 195mPt in causing high-LET radiation type biological effects. The extreme radiotoxicity of intranuclearly localized 195mPt, in conjunction with the proclivity of platinum chemotherapy agents to bind to DNA in the cell nucleus, suggests that the combination of chemical effects and the effects of Auger electrons that can be obtained with radioplatinum coordination complexes may have potential in the treatment of cancer.

Testicular and plasma ascorbic acid levels in mice following dietary intake: a high-performance liquid chromatographic analysis.

A modified buffer system is reported for the analysis of vitamin C in mouse plasma and testes, on a reversed-phase high-performance liquid chromatographic column with ultraviolet detection. The buffer, consisting of 0.1 M NaH2PO4 and 0.2 mM Na2EDTA adjusted to pH 3.1 with orthophosphoric acid, resolved the ascorbic acid (AA) peak allowing clear quantitation of the chemical. The method is also applicable to the assay of dehydroascorbic acid after its reduction to ascorbic acid, and overcomes problems of AA stability encountered in previously reported procedures. Using the present technique, variations in the vitamin levels of plasma and testes are studied from 3 to 29 days after the commencement of feeding a vitamin C-rich diet (1%, w/w) in mice. The plasma AA levels were elevated above the controls by a factor of 2.5 by day 8. Contrary to this, testicular AA levels increased marginally (1.2-fold) by day 12 and were maintained at levels close to the control values thereafter. It appears that the feedback inhibition mechanism which is effective in plasma is not operational in the testes. The findings are of clinical significance in that testicular AA levels do not change significantly as a result of dietary intake of vitamin C, whereas plasma AA levels do show an increase.

Vitamins as Radioprotectors In Vivo. I. Protection by Vitamin C against Internal Radionuclides in Mouse Testes: Implications to the Mechanism of Damage Caused by the Auger Effect

The potential of vitamin C, an antioxidant, to protect the radiosensitive spermatogonial cells in mouse testes against the effects of chronic irradiation by radionuclides incorporated into tissue was investigated. Interestingly, when injected intratesticularly, a small and nontoxic amount of vitamin C (1.5 microgram in 3 microliters saline) protected the spermatogonia against the damage associated with high-LET radiation caused by Auger electrons from similarly administered 5-(125I)-iodo-2-deoxyuridine (125IdU). A dose modification factor (DMF) of 2.3 was obtained. In contrast, no protection was observed when 210Po, an alpha-particle emitter, was administered similarly. These findings suggest that the mechanism of action of the Auger effect is of an indirect nature, which is in contrast to the direct action generally believed to be responsible for biological damage caused by high-LET radiations.

Relative biological effectiveness of 99mTc radiopharmaceuticals

The radiotoxicity of three 99mTc-labeled compounds is investigated using spermatogenesis in mouse testis as the experimental model, and spermatogonial cell survival as the biological end point. The radiopharmaceuticals studied are pertechnetate (99mTcO4-), pyrophosphate (99mTc-PYP), and hydroxyethylene diphosphate (99mTc-HDP). The mean lethal doses at 37% survival (D37) are 0.70 +/- 0.06, 0.84 +/- 0.13, and 0.59 +/- 0.08 Gy for 99mTcO4-, 99mTc-PYP, and 99mTc-HDP, respectively. When these results are compared with the D37 value obtained with external x rays or internal gamma rays, the relative biological effectiveness (RBE) of these compounds are 0.94 +/- 0.09, 0.79 +/- 0.13, and 1.1 +/- 0.16, respectively. These results show that the radiotoxicity of 99mTc in mouse testis is essentially similar to that of low-LET radiations (i.e., RBE approximately 1). To understand these results, the distribution of these radiocompounds in the testis is determined and correlated with the observed RBE values. The expected range of RBE values for 99mTc radiopharmaceuticals in organs is 0.95 to 1.5, depending on the fraction of organ activity that is bound to DNA. This suggests that the Auger electrons emitted in the decay of 99mTc are not capable of causing extreme toxicity in vivo. These results provide further support for 99mTc as the radionuclide of choice for imaging in nuclear medicine.

Relative Biological Effectiveness of Alpha-Particle Emitters In Vivo at Low Doses

The therapeutic potential of radionuclides that emit alpha particles, as well as their associated health hazards, have attracted considerable attention. The 224Ra daughters 212Pb and 212Bi, by virtue of their radiation properties which involve emission of alpha and beta particles in their decay to stable 208Pb, have been proposed as candidates for radioimmunotherapy. Using mouse testes as the experimental model and testicular spermhead survival as the biological end point, the present work examines the radiotoxicity of 212Pb and its daughters. When 212Pb, in equilibrium with its daughters 212Bi, 212Po and 208Tl, was administered directly into the testis, the dose required to achieve 37% survival (D37) was 0.143 +/- 0.014 Gy and the corresponding RBE of the mixed radiation field was 4.7 when compared to the D37 for acute external 120 kVp X rays. This datum, in conjunction with our earlier results for 210Po, was used to obtain an RBE-LET relationship for alpha particles emitted by tissue-incorporated radionuclides: RBE alpha = 4.8 - 6.1 x 10(-2) LET + 1.0 x 10(-3) LET2. Similarly, the dependence of RBE on alpha-particle energy E alpha was given by RBE alpha = 22 E(-0.73) alpha. These relationships, based on in vivo experimental data, may be valuable in predicting biological effects of alpha-particle emitters.

bis-benzimidazole dyes, Hoechst 33258 and Hoechst 33342: Radioiodination, facile purification and subcellular distribution

A simple HPLC method is presented for the purification of DNA binding bis-benzimidazole dyes Hoechst 33258, Hoechst 33342 and 131I-iodoHoechst 33258. The mobile phase, consisting of methanol and aqueous ammonia (0.2%) in the ratio 2:3, resolved and separated the radiochemical from unlabeled ligand and other reagents used in the reaction, thereby resulting in high radiochemical purity and yield. The iodinated Hoechst 33258 did not show any selective binding to nuclear DNA when cell fractionation studies were performed with cultured mammalian cells as well as in mice testes. Fluorescence microscopy studies with V79 cells stained with these dyes, showed the superiority of Hoechst 33342 in selective localization in nuclear DNA compared to Hoechst 33258. The difference in behavior of these two dyes in terms of binding to nuclear DNA, and hence their ability to provide protection against damage caused by ionizing radiation, may be explained on the basis of the molecular charge. The high chemotoxicity of Hoechst 33342 observed in the present studies suggests that its usefulness as a radioprotector against chronic irradiation of tissue by incorporated radionuclides may be limited.

CALCULATION OF EQUIVALENT DOSE FOR AUGER ELECTRON EMITTING RADIONUCLIDES DISTRIBUTED IN HUMAN ORGANS

Radionuclides that emit Auger electrons can be extremely radiotoxic depending on the subcellular distribution of the radiochemical. Despite this, ICRP 60 provides no guidance in the calculation of equivalent dose H(T) for Auger electrons. The recent report by the American Association of Physicists in Medicine recommends a radiation weighting factor wR of 20 for stochastic effects caused by Auger electrons, along with a method of calculating the equivalent dose that takes into account the subcellular distribution of the radionuclide. In view of these recommendations, it is important to reevaluate equivalent doses from Auger electron emitters. The mean absorbed dose per unit cumulated activity (S-value) from Auger electrons and other radiations is calculated for ninety Auger-electron-emitting radionuclides distributed in human ovaries, testes and liver. Using these S-values, and the formalism given in the recent AAPM report, the dependence of the organ equivalent doses on subcellular distribution of the Auger electron emitters is examined. The results show an increase in the mean equivalent dose for Auger electron emitters when a significant fraction of the organ activity localizes in the DNA.