Multicenter ProstaScint imaging findings in 2154 patients with prostate cancer. The ProstaScint Imaging Centers.

OBJECTIVES: To report the results of a retrospective study of 2290 ProstaScint scans of 2154 patients with prostate carcinoma done at 15 institutions. METHODS: The results were analyzed by logistic regression after stratification of the patients into four groups: group 1, newly diagnosed; group 2, after radical prostatectomy with a rising prostate-specific antigen (PSA) level; group 3, after radiation therapy with a rising PSA level; and group 4, after hormonal therapy. RESULTS: The PSA level and ProstaScint scans positive in the prostate bed (P <0.001) and for pelvic metastases (P <0.001), but not for extrapelvic metastases, correlated significantly in group 1 patients. In group 2, the association for detecting fossa recurrence was weaker (P = 0.033) and was insignificant for pelvic and extrapelvic metastases. Patients in group 3 also exhibited a weak PSA-ProstaScint association for detecting fossa recurrence (P = 0.038), and was insignificant for pelvic and extrapelvic metastases. No significant PSA-ProstaScint correlation was found in patients in group 4 for fossa recurrence, pelvic or extrapelvic metastases. The distribution of positive ProstaScint results among the prostate/prostate bed, pelvic nodes, and extrapelvic nodes was nearly equal for all groups, except that a significantly greater percentage of extrapelvic metastases was found in the hormonal group (group 4). The ProstaScint results were independent of the Gleason score for 260 patients before and 285 patients after therapy. CONCLUSIONS: The results of this study underscore the complementary diagnostic value of ProstaScint to PSA level and Gleason score as an independent indicator of prostate cancer recurrence and metastases and in identifying extrapelvic metastases in both newly diagnosed and recurrent prostate cancer.

October 18, 2000 8:30-8:45. Problem Solving with FDG Positron Emission Mammography.

Purpose: To examine one potential clinical application of dedicated devices for positron emission mammography.Background: Dedicated devices for breast imaging with FDG have technical specifications that appear well-suited for detecting small breast cancers, including registration with x-rays, high count sensitivity, and spatial resolution in the 2-3 mm range.Methods: In IRB-approved clinical trials, patients who were scheduled for biopsy for suspicious findings on x-ray mammograms were injected with 10-20 mCi FDG intravenously one to two hours prior to x-ray guided core biopsy or lumpectomy. Positron emission mammograms using a dedicated device were performed just before biopsy, and results compared to final histopathology.Results: Three patients whose cancers were missed on initial x-ray mammograms or whose mammograms contained subtle abnormalities that were not strongly suspicious for cancer, were shown to have very suspicious hot spots on positron emission mammograms. These patients were later confirmed to have cancer.Discussion: The role for dedicated devices for positron emission mammography is evolving. Finding cancers that are either mammographically occult or have low indices of suspicion is a potential application for this technique. Further clinical and technical work will be needed to better define the strengths and weaknesses of the device in this application.

Innovations in breast cancer imaging: PET for diagnosis and follow-up.

Could positron emission tomography (PET), which identifies the higher metabolism of malignant tumors, become the noninvasive test needed to assess the thousands of falsely positive mammograms? Could serial scans be used to evaluate response to chemotherapy? This team examines the data on current and potential uses of PET.

Prediction of Salmonella mutagenicity.

The ability of a number of prediction systems was examined to determine how well they could predict Salmonella mutagenicity. The prediction systems included two computer-based systems (CASE and TOPKAT), the measurement of a physiochemical parameter (ke) and the use of structural alerts by an expert chemist. The computer-based systems operators and the chemist were supplied with the structures of 100 chemicals that had been tested for mutagenicity in the Salmonella test; the actual chemicals were needed for the physiochemical measurement. None of the participants was provided with the chemical names or Salmonella test results prior to submitting their predictions. The three systems that predicted the mutagenicity from the structure of the chemicals produced equivalent results (71-76% concordance with the Salmonella results); the physiochemical system produced a lower (60-61%) concordance.

Interlaboratory comparison of the effects of radon on L5178Y cells: dose contribution of radon daughter association with cells.

The cytotoxic and mutagenic effects of radon and its progeny were compared in murine lymphoblast L5178Y-R16 cells after exposure at three institutions. The cells were exposed to 222Rn at Case Western Reserve University (CWRU) and Pacific Northwest Laboratories (PNL) and to 212Bi, a decay product of 220Rn, at the University of Chicago (UC). The dose to the cell nucleus was calculated using a dosimetric model which addressed both the contribution of the dose from the radioactivity in the medium and that associated with the cells. The dose-response curves for cell survival showed D0's of 0.30 Gy at CWRU, 0.20 Gy at PNL, 0.37 Gy for chelated 212Bi, and 0.13 Gy for unchelated 212Bi. Induced mutant frequencies at the thymidine kinase locus at the 37% survival level were 1470 x 10(-6) at CWRU, 1518 at PNL, and 2414 x 10(-6) at UC using combined results for chelated and unchelated 212Bi. The variation between institutions was greater than obtained in a previous interlaboratory comparison of the effects of radon on CHO cells. Since less radioactivity was associated with CHO cells than L5178Y cells, we have concluded that the variation between institutions in the case of L5178Y cells is caused by the differences in cell-associated radioactivity and errors related to the measurement of this parameter.

Cytotoxic and mutagenic effects of radon and radon daughters on murine L5178Y lines differing in DNA repair.

The effects of 222Rn were measured in mouse L5178Y (LY) lymphoblasts that differ in repair capabilities. Line LY-S1 is deficient in the repair of X-radiation-induced DNA doublestrand breaks, while lines LY-R16 and LY-R83 are presumed to be deficient in the excision of UV-radiation-induced pyrimidine dimers. Line LY-R83 is hemizygous while the other two lines are heterozygous at the thymidine kinase (tk) locus. After exposure to radon the D0's were found to be very similar for the three lines (0.30-0.31 Gy), whereas for X radiation the D0 for line LY-S1 is lower (0.7 Gy) than that for the two LY-R lines (1.3 Gy). Mutant frequencies at the tk locus were higher per gray after treatment with radon than X radiation, but at equitoxic doses the mutant frequencies were similar for X and alpha-particle radiation. A low radon-induced mutant frequency was observed for the hemizygous line, in agreement with the hypothesis that multilocus lesions were induced by the alpha-particle radiation and that mutants bearing intergenic lesions were not recovered in the TK+/- line. The entire active tk allele was lost by 81% of the TK-/- mutants of line LY-R16. In lines LY-S1 and LY-R16, 39-43% of the TK-/- mutants exhibited loss of galactokinase activity, indicating that the mutational lesion inactivating the tk gene frequently extended to the neighboring galactokinase gene.

A radon generator/delivery system.

A radon-generating system is described in which 222Rn, emanating from 226Ra stored in an aluminum containment vessel, may be pumped into a syringe for subsequent injection into a standard spinner flask containing tissue culture medium. The radium-containment vessel is sealed by an indium gasket and three metal bellows valves, one of which was used to fracture the glass capsule that contained 2.9 GBq of radium salt. A rotating piston pump transfers radon-enriched air from the radium-containment vessel to a delivery loop that includes a transfer syringe. The flow of air and radon through the loop is manipulated by three crossover ball valves, one of which may be set to fill the syringe. A charcoal trap is provided to collect residual radon left in the delivery loop after the transfer syringe has been filled. The protocol used to expose cells to radon and its progeny is described as well as the dosimetry that is used to estimate the dose delivered to the cells. A description of safety precautions taken in fabricating the generator and in conducting radiobiological studies is also presented.

Response of the ke test to NCI/NTP-screened chemicals. III. Complementary value of ke in screening for carcinogens.

The value of using a physico-chemical carcinogen-screening test, the ke test, in conjunction with the Salmonella typhimurium/microsome assay (the Ames test) and/or structural alerts of reactivity (the S/A test), is analyzed on the basis of the response of the three tests to 171 chemicals of known rodent carcinogenicity. The Ames test is widely used to screen chemicals for potential carcinogenicity; however, its relatively low sensitivity (proportion of true positives among carcinogens tested) has prompted a search for complementary tests that increase sensitivity without an unacceptable decrease in specificity (proportion of true negatives among non-carcinogens tested). The S/A test is a structural analysis based on recognition of chemicals groups likely to react with DNA. The S/A test does not complement the Ames test well, because of the high similarity of responses (dependence) between these two tests. The ke test measures the affinity of a test chemical for electrons, and has a sensitivity and specificity comparable to the Ames test. The ke test is shown in this work to complement both the Ames test and the S/A test. Addition of the ke test to either the Ames test or the S/A test results in a substantial decrease in false negatives and an approximately equal increase in false positives, which is a trade-off that would be desirable in all but the least risk averse situations. The S/A and ke battery has a sensitivity of > 0.9, and could be applied to untested chemicals without any biological testing. In view of these observations, it is proposed that the ke test be considered in developing future strategies to optimize the screening of potential carcinogens in the most cost-effective manner.

Response of the ke test to NCI/NTP-screened chemicals. II. Genotoxic carcinogens and non-genotoxic non-carcinogens.

A physico-chemical carcinogen-screening test was used to measure the rate constants of electron attachment, kes, of 105 chemicals that had been screened in long-term rodent bioassays and short-term in vitro tests by the NCI/NTP. In the ke test, a pulse-conductivity technique is used to generate and monitor the decay of excess electrons that serve as nucleophilic surrogates for the target tissue of rodents. Of the 61 chemicals that had been found to be rodent carcinogens as well as Salmonella mutagens, 36 yield kes that are equal to or greater than the diffusion-controlled ke of carbon tetrachloride and are considered to be positive ke test responses. In contrast, 29 of the remaining 44 chemicals that are putative non-carcinogens and non-mutagens yield kes that are negative ke test responses. These results are combined with the ke responses of 46 non-mutagenic carcinogens and 20 mutagenic non-carcinogens that were reported earlier and are evaluated to determine the degree to which the measure of electron-accepting capacity that ke provides complements or overlaps the electrophilicity or DNA reactivity of chemicals that is indicated by positive mutagenicity responses in the Ames Salmonella tester strains or by positive structural alerts, S/As, of the chemicals. The combined ke test results indicate that the overall predictivity of the ke test is comparable to and complements the Ames Salmonella test and S/As in identifying rodent carcinogens. Moreover, the electrons serve as non-discriminate nucleophilic targets for both genotoxic and non-genotoxic electron-accepting molecules and appear to attach with equal efficiency to carcinogens that are active in various tissues of rodents. This property of excess electrons suggests that the predictivity of the ke test could be enhanced by combining the measured ke with an appropriate lipophilicity or pharmacokinetic parameter. A pre-chemical electron-transfer step that had been proposed to precede chemical interactions between the carcinogen and target tissue is discussed in light of recent developments in electron-donor/-acceptor chemistry and in the application of structure--activity relationships to identify carcinogens.

Prospective ke screening of potential carcinogens being tested in rodent bioassays by the US National Toxicology Program.

Values of ke, the rate constant of electron attachment, were measured in cyclohexane for 31 of 44 chemicals now being screened for carcinogenicity in rodent bioassays conducted by the US National Toxicology Program (NTP). These kes provide a physico-chemical measure of electron-solute interaction in a non-polar medium which had been found to be correlated with solute carcinogenicity in our earlier studies of the kes of chemical carcinogens and putative noncarcinogens. The ke test yields 15 negative and 16 positive responses for the 31 chemicals that were screened in this study. These ke results are compared with the predictions of other carcinogen-screening methods that have been applied to the same chemicals and also reported in this Discussion Forum.

In vitro exposure of mammalian cells to radon: dosimetric considerations.

We have developed a model to calculate the dose to the cell nucleus in cells exposed in suspension to radon and/or radon progeny. The model addresses the influence of (1) different radiation qualities and energies in the irradiation milieu; (2) the contribution to dose from radioactivity in the medium surrounding the cell after exposure to the radon gas as well as that from excess radon progeny associated with the cell; (3) the geometry of the cell and of the radiosensitive target, the cell nucleus; (4) the intracellular localization of the radionuclides; (5) attenuation of the alpha particles by the cytoplasm; (6) the radionuclide concentrations in the medium; and (7) the length of exposure. Investigation of the influence of these various parameters was made using an irradiation system in which cells were exposed to 212Bi, which decays to stability with the emission of an alpha particle (either 6.05 or 8.78 MeV). The information from these studies was then used to develop the system further for more complex systems in which 222Rn and its progeny are present. The model takes into account the contribution of dose from different radiation sources using scintillation counts of the medium and the cells, and it is useful for calculations of dose in situations where cells are exposed in suspension culture.

Response of the ke test to NCI/NTP-screened chemicals. I. Non-genotoxic carcinogens and genotoxic non-carcinogens.

The response of a physico-chemical carcinogen-screening test, the k(e) test, to 46 rodent carcinogens and 20 putative non-carcinogens that had been screened in long-term two-species bioassays by the National Cancer Institute/National Toxicology Program are reported. All of the chemicals screened are those that yield mutagenicity responses in the Ames Salmonella/microsome test that are either equivocal or contrary to the rodent carcinogenicity responses. The electron attachment rate constants, k(e)S, of the test chemicals in cyclohexane at 21 degrees C were measured using a pulse-conductivity technique. The k(e)S of 27 of the 46 rodent carcinogens (59%) are equal or greater than the diffusion-controlled k(e) of carbon tetrachloride, which is regarded as the boundary between a positive and negative response; the k(e)S of 8 of the 20 mutagenic non-carcinogens (40%) are less than diffusion-controlled. If the boundary between positive and negative k(e) responses is decreased to half the diffusion-controlled k(e), six additional carcinogens yield a positive ke response which increases the k(e) test sensitivity to 72% while the specificity to non-carcinogens remains at 40%. Comparison of these k(e)S with measures of the chemicals' electrophilicity that had been inferred from chemical structure indicates that k(e) provides a markedly different measure of electrophilicity and one that complements the Ames Salmonella assay. The use of the k(e) test as an analytical tool to indicate the presence of electron-attaching impurities in solvents such as benzene is discussed, as is the sensitivity of the k(e) test to rodent-liver carcinogens.

A physico-chemical screening test for chemical carcinogens: the ke test.

A pulse-conductivity technique was used to measure the rate at which excess electrons in liquid cyclohexane attach to carcinogens and non-carcinogens in order to determine if the electron attachment rate constant, ke, could be used to screen potential carcinogens. The keS of 114 chemicals are reported; these chemicals are among 182 that had previously been tested in a validation study of several short-term carcinogen-screening bioassays. The remaining 68 chemicals for which keS were not measured include chemicals that were unavailable, were not sufficiently stable or soluble in cyclohexane, or did not have a well-defined mol. wt. For the 114 chemicals that were tested, 35 are carcinogens, 50 are putative non-carcinogens and 29 have not been adequately tested or yielded equivocal responses in animal-test studies. Diffusion-controlled keS were measured for 27 of the 35 carcinogens tested whereas the keS of 45 of the 50 non-carcinogens were less than diffusion controlled. From these results, several measures of the predictive performance of using a diffusion-controlled ke to indicate a positive response to a carcinogen were calculated and compared with the Ames-test predictiveness in screening the same chemicals. The predictive criteria calculated were sensitivity, specificity, accuracy and predictive value, all of which were greater for the ke test than for the Ames test. Comparisons of the chemicals that yielded false-negative responses in the ke and Ames tests indicate a high degree of independence between the two which implies that the tests could be efficaciously used in a battery of short-term tests. Rationales are offered concerning the observed ke--carcinogenicity correlation and the apparent lack of the need for procarcinogens to be metabolically activated to yield a positive ke response.

Comparative lethal effects of uv and ionizing radiation in Ames tester strains of Salmonella.

In the three (parent-daughter) pairs of Ames Salmonella tester strains TA1535-TA100, TA1537-TA2637, and TA1538-TA98 in which the daughter strains carry the pKM101 plasmid but the parent strains do not, the pKM101 plasmid uniformly confers resistance of the host to uv radiation which indicates that the muc genes of the plasmid are present and function correctly in all three daughter strains. This uniform protection against killing by uv contrasts with the lethality responses of the same parent-daughter pairs to ionizing radiation (ir) where pKM101 again confers lethality protection to TA100 and TA2637 but sensitizes TA98 toward the lethal effects of ir. From these results we conclude that the pathways for error-prone repair of lethal lesions induced by uv and by ionizing radiation are not the same and that the muc genes of the plasmid alone are not sufficient to carry out error-prone repair of lethal lesions induced by ionizing radiation. We infer that a segment of plasmid DNA that is present in TA100 and TA2637 and is required to repair potentially lethal damage induced by ir is deleted in TA98.

Radiation-induced mutagenicity and lethality in ames tester strains of salmonella.

Mutation and killing induced by X radiation and 60CO gamma radiation were studied in six different histidine-requiring auxotrophs of Salmonella typhimurium. Strain TA100, which is sensitive to base-pair substitutions, and strains TA2637 and TA98, which are sensitive to frameshifts, carry the pKM101 plasmid and exhibit significantly higher radiation-induced mutations compared to their plasmidless parent strains TA1535, TA1537, and TA1538, respectively. Among the plasmid-containing strains, TA98 and TA2637 are much more sensitive to the mutagenic action of radiation than is TA100 based on a comparison with their respective spontaneous mutation rates; however, no uniformity was observed in the responses of the strains to the lethal action of ionizing radiation. The pKM101 plasmid provides partial protection against lethality in TA100 and TA2637, whereas the same plasmid enhances the lethal action of ionizing radiation in TA98. The following conclusions are consistent with these observations: (1) the standard Ames Salmonella assay correctly identifies ionizing radiation as a mutagenic agent; (2) frameshift-sensitive parent strains are more sensitive to the mutagenic effects of ionizing radiation than is the only strain studied that is sensitive to base-pair substitutions; and (3) enhancement of mutagenesis and survival is related to plasmid-mediated repair of DNA damage induced by ionizing radiation and does not involve damage induced by Cerenkov-generated uv radiation which is negligible for our irradiation conditions.

Quasifree electron attachment to carcinogens in liquid cyclohexane.

The quasifree electron attachment rate constant, kappa e, was measured in liquid cyclohexane at 20 degrees C for 76 chemicals that had been tested for bacterial mutagenicity in at least one of five earlier studies designed to determine the correlation between bacterial mutagenicity and animal carcinogenicity. A millionfold range of kappa e's was observed with the kappa e of 37 of 42 carcinogens being equal to or greater than the diffusion-controlled electron attachment rate constant, kappa d, of 2.9 x 10(12) M-1 s-1, whereas kappa e was less than kappa d for 30 of 34 noncarcinogens. These results yield a sensitivity of 88 percent for kappa e greater than or equal to kappa d indicating a chemical's carcinogenicity and a specificity also of 88% for kappa less than kappa d indicating a noncarcinogenic chemical. The observed quasifree electron attachment sensitivity and specificity compare favorably with previously observed Ames-test mutagenicity/carcinogenicity correlations which suggests that kappa e measurements also may be of value in a carcinogen-screening program.

Conjecture on the role of dry charges in radiosensitization.

A model of cellular radiation damage that involves dry electrons and holes is proposed and the implications of this model to hypoxic radiosenstization are discussed. The transport and reaction properties of dry charges in the structured water surrounding polar biomolecules are considered and a solvation time of 600 picoseconds in this medium is derived. Pre-solvation charge migration, attachment and recombination and the relevance of these processes to sensitization are discussed. DNA damage by physico-chemical interactions with dry holes is proposed and is shown to be modified by oxygen and other electron affinic sensitizers.