@article{thrall_rosner_azuma_mcentee_raleigh_1997, title={Hypoxia marker labeling in tumor biopsies: quantification of labeling variation and criteria for biopsy sectioning}, volume={44}, ISSN={["0167-8140"]}, DOI={10.1016/s0167-8140(97)01931-2}, abstractNote={The error associated with using biopsy-based methods for assessing parameters reflective of the tumor microenvironment depends on the variability in distribution of the parameter throughout the tumor and the biopsy sample. Some attention has been given to intratumoral distribution of parameters, but little attention has been given to their intrabiopsy distribution. We evaluated the intrabiopsy distribution of CCI-103F, a 2-nitroimidazole hypoxia marker.The hypoxia marker CCI-103F was studied in dogs bearing spontaneous solid tumors. Two biopsies were taken from each of seven tumors, for a total of 14 biopsies. Biopsies were serially sectioned and four to six contiguous slides from each 100-150 microm of the biopsy were used to formulate the best estimate of CCI-103F labeled area throughout the biopsy sample. One, two or four slides were then randomly selected from each biopsy and the labeled area, based on this limited sample, was compared to the estimate obtained from counting all available slides. Random sampling of slides was repeated 1000 times for each biopsy sample.CCI-103F labeling variance throughout the biopsy decreased as the estimated overall labeled area in the biopsy decreased. The error associated with estimating the overall labeled area in a biopsy from a randomly selected subset of slides decreased as the number of slides increased, and as the overall labeled area in the biopsy decreased. No minimally labeled biopsy was classified as unlabeled based on limited sampling.With regard to CCI-103F labeling, quantification of the labeled area in four randomly selected slides from a biopsy can provide, in most biopsies, an estimate of the labeled area in the biopsy within an absolute range of +/-0.05.}, number={2}, journal={RADIOTHERAPY AND ONCOLOGY}, author={Thrall, DE and Rosner, GL and Azuma, C and McEntee, MC and Raleigh, JA}, year={1997}, month={Aug}, pages={171–176} }
 @article{azuma_raleigh_thrall_1997, title={Longevity of pimonidazole adducts in spontaneous canine tumors as an estimate of hypoxic cell lifetime}, volume={148}, ISSN={["1938-5404"]}, DOI={10.2307/3579536}, abstractNote={The longevity of pimonidazole adducts in tumors was quantified as an estimate of the lifetime of hypoxic cells. Pimonidazole was given before irradiation to 12 dogs bearing spontaneous tumors, and tumors were biopsied 24, 48 and 72 h later. Pimonidazole antigen was quantified in the biopsies using ELISA and immunohistochemistry. Pimonidazole antigen was detectable in the initial biopsy in all dogs. In 5 dogs the amount of detectable antigen decreased to less than 50% of the initial amount, in 5 other dogs the amount of detectable antigen decreased to an amount between 50 and 100% of the initial amount, and in 2 dogs the amount of antigen appeared to increase relative to the initial amount. Tumors with high initial adduct concentration were characterized by greater decreases in adduct concentration than tumors with low initial adduct concentration. Immunohistochemically, labeled cells were present in 11 of 12 tumors. The geographic area in tumor biopsies labeled immunohistochemically with pimonidazole adducts (labeled area fraction) tended to decrease over time in 6 dogs, remain stable in 4 dogs and seemingly increase in 1 dog. There was no relationship in individual tumors between the relative change in antigen concentration and the relative change in labeled area fraction. Hypoxic cells which bind pimonidazole may persist for days during fractionated radiation therapy, and the potential exists for them to exert a negative effect on the host.}, number={1}, journal={RADIATION RESEARCH}, author={Azuma, C and Raleigh, JA and Thrall, DE}, year={1997}, month={Jul}, pages={35–42} }
 @article{kennedy_raleigh_perez_calkins_thrall_novotny_varia_1997, title={Proliferation and hypoxia in human squamous cell carcinoma of the cervix: First report of combined immunohistochemical assays}, volume={37}, ISSN={["0360-3016"]}, DOI={10.1016/S0360-3016(96)00539-1}, abstractNote={Preliminary data establishes pimonidazole as a useful probe for human tumor hypoxia. Although different in chemical structure and properties, pimonidazole is indistinguishable from other markers in its binding to hypoxic cells in human tumors. Pimonidazole hydrochloride is convenient to administer and is well tolerated when given as single intravenous infusion of 0.5 g/m2. Comparison of immunostaining for pimonidazole binding and for PCNA or MIB-1 shows an inverse relationship between proliferation and hypoxia.}, number={4}, journal={INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS}, author={Kennedy, AS and Raleigh, JA and Perez, GM and Calkins, DP and Thrall, DE and Novotny, DB and Varia, MA}, year={1997}, month={Mar}, pages={897–905} }
 @article{cline_rosner_raleigh_thrall_1997, title={Quantification of CCI-103F labeling heterogeneity in canine solid tumors}, volume={37}, ISSN={["0360-3016"]}, DOI={10.1016/S0360-3016(96)00559-7}, abstractNote={The purposes of this study were to assess sources of variation in the distribution of nitroimidazole-labeled hypoxic cells in canine tumors and to quantify the reliability of estimating overall nitroimidazole-labeled area fraction from biopsies.Hypoxic cells were labeled in 24 canine tumors by immunostaining of the nitroimidazole hypoxia marker CCI-103F. In tumors with a volume < 100 cm3, each cubic centimeter of tumor was examined; in larger tumors 100 randomly selected 1 cm3 samples were examined. These data were used to estimate the overall CCI-103F-labeled area fraction in the tumor. A variance components model was used to quantify intertumoral, intratumoral, and within slide (residual) sources of variation. The ability to estimate intratumoral CCI-103F-labeled area fraction based on information obtained from biopsies was assessed by randomly selecting two or four samples from the dataset for each tumor and comparing the mean CCI-103F-labeled area fraction from this limited sample to the labeled area fraction based on each cubic centimeter; this simulation process was repeated 1000 times.Intratumoral (27% of total) and intertumoral (30% of total) variation in CCI-103F-labeled area fraction were similar. Residual variation (variation at the microscopic level) accounted for 43% of total variation in CCI-103F labeling. Intratumoral variation in labeling decreased as the intratumoral CCI-103F mean labeled area fraction decreased. The accuracy of estimating the intratumoral CCI-103F-labeled area fraction in a tumor from limited sampling increased as the number of samples increased or the intratumoral labeled area fraction decreased. When four random samples were used to estimate overall CCI-103F-labeled area fraction in the tumor, estimates from approximately 90% of the 1000 simulations were within 0.10 of the intratumoral CCI-103F-labeled area fraction. Classifying a minimally labeled tumor as unlabeled based on limited sampling was unlikely.Despite intratumor variation, acceptable estimates of nitroimidazole-labeled cells in a tumor may be obtained from a clinically feasible number of biopsies.}, number={3}, journal={INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS}, author={Cline, JM and Rosner, GL and Raleigh, JA and Thrall, DE}, year={1997}, month={Feb}, pages={655–662} }