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Lethal member of complementation group A, gives homozygous and hemizygous flies which are clearly ras in eye color, suggesting that the remaining members of this complementation group are ras-Z's. All 24 mutations were tested in double heterozygotes with ras' in a v background to facilitate observation of the ras phenotype. Six members of group A including J5 ; gave raseyed flies and are therefore ras-1's; none of the remaining 18 mutants were obviously different in phenotype from v v . two group A mutations, F1 and F19, gave ras + eye-color in double heterozygotes with ras', so they were not classified as ras-Z on the basis of this test. T h e allelic variation implied by this observation was further investigated. In order to differentiate possible allelic variation from the effects of modifiers and of experimental fluctuations, each group A mutation was placed in a different genetic background, as described in MATERIALS AND METHODS. T h e original and the substituted stocks were then used to investigate the phenotype of each mutant in heteroallelic combination with gualts, p u r l purl' and ras'. In the cases of the auxotrophic mutations, survival of the heterozygotes on axenic yeast-sucrose and SANG'S nucleoside-free media was examined at 25 " and 29 . T rus' combinations were examined chromatographically for pteridine content. Sample alleles from the remaining seven complementation groups gave no evidence of allelism with any of the four nonlethals results not shown ; . Survival of gual * ' ras-l combinations: guul" homozygotes and hemizygotes show about 5% survival at 29" and 50% survival at 25" when grown on nucleoside-free defined medium FALK1973 ; . An approximately similar temperature-dependent differential is found when this mutant is exposed by a deficiency JOHNSON, WOLOSHYN and NASH1979 ; . T h same genotypes survive as well as controls on yeast-sucrose medium at either temperature. Table 3 shows the survival of the new ras-1 mutations in combination with p a l yeast-sucrose medium the gual' * ras-1 flies show high levels of survival in all cases. In only one case Zz F1 m ; survival significantly, although marginally, depressed. On defined medium, all 32 crosses demonstrate significant depression of survival. Seven mutations D12, F I , F19, H6, H25, K27 and N23 ; show low survival less than 6% ; at 29" and seem to be at least as extreme as guul" itself. T h e eighth mutation J5 ; survives considerably better and would therefore appear to be less defective than gual". T h e result is reflected in the results at 25". At 25", K27 also survives relatively well in both backgrounds; the remainder all show severely depressed survival compared with guall" Df JOHNSON, WOLOSHYN and NASH1979 ; . This latter phenomenon, in which the heteroallelic combination has a more extreme phenotype than the deficiency heterozygote, was previously described by NASHet al. 1981 ; . Survival of purl' ras-l combinations: p u r mutation that shows less than 5% survival on nucleoside-free medium FALKand NASH 1974a its supplementation and complementation behavior suggest a main defect in p u function and a minor defect in g u function FALK1973 ; . Deficiency heterozygotes survive poorly on yeast-sucrose medium JOHNSON, WOLOSHYN and.

Pulmonary fibrosis is still unknown despite the fact that pulmonary fibrosis in rodents after intratracheal i.t. ; administration of a single dose of BLM has been widely used as a model to study the mechanisms of fibrotic lung disease [4]. These studies have revealed an acute cellular infiltrate after BLM treatment, which was comprised of macrophages, granulocytes, and lymphocytes in the alveolar spaces and interstitial tissue [57]. That lymphocytes may play a role in BLM-induced pulmonary fibrosis has been suggested by studies in which inhibition or depletion of lymphocytes by either antilymphocyte antibody, mAb to T cell subsets, or steroids suppressed BLM-induced pulmonary fibrosis [810]. However, results from experiments using nude mice that lacked T cells have conflicted, showing either a reduction or no change in BLM-induced fibrosis [11, 12]. More recently, studies have focused on the role of fibrogenic cytokines in the development of BLM-induced pulmonary fibrosis. A variety of cytokines and chemokines have been implicated, such as interleukin-1 IL-1 ; [13, 14], tumor necrosis factor- TNF- ; [15, 16], macrophage inflammatory protein-1 MIP-1 ; [17], interleukin-5 IL-5 ; [18], and transforming growth factor TGF- ; [1922]. Lung macrophages have been reported as the primary source of these cytokines [1322] but other cell types such as fibroblasts [23], endothelial cells [24], and eosinophils [18 ] also have been described to secrete TGF- after BLM stimulation. Antibodies against TGF- [25] or TNF- [15] and human recombinant soluble TNF receptor [16] have also been shown to markedly inhibit the development of BLM-induced pulmonary fibrosis in mice. The purpose of this study was to investigate the role of lung T cells and cytokines in the pathogenesis of BLM-induced pulmonary fibrosis. Based on previous studies we hypothesized that T cell activation and cytokine secretion early after BLM treatment is an initial step in triggering the process of pulmonary fibrosis by activating macrophages to secrete fibrogenic factors. To test this hypothesis, BLM-induced fibrosis was analyzed in C57BL 6 SCID, CB.17 SCID C57BL 6 SCID CB.17 SCID ; F1 mice, and their wild-type counterparts. The results demonstrated that C57BL 6 and C57BL 6 BALB c ; F1, as well as their SCID counterparts, were equally susceptible to. Sample represents -0.1% of the blood volume. Thus, with 1000 cancer cells in the circulation there would be a 70% chance of getting a positive result according to the Poisson distribution ; if the method is able to detect one cell. By the present method we detected 10 LNCap cells 6 X 10 blood leukocytes, corresponding to 1.6 LNCap cells l06 blood cells. A recently described RT-PCR had similar sensitivity, i.e., 1 LNCap cell in 106 peripheral blood mononuclear cells 17 ; . The sensitivity could be increased by taking a larger volume of blood, but the possibilities of this approach and pindolol.

Middot; it is not known whether photofrin passes into breast milk. Table 2. Comparison of blood, early secretion and late secretion and pram Cure of xenografted human carcinomas by BR96-doxorubicin immunoconjugates. Science Washington DC ; , 261: 212215, 1993. Lu, Z-R., Kopeckova, P., and Kopecek, J. Polymerizable Fab antibody fragments for targeting of anticancer drugs. Nat. Biotechnol., 17: 11011104, 1999. Tolcher, A. W., Sugarman, S., Gelmon, K. A., Cohen, R., Saleh, M., Isaacs, C., Young, L., Healey, D., Onetto, N., and Slichenmyer, W. Randomized Phase II study of BR96-doxorubicin conjugate in patients with metastatic breast cancer. J. Clin. Oncol., 17: 478 484, Markman, M. Basic Cancer Medicine. Philadelphia, PA: W. B. Saunders Company, 1997. 18. Meirow, D. Ovarian injury and modern options to preserve fertility in female cancer patients treated with high dose radio-chemotherapy for hemato-oncological neoplasias and other cancers. Leuk. Lymphoma, 33: 6576, 1999. Lidor, Y. L., O'Briant, K. C., Xu, F. J., Hamilton, T. C., Ozols, R. F., and Bast, R. C., Jr. Alkylating agents and immunotoxins exert synergistic cytotoxicity against ovarian cancer cells. J. Clin. Investig., 92: 2440 2447, Canti, G., Nicolin, A., Cubeddu, R., Taroni, P., Bandieramonte, G., and Valentini, G. Antitumor efficacy of the combination of photodynamic therapy and chemotherapy in murine tumors. Cancer Lett., 125: 39 44, Brophy, P. F., and Keller, S. M. Adriamycin enhanced in vitro and in vivo photodynamic therapy of mesothelioma. J. Surg. Res., 52: 631 634, Krinick, N. L., Sun, Y., Joyner, D., Spikes, J. D., Straight, R. C., and Kopecek, J. A polymeric drug delivery system for the simultaneous delivery of drugs activatable by enzymes and or light. J. Biomater. Sci. Polym. Ed., 5: 303324, 1994. Peterson, C. M., Lu, J. M., Sun, Y., Peterson, C. A., Shiah, J. G., Straight, R. C., and Kopecek, J. Combination chemotherapy and pho todynamic therapy with N- 2-hydroxypropyl ; methacrylamide copolymer-bound anticancer drugs inhibit human ovarian carcinoma heterotransplanted in nude mice. Cancer Res., 56: 3980 3985, Peterson, C. M., Lu, J. M., Gu, Z. W., Shiah, J. G., Lythgoe, K., Peterson, C. A., Straight, R. C., and Kopecek, J. Cooperativity between free and N- 2-hydroxypropyl ; methacrylamide copolymer bound Adriamycin and mesochlorin e6 monoethylene diamine induced photodynamic therapy in human epithelial ovarian carcinoma in vitro. Int. J. Oncol., 15: 516, 1999. Shiah, J-G., Sun, Y., Peterson, C. M., and Kopecek, J. Biodistribu tion of free and N- 2-hydroxypropyl ; methacrylamide copolymer-bound mesochlorin e6 and Adriamycin in nude mice bearing human ovarian carcinoma OVCAR-3 xenografts. J. Controlled Release, 61: 145157, 1999. Omelyanenko, V., Kopeckova, P., Gentry, C., Shiah, J. G., and Kopecek, J. HPMA copolymer-anticancer drug-OV-TL 16 antibody conjugates. I. Influence of the method of synthesis on the binding affinity to OVCAR-3 ovarian carcinoma cells in vitro. J. Drug Targeting, 3: 357373, 1996. Hashimoto, Y., Hirano, T., and Yamaguchi, N. Novel after-loading interstitial photodynamic therapy of canine transmissible sarcoma with photofrin II and excimer dye laser. Jpn. J. Cancer Res., 86: 239 244, Takakura, Y., and Hashida, M. Macromolecular carrier systems for targeted drug delivery: pharmacokinetic considerations on biodistribution. Pharm. Res. NY ; , 13: 820 831, Pimm, M. V., Perkins, A. C., Strohalm, J., Ulbrich, K., and Duncan, R. scintigraphy of the biodistribution of 123I-labelled N- 2-hydroxypropyl ; methacrylamide copolymer-doxorubicin conjugates in and photofrin.

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