[18]F-fluoroethyl-l-tyrosine positron emission tomography for radiotherapy target delineation: Results from a Radiation Oncology credentialing program.

Background and purpose: The [18]F-fluoroethyl-l-tyrosine (FET) PET in Glioblastoma (FIG) study is an Australian prospective, multi-centre trial evaluating FET PET for newly diagnosed glioblastoma management. The Radiation Oncology credentialing program aimed to assess the feasibility in Radiation Oncologist (RO) derivation of standard-of-care target volumes (TVMR) and hybrid target volumes (TVMR+FET) incorporating pre-defined FET PET biological tumour volumes (BTVs). Materials and methods: Central review and analysis of TVMR and TVMR+FET was undertaken across three benchmarking cases. BTVs were pre-defined by a sole nuclear medicine expert. Intraclass correlation coefficient (ICC) confidence intervals (CIs) evaluated volume agreement. RO contour spatial and boundary agreement were evaluated (Dice similarity coefficient [DSC], Jaccard index [JAC], overlap volume [OV], Hausdorff distance [HD] and mean absolute surface distance [MASD]). Dose plan generation (one case per site) was assessed. Results: Data from 19 ROs across 10 trial sites (54 initial submissions, 8 resubmissions requested, 4 conditional passes) was assessed with an initial pass rate of 77.8 %; all resubmissions passed. TVMR+FET were significantly larger than TVMR (p < 0.001) for all cases. RO gross tumour volume (GTV) agreement was moderate-to-excellent for GTVMR (ICC = 0.910; 95 % CI, 0.708-0.997) and good-to-excellent for GTVMR+FET (ICC = 0.965; 95 % CI, 0.871-0.999). GTVMR+FET showed greater spatial overlap and boundary agreement compared to GTVMR. For the clinical target volume (CTV), CTVMR+FET showed lower average boundary agreement versus CTVMR (MASD: 1.73 mm vs. 1.61 mm, p = 0.042). All sites passed the planning exercise. Conclusions: The credentialing program demonstrated feasibility in successful credentialing of 19 ROs across 10 sites, increasing national expertise in TVMR+FET delineation.

Genomic testing for copy number and single nucleotide variants in spermatogenic failure.

PURPOSE: To identify clinically significant genomic copy number (CNV) and single nucleotide variants (SNV) in males with unexplained spermatogenic failure (SPGF). MATERIALS AND METHODS: Peripheral blood DNA from 97/102 study participants diagnosed with oligozoospermia, severe oligozoospermia, or non-obstructive azoospermia (NOA) was analyzed for CNVs via array comparative genomic hybridization (aCGH) and SNVs using whole-exome sequencing (WES). RESULTS: Of the 2544 CNVs identified in individuals with SPGF, > 90% were small, ranging from 0.6 to 75 kb. Thirty, clinically relevant genomic aberrations, were detected in 28 patients (~ 29%). These included likely diagnostic CNVs in 3/41 NOA patients (~ 7%): 1 hemizygous, intragenic TEX11 deletion, 1 hemizygous DDX53 full gene deletion, and 1 homozygous, intragenic STK11 deletion. High-level mosaicism for X chromosome disomy (~ 10% 46,XY and ~ 90% 47,XXY) was also identified in 3 of 41 NOA patients who previously tested normal with conventional karyotyping. The remaining 24 CNVs detected were heterozygous, autosomal recessive carrier variants. Follow-up WES analysis confirmed 8 of 27 (30%) CNVs (X chromosome disomy excluded). WES analysis additionally identified 13 significant SNVs and/or indels in 9 patients (~ 9%) including X-linked AR, KAL1, and NR0B1 variants. CONCLUSION: Using a combined genome-wide aCGH/WES approach, we identified pathogenic and likely pathogenic SNVs and CNVs in 15 patients (15%) with unexplained SPGF. This value equals the detection rate of conventional testing for aneuploidies and is considerably higher than the prevalence of Y chromosome microdeletions. Our results underscore the importance of comprehensive genomic analysis in emerging diagnostic testing of complex conditions like male infertility.

A prospective evaluation of health-related quality of life after skull base re-irradiation.

PURPOSE: To report cancer control outcomes and health-related quality of life (HRQoL) outcomes after highly conformal skull-based re-irradiation (re-RT). METHODS: Patients planned for curative intent re-RT to a recurrent or new skull base tumor were enrolled. HRQoL were assessed using the MD Anderson Symptom Inventory Brain Tumor (MDASI-BT) and the anterior skull base surgery quality of life (ASBQ) questionnaires. RESULTS: Thirty-nine patients were treated with stereotactic body RT or intensity modulated RT. Median follow-up was 14 months. Progression free survival was 71% at 1-year. There was mild clinically significant worsening of fatigue, lack of appetite and drowsiness (MDASI-BT), and physical function (ASBQ) at the end of RT, followed by recovery to baseline on subsequent follow-ups. Subjective emotions were clinically improved at 12 months, with patients reporting feeling less tense/nervous. CONCLUSION: Conformal skull base re-RT is associated with mild immediate deterioration in physical function followed by rapid and sustained recovery.

Radiographic retropharyngeal lymph node involvement in HPV-associated oropharyngeal carcinoma: Patterns of involvement and impact on patient outcomes.

BACKGROUND: The objective of the current study was to characterize the incidence, pattern, and impact on oncologic outcomes of retropharyngeal lymph node (RPLN) involvement in HPV-associated oropharyngeal cancer (OPC). METHODS: Data regarding patients with HPV-associated OPC who were treated at The University of Texas MD Anderson Cancer Center with intensity-modulated radiotherapy from 2004 through 2013 were analyzed retrospectively. RPLN status was determined by reviewing pretreatment imaging and/or reports. Outcomes analysis was restricted to patients with lymph node-positive (+) disease. Kaplan-Meier survival estimates were generated and survival curves were compared using the log-rank test. Bayesian information criterion assessed model performance changes with the addition of RPLN status to current American Joint Committee on Cancer staging. Competing risk analysis compared modes of disease recurrence. RESULTS: The incidence of radiographic RPLN involvement was 9% (73 of 796 patients) and was found to vary by primary tumor site. The 5-year rates of freedom from distant metastases (FDM) and overall survival were lower in patients with RPLN(+) status compared with those with RPLN-negative (-) status (84% vs 93% [P = .0327] and 74% vs 87% [P = .0078], respectively). RPLN(+) status was not found to be associated with outcomes on multivariate analysis. Bayesian information criterion analysis demonstrated that current American Joint Committee on Cancer staging was not improved with the inclusion of RPLN. Locoregional and distant disease recurrence probabilities for those patients with RPLN(+) status were 8% and 13%, respectively, compared with 10% and 6%, respectively, for those with RPLN(-) status. RPLN(+) status portended worse 5-year FDM in the low-risk subgroup (smoking history of <10 pack-years) and among patients who received concurrent chemotherapy but not induction chemotherapy. CONCLUSIONS: RPLN(+) status was associated with worse overall survival and FDM on univariate but not multivariate analysis. In subgroup analyses, RPLN(+) status was associated with poorer FDM in both patients with a smoking history of <10 pack-years and those who received concurrent chemotherapy, suggesting that RPLN(+) status could be considered an exclusion criteria in treatment deintensification efforts seeking to omit chemotherapy.

Spiral brush cervical biopsy experience in a community clinic.

OBJECTIVE: This article seeks to demonstrate the experience of implementing the spiral brush in several community clinic locales. Before the introduction of the spiral brush cervical biopsy in 2002 there were few alternatives to colposcopy directed punch biopsy when evaluating and managing abnormal dysplastic Papanicolaou (pap) smear or a visually abnormal cervix. Subsequent investigations validated the spiral brush usage but there are limited reports for its implementation in primary care colposcopy. PATIENTS AND METHODS: Over a two year period (2004-2006) patients with internal referrals for colposcopy received the spiral brush cervical biopsy. Those that resulted in the diagnoses of high grade squamous intraepithelial lesion (HSIL) diagnoses (CIN2-3) were compared to the final pathology diagnosis from the loop excision specimen. RESULTS: 15 cases of HSIL were identified with subsequent loop excision. Comparison of the pathology diagnosis from the loop excision and the spiral brush biopsy resulted in 13.3% (n=2) of cases differing. In both cases, low grade squamous intraepithelial lesion (LSIL) diagnoses (CIN1) were found whereas the remainder had the same diagnosis of HSIL. CONCLUSIONS: These results showed acceptable rates of concordance with traditional pathology specimens which supports the use of this Food and Drug Administration (FDA) approved device within a primary care setting.

Etoposide induction of tumor immunity in Lewis lung cancer.

PURPOSE: To determine if the antineoplastic effect of etoposide includes alteration in Lewis lung cancer cells which evoke an immunologic response in C57B1/6 host mice. METHODS AND RESULTS: Of C57B1/6 mice injected with 10(6) Lewis lung cancer (3LL) cells followed by treatment with a single 50 mg/kg dose of etoposide (VP-16), 60% survived over 60 days, in contrast to untreated control mice which died within 30 days. Approximately 40% of surviving mice rejected a subsequent challenge with 3LL. Their splenocytes protected naive mice injected with 3LL. To test if VP-16 treatment produced alterations in 3LL cells, which induce host immunity, leading to tumor rejection, C57B1/6 mice were injected with 3LL cells that had survived an 80-90% lethal concentration of VP-16 in vitro. These cells killed 75% of recipient mice but 60% of the surviving mice rejected challenge with 3LL. Splenocytes harvested from tumor-rejecting mice protected naive mice injected with 3LL. CONCLUSION: These results support the hypothesis that in addition to its antineoplastic cytotoxic effect, VP-16 induces changes in 3LL cells which are recognized by the host immune system resulting in immune rejection of 3LL. often immunosuppressive and therapeutic advantage is generally based on the tumor cytotoxicity of individual drugs or combinations of drugs [13]. Our earlier work showed a link between the use of cytotoxic chemotherapy with etoposide (VP-16) and the induction of an immune response against syngeneic murine leukemia in the intact host [16]. VP-16 is an immunosuppressive topoisomerase II-inhibiting drug which induces tumor cell apoptosis and is frequently used clinically to treat a variety of tumors [1, 3, 9, 10]. We have noted that the addition of cyclosporin A to VP-16 produces CD8 T lymphocyte-mediated tumor-specific immunity in mice bearing L1210 leukemia [17]. We have extended these experiments to a spontaneously arising non-carcinogen-induced neoplasm, Lewis lung cancer (3LL), and now report that surviving mice successfully treated with VP-16, in the absence of cyclosporin A, reject challenge with 3LL. In addition, results are presented to show that VP-16 modifies 3LL cells rendering them immunogenic. These findings are submitted to support the hypothesis that VP-16-induced cytotoxic changes include cellular membrane alterations in 3LL cells which are recognized by the immune system and cause rejection of this syngeneic lung tumor.

Re-epithelialization in cornea organ culture after chemical burns and excimer laser treatment.

OBJECTIVE: To describe the epithelial healing rates observed in freshly cultured rabbit corneas chemically burned with high-concentration hydrochloric acid (HCl) and sodium hydroxide (NaOH) and subsequently treated with phototherapeutic keratectomy (PTK). METHODS: We obtained 126 fresh corneoscleral rims from cadaveric New Zealand white rabbits. Each cornea was exposed to 4-mm cellulose sponges soaked in a solution of topical 0.9% isotonic sodium chloride solution, 2M HCl, or 0.5M NaOH. A transepithelial PTK (6-mm zone; 100-microm ablation depth) was then performed using the excimer laser (150-mJ/cm(2) energy pulse; 20 nanosecond duration; and 10-Hz frequency). Corneas were placed in tissue culture, and 1 cornea from each group was taken out of culture each day after treatment. Re-epithelialization was monitored by means of fluorescein staining, slitlamp photography, and histopathological analysis. RESULTS: Corneas treated with HCl and NaOH exhibited immediate epithelial defects that slowly healed over time. In PTK-treated corneas, the re-epithelialization rate was accelerated compared with that of controls (P =.003 for the HCl group, and P<.001 for the NaOH group). The new epithelial layers were smoother in PTK-treated corneas, as confirmed by results of histopathological analysis. CONCLUSION: Corneal damage caused by HCl and NaOH may be modulated in vitro by PTK in this rabbit model. CLINICAL RELEVANCE: After corneal chemical damage, 193-nm excimer laser PTK accelerates epithelial wound healing.

Simple organ cornea culture model for re-epithelialization after in vitro excimer laser ablation.

BACKGROUND AND OBJECTIVE: Most of the in vitro work to characterize the effects of clinical laser surgery on corneal tissues has concentrated on the effects on stromal keratocytes and endothelium with little attention being paid to corneal epithelium. Our purpose is to describe the epithelial healing rates observed in freshly cultured rabbit corneas treated with phototherapeutic keratectomy (PTK). STUDY DESIGN/MATERIALS AND METHODS: Corneas were placed in a simple organ culture system, with media change every 2 days. A clinical excimer laser was used to perform a 6 mm diameter, 100 microm depth transepithelial PTK on 24 cultured rabbit corneas, 1 day after culture initiation. For each post-treatment day, one experimental and one control cornea were removed from culture and stained with fluorescein, photographed, and fixed for histology. Epithelial defect area was measured with digital imaging software and analyzed statistically to assess the re-epithelialization rate. RESULTS: Control corneas, maintained in culture for 1-4 days, had no epithelial defects. Those corneas treated with PTK exhibited an immediate epithelial defect that slowly healed over 3 days. This was confirmed on histopathological analysis. A significant linear trend in re-epithelialization across the time points studied was found (F = 80.48, P = 0.0029). The slope of the linear regression model showed an estimate rate of re-epithelialization of -6.70 over the 3 days. CONCLUSION: We have described the development of a simple, whole organ, rabbit cornea culture model for re-epithelialization after PTK. Our rates of epithelial healing resemble those found in the literature in live rabbit models. Therefore, this model may possibly be used to monitor epithelial wound healing in different corneal diseases or injuries.

Assessment of analgesia in human chronic pain. Randomized double-blind crossover study of once daily repro-dose morphine versus MST continus.

OBJECTIVE: This study evaluated Repro-Dose morphine (RDM; Reliadol from Nycomed Pharma), a new once daily controlled-release morphine formulation, against twice daily MST Continuous (MST) at steady state in patients with chronic opioid responsive pain. METHODS: A randomized double-blind two-way crossover design was used to evaluate the efficacy and adverse effects of RDM once daily or MST twice daily, at the same total daily doses, in patients with chronic stable pain (dose range 20-120 mg per day). During the RDM limb of the study active drug was administered in the evening and placebo in the morning. Dextromoramide was provided as escape analgesia throughout the study. Following a 5-day screening period, during which stability of oral opioid dose was verified, patients underwent two 5-day treatment periods, (one MST, one RDM) in random sequence. Pain scores, escape analgesia requirements and side-effects were compared using data from days 3, 4 and 5 of each treatment period. Any events or medication changes occurring during the study period thought liable to influence analgesia were regarded as protocol violations. Overall assessment and period preference was assessed by direct questioning. RDM treatment was regarded as successful if the amount of escape medication required during the RDM period was equal to or less than that required during the MST period. RESULTS: Forty-seven patients were included in the study, of whom 40 completed both periods [the intention to treat (ITT) population], 31 in strict accordance with the protocol [the per protocol (PP) population]. Results were similar for both populations. There was no significant difference in pain scores or incidence of adverse events occurring during the MST and RDM periods. For the ITT population, requirements for escape medication during the RDM period were less than, equal to or greater than those recorded during the MST period for 14, 15, and 11 patients, respectively. Twenty-nine of 40 patients (72.5%) were therefore RDM treatment successes (95% confidence interval 56.1-85.4%). The percentage of patients preferring RDM (45%) combined with those with no preference (32.5%) was significantly higher than those preferring MST (22.5%; P = 0. 0003). CONCLUSIONS: Oral morphine administered as RDM once daily is at least as effective and well tolerated as MST twice daily, with over 70% of patients in this double-blind crossover study reporting that RDM was equal or superior to MST.

Thermal sensitivity of endodontically treated teeth.

CASE REPORTS: The problem of thermal sensitivity following non-surgical root-canal treatment is explored and case reports are presented. Possible causes for post-treatment discomfort from endodontic and restorative aetiologies are discussed, as are the mechanisms to explain the patients' painful experiences. Treatment of this problem may vary from the simple replacement of a defective restoration to a more extensive non-surgical retreatment of the case, despite radiographic evidence of an acceptable root filling and normal periradicular tissues.

Superiority of cyclosporin A over PSC-833 in enhancement of VP-16 efficacy in murine tumors in vivo.

PSC-833, a non immunosuppressive analogue of cyclosporin A, is an effective modulator of the multidrug-resistant tumor phenotype. Since both PSC-833 and cyclosporin A also enhance the cytotoxicity of VP-16 against drug sensitive L1210 leukemia cells in vitro we compared these agents as modulators of VP-16 efficacy in vivo. Compared to VP-16 treatment alone both PSC-833 and cyclosporin A significantly altered the survival of L1210 leukemia-bearing BDF/1 mice and Lewis lung carcinoma-bearing C57/B1 mice. Cyclosporin A enhanced VP-16 efficacy whereas PSC-833 impaired VP-16 efficacy against these murine tumors. Possible reasons for these disparate effects are discussed.

Microtubule-dependent multilobular organization of the nucleus in sensitive and multidrug-resistant L0 leukemia cells.

The relationship between the nuclear morphology and the microtubular organization of L100 and L1000 cells, two vincristine-induced multidrug resistant human acute lymphocytic leukemia cell lines, was examined and compared to that of L0 parental cells. The L0 parental cells contained a round nucleus and the microtubules were evenly distributed throughout the cytoplasm. In contrast, the microtubules of the L100 and L1000 cells were localized between the lobular structures of a multilobulated nucleus. Disassembly of microtubules in L100 and L1000 cells by colchicine resulted in the loss of the multilobulated morphology of the nucleus. While the total cellular content of tubulin of L0 and L100 cells was similar, the content of microtubules of L100 cells was only 55% of that observed in L0 cells. Two, 28 kDa (pI 6.9) and 31 kDa (pI 4.4), microtubule-associated proteins were found to be overexpressed in L100 and L1000 cells. The results indicate that the multilobulated nuclear morphology of L100 and L1000 cells is dependent upon the unique and intact organization of the microtubules; the distinct organization of the microtubules and the multilobular nuclear morphology of the two resistant cell lines may be due to the differential expression of specific microtubule-associated proteins.

Comparison of cyclosporin A, verapamil, PSC-833 and cremophor EL as enhancing agents of VP-16 in murine lymphoid leukemias.

Although verapamil, cyclosporin A. cremophor EL and PSC-833 are active as multidrug resistance modulators, there has been limited study of these compounds as possible chemotherapy enhancing agents against drug-sensitive tumors. We compared these agents as modifiers of VP-16 cytotoxicity in vitro and modifiers of VP-16 efficacy in vivo against drug-sensitive P388 and L1210 leukemias. Our study indicates that cyclosporin A enhances VP-16 cytotoxicity to a significantly greater extent than equimolar concentrations of verapamil or PSC-833. Although cremophor EL shows significantly greater activity than verapamil in VP-16 cytotoxicity enhancement in vitro, it is ineffective when added to VP-16 therapy of mice bearing L1210 leukemia.

Cyclosporin A/VP-16 produced immunity to L1210 leukemia: the participation of cytotoxic CD8 T-lymphocytes.

The role of the immune response in the chemotherapeutic cure of an intact host with neoplasia is not well defined. We have previously shown that the addition of cyclosporin A to VP-16 therapy of BDF/1 mice with L1210 leukemia produces immunity to leukemia in long-surviving host animals. We now characterize this immunity as being tumor specific and related to the participation of CD8 T-lymphocytes. Splenocytes derived from L1210 leukemia immune mice are cytotoxic to L1210 cells after in vitro restimulation, compared to splenocytes harvested from nonimmune control mice. This cytotoxicity is lost by CD8 T-lymphocyte depletion and persists in Ia antigen blocking experiments. Cytotoxicity is selective for L1210 leukemia compared to P388 leukemia, an alternate Ia antigen expressing methylcholanthrine-induced acute lymphoid leukemia, and L1210 leukemia-immune mice remain susceptible to P388 leukemia in vivo demonstrating specificity of the immune response generated by cyclosporin A/VP-16 therapy.

Synergistic interaction of cyclosporin A and verapamil on vincristine and daunorubicin resistance in multidrug-resistant human leukemia cells in vitro.

We studied the effects of cyclosporin A and verapamil on the modulation of vincristine and daunorubicin resistance in a multidrug-resistant subline of human T-cell acute lymphatic leukemia GM3639. Our results show that cyclosporin A is more effective than verapamil as a modulator of the high degree of primary vincristine resistance and the low degree of daunorubicin cross-resistance expressed by this cell line. Isobologram analysis revealed that the combined modulators act synergistically in correcting both vincristine and daunorubicin resistance.

Novel nucleolar and nuclear morphology in a vincristine-dependent human leukemia cell line (L100).

Acquired resistance to chemotherapeutic drugs by tumor cells is an important obstacle to effective therapy of human malignancy. We now describe a vincristine (VCR)-induced multidrug-resistant (MDR) human acute lymphatic leukemia cell line, the sustained in vitro growth of which is dependent on vincristine. The doubling time for parental drug-sensitive cells (L0) is 40.2 +/- 13.2 h and for the MDR subline (L100) 62.5 +/- 11.3 h. L100 cells have similar G2 and mitotic phase to parental cells, express the MDR phenotype, and are characterized by novel morphologic features with multilobulated nuclei and multiple small nucleoli. Compared with L0 cells which have 2-3 nucleoli per cell, L100 cells have 7-8 nucleoli per cell. Average nucleolar area is 11.3 +/- 7.3 microns 2 for L0 and 2.5 +/- 2.4 microns 2 for L100 cells determined by the laser scanning method. The striking morphologic abnormalities of L100 cells suggest a drug-induced cytoskeletal abnormality. The relationship of these abnormalities to the VCR growth dependence of L100 cells is discussed.

Development of cyclosporin A mediated immunity in L1210 leukaemia.

Cyclosporin A (CsA) is an effective modulator of multidrug resistance (MDR) in vitro and in murine tumour systems in vivo. We now report the production of immunity to L1210 leukaemia by the addition of CsA to VP-16 therapy of leukaemic BDF/1 mice. VP-16/cyclosporin A tumour immunity induction arises as a consequence of active therapy independently of immunisation with modified tumour cells. The addition of CsA to VP-16 prolongs survival of BDF/1 host mice bearing L1210 leukaemia beyond that produced by equivalent dose VP-16 alone. A subpopulation of 60-day surviving mice after combined VP-16/CsA are immune to rechallenge with the same leukaemia inoculum to which they were originally exposed. Spleen cells from immune mice adoptively transfer anti-L1210 leukaemia immunity to untreated BDF/1 mice in a dose dependent, statistically significant manner. Adoptive transfer experiments additionally suggest active recruitment of immunologic response in recipient animals: (1) We have been able to perpetuate leukaemia immunity in four sequential cohorts of naive recipient mice. This propogation of adoptive immunity is accomplished by use of spleen cells harvested from each preceeding passively-protected animal cohort; (2) Cyclophosphamide pretreatment of adoptive transfer recipient mice abrogates the ability of their splenocytes to perpetuate passive protection in sequential adoptive transfer experiments.

Cyclosporin A and verapamil enhancement of daunorubicin-produced nucleolar protein B23 translocation in daunorubicin-resistant and -sensitive human and murine tumor cells.

It has recently been shown that anthracycline antibiotic-resistant tumor cells are less responsive to daunorubicin-stimulated B23 nucleolar phosphoprotein translocation than drug-sensitive cells. Since cyclosporin A and verapamil reverse primary acquired and secondary cross-resistance to daunorubicin, we investigated the effect of these agents on nucleolar B23 translocation in sensitive and resistant tumors. We compared modified to baseline B23 phosphoprotein distribution between predominantly nucleolar, mixed nucleolar-nuclear, or nuclear immunofluorescence using a monoclonal anti-B23 antibody in parental drug-sensitive and multidrug-resistant acute lymphatic leukemia and in daunorubicin-sensitive and -resistant murine hepatoma. Our experiments show that cyclosporin A and verapamil alone have no effect on B23 phosphoprotein translocation, but that the addition of either agent to sensitive parental or resistant tumor sublines markedly enhances daunorubicin-stimulated translocation. This effect correlates with the correction of impaired daunorubicin inhibition of RNA synthesis by cyclosporin A and verapamil in the resistant sublines. Our observations suggest that nucleolar B23 phosphoprotein is an important site in the modulation of anthracycline antibiotic antitumor activity.

Enhancement by cyclosporin A of daunorubicin efficacy in Ehrlich ascites carcinoma and murine hepatoma 129.

Cyclosporin A abrogates pleiotropic drug resistance in certain experimental tumors. Its impact on drug-sensitive tumors has not been investigated. Our studies show that in drug-sensitive Ehrlich ascites carcinoma and hepatoma 129 cyclosporin A enhances daunorubicin inhibition of DNA synthesis in vitro and prolongs survival of host mice in vivo. Of particular interest is that cyclosporin A converts ineffective daunorubicin regimens into those which result in prolongation of host mice survival. Other agents known to reverse pleiotropic drug resistance are reported to exert their effects by increasing intracellular drug accumulation. In contrast, our studies of drug transport in drug-sensitive Ehrlich ascites carcinoma and hepatoma 129 show that cyclosporin A causes minimal enhancement of [3H]daunorubicin uptake without inhibition of [3H]daunorubicin efflux in both the presence and absence of interrupted active daunorubicin efflux. This suggests that the mechanism of action of daunorubicin enhancement by cyclosporin A in drug-sensitive tumors is not simply the result of increased intracellular daunorubicin accumulation. In vivo dosages of cyclosporin A in the current study are comparable to those which can be used with reasonable safety in humans. We conclude that cyclosporin A may be useful in the potentiation of anthracycline antibiotic therapy directed against drug-sensitive as well as drug-resistant tumors.