Deutscher et al. BMC Cancer 2010, 10:519 http://www.biomedcentral.com/1471-2407/10/519 similar to women with more advanced disease than in women with benign pathology. Figure 1 also suggests that a high Tn concentration is consistent with AH or more advanced disease. The study had limitations. First, the ability of TF or Tn to determine if a woman had AH was limited by the small number of women in the study with AH. Second, since only one ND sample collection was feasible in the women who soon after sample collection had surgery on the breast, we do not know the variability in TF or Tn expression independent of disease. A study which col- lects sequential ND samples from women at increased breast cancer risk, including women with a biopsy demonstrating AH, may help to address these limitations. Conclusions TF and Tn are unique as biomarkers in that they are carbohydrates, whereas most disease markers both in use and undergoing evaluation are proteins. The expres- sion of TF correlated with the presence of breast atypia and cancer. A multicenter study should be conducted to confirm our findings, and to determine the usefulness of these markers in disease detection. Abbreviations Abs: absorbance; AH: atypical hyperplasia; CV: coefficient of variation; DCIS: ductal carcinoma in situ; H: usual hyperplasia; ICC: intraclass correlation coefficient; NAF: nipple aspirate fluid; ND: nipple discharge; OR: odds ratio; P: pathologic; ROC: receiver operating characteristic Acknowledgements Partial funding for this research was provided by NIH grant CA119095 and a grant from Allied Minds Author details 1University of Missouri Departments of Biochemistry, One Hospital Drive, Columbia, MO 65212, USA. 2University of Missouri, Department of Biostatistics, One Hospital Drive, Columbia, MO 65212, USA. 3The Harry S. Truman Veteran’s Administration, 800 Hospital Drive, Columbia MO 65201, USA. 4Royal Marsden NHS Foundation Trust, Fulham Road, London SW3 6JJ, UK. 5University of North Dakota, Center for Health Promotion and Prevention Research 501 N. Columbia Rd., Grand Forks, ND 58202, USA. 6University of North Dakota, Department of Surgery, 501 N. Columbia Rd., Grand Forks, ND 58202, USA. Authors’ contributions SLD helped design the study, supervised the TF and Tn analyses, and participated in manuscript review. MD, JN and SK conducted the TF and Tn analyses. GG assisted in participant recruitment, sample collection, and manuscript review. JH, NV-H and JH conducted statistical analyses for the manuscript. JH and NV-H also prepared figures for the manuscript. TQ helped design the study and assisted in manuscript review. ERS helped design the study, recruited participants and collected samples that were analyzed, oversaw the entire project, and wrote the manuscript. All authors read and approved the final manuscript. Competing interests ERS is a member of the Scientific Advisory Board of Atossa Genetics, Inc. He does not hold stock nor shares in the company. Atossa Genetics did not assist in the design of the study, did not finance the study, and did not review the manuscript prior to its submission. Page 7 of 7 Received: 1 April 2010 Accepted: 1 October 2010 Published: 1 October 2010 References 1. Springer GF: T and Tn, general carcinoma autoantigens. Science 1984, 224(4654):1198-1206. 2. Imai J, Ghazizadeh M, Naito Z, Asano G: Immunohistochemical expression of T, Tn and sialyl-Tn antigens and clinical outcome in human breast carcinoma. Anticancer Res 2001, 21(2B):1327-1334. 3. Springer GF, Desai PR, Banatwala I: Blood group MN antigens and precursors in normal and malignant human breast glandular tissue. J Natl Cancer Inst 1975, 54(2):335-339. 4. Sauter ER, Klein-Szanto A, Macgibbon B, Ehya H: Nipple aspirate fluid and ductoscopy to detect breast cancer. Diagn Cytopathol 2010, 38:244-251. 5. Kumar SR, Sauter ER, Quinn TP, Deutscher SL: Thomsen-Friedenreich and Tn antigens in nipple fluid: carbohydrate biomarkers for breast cancer detection. Clin Cancer Res 2005, 11(19 Pt 1):6868-6871. 6. Sauter ER, Ross E, Daly M, Klein-Szanto A, Engstrom PF, Sorling A, Malick J, Ehya H: Nipple aspirate fluid: a promising non-invasive method to identify cellular markers of breast cancer risk. Br J Cancer 1997, 76(4):494-501. 7. Smith GP: Kinetics of amine modification of proteins. Bioconjug Chem 2006, 17(2):501-506. 8. Sauter ER, Wagner-Mann C, Ehya H, Klein-Szanto A: Biologic markers of breast cancer in nipple aspirate fluid and nipple discharge are associated with clinical findings. Cancer Detect Prev 2007, 31(1):50-58. 9. Babino A, Oppezzo P, Bianco S, Barrios E, Berois N, Navarrete H, Osinaga E: Tn antigen is a pre-cancerous biomarker in breast tissue and serum in n-nitrosomethylurea-induced rat mammary carcinogenesis. Int J Cancer 2000, 86(6):753-759. 10. Vogel VG, Costantino JP, Wickerham DL, Cronin WM, Cecchini RS, Atkins JN, Bevers TB, Fehrenbacher L, Pajon ER Jr, Wade JL, et al: Effects of tamoxifen vs raloxifene on the risk of developing invasive breast cancer and other disease outcomes: the NSABP Study of Tamoxifen and Raloxifene (STAR) P-2 trial. JAMA 2006, 295(23):2727-2741. 11. Khan A, Newman LA: Diagnosis and management of ductal carcinoma in situ. Curr Treat Options Oncol 2004, 5(2):131-144. Pre-publication history The pre-publication history for this paper can be accessed here: http://www.biomedcentral.com/1471-2407/10/519/prepub doi:10.1186/1471-2407-10-519 Cite this article as: Deutscher et al.: Carbohydrate antigens in nipple aspirate fluid predict the presence of atypia and cancer in women requiring diagnostic breast biopsy. BMC Cancer 2010 10:519. Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit