Eileen B. King

Factors associated with obtaining nipple aspirate fluid: Analysis of 1428 women and literature review

SummaryStudies of cytologic and biochemical constituents of nipple aspirates of breast fluid have contributed to understanding the natural history of benign and malignant breast disease. We conducted multivariate analyses using 1428 women from a recent case-control study of breast disease to determine which factors were independently associated with the ability to obtain breast fluid from nonlactating women. We then compared results from these analyses to the results from five previous studies that also used the aspiration technique of Sartorius. Four factors were consistently associated across studies with increased ability to obtain breast fluid: 1) age up to 35 to 50 years; 2) earlier age at menarche; 3) non-Asian compared to Asian ethnicity; and 4) history of lactation. Exogenous estrogen use, endogenous estrogen concentrations, phase of menstrual cycle, family history of breast cancer, type of menopause, and less than full-term pregnancy consistently did not influence ability to obtain fluid.New findings from this study shed light on some apparently contradictory findings from the previous studies. In particular, this study showed that the effects of age on ability to obtain fluid appeared to be independent of the effects of menopause. Furthermore, discrepancies in previous findings on the effects of parity on ability to obtain fluid may be explained by our finding that the increased ability to obtain fluid from parous compared to nulliparous women applied only to parous women who had breastfed.

Image cytometric classification of premalignant breast disease in fine needle aspirates

Image cytometry for the classification of fine needle aspirate (FNA) biopsies was evaluated in samples from 39 women. Eighteen of them had benign lesions, seven had premalignant lesions, nine had carcinoma in situ and five had carcinoma. The term, premalignant, here refers to lesions with an increased risk of developing into breast cancer (atypical hyperplasia and, to a lesser extent, moderate or florid hyperplasia). The classifications by cytometry were compared with the microscopic diagnoses of the same FNA samples and of tissue from a subsequent surgical biopsy of the same area. One slide from each breast FNA sample was restained in Azure‐A Feulgen. Breast epithelial cells were measured using a texture analysis program on the Leitz TAS‐plus. The mean, standard deviation (SD), and interquartile range were calculated for each of 12 nuclear parameters from 200 cells per slide. A discriminant analysis was used to develop a statistical model for classifying individual samples. Six of seven atypical proliferative lesions (atypical hyperplasia and moderate hyperplasia) were identified by image cytometry, but were unrecognized by conventional microscopic examination.

Cerumen phenotype and proliferative epithelium in breast fluids of U.S.-born vs. immigrant Asian women: a possible genetic-environmental interaction.

SummaryTo determine whether a genetic-environmental interaction exists between the breast, a modified apocrine gland, its secretions, and the genetic polymorphic phenotypes of wet and dry cerumen, we examined nipple aspirate fluid (NAF) for proliferative disease in 172 U.S.-born and immigrant Chinese and Japanese women.Cytologic evidence of proliferative epithelial cells (benign hyperplasia and/or atypical hyperplasia) was found in the NAF of 36 women (20.9%). A significantly higher incidence of proliferative epithelial cells was present in the NAF of U.S.-born than in immigrant Asian women (28.6% vs. 16.5%) (p = 0.05).A higher proportion of U.S.-born Asian women with wet cerumen than women with dry cerumen had proliferative epithelial cells in NAF: 39.3% vs. 20.0% (p = 0.08). No significant difference in NAF proliferative cells was found between immigrant women with wet and dry cerumen: 15.8% vs. 17.3%; p = 0.50. A strong association of proliferative epithelial cells and cerumen phenotype was found in parous U.S.-born women (wet = 47.6% vs. dry = 16.0%; p = 0.002). No significant association with wet and dry cerumen phenotype was found in parous immigrant women (wet = 12.2% vs. dry = 20%).These findings support the hypothesis that an apocrine genetic polymorphic trait differentially influences susceptibility of the breast to proliferative disease in Asian women born in environments presumed to be of high risk for breast cancer compared to women from low risk environments.

Association of breast fluid coloration with age, ethnicity, and cigarette smoking

SummaryNipple aspirates of breast fluid (NAF) occur with different colorations (colorless, white, pale yellow, dark yellow, brown, green, and black). Increasing concentrations of cholesterol, cholesterol 5,6-epoxides, estrogens, and fluorescent products of lipid peroxidation have been positively associated with the dark colorations (dark yellow, brown, green, and black). Because of the absence of data on these variations in breast fluid coloration, we made an exploratory study of their possible associations with age, ethnicity, clinical breast status, and breast cancer risk factors. Dark NAF colorations increased with age among white women from 22.5% at 20–29 years to 49.2% at 50–59 years. Among Chinese and Japanese women, the overall proportion of dark breast fluids was significantly lower (highest proportion 23.5%).A positive association of dark NAF coloration was found with current cigarette smoking (odds ratio = 1.64 [1.04–2.59]). A dose response between amount smoked and dark coloration was found in women <50 years of age, with women who smoked more than one pack per day having an odds ratio of 2.31 (1.30–4.67). No significant association of dark NAF was found with the major breast cancer risk factors or with actual benign or malignant breast disease.The dark colorations may represent pigmented products of apocrine gland secretion, lipofuscin complexes of peroxidated lipoprotein, breakdown products of hemoglobin, and possibly, diet-related secretory products.

Multiple Sampling for Increasing the Diagnostic Sensitivity of Nipple Aspirate Fluid for Atypical Cytology

OBJECTIVE To determine if repeated collection of nipple aspirate fluid (NAF) can improve the diagnostic sensitivity for cytologic atypia, a marker of increased risk of breast cancer. STUDY DESIGN Two hundred sixty-seven women without known breast disease volunteered for NAF cytology at 5 6-month intervals over 2 years. NAF samples were prepared on Millipore filters (Millipore Filter Corp., Bedford, Massachusetts, U.S.A.) and stained with a modified Papanicolaou method. Fluid availability and cellular abnormalities were evaluated for each collection attempt. Cellular findings were classified as benign, hyperplasia or atypia. RESULTS NAF was obtained from 178 women (66.6%) at the first visit and from an additional 15, 10, 2 and 4 women at visits 2, 3, 4 and 5, respectively, for a cumulative total of 78.2% by visit 5. The number of women yielding NAF containing hyperplastic or atypical epithelial cells was determined at each visit. Hyperplastic cells were found in 34 (19.1%) at visit 1 and in an additional 20, 10, 5 and 4 women at visits 2, 3, 4 and 5, respectively. Atypical epithelial cells were present in 12 (6.7%) women at the initial visit and in an additional 11, 7, 5 and 1 women at visits 2, 3, 4 and 5, respectively, for a cumulative percent of 18.2 at visit 5. NAF could not be obtained from 58 women at any visit. CONCLUSION These findings suggest that an optimum collection method for NAF cytology should consist of at least 3 or 4 separate fluid aspiration attempts. Reviewing repeated multiple samples instead of 1 increases the number of women who can be evaluated and the likelihood of detecting cytologic atypia.

Flow cytometry of human gynecologic specimens using log chromomycin A3 fluorescence and log 90 degrees light scatter.

Flow cytometry and electronic cell sorting are being investigated to screen gynecologic specimens for cervical neoplasia. Cellular DNA content is quantitated by Chromomycin A3 fluorescence and cell size is quantitated by 90 degrees light scatter; the logarithms of the measured intensities are used to produce a two parameter histogram. To determine the cell types responsible for signals in various histogram regions, systematic electronic cell sorting is performed. The sorted fractions are sedimented into microscope slides and stained by the Papanicolaou technique. The cells in each fraction are identified by conventional cytomorphologic criteria. Morphologic analysis of sorted cells reveals histogram regions corresponding to specific cell types. One very important region contains the highest concentration of signals from abnormal cells and is therefore the best region to analyze for specimen abnormality. However, because a significant number of signals in this region are from normal cells, specimens cannot be diagnosed by their analysis. Another important histogram region is composed primarily of signals from endocervical columnar and metaplastic cells. The presence of such cells is a good criterion for specimen adequacy, therefore analysis of signals in this region is essential to assess specimen adequacy for automatic screening.