Nipple Aspirate Research Articles

Biomarkers associated with breast cancer are associated with obesity

Background: Obesity is linked to the development of postmenopausal breast cancer, and some studies indicate obesity predicts a worse prognosis for premenopausal women who develop the disease. It was our hypothesis that proteins associated with breast cancer would be associated with body mass index (BMI). Methods: We searched our database of women enrolled in breast health translational research trials for information on BMI and markers predictive of breast cancer (basic fibroblast growth factor (bFGF), prostate-specific antigen (PSA), human kallikrein (hK)2, and urinary plasminogen activator (uPA). Information on BMI and one or more nipple aspirate fluid (NAF) or serum biomarkers was available from 382 women. Results: In this data set, NAF and serum levels of PSA (nPSA and sPSA), and NAF levels hK2, bFGF and uPA were each associated with pre- and/or postmenopausal breast cancer. sPSA was inversely associated with BMI in both pre- ( r = −.56, p = .001) and postmenopausal women ( r = −.62, p = .0035) without breast cancer. This association was lost when controlling for plasma volume. In women without breast cancer, NAF bFGF ( p = .07, premenopausal subjects) and NAF hK2 ( p = .09, postmenopausal subjects) were borderline associated with BMI. In women with breast cancer, nPSA was inversely ( r = −.53, p = .049) associated with BMI in premenopausal women and directly associated with BMI in postmenopausal women ( r = .37, p = .017). nPSA trended higher in hormone sensitive cancers, especially those that expressed progesterone receptor ( p = .059). Conclusions: sPSA was inversely associated with BMI in all pre- and postmenopausal women and specifically in pre- and postmenopausal women without breast cancer. NAF PSA was associated with BMI in pre- and postmenopausal women with breast cancer. Evaluating the change in PSA with changes in weight may provide clues regarding a subject's breast cancer risk.

Black Cohosh Does Not Exert an Estrogenic Effect on the Breast

Women's Health Initiative findings indicate that hormone replacement therapy may increase breast cancer and cardiovascular disease risk. Black cohosh extract (BCE) is a popular alternative that reduced menopausal symptoms in several clinical trials. Preclinical studies have addressed the estrogenic properties of BCE, with conflicting results. The estrogenic influence of BCE on the breast has not been investigated. Black cohosh is standardized to triterpenes, but the activity and mechanism of action of these compounds are unknown. The study goals were to determine 1) triterpene content of 2 commercially available BCE preparations and 2) the effect of BCE on circulating and breast-specific estrogenic markers. Two black cohosh preparations were analyzed for triterpene content. Postmenopausal women took BCE for 12 wk followed by a 12-wk washout. One BCE preparation contained trace amounts and another contained 2.5% triterpenes. Women taking BCE with 2.5% triterpenes experienced relief of menopausal symptoms, with reversion toward baseline after washout. BCE had no effect on estrogenic markers in serum and no effect on pS2 or cellular morphology in nipple aspirate fluid. Triterpene content in commercially available black cohosh preparations varies. BCE standardized to 2.5% triterpenes relieved menopausal symptoms without systemic or breast-specific estrogenic effects.

Nipple aspirate fluids from women with breast cancer contain increased levels of group IIa secretory phospholipase A2

Arachidonic acid, a bioactive molecule metabolized into prostaglandins and leukotrienes, contributes to cellular proliferation and tumor progression. Group IIa secretory phospholipase A2 (sPLA2-IIa) can facilitate arachidonate release from cellular phospholipids, suggesting its involvement in tumor evolution. Analysis of breast nipple aspirate fluid (NAF), a noninvasive research tool, allows the identification of biomarkers of breast cancer. We sought to determine whether sPLA2-IIa expression might be related to breast cancer development or progression. sPLA2-IIa expression was evaluated in NAF samples from 110 women (57 women with and 53 without breast cancer) using ELISA and western blotting; ultrastructural immunolocalization was performed in epithelial cells floating in NAF. Immunocytochemistry revealed that sPLA2-IIa is a constitutive intracellular protein suggesting that breast ductal cells synthesize and secrete the 14 kDa protein in NAF. Among all 110 subjects, sPLA2-IIa expression was significantly increased both in NAF and within ductal epithelial cells from cancer containing breasts. While in healthy women menopausal status did not influence sPLA2-IIa expression (P = 0.457), among patients with breast cancer there was a significant down-regulation in postmenopausal subjects (P < 0.0001). Moreover, sPLA2-IIa concentration in NAF from breast cancer patients was positively correlated with tumor stage (r (2) = 0.979, P = 0.0012), suggesting an active secretion/accumulation of the enzyme in NAF based on tumor burden. sPLA2-IIa activity may serve a dual role in breast carcinogenesis, beneficial in its release of arachidonate and detrimental in the metabolic conversion of arachadonic acid into prostaglandins and leukotrienes.

Breast Ductal Secretions: Clinical Features, Potential Uses, and Possible Applications

Nipple discharge accounts for approximately 5% of visits to a breast specialist surgical practice and may be encountered as the chief complaint by many other types of physicians. The vast majority of breast cancers originate in the ductal system, which prompted interest in the evaluation of the intraductal approach to breast cancer. Ductoscopy, nipple aspiration, and ductal lavage have emerged as innovative fields of study that may have clinical applications. We performed a literature search of published manuscripts using the keywords nipple discharge, breast ductal secretions, and intraductal approach. We also report our single-institution experience in managing nipple discharge. We present our institutional algorithm for the management of nipple discharge. The possible etiologies of nipple discharge and the appropriate workup are reviewed. Three evolving minimally invasive techniques for the evaluation of high-risk patients include ductoscopy, nipple aspiration, and ductal lavage. Nipple aspiration and ductal lavage fluid may be assayed for cytology, genomic, gene expression, and proteomic studies. Several different translational approaches are being undertaken to investigate the local microenvironment associated with the development and progression of breast carcinoma. Nipple aspiration fluid and ductal lavage offer the opportunity to study the local microenvironment of the ductal system, which is where most breast cancers originate. These powerful approaches to biomarker analysis could be applied to the prevention and treatment of breast cancer.

C-reactive protein in nipple aspirate fluid associated with Gail model factors.

The majority of breast cancers originate in the epithelial lining of the breast ductal system. Premalignant cell damage in this lining may produce biochemical signals that deliver inflammatory proteins to the site. The presence of C-reactive protein (CRP) in nipple aspirate fluid (NAF) may reflect an inflammatory state indicative of a premalignant breast microenvironment. This study ascertained CRP's presence in NAF and evaluated if risk factors, as identified by the Gail model, were associated with NAF CRP levels among healthy women. NAF CRP levels were assayed in 59 women. CRP was present in NAF and significantly (p = .04) and positively related to breast cancer risk as predicted by the Gail model. CRP is differentially present in NAF and varies by Gail model risk factors. CRP in NAF holds promise as a noninvasive biomarker that detects a precarcinogenic breast ductal microenvironment and may contribute to the diagnosis of breast cancer early in the course of the disease when prognosis is most favorable.

A comparative proteinomic analysis of nipple aspiration fluid from healthy women and women with breast cancer

This pilot study examines the feasibility of nipple aspiration to distinguish women with breast cancer from healthy women using surface-enhanced laser desorption ionisation time-of-flight mass spectrometry (SELDI-TOF/MS). Nipple aspiration fluid (NAF) was collected from each breast in 21 women newly diagnosed with unilateral breast cancer and 44 healthy women. No differences were found when proteomic profiles of NAF from the cancer-bearing breast and the contralateral non-cancerous breast were compared. In contrast, 9 protein peaks were significantly different between the cancer-bearing breast compared with healthy women and 10 peaks were significantly different between the contralateral healthy breast and healthy women ( P < 0.05). These data suggest that invasive breast cancer may result in a field change across both breasts and that proteomic profiling of NAF may have more value in breast cancer risk assessment than as a diagnostic or screening tool.

Comparison of Ductal Lavage and Random Periareolar Fine Needle Aspiration as Tissue Acquisition Methods in Early Breast Cancer Prevention Trials

Short-term phase I and phase II breast cancer prevention trials require tissue acquisition at baseline and after intervention to evaluate modulation of potential biomarkers. Currently used tissue acquisition methods include ductal lavage (DL), random periareolar fine needle aspiration (RPFNA), and core needle biopsy. The optimum method to retrieve adequate samples and the most accepted method by study participants is not known. We compared RPFNA and DL as breast tissue acquisition methods for short-term breast cancer prevention trials by evaluating sample adequacy and tolerability in subjects who participated in two prospective phase II breast cancer prevention trials. Eighty-six women at increased risk for breast cancer were included in this study and underwent baseline DL and RPFNA. High risk was defined as having a 5-year Gail score of >1.67% or a history of atypical hyperplasia (AH), lobular carcinoma, or breast cancer. Median age was 54.5 years (range, 39-75 years); 75% of the women were postmenopausal. About 51% of the women yielded nipple aspiration fluid, and breast fluid samples via DL were retrieved in 73% of these subjects. Of these samples, 71% were adequate samples (greater than 10 epithelial cells). However, when the entire cohort was considered, only 31% of the subjects had adequate samples. RPFNA was also attempted in all subjects, and sample retrieval rate was 100%. Out of these, 96% of the subjects had adequate samples. In DL samples, AH rate was 3.7% was and hyperplasia (H) rate was 11.1%. In RPFNA samples, AH rate was 12.9%, and H rate was 24.7%. Cytology findings in RPFNA samples correlated with age, menopausal status, and breast cancer risk category (previous history of lobular carcinoma in situ). Both procedures were well tolerated, and no complications occurred among participants. Considering that the main end point for short-term prevention trials is the modulation of biomarkers, it is important to optimize adequate sample acquisition; therefore, RPFNA is a more practical option for future phase I and II breast cancer prevention trials compared with DL.

Proteomic Analysis of Breast Tissue and Nipple Aspirate Fluid for Breast Cancer Detection

Clinical proteomics has great promise, but many hurdles. Proteomic approaches may prove to be the answer to decades of frustrating searches for markers that are specific and selective for breast cancer. In order to achieve this goal, the challenge arises that breast cancer is a heterogeneous disease and individual variations are enormous. To identify sensitive and specific breast cancer markers will require standardization of every method, from tissue collection through statistical analysis and diligence. Small pilot studies of tumors with varied characteristics will not suffice; large, homogeneous sample sets will be the most efficient route to identify breast cancer markers. Proteomic analysis of nipple aspirate fluid may identify markers that can be used for screening with this noninvasive technique. Proteomic analysis of breast biopsies will allow more detailed characterization of the individual's cancer, with therapy tailored to the disease based on this information.

Proteomics of nipple aspirate fluid, breast cyst fluid, milk, and colostrum

Proteomic analysis of breast fluids has wide-ranging clinical applications. Protein expression in nipple aspirate fluid and breast cyst fluid may prove valuable for the detection and monitoring of breast cancer, but has been hampered by the lack of a single marker with sufficient breast cancer sensitivity and specificity to be clinically useful. The assessment of multiple proteins may offer a more powerful cancer detection tool. Breast cancer is particularly difficult to detect in women who are lactating. The identification of cancer predicting proteins in milk may prove very helpful in an early cancer detection in this group of women. A better understanding of the protein composition of milk and colostrum should improve infant care and nutrition, and lead to alternatives for individuals with milk allergies.

Effects of High Fruit-Vegetable and/or Low-Fat Intervention on Breast Nipple Aspirate Fluid Micronutrient Levels

A change in diet is known to affect micronutrient levels in blood but to what extent diet can affect micronutrient levels in the breast is not yet well established. Healthy, premenopausal women with a family history of breast cancer were randomized across four diet arms for 1 year in a 2 x 2 factorial design study: control, low-fat, high fruit-vegetable, and combination low-fat/high fruit-vegetable diets. Subjects were asked to collect breast nipple aspirate fluid (NAF) at 0, 6, and 12 months, and levels of micronutrients were measured in the fluid. A total of 122 women were enrolled, 97 were retained for 12 months, and sufficient NAF for analysis was available from 59 women at baseline, 49 at 6 months, and 50 at 12 months. Repeated measures mixed-model ANOVA was used to model the data using cholesterol levels and lactation duration as covariates, where appropriate. The high fruit-vegetable intervention, regardless of fat intake, significantly increased total carotenoid levels in NAF. In the low-fat arm, levels of total carotenoids decreased over time relative to control. Levels of total tocopherols and retinol did not change significantly. Levels of 15-F(2t)-isoprostane, a marker of lipid peroxidation, also did not change significantly over time, although there was a decrease observed in the combination arm. These results indicate that total carotenoid levels in NAF can be significantly increased in the breast NAF with a high fruit-vegetable diet. A low-fat diet that was achieved with little increase in fruit and vegetable intake, however, decreased NAF carotenoid levels.

Increased expression of the inflammatory protein YKL‐40 in precancers of the breast

Serum levels of YKL-40 have been associated with inflammatory diseases and breast cancer. Our purpose was to determine if YKL-40 in breast tissue, nipple aspirate fluid (NAF) and serum is (i) concentrated in NAF compared to matched serum, (ii) increased in the NAF, serum or tissue of women with biopsy proven precancer or cancer compared to healthy women and (iii) influenced by menopausal status. 118 women (61 healthy subjects, 10 with precancer and 47 with breast cancer) aged 17-95 years provided NAF with or without serum samples for analysis. Matched tissue was analyzed from a subset of subjects who underwent breast biopsy. All NAF and serum samples had detectable levels of YKL-40. Median YKL-40 levels for the entire cohort were 683 fold higher in NAF than serum. Premenopausal subjects had higher NAF and lower serum levels of YKL-40 than postmenopausal subjects. YKL-40 levels in NAF but not serum were higher in women with precancer (atypical hyperplasia and lobular carcinoma in situ) than in either healthy subjects (p = 0.025) or subjects with breast cancer (p = 0.015). In women with precancer, YKL-40 distribution in tissue correlated with YKL serum level (p = 0.043). YKL-40 is concentrated in NAF, with the highest concentrations in premenopausal women. NAF levels of YKL-40 are significantly higher in women with precancers than healthy subjects, suggesting that measuring YKL-40 in NAF may improve the identification of women at increased breast cancer risk.

The Reliability of Nipple Aspirate and Ductal Lavage in Women at Increased Risk for Breast Cancer—a Potential Tool for Breast Cancer Risk Assessment and Biomarker Evaluation

Ductal lavage, a technique used to sample epithelial cells from breast ducts, has potential use in risk assessment and biomarker evaluation among women at increased risk for breast cancer. However, little is known about the reliability of the procedure. We evaluated the reliability of nipple aspirate (NAF) and ductal lavage at two time points 6 months apart in women at increased risk for breast cancer. Eligible women had a 5-year Gail risk >or=1.66% or lifetime risk of >20%, and/or a family history or personal history of breast cancer. All ducts that produced NAF were cannulated. The kappa statistic was used to evaluate reliability of NAF production, cellular yield, and cytologic diagnosis. Sixty-nine women (mean age, 47 years) were enrolled over 35 months. Forty-seven returned for a second visit. At baseline, 65% of premenopausal and 41% of postmenopausal women produced NAF (P = 0.05), of which 72% underwent successful lavage of at least one duct. Samples of inadequate cellular material for diagnosis were significantly more likely in postmenopausal women than in premenopausal women (P = 0.04). Of the women who returned for a second visit, 18 of 24 who produced NAF had at least one duct successfully cannulated. Twenty-four ducts in 14 women were lavaged twice. Among these ducts, cellular yield for the two time points was inconsistent (kappa = 0.33 +/- 0.13), and only fair cytologic agreement was observed (kappa = 0.32 +/- 0.15). Ductal lavage was associated with moderate discomfort. Currently, the use of ductal lavage is limited by technical challenges in duct cannulation, inconsistent NAF production, a high rate of inadequate cellular material for diagnosis, fair cytologic reproducibility, and low participant return rates.

The relation of soy intake and isoflavone levels in nipple aspirate fluid

Given the possible breast cancer preventive effects of isoflavones, substances with an estrogen-like structure occurring in soybeans, it is of interest to assess levels directly in the breast. The objective of this pilot study was to measure isoflavones in nipple aspirate fluid. In a pilot study with 11 women, nipple aspirate fluid and urine samples were collected at baseline and after 1 month of consuming two servings of soymilk per day. In a parallel investigation, one woman donated nipple aspirate fluid, blood, and urine on the same day after consuming different amounts of soy for 9 days. Nipple aspirate fluid was collected with an aspirator, diluted 1 : 11, and stored at -20 degrees C. Isoflavonoids in nipple aspirate fluid, serum, and urine were liquid-liquid extracted after enzymatic hydrolysis followed by liquid chromatography electrospray ionization high resolution tandem mass spectrometry analysis using triply C labeled internal isoflavonoid standards. After the 30-day soy intervention, median nipple aspirate fluid yield changed very little (P=0.47), whereas urinary isoflavonoid excretion (P=0.04) and isoflavonoid levels in nipple aspirate fluid (P=0.12) increased substantially. For the nine samples collected in one participant, isoflavonoids measured in nipple aspirate fluid, plasma, and urine were highly correlated with the number of daily soy servings and with each other. This pilot study indicates that isoflavonoids in nipple aspirate fluid are 10 times lower than in plasma and are closely related to levels of isoflavonoids in urine and plasma. These findings show that isoflavonoids are present in breast fluid and may act directly on breast tissue.

Human breast biomonitoring and environmental chemicals: use of breast tissues and fluids in breast cancer etiologic research

Extensive research indicates that the etiology of breast cancer is complex and multifactorial and may include environmental risk factors. Breast cancer etiology and exposure to xenobiotic compounds, diet, electromagnetic fields, and lifestyle have been the subject of numerous scientific inquiries, but research has yielded inconsistent results. Biomonitoring has been used to explore associations between breast cancer and levels of environmental chemicals in the breast. Research using breast tissues and fluids to cast light on the etiology of breast cancer is, for the most part, predicated on the assumption that the tissue or fluid samples either contain measurable traces of the environmental agent(s) associated with the cancer or that they retain biological changes that are biomarkers of such exposure or precursors of carcinogenic effect. In this paper, we review breast cancer etiology research utilizing breast biomonitoring. We first provide a brief synopsis of the current state of understanding of associations between exposure to environmental chemicals and breast cancer etiology. We then describe the published breast cancer research on tissues and fluids, which have been used for biomonitoring, specifically human milk and its components, malignant and benign breast tissue, nipple aspirate fluid (NAF) and breast cyst fluid. We conclude with a discussion on recommendations for biomonitoring of breast tissues and fluids in future breast cancer etiology research. Both human milk and NAF fluids, and the cells contained therein, hold promise for future biomonitoring research into breast cancer etiology, but must be conducted with carefully delineated hypotheses and a scientifically supportable epidemiological approach.

Patient variables that predict atypical cytology by nipple duct lavage

Nipple duct lavage (NDL) may be useful for breast cancer risk stratification. Published guidelines recommend that women with a 5-year Gail risk >/=1.7% should consider undergoing lavage for any fluid-yielding ducts, but it is not known whether increased breast cancer risk or nipple fluid production predict lavage atypia. One hundred fifty women unselected for breast cancer risk underwent NDL with cannulation of all nipple aspirate fluid (NAF)-producing ducts and at least 1 dry duct. This resulted in 516 lavage samples. The rate of cytologic atypia was compared for NAF-positive ducts and NAF-negative ducts and for women with 5-year Gail risks >/=1.7% and <1.7%. Ducts from breasts with cancer (N = 113) were excluded from the analysis. The cytologic atypia rate was similar for the 240 NAF-producing ducts (19%) and the 163 dry ducts (15%; P = .36). No significant differences were observed when atypia was categorized as mild (13% vs 10%; P = .63) or marked (6% vs 4%; P = .53). Among the 83 patients who were unaffected by breast cancer, atypia was diagnosed in 15 of 44 patients (34%) with a 5-year Gail risk <1.7% and in 11 of 39 patients (28%) with a 5-year Gail risk >/=1.7% (P = .74). Neither NAF production nor 5-year Gail risk predicted lavage atypia. Limiting NDL to fluid-producing ducts in women with a 5-year Gail risk >/=1.7% significantly reduced the sensitivity of the test for population screening.

Breast carcinogenesis: can the examination of ductal fluid enhance our understanding?

To explore current breast carcinogenesis theories and the possibility of examining breast epithelial cells to confirm steps in the carcinogenic process and the relationship between intraductal sampling techniques and their role in enhanced risk prediction. Published articles, textbooks, and conference proceedings. Examining breast epithelial cells may provide insight into the carcinogenic process while it is occurring. Methods of extracting breast epithelial cells include nipple aspiration, ductal lavage, and periareolar fine-needle aspiration. Nipple aspiration, ductal lavage, and periareolar fine-needle aspiration are viable means of examining possible precursors to breast tumors. Differentiating between true precursors and benign changes is an important step in breast cancer risk assessment. Nipple aspiration and ductal lavage may be performed in an outpatient setting. RNs and advanced practice nurses may perform these procedures and discuss results with patients.

The 8‐epimer of prostaglandin F2α, a marker of lipid peroxidation and oxidative stress, is decreased in the nipple aspirate fluid of women with breast cancer

Breast cancer (BC), a worldwide disease with increasing incidence, develops from ductal/lobular epithelium. Nipple aspirate fluid (NAF), secreted from the breast ducts and lobules, can be analyzed to assess breast metabolic activity. Whether lipid peroxidation in the mammary gland promotes or prevents tumorigenesis is unclear. Malondialdehyde (MDA) and the 8-epimer of Prostaglandin F(2alpha) (8-iso-PGF(2alpha)), two lipid peroxidation markers, were studied in milk (n = 10), NAF (n = 140) and plasma (n = 35) samples. MDA was detected in all plasma, in 80% of milk samples and in 95% of NAF samples. MDA levels in NAF and plasma were significantly higher than in milk (p = 0.016 and p = 0.029, respectively). We found no significant difference between levels of MDA in NAF samples from BC patients compared to healthy controls. 8-iso-PGF(2alpha) was detectable in all samples. 8-iso-PGF(2alpha) median levels in NAF were significantly higher than in both milk and plasma (p < 0.0001). The highest 8-iso-PGF(2alpha) levels were found in NAF from healthy women, significantly higher than in women with BC (p < 0.0001). No significant differences were found in both markers after the age-adjustment. High levels of lipid peroxidation products in NAF suggest their in situ production in the nonlactating breast. Active lipid peroxidation may have a physiologic role in the normal mammary gland. Lower levels of 8-iso-PGF(2alpha) in NAF from BC patients suggest altered production of arachidonic acid metabolites during breast carcinogenesis.

Detection of breast cancer biomarkers in nipple aspirate fluid by SELDI-TOF and their identification by combined liquid chromatography-tandem mass spectrometry

Screening mammography is the most effective tool available for breast cancer detection. While screening mammography saves lives, it has intrinsic problems that limit further improvement. We hypothesize that protein biomarkers in nipple aspirate fluid (NAF) may separate the cancer from the non-cancer state, and therefore can be used for breast cancer detection. In this study the proteins in NAF were analyzed by surface-enhanced laser desorption ionization coupled to time-of-flight mass spectrometry (SELDI-TOF) in the m/z 5,000-85,000 range. Two methods were used to normalize spectra. Then differentially expressed signals that separate cancer from non-cancer conditions were selected by two specifically developed statistical algorithms. Proteins of interest were identified by combined liquid chromatography-tandem mass spectrometry. A set of 8 markers were identified which collectively gave 63% sensitivity, 89% specificity and 76% accuracy for distinguishing cancer from non-cancer. Further improvements in the specificity and sensitivity of this strategy could come from the development of methods for more precise quantification of the biomarkers of interest and also from focusing on the low abundant components that are not evident when unfractionated NAF is analyzed directly.

Biologic markers of breast cancer in nipple aspirate fluid and nipple discharge are associated with clinical findings

Background: The aim of this prospective study was to assess predictive markers in nipple aspirate fluid (NAF) and pathologic nipple discharge (PND) collected prior to excisional breast biopsy, as well as clinical factors available prior to biopsy, with histopathologic results in women with a radiographically suspicious and/or palpable breast lesion. Methods: 208 NAF samples from 191 women were evaluated for the following candidate predictive proteins and cellular markers: prostate-specific antigen (PSA), human glandular kallikrein 2 (hK2), basic fibroblast growth factor (bFGF), S phase fraction (SPF), DNA index, and cytology. Clinical factors included whether or not the lesion was palpable, menopausal status, history of pregnancy, history of birth control or hormone replacement use, and PND. Results: Considering all women, bFGF ( p = 0.005) and SPF (0.031) were associated, and abnormal cytology approached an association ( p = 0.056) with the presence of breast cancer. Women with PND were less likely to have breast cancer (4 vs. 37%, p < 0.001) or palpable lesions (10 vs.43%, p < 0.001), were younger, had lower PSA levels ( p = 0.046), and were more likely to have atypical NAF cytology ( p = 0.002). Excluding PND, increased age, postmenopause (both p < 0.01), high bFGF ( p = 0.004) and low PSA ( p = 0.05) were associated with cancer. The best breast cancer predictive model included cytology, bFGF, and age (88% sensitive and 57% specific). When the data were divided by menopausal status, the optimal models to predict breast cancer, which included NAF hK2 or PSA and age, were 100% sensitive and 41% specific in pre- vs. 93% sensitive and 12% specific in postmenopausal women. Conclusion: NAF and clinical biomarkers are sensitive predictors of whether a breast contains cancer, and may ultimately help guide treatment. Future studies to determine the optimal combination of predictive markers are warranted.

Human body fluid proteome analysis.

The focus of this article is to review the recent advances in proteome analysis of human body fluids, including plasma/serum, urine, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, synovial fluid, nipple aspirate fluid, tear fluid, and amniotic fluid, as well as its applications to human disease biomarker discovery. We aim to summarize the proteomics technologies currently used for global identification and quantification of body fluid proteins, and elaborate the putative biomarkers discovered for a variety of human diseases through human body fluid proteome (HBFP) analysis. Some critical concerns and perspectives in this emerging field are also discussed. With the advances made in proteomics technologies, the impact of HBFP analysis in the search for clinically relevant disease biomarkers would be realized in the future.