Maternal areola pH: A chemical basis for mother-infant recognition.

BACKGROUND: The human female's areolar region is the point of arrival of a natural progression from birth to breastfeeding continuum, supported by numerous functional features. AIMS: The aim of this study was to look more closely into the areola' skin chemical signals that are uniquely important for mother-infant chemoemission, chemoreception, and breastfeeding continuum. STUDY DESIGN: A dermatological study of the areolae and corresponding breast quadrants was undertaken on 71 healthy, at-term women. Areolar and corresponding breast quadrant pH were assessed pre-partum and 1 and 2 days post-partum using a corneometer (Soft Plus 5.5; Callegari S.P.A., Parma, Italy). RESULTS: Pre-partum, the pH of the areola was significantly higher than the pH of the breast quadrant (4.25 ±â€¯0.26 vs. 4.06 ±â€¯0.44, p < 0.015). Post-partum, the pH of the areola was significantly higher that the pH of the breast quadrant both on day one (4.28 ±â€¯0.31 vs 4.01 ±â€¯0.25, p < 0.001) and on day two (4.39 ±â€¯0.19 vs 4.01 ±â€¯0.16, p < 0.001), respectively. In addition, pre-labor pH of the areola increased on day one post-partum and significantly on day two postpartum (4.25 ±â€¯0.26 vs 4.39 ±â€¯0.19, p 0.004). Moreover, the pH of the areola increased significantly from day one to day two post-partum (4.28 ±â€¯0.31 v s 4.39 ±â€¯0.19, p < 0.041). CONCLUSIONS: Our findings show, for the first time, that the areola has a higher pH than the surrounding breast skin, and this increases from day one to day two postpartum. We believe that the pH changes of the areolar region may act as a unique chemical signal to guide the infant directly to the nipple.

Zinc delivery from non-woven fibres within a therapeutic nipple shield.

A Therapeutic Nipple Shield (TNS) was previously developed to respond to the global need for new infant therapeutic delivery technologies. However, the release efficiency for the same Active Pharmaceutical Ingredient (API) from different therapeutic matrices within the TNS formulation has not yet been investigated. To address this, in-vitro release of elemental zinc into human milk from two types of Texel non-woven fibre mats of varying thickness and different gram per square meter values, placed inside the TNS was explored and compared to the release from zinc-containing rapidly disintegrating tablets. In-vitro delivery was performed by means of a breastfeeding simulation apparatus, with human milk flow rates and suction pressure adjusted to physiologically relevant values, and release was quantified using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). It was found that a total recovery of 62-64 % elemental zinc was obtained after the human milk had passed through the fibre insert, amounting to a 20-48% increase compared to previous zinc delivery studies using rapidly disintegrating tablets within the TNS. This indicates that non-woven Texel fibre mats were identified as the superior dosage form for oral zinc delivery into human milk using a TNS.

Solid papillary carcinoma of the nipple: an in situ carcinoma or an expansive growth tumor?

Papillary breast lesions are a heterogeneous group of tumors which mainly arise in the central mammary region, ranging from benign to malignant. Among them, solid papillary carcinoma (SPC) represents a very uncommon variant with indolent clinical behavior and excellent prognosis. The categorization of papillary lesions as benign, atypical or malignant is often difficult even for experienced pathologists. Furthermore, for prognostic purposes, to decide whether to consider a lesions as in situ when it is not associated with frank invasive foci of carcinoma may be problematic. We present a case of solid papillary carcinoma arising in the nipple with an expansive and circumscribed growth, mimicking an in situ lesion of the breast on the hematoxylin and eosin stained sections, but in which a myoepithelial layer around neoplastic nodules could not be detected by using immunohistochemistry. To the best of our knowledge, primary origin in the nipple is very rare for SPCs and it has been described only once in the literature. The case we herein illustrate is of interest not only because of its origin in the nipple, but also because of its not in situ, but invasive, although expansive and not infiltrative growth. In the differential diagnosis, nipple disorders as adenoma and syringomatous adenoma, usual ductal hyperplasia (UDH), papilloma, intracystic papillary carcinoma, lobular carcinoma in situ, ductal carcinoma in situ and skin adnexal tumors are considered.

Pigmented Paget's disease of the nipple.

Pigmented Paget's disease of the nipple (PPD) is an uncommon variant of Paget's disease. An accumulation of melanin within the lesion imparts a brow color to the affected area, so the lesion might clinically as well as histologically mimic melanoma. We present a case of PPD in a 60-year-old woman.

Nipple eczema, an indicative manifestation of atopic dermatitis? A clinical, histological, and immunohistochemical study.

Nipple eczema exhibits as a minor manifestation of atopic dermatitis (AD) or occurs as a single skin symptom on the nipple. To characterize the relationship between nipple eczema and AD, a clinical evaluation and an immunohistochemical study were performed. All cases of nipple eczema were confirmed histopathologically. We divided the patients with nipple eczema into 2 groups, namely, those with AD and those without AD, and compared several clinical features. Upon histological examination, the degree of inflammation was subjectively graded as mild, moderate, or severe by 2 separate investigators. Immunohistochemical stainings were performed by using antiinterleukin (IL)-4, anti-IL-13, anti-CD4, and anti-CD8 antibodies, and the results were scored semiquantitatively. In 43 cases evaluated, 12 were nipple eczema with AD. The clinical analysis and histological examination showed no significant differences between the groups. There were consistent findings of IL-4 expressions throughout the epidermis and IL-13 expression mainly in the perivascular area of the dermis. Although CD4 and CD8 were expressed in the cells in the dermis, CD8 expression was detected in the serocrusts of the epidermis. Expression levels of IL-4, IL-13, CD4, and CD8 exhibited no significant differences between the nipple eczema group with AD and the nipple eczema group without AD. Although nipple eczema may accompany AD, we found no definite differences in the degree or pattern of inflammation and cytokine expression level regardless of whether AD was present or not. Serocrust formation seemed to be mainly a collection of CD8-positive cells.

Toker cells of the nipple are commonly associated with underlying sebaceous glands but not with lactiferous ducts.

AIMS: Toker cells are clear cells present in the squamous epithelium of the nipple of some women. In contrast to squamous epithelium, they are cytokeratin 7 (CK7) positive. The origin of these cells is not completely understood. It has been suggested that they may represent abortive glands or migratory ductal cells; and may be precursors of Paget's disease of the nipple. Our aim was to investigate the incidence and distribution of Toker cells and their relationship with lactiferous ducts. METHODS: We examined nipple sections from 100 consecutive mastectomies performed at Charing Cross hospital. New sections were stained for CK7 using the immunoperoxidase technique. RESULTS: Toker cells were identified in 11 cases. They were always clustered within the squamous epithelium superficial to sebaceous glands with no relationship with lactiferous ducts. Two cases in the study had Paget's disease and these were not associated with underlying sebaceous glands. CONCLUSIONS: This study suggests that Toker cells are more likely to be developmentally related to sebaceous glands rather than lactiferous ducts. This raises doubts about the presence of a relationship between Toker cells and the common forms of Paget's disease, as the latter are commonly seen in association with ductal carcinoma in situ (DCIS) involving underlying lactiferous ducts. Toker cells, however, may be related to a less common form of Paget's disease which is not associated with underlying DCIS.

Metabolomic characterization of nipple aspirate fluid by (1)H NMR spectroscopy and GC-MS.

Nipple aspirate fluid (NAF) is a noninvasively obtained biofluid from the duct openings of the breast. NAF components are constantly secreted, metabolized, and reabsorbed by the epithelial lining of the lactiferous ducts of the breast. NAF has been studied as a potential breast tissue surrogate for the discovery of novel breast cancer risk, early detection, and treatment response biomarkers. We report the first unsupervised metabolite characterization of nipple aspirate fluid using NMR and GC-MS using convenience samples previously collected from four premenopausal and four postmenopausal women. A total of 38 metabolites were identified using the two analytical techniques, including amino acids, organic acids, fatty acids, and carbohydrates. Analytical reproducibility of metabolites in NAF by GC-MS was high across different extraction and analysis days. Overall, 31 metabolites had a coefficient of variation below 20%. By GC-MS, there were eight metabolites unique to NAF, 19 unique to plasma, and 24 shared metabolites. Correlative analysis of shared metabolites between matched NAF and plasma samples from pre- and postmenopausal women shows almost no correlations, with the exception being lactic acid, which was significantly negatively correlated (R(2) = 0.57; P = 0.03). These results suggest that NAF is metabolically distinct from plasma and that the application of metabolomic strategies may be useful for future studies investigating breast cancer risk and intervention response biomarkers.

Pl6 expression in Paget's disease of the breast.

BACKGROUND: Paget's disease of the nipple is generally associated with an underlying invasive cancer or an underlying ductal carcinoma in situ. Epidermotropic theory maintains that Paget's cells are derived from an underlying mammary in situ adenocarcinoma. Because p16 protein plays a major role in cell-cycle control and in tumoral cell mobility, we analyzed p16 expression in Paget's disease of the nipple and in associated underlying ductal carcinoma in situ. METHODS: The expression of p16 protein was analyzed by immunohistochemistry in eight cases of Paget's disease of the nipple with associated underlying ductal carcinoma in situ. The Student's t-test (2-tailed) was used to establish the equality of means. RESULTS: The expression of p16 protein was observed in 87.5% (7/8 cases) both in the nipple disease and in the associated underlying ductal carcinoma in situ. The difference between the two populations was not statistically significant. In normal breast tissue, no expression of the protein was observed. CONCLUSION: The positive p16 expression in Paget's disease of the nipple and the underlined ductal carcinoma in situ and its role in cell motility lead us to propose a role of p16 in the spread of this disease.

uPA is upregulated by high dose celecoxib in women at increased risk of developing breast cancer.

BACKGROUND: While increased urokinase-type plasminogen activator (uPA) expression in breast cancer tissue is directly associated with poor prognosis, recent evidence suggests that uPA overexpression may suppress tumor growth and prolong survival. Celecoxib has been shown to have antiangiogenic and antiproliferative properties. We sought to determine if uPA, PA inhibitor (PAI)-1 and prostaglandin (PG)E2 expression in nipple aspirate fluid (NAF) and uPA and PGE2 expression in plasma were altered by celecoxib dose and concentration in women at increased breast cancer risk. METHODS: NAF and plasma samples were collected in women at increased breast cancer risk before and 2 weeks after taking celecoxib 200 or 400 mg twice daily (bid). uPA, PAI-1 and PGE2 were measured before and after intervention. RESULTS: Celecoxib concentrations trended higher in women taking 400 mg (median 1025.0 ng/mL) compared to 200 mg bid (median 227.3 ng/mL), and in post- (534.6 ng/mL) compared to premenopausal (227.3 ng/mL) women. In postmenopausal women treated with the higher (400 mg bid) celecoxib dose, uPA concentrations increased, while PAI-1 and PGE2 decreased. In women taking the higher dose, both PAI-1 (r = -.97, p = .0048) and PGE2 (r = -.69, p = .019) in NAF and uPA in plasma (r = .45, p = .023) were correlated with celecoxib concentrations. CONCLUSION: Celecoxib concentrations after treatment correlate inversely with the change in PAI-1 and PGE2 in the breast and directly with the change in uPA in the circulation. uPA upregulation, in concert with PAI-1 and PGE2 downregulation, may have a cancer preventive effect.

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.

The 8-epimer of prostaglandin F(2alpha), 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.

Identification of markers for nipple epidermis: changes in expression during pregnancy and lactation.

In vertebrates, specific regions of skin crucial for interaction with and manipulation of elements in the environment are characterized by specialized epidermis. Regions of specialized epidermis show distinct patterns of cellular differentiation and express specific keratins that provide an increased ability to withstand mechanical strain. The nipple, which must endure the mechanical strain of nursing, is a type of specialized epidermis. The entire ventral skin of the keratin 14 promoter driven PTHrP mouse provides a model for nipple development. To identify novel markers for this specialized epidermis, we have used two-dimensional (2-D) gels, mass spectrometric protein identification, Western blotting and immunohistochemistry to compare intermediate filament preparations from the nipple-like K14-PTHrP ventral skin to that of wild-type littermates. We identified 64 spots on 2-D gels that were increased in expression in the nipple-like skin of the female K14-PTHrP mouse and 11 spots that were elevated in the wild type. Microsequencing suggested that K17 and epiplakin were among the proteins with the greatest increase in expression in the K14-PTHrP ventral skin. Using Western blots and immunohistochemistry, we evaluated the expression of these proteins as well as K6 in the wild-type nipple, K14-PTHrP ventral skin and wild-type ventral skin. In addition, we found that the expression of K6 was minimally changed in the pregnant and lactating nipple, but the expression of a previously identified marker, K2e, was reduced during lactation. Using a model of the mechanical strain induced by nursing, we found that K2e but not K6 expression was responsive to this condition. The identification of epidermal markers and their expression patterns will provide insight into the cellular differentiation patterns of the nipple and the underlying epidermal-mesenchymal interactions that direct this differentiation.

Proteomic analysis of nipple aspirate fluid throughout the menstrual cycle in healthy pre-menopausal women.

A proteomic approach to nipple aspiration fluid (NAF) has been used in a number of studies comparing women with breast cancer and healthy women. However, to make useful comparisons between women with breast cancer and healthy women it is necessary to establish whether there is physiological variation in the proteomic profiles of NAF. The purpose of this study was, for the first time, to examine how the proteomic profile of NAF using surface-enhanced laser desorption ionisation time-of-flight mass spectrometry varies across the menstrual cycle in healthy pre-menopausal women. Twelve women were recruited and nipple aspiration was carried out weekly from both breasts of each subject for two menstrual cycles. Matching serum samples for luteinising hormone, follicle stimulating hormone and oestradiol were obtained at each aspiration attempt. Statistically significant peaks were found for three healthy volunteers (p < 0.05). However, the peaks that varied across the menstrual cycle were different from one healthy volunteer to another and the differences were small compared with the large variation in proteomic profiles between healthy volunteers. This study provides proof of concept that the NAF proteomic profile does not vary substantially during the menstrual cycle and that therefore it is valid to compare NAF profiles from pre-menopausal women that have been taken at different stages in the menstrual cycle.

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.

Microbiological and biochemical assessment of the surface area of breast nipples and breast milk of lactating women in Calabar, southeastern Nigeria.

Microbiological and biochemical analyses of 59 breast nipple swab samples and 15 manually expressed breast milk samples of lactating mothers aged 15 to 40 years, was carried out using standard procedures. The incidence of bacterial species in swab samples was Staphylococus aureus (63.8%), Streptococcus sp (25.5%), Escherichia coli (6.4%) and Klebsiella sp (4.26%), indicating the poor sanitary status of the lactating mothers. S. aureus was recovered from only 1 (6.7%) of the milk samples, indicating that breast milk is relatively sterile. The nutritional values for the breast milk were 22.5 mg/ml (protein), 0.3 mg/ml (calcium), 3.5 mg/ml (sugar) and 300 microg/ml (vitamin A) in age group 15-20 years, and 16.4 mg/ml (protein), 0.16 mg/ml (calcium), 1.8 mg/ml (sugar) and 100 microg/ml (vitamin A) in the age group 36-40 years. In conclusion, the nutritive and antimicrobial properties of breast milk decrease with increasing age of lactating mothers. The need for public health enlightenment of lactating mothers regarding hygiene, and the provision of oral vitamin A supplement to infants, is discussed.

Fibrous Histiocytoma-like Spindle-Cell Proliferation in the Nipple After Body-Piercing.

We report the case of a 19-year-old pregnant woman who presented with a nipple tumor. The lesion consisted in a spindle-cell proliferation with histologic features similar to those of fibrous histiocytoma, with a highly vascularized stroma. Although it showed low mitotic activity, scattered marked atypical cells with prominent nucleoli were identified, thus raising concern about the benign nature of the tumor. Immunohistochemical evaluation revealed that the spindle cells were diffusely positive for vimentin, focally positive for CD68, and negative for all the other tested antibodies. The patient had a total excision of the lesion and she is free of disease after 30 months. To our knowledge this is the first reported case of a lesion of this type in the nipple after body-piercing.