Cellular composition of the nipple aspirate specimen of breast fluid. I. The benign cells.

A nipple aspirator device was used to obtain breast secretions for cytologic examination, as well as for viral and biochemical analysis. Examination of the first 1,456 specimens from 796 women revealed ductal epithelium in 54%. Ductal epithelial cells were often absent in specimens from normal women; however, 78.5% of women with benign breast disease on tissue biopsy had specimens containing ductal epithelium. Apocrine metaplastic cells were a further indication of the presence of breast disease, and were rarely found in specimens from asymptomatic women. Foam cells were often abundant in specimens from normal breasts, but were found in decreased numbers in specimens from women benign breast disease. Differences in the occurrence of ductal epithelial cells, apocrine metaplastic cells, and foam cells suggest an alteration in the rate of maturation of ductal epithelium in women with both benign and malignant breast disease. The finding of a relative abundance of cells in nipple aspirate specimens from women with breast disease and few or no cells in specimens from women with normal breasts is believed to be of great importance in the cytologic evaluation of nipple aspirate specimens.     

Characterization of foam cells in nipple aspirate fluid

Foam cells with abundant vacuolated cytoplasm are prominent in most samples of spontaneous nipple discharge, nipple aspirate fluid, and ductal lavage. Although several investigators have attempted to characterize these cells, there is no consensus about whether these cells are derived entirely from macrophages or from both ductal epithelial cells and macrophages. Using immunocytochemical methods, we studied 20 paired specimens of nipple aspirate fluid containing abundant foam cells obtained from the involved breast of women with in situ or invasive carcinoma and from the contralateral normal breast. We used a cocktail of anticytokeratin antibodies including AE1, AE3, and CAM5.2 and the macrophage marker KP1 (CD68). In addition, we examined samples by electron microscopy. The foam cells were consistently negative for cytokeratin and positive for CD68. In every case electron microscopy of these cells revealed irregular outlines with short cytoplasmic processes. The cytoplasm was abundant and contained numerous lysosomes, a small Golgi complex, lipid droplets, mitochondria, and short profiles of rough endoplasmic reticulum. There was no evidence, however, of cell junctions or tonofilaments. The immunocytochemical and electron microscopic findings of our study together clearly support a macrophage derivation for foam cells in nipple aspirate fluid.     

Increased levels of erythropoietin in nipple aspirate fluid and in ductal cells from breast cancer patients.

BACKGROUND: Erythropoietin (Epo) is an important regulator of erythropoiesis, and controls proliferation and differentiation of both erythroid and non-erythroid tissues. Epo is actively synthesized by breast cells during lactation, and also plays a role in breast tissues promoting hypoxia-induced cancer initiation. Our aims are to perform an exploratory investigation on the Epo accumulation in breast secretions from healthy and cancer patients and its localization in breast cancer cells. METHODS: Epo was determined by ELISA, immunoprecipitation, western blot and immunocytochemical analyses in 130 Nipple Aspirate Fluids (NAF) from 102 NoCancer and 28 Breast Cancer (BC) patients, comparing results with those found in 10 milk, 45 serum samples and breast cancer cell lines. RESULTS: Epo levels in NAFs were significantly higher than those in milk and serum. No difference in Epo electrophoretic mobility was found among NAF, milk and serum samples, and conditioned cell culture medium. Immunolocalization of intracellular Epo in ductal cells floating in BC NAFs was similar to those of cancer cell lines. No significant correlation between TNM classification and Epo in NAFs from BC patients was found. Significantly higher Epo concentration was found in NAF from BC patients compared to NoCancer. CONCLUSION: We demonstrate that breast epithelial cells are a source of Epo in breast microenvironment, suggesting the presence of a paracrine/autocrine Epo function in NAFs, triggering off intracellular signaling cascade with subsequent BC initiation.     

Nipple aspirate fluid and ductoscopy to detect breast cancer

We prospectively performed cytologic assessment and image analysis (IA) on matched nipple aspirate fluid (NAF) and mammary ductoscopy (MD) specimens to determine (1) the accuracy of these methods in cancer detection and (2) whether the two collection methods provide complementary information. NAF and MD specimens were collected from 84 breasts from 75 women (nine bilateral samples) who underwent breast surgery. Cytologic evaluation was performed on all samples. IA was performed on slides with sufficient epithelial cells. Cytologic evaluation proved more accurate in patients without pathologic spontaneous nipple discharge (PND) than those with PND, mainly because of the potential false positive diagnosis in the latter. While the sensitivity of NAF and MD cytology was low (10% and 14%, respectively), both were 100% specific in cancer detection in the non‐PND cohort. Combining NAF and MD cytology information improved sensitivity (24%) without sacrificing specificity. Similar to cytology, IA was more accurate in patients without PND having high specificity (100% for aneuploid IA), but relatively low sensitivity (36%). Combining NAF and MD cytology with aneuploid IA improved the sensitivity (45%) while maintaining high specificity (100%). The best predictive model was positive NAF cytology and/or MD cytology combined with IA aneuploidy, which resulted in 55% sensitivity and 100% specificity in breast cancer detection. Cytologic evaluation and IA of NAF and MD specimens are complementary. The presence of atypical cells arising from an intraductal papilloma in ductoscopic specimens is a potential source of false positive diagnosis in patients with nipple discharge. Diagn. Cytopathol. 2010 © 2009 Wiley‐Liss, Inc.     

Computer assisted diagnosis of fine needle aspirate of the breast.

The development of the National Breast Screening Programme has created a demand for the widespread availability of fine needle aspiration cytology services. To meet this demand there must be a rapid increase in the number of pathologists and laboratories able to offer this service. In turn there is a need for improved training methods. The technique of fine needle aspiration cytology is not inherently complicated. The number of possible conclusions is essentially limited to four: unsatisfactory, benign, suspicious and malignant. A computer based expert system, designed to assist pathologists in the diagnosis of fine needle aspirates of the breast, has been developed. The system prompts pathologists to categorize a number of variables in the aspirate including nuclear and cytoplasmic features, and the degree of cellular cohesion, and uses these data to reason about possible conclusions. The final diagnosis is displayed with a detailed explanation listing the factors supporting it. Initial trials with this system have been encouraging and it is envisaged that this system will be of value both in training and as an aid to routine diagnosis.     

Immunoglobulin profile of tracheal aspirate fluid in intubated children.

Pulmonary lavage immunoglobulins IgG, IgA and IgM were measured in intubated ventilated neonates during their period of intubation (range 1-64 days, mean 12). The neonates were divided into two groups based on gestational age (group 1 26-32 weeks, group 2 33-40 weeks). IgG levels were high at birth, and decreased exponentially throughout the period of intubation. There was no statistical difference in IgG levels between the two groups. IgA and IgM levels were low at birth, and increased linearly, there being a significantly greater increase with age in Group 2 (the more mature gestationally) for both immunoglobulins. Two groups of older children were also studied (2-4 year olds, and 8-10 year olds). In the 2-4 year age group, IgG levels were similar to those seen in the immediate newborn period, were quantitatively greater than IgA and IgM, and were not significantly different from levels in the 8-10 year olds. IgA and IgM levels were also not significantly different between the two groups.     

How the neonatal rat gets to the nipple. II. Changes during development

In the course of postnatal development, the motor sequence executed by pups in order to attach to the dam's nipple undergoes extensive changes. During the 1st postnatal week, the pup performs a rotation along the longitudinal axis of its trunk to achieve a supine posture under the mother. The pup then crawls on the maternal ventrum while in the supine posture, searching for, finding, and attaching to a nipple. During the 2nd postnatal week, this sequence is modified and the pup first searches and establishes contact with a nipple before rotating to the supine posture. This sequence of movements is then truncated. By postnatal Day 11, pups may attach to a nipple while in a prone posture. Developmental changes in supination before attaching to the nipple are reminiscent of changes in righting during a similar period of development. These observations support the idea that both righting and postural adjustments involved in attachment to the nipple derive from common motor modules, with righting executed in the direction of gravity and rotation to the nipple executed against the force of gravity. The parallel structure of these behaviors is consistent with a common origin and similar control mechanisms for these distinct motor behaviors that are expressed early in postnatal development.     

Cellular fatty acid composition of Plesiomonas shigelloides.

The cellular fatty acid compositions of 29 strains of Plesiomonas shigelloides and 5 strains of Aeromonas hydrophila were studied. The cellular fatty acid compositions of all the Plesiomonas strains were identical and characterized by the presence of hexadecanoate (16:0) (33%), hexadecenoate (16:1) (28%), octadecenoate (18:1) (9%), and octadecanoate (18:0) (6%). The cellular fatty acid composition of A. hydrophila was similar to that of the Plesiomonas strains, except that the former contained an average of 25% 16:0, 29% 16:1, 12% 18:1, and 2% 18:0 acids compared with 33, 28, 9, and 6%, respectively, for the latter. The percentage ratios of 16:1 to 16:0 and 18:1 to 18:0 could be used to differentiate P. shigelloides from A. hydrophila. These ratios were 0.8 and 1.5 for the former and 1.2 and 6.0 for the latter.     

Cellular fatty acid composition of Campylobacter fetus.

The cellular fatty acid composition of 29 human and animal isolates of Campylobacter fetus was determined with gas-liquid chromatography. All strains contained small amounts of 3-hydroxytetradecanoic acid, suggesting the presence of lipid A. Each of 13 different blood or stool isolates of C. fetus subsp. jejuni obtained from humans or fowl contained a 19-carbon cyclopropane acid which was not present in C. fetus subsp. fetus (6 strains from cattle) or C. fetus subsp. intestinalis (10 strains from humans and cattle). These findings indicate that C. fetus subsp. jejuni can be rapidly differentiated from other C. fetus by gas-liquid chromatography analysis of cellular fatty acids.     

Cellular fatty acid composition of Lautropia mirabilis.

Ten strains of Lautropia mirabilis (ATCC 51599(T) and nine phenotypically similar clinical isolates) were examined for cellular fatty acid (CFA) composition to evaluate their chemical relatedness to known bacterial species and groups. The CFAs were liberated from whole cells by base hydrolysis, methylated, and analyzed by gas-liquid chromatography. CFA profiles were generated by using a commericial software package (MIDI, Newark, Del.). All strains tested had an identical CFA profile characterized by major amounts of 16:1omega7c (41%) and 16:0 (44%); smaller amounts (1 to 4%) of 3-OH-10:0, 12:0, 14:0, 15:0, and 18:1 omega7c; trace amounts (<1%) of 10:0, 18:2 and 18:0; and no cyclopropane acids. This profile was similar to the CFA profiles of Acidovorax delafieldii, Comamonas terrigena, and strains of an unclassified Centers for Disease Control group designated weak oxidizer group 1. CFA analysis, when supplemented by phenotypic characterization, is useful for the identification of L. mirabilis isolates.     

Cellular fatty acid composition of Haemophilus equigenitalis.

The cellular fatty acid composition of eight Haemophilus equigenitalis strains was determined by gas-liquid chromatography. All strains showed a grossly similar pattern characterized by large amounts of 18:1 and 16:0. The amounts of 16:1, 18:2, 18:0, 3-OH 14:0, 3-OH 16:0, and 3-OH 18:1 were relatively small.     

Cellular fatty acid composition of Francisella tularensis.

Several unusual fatty acids characterized strains of Francisella tularensis. Long-chain (C20-C26) acids and the hydroxy acids 2-hydroxy-decanoate, 3-hydroxy-hexadecanoate, and 3-hydroxy-octadecanoate appeared to be of special diagnostic value.     

Cellular homeostasis and the breast

Increasing our knowledge regarding the control of cellular homeostasis in the normal human breast is important in understanding how breast tumours arise and whether compounds used for hormone replacement therapy are able to promote tumour formation In our studies on tissue from pre-menopausal, non-pregnant, non-lactating women, oestradiol is the main ovarian steroid mitogen for the breast epithelium whereas progesterone has little or no effect. Oestradiol appears to influence proliferative activity indirectly via oestrogen-receptor positive cells which control the activity of adjacent division competent cells by means of paracrine or juxtacrine growth factors. After the menopause, however, a mitogenic effect of progesterone becomes apparent which may be due to the reduction in endogenous oestradiol levels or, possibly, to tissue specific alterations in steroid sensitivity. Whatever the mechanism, the proliferative effects of progesterone on the post-menopausal breast have implications for the use of HRT.     

Imaging findings of metastatic disease to the breast.

The purpose of this study was to evaluate imaging findings of metastatic disease to the breast. We analyzed 15 cases that had been previously examined by radiographic study (comprising a total of 11 mammograms, 13 ultrasounds and 3 chest CT). Primary malignancies included the following: 6 breast cancers, 5 stomach cancers, 2 melanomas, 1 cervical cancer, and 1 lung cancer. Radiologic findings were analyzed and the 15 cases were divided into two groups: hematogeneous and lymphangitic metastasis. Eleven cases were classified as lymphangitic metastasis (73.3%) and 4 cases were hematogeneous metastasis (26.7%). Multiple masses were shown in 13 cases (86.7%), and 12 cases displayed unilateral lesions (80%). There was no evidence of calcification. In the 11 cases with available mammogram, 7 cases (63.6%) revealed multiple masses with well-defined (2 cases, 28.6%) or ill-defined margin (5 cases, 71.4%) and 6 cases (54.5%) showed diffuse increased density and skin thickening. Among the 11 cases that revealed mass lesions on ultrasound, 2 cases showed a well-defined margin (18.2%) and 9 cases showed an ill-defined margin (81.8%). Posterior shadowing of the mass was seen in only 1 case. Four cases (36.3%) showed a minimal echogenic boundary around the mass. Eight cases showed diffuse skin thickening (53.3%). Three cases with chest CT showed multiple masses and/or diffuse skin and trabecular thickening of the breast. In conclusion, the characteristic imaging findings of metastatic disease to the breast are multiple ill-defined masses with a lack of desmoplastic reaction and no calcification, as compared to primary breast carcinoma. The diffuse lesions without evidence of breast mass on mammogram and US could be distinguishable from inflammatory carcinoma of breast origin.     

Neonatal thymus grafts. II. Cellular events.

Neonatal thymus undergoes sequential necrosis and cell regeneration after grafting under the kidney capsule in normal and thymectomized recipients. This regeneration assessed in terms of cell number, response to mitogens, in vivo steroid sensitivity, level of antigen-binding cells, and cellular distribution on Ficoll gradient is not influenced by the presence of host thymus or by FTS injections. The grafted thymus shows a similar pattern of development as control thymus of the same age followed up in situ, with the exception of age-related involution the onset of which takes place earlier in the grafted thymus.     

Cellular composition of the bone marrow in the chicken. I. Identification of cells.

Incorporation of Fe55 in vivo was used for verifying on radioautographs the identity of chicken bone marrow cells that are in the process of hemesyntheses. Under the experimental conditions all labeled cells may be considered to be linked with erythroid differentiation. They were classified into five maturational stages according to their morphology and capacity for DNA synthesis. Granulocytes were identified by the presence of specific granules. All mononuclear cells were classified as lymphocytes which had a pachychromatic nucleus, a high nucleocytoplasmic ratio, lacked the capacity for DNA synthesis, and resembled small lymphocytes of the bursa, spleen and bone marrow that bound, in vitro, anti-chicken gamma globulins labeled with I125. Radioautography with H3TdR was used to identify proliferating and non-proliferating members of each cell population.     

Cellular composition of the bone marrow in the chicken. I. identification of cells

Incorporation of Fe⁵⁵ in vivo was used for verifying on radioautographs the identity of chicken bone marrow cells that are in the process of heme syntheses. Under the experimental conditions all labeled cells may be considered to be linked with erythroid differentiation. They were classified into five maturational stages according to their morphology and capacity for DNA synthesis. Granulocytes were identified by the presence of specific granules. All mononuclear cells were classified as lymphocytes which had a pachychromatic nucleus, a high nucleocytoplasmic ratio, lacked the capacity for DNA synthesis, and resembled small lymphocytes of the bursa, spleen and bone marrow that bound, in vitro, anti-chicken gamma globulins labeled with I¹²⁵. Radioautography with H³TdR was used to identify proliferating and non-proliferating members of each cell population. With the use of the morphological criteria thus established, reproducible differential counts could be performed on chicken bone marrow smears also in the absence of specific labeling. In normal, 3, 4, and 8 week old White Leghorn chicks, such counts revealed the bone marrow as a member of the lymphomyeloid complex preferentially adapted to the production and maturation of erythroid cells, while the reserve of granulocytes was found to be small compared to that of mammalian marrow. The percentage of lymphocytes appears to increase with age but, in contrast to rodent marrow, a proliferating precursor cell pool for lymphocytes could not be identified in chicken marrow up to the age of eight weeks.     

Cellular fatty acid composition of isolates from Legionnaires disease.

The cellular fatty acids of four isolates from Legionnaires disease and two antigenically related isolates were identified by gas chromatography, mass spectrometry, and associated techniques. The six isolates had essentially the same fatty acid composition, which was characterized by large amounts (greater than 80%) of branched-chain acids.