Toward the first definition of taste rosea in female breasts: histological analysis / La primera definición del sabor rosea en mamas femeninas: análisis histológico

Although various neuropsychochemical theories have been established about why breastfeeding mothers feel hedonic sensation, the underlying neural mechanism has not been adequately clarified. We aimed to investigate if there is hedonic sensation-initiated taste-bud like structures stimulated by sugars in the milk-secreting lactiferous ducts of mammary glands of breastfeeding female rats. In this study, twenty-two female rats were chosen which six of the virgin (n=6), six of pregnant (n=6) and ten of breastfeeding (n=10). We examined lactiferous ducts/nipples of mammary glands of all animals. They were sacrificed following intracardiac formalin injection, and their breast tissues were removed with covering tissues and fixed with 10 % of formalin solution. After current histological procedures, the tissues were examined by light microscope to assess taste-bud like structures, and their numerical densities were calculated by using stereological methods. Results were analyzed statistically. Taste-buds like structures with neuron-like appendages at the apical ends were discovered in lactiferous ducts. The taste rosea numbers were estimated as 3±1/mm3 in virgins, 167±27/mm3 in pregnant and 375±63/mm3 in breastfeeding animals. The taste rosea numbers were greater in breastfeeding rats than those of virgins and pregnant rats. They named as taste rosea resembling flower bucket which has not been mentioned in the literature so far.

Existen varias teorías neuropsicoquímicas, referente a la sensación hedónica que sienten las mujeres al amamantar, y el mecanismo neural subyacente. No obstante, estas aún no se aclaran adecuadamente. El objetivo de este estudio, fue investigar si existen estructuras hedónicas iniciadas por la sensación gustativa estimuladas por los azúcares en los conductos mamarios secretores de leche, de las glándulas mamarias de las ratas durante el período de lactancia. En este estudio, se eligieron 22 ratas hembras, seis de estas no preñadas como grupo control, seis preñadas y diez en período de lactancia. Examinamos los conductos lactíferos / pezones de las glándulas mamarias de los tres grupos. Los animales fueron sacrificados por medio de inyección intracardíaca de formalina. El tejido mamario se fijó en solución de formalina al 10 %. La muestras histólogicas fueron examinadas a través microscopía óptica con la finalidad de evaluar estructuras con características morfológicas similares a las papilas gustativas. Su densidad de número se calculó utilizando métodos estereológicos. Los resultados fueron analizados estadísticamente. En los conductos mamarios se observaron dos estructuras con con características morfológicas tipo papilas gustativas con apéndices neuronales en los extremos apicales. Los números se estimaron en 3±1/mm3 en el grupo control, 167±27/mm3 en gestantes y 375±63/mm3 en animales lactantes. El número de estructuras características morfológicas similares a las papilas gustativas fue mayor en las ratas amamantando que en el grupo control y que en las ratas preñadas. Conocido como sabor rosea debido a que se asemeja a un ramo de flores, lo que hasta ahora no se ha mencionado en la literatura.

La relación núcleo-citoplasmática en la diferenciación y transformación celular del epitelio mamario / The nucleo-cytoplasmic relationship in the differentiation and cellular transformation of the mammary epithelium

ANTECEDENTES: Las células pertenecientes a la línea HC11 son estimuladas tanto a proliferar mediante el Factor de Crecimiento Epidérmico, como a diferenciar estimuladas con dexametasona, insulina y prolactina, dando origen a los tipos celulares HC11 GM y HC11 IM, respectivamente. De igual manera cuando ellas son transfectadas con el oncogén ras generan Q6 GM y Q6 IM, células transformadas. Producto de ambos mecanismos estas células asumen distintas propiedades modificando tanto sus componentes como sus funciones, los cuales pueden ser cuantificados mediante técnicas morfométricas. OBJETIVO: Evidenciar en términos cuantitativos y morfológicos las variaciones experimentadas tanto por los núcleos como los citoplasmas y su correspondiente relación núcleo-citoplasmática (N/C) pertenecientes a células mamarias de la línea HC11 con el decorrer de los mecanismos de diferenciación y de transformación celular. MÉTODO: Se estudió a nivel de microscopia electrónica de transmisión los tipos celulares en proceso de diferenciación y transformación, cuantificando variaciones de la relación núcleo-citoplasmática y su respectiva funcionalidad. RESULTADOS: Se evidencian diferencias estadísticamente significativas referentes a las áreas nucleares y citoplasmáticas pertenecientes a estos tipos celulares. CONCLUSIÓN: Las células del epitelio mamario en proceso de diferenciación como de transformación, presentan diferentes valores en su relación N/C hecho que responde a funcionalidades específicas en cada tipo celular.

BACKGROUND: Cells of the HC11 line are stimulated to proliferate using the Epidermic Growth Factor, and to differentiate with dexamethasone, insulin and prolactin, giving rise to cell types HC11 GM and HC11 IM, respectively. Likewise when they are transfected with the ras oncogen they generate Q6 GM and Q6 IM transformed cells. As a result of these two mechanisms, these cells assume different properties, in which both their components and their functions are modified. The modifications can be quantified by morphometric techniques. OBJECTIVE: To show in quantitative and morphological terms the variations effected in both the nuclei and the cytoplasms, and the corresponding nuclear-cytoplasmic ratio of mammary cells of the HC11 line, under the effects of cellular differentiation and transformation mechanisms. METHOD: The cell types undergoing differentiation and transformation processes were studied by transmission electron microscope, permitting quantification of variations in the nuclear-cytoplasmic ratio and its relation with the respective functions. RESULTS: Statistically significant differences were found in the nuclear and cytoplasmic areas of these cell types. CONCLUSION: cells of the mammary epithelium undergoing differentiation and transformation processes present different values for their nuclear-cytoplasmic ratio, and this responds specific functions in each cell-type.

Las mitocondrias en la transformación celular del epitelio mamario / The mitochondria in the mammary epithelial cells transformation

Antecedentes: La adquisición del fenotipo metastático es el resultado de la potente acción generada por oncogenes transformantes sobre una célula normal los cuales con la consiguiente expresión de oncoproteínas determinan drásticos cambios tanto en la morfología como en los volúmenes de los componentes celulares, generando una célula con diferente funcionalidad. Este mecanismo corresponde a la transformación celular. Objetivo: Precisar las modificaciones que caracterizan al mecanismo transformante en células de epitelio mamario transfectado con el oncogén ras (HC11 ras) en comparación con su tipo celular normal (HC11GM). Método: Se estudió con microscopia electrónica de transmisión aplicando técnicas morfométricas a estos tipos celulares con énfasis en las mitocondrias, cuantificando variaciones del organelo generador de energía. Resultados: Todos los parámetros mitocondriales evaluados en el tipo celular transformado presentan diferencias significativas con respecto a la célula normal. Conclusión: Las drásticas modificaciones experimentadas por las mitocondrias se reflejan en la adquisición de nuevos requerimientos energéticos y metabólicos en la célula transformada.

Background: The acquisition of the metastatic phenotype is the result of the transforming oncogene powerful action over a normal cell which with the subsequent oncoprotein expression leads to drastic changes in morphology as well as in cell components volumes, generating a cell with a different function. This mechanism relates to the cell transformation. Objective: To specify the modifications that characterize the transforming mechanism in mammary epithelial cells transfected with the ras oncogene comparing them with its normal cell type. Method: Transmission electronic microscopy using morphometric techniques was applied to this cell types, emphasizing mitochondria variations, trying to clarify its role in each cell type metabolism. Results: Everyone mitochondrial parameters examined in transformed cell type present significant differences regarding to the normal cells. Conclusión: The drastic changes in mitochondria are reflected in the acquisition of new energy requirements and metabolism in the transformed cell.

Differential subcellular distribution of glucose transporters GLUT1-6 and GLUT9 in human cancer: ultrastructural localization of GLUT1 and GLUT5 in breast tumor tissues.

It has been proposed that the enhanced metabolic activity of tumor cells is accompanied by an increased expression of facilitative hexose transporters (GLUTs). However, a previous immunohistochemical analysis of GLUT1 expression in 154 malignant human neoplasms failed to detect the GLUT1 isoform in 87 tumors. We used 146 normal human tissues and 215 tumor samples to reassess GLUT1 expression. A similar number of samples were used to compare the expression of GLUT2-6 and 9. The classical expression of GLUT1-5 in different normal human tissues was confirmed, however, we were unable to detect GLUT2 in human pancreatic islet cells. GLUT6 was principally detected in testis germinal cells and GLUT9 was localized in kidney, liver, heart, and adrenal. In tumor samples, GLUT1, 2, and 5 were the main transporters detected. GLUT1 was the most widely expressed transporter, however, 42% of the samples had very low-to-negative expression levels. GLUT2 was detected in 31% of the samples, being mainly expressed in breast, colon, and liver carcinoma. GLUT5 was detected in 27% of breast and colon adenocarcinoma, liver carcinoma, lymphomas, and testis seminoma samples. In situ RT-PCR and ultrastructural immunohistochemistry confirmed GLUT5 expression in breast cancer. GLUT6 and 9 are not clearly over-expressed in human cancer. The extensive expression of GLUT2 and 5 (glucose/fructose and fructose transporters, respectively) in malignant human tissues indicates that fructose may be a good energy substrate in tumor cells. Our functional data obtained in vitro in different tumor cells support this hypothesis. Additionally, these results suggest that fructose uptake could be used for positron emission tomography imaging and, may possibly represent a novel target for the development of therapeutic agents in different human cancers.

Nuclear and nucleolar image analysis of human breast epithelial cells transformed by benzo[a]pyrene and transfected with the c-Ha-ras oncogene.

Changes in nuclear and nucleolar morphometric parameters were investigated by image analysis procedures in human breast MCF-10F epithelial cells expressing different stages of the tumourigenic progression after benzo[a]pyrene (BP) transformation (BP1, BP1-E, and BP1-E1 cell lines), and additionally transfected with the c-Ha-ras oncogene (BP1-Tras cell line). Nuclear pleomorphism was evident in all the transformed cells. The analysis of different morphometric parameters did not show a clear relationship between specific nuclear and nucleolar changes and the expression of the different stages of the tumourigenesis, with the exception of the nucleolar size, which could be associated to the expression of the tumourigenic phenotype, and a nucleolar area/nuclear area ratio, which discriminated the immortalized, the transformed, and the tumourigenic phenotypes from one another. The nuclear morphometric data established for the BP-transformed cells and for the cells additionally transfected with the c-Ha-ras oncogene were suggestive of complex and distinct morphofunctional mechanisms involving the in vitro transformation of the MCF-10F cells. The nuclear changes found in the BP1-Tras cell line were assumed to be related to the additional effects and/or enhanced genomic instability induced by transfection with the ras oncogene.

Estudo das fibras elasticas em glandulas mamárias de mulheres fumantes e näo fumantes / Morphometrical study of elastic tissue in breast of smoking and non-smoking women

Foram estudadas 51 mamas de mulheres normais. As mamas foram coradas pela hematoxilina-eosina e resorcina de Weigert para estudo morfometrico dos componentes histologicos das glandulas mamárias e do sistema elastico nessas estruturas, respectivamente. Os resultados morfometricos correspondentes aos componentes da glândula mamária revelaram maiores áreas de tecido conjuntivo e menores de tecido adiposo na regiäo retroareolar. Com relaçäo a porçäo glandular propriamente dita, encontramos maiores áreas ocupadas por acinos. Através da coloraçäo especifica para o sistema elastico, verificamos que a maior concentraçäo de fibras do sistema elastico aconteceu ao redor dos ductos. O estroma apresentou fibras do sistema elastico em quantidade menor do que as demais regiöes. A proporçäo de ductos foi maior nas mulheres mais velhas, assim como a proporçäo de acinos teve comportamento inverso...

Study of the action of tamoxifen on the mammary gland epithelium of premenopausal patients by lysosome quantification

Tamoxifen is an antiestrogen drug widely utilized for the adjuvant hormonal treatment of breast carcinoma. Its use in the primary prophylaxis of this disease is currently being proposed. Although the drug has few side effects, its precise action on breast tissue that has not undergone neoplastic transformation has not been fully elucidated. This prospective, randomized study assessed the estrogen activity of tamoxifen on the mammary gland epithelium of premenopausal patients using a quantitative analysis of mammary epithelium lysosomes identified by the cytochemical technique of GOMORI for acid phosphatase and by light microscopy. Tamoxifen significantly increased the number of lysosomes only during the secretory phase of the menstrual cycle. We concluded that the early effect of the drug on normal mammary tissue is synergistic with the effect of estrogen during the premenopausal period.

Study of the action of tamoxifen on the mammary gland epithelium of premenopausal patients by lysosome quantification.

Tamoxifen is an antiestrogen drug widely utilized for the adjuvant hormonal treatment of breast carcinoma. Its use in the primary prophylaxis of this disease is currently being proposed. Although the drug has few side effects, its precise action on breast tissue that has not undergone neoplastic transformation has not been fully elucidated. This prospective, randomized study assessed the estrogen activity of tamoxifen on the mammary gland epithelium of premenopausal patients using a quantitative analysis of mammary epithelium lysosome identified by the cytochemical technique of GOMORI for acid phosphatase and by light microscopy. Tamoxifen significantly increased the number of lysosomes only during the secretory phase of the menstrual cycle. We concluded that the early effect of the drug on normal mammary tissue is synergistic with the effect of estrogen during the premenopausal period.

Actividad proteín-quinasa endógena y fosforilación de protaínas intracelulares en cáncer de mama: relación con elementos hsitológicos de pronóstico / Endogenous protein-kinase activity and intracellular protein phosphorylation in breast cancer: relationship with histological elements of prognostic

Existen evidencias previas que demuestran un aumento de la actividad global de proteín-quinasa en tejido mamario canceroso, lo que se traduce en patrones de fosforilación proteica alterados, tanto para proteínas de membrana (plasmática y nuclear) como citoplásmicas. La fosforilación proteica es un importante mecanismo de regulación de ciclo celular, de modo que el hallazgo de fosforilaciónes anormales dentro de las células patológicas puede constituir un indicio de la alteración que induce la neoplasia, y en la práctica podría relacionarse con la evolución del tumor. Con el objeto de cuantificar los niveles de fosforilación endógena de proteínas y de demostrar que los patrones de fosforilación en las células neoplásicas son diferentes de los de la células no neoplásicoas, se realizó un estudio en homegenizado de tejido mamario obtenido de 18 pacientes, en cada una de las cuales se extrajó una muestra de tejido canceroso y otra de parénquima mamario normal. En ellas se determinó la actividad PK endógena con técnica de Roskoski modificada y se determinó los patrones de fosforilación mediante análisis electroforético y posterior exposición en placas de autorradiografía.En 17 de los 18 casos, se detectó una diferencia entre los tejidos normales y los neoplásicos reflejado en los patrones de fosforilación respectivos. Las diferencias más notables se encontraron en la fosforilación endógena de la proteína de 59,5 kd que se fosforila en el 88,9 por ciento de los cánceres y en el 16,7 por ciento de los tejidos normales; la proteína de 13 kd que se fosforila en el 72,2 por ciento de los cánceres y en el 22,2 por ciento de los tejidos normales y en la proteína de 150 kd que se fosforila en el 94,4 por ciento de los cánceres y en el 16,7 por ciento de los tejidos normales. Las diferencias observadas demuestran que existe una notable alteración en los mecanismos de regularización de la fosforilación proteíca dentro de la célula de cáncer de mama

Diferenciación celular del epitelio mamario en cultivo / Cellular differentiation of the mammary epitelium in culture

The functional differentiation of mammary epithelial cells requires specific hormones, growth factors and chemical signals of the cellular environment. These signals consist in cell communication with the extracellular matirx and cell-cell interactions whic though an intrincated mechanism lead to the events that trigger milk secretion. The mammary epithelial cells HC!! from the cellular line COMMA-1D were cultured in the presence of lactogenic hormones such as insuline, dexametazone and prolactin and further stimulated with the epidermal growth factor. In these conditions the cell differentiated, presenting quantitative changes in shape and volume on their organelle cell structure. These cells synthesize betha casein and the intermediary filaments of keratin demonstrated by immunoblotting as well as electronic micrographies

Mammary epithelial hyperplasias: alterations related solely to proliferation?

The cytogenetic analysis of five mammary epithelial hyperplasias is reported. Only two cases had clonal chromosome alterations. Structural alterations involved chromosomes 1 and 5. Numerical alterations involved chromosomes X, 1, 6, 9, and 19. In four cases, the modal number was in the diploid range, and one case was tetraploid. The study of benign proliferations and their comparison with the chomosome alterations of their malignant counterparts could lead to a better understanding of the genes acting solely in cell proliferation and those that cause these cells to undergo malignant transformation and to become invasive.

Quantitative study of the lysosomes of epithelial cells from human mammary glands and fibroadenoma during proliferative and secretory phase of the menstrual cycle.

The cyclic lysosome number variation in fibroadenoma and normal breast epithelial cells in 13 women, using light microscopy and Gomory acid phosphatase method, was studied. The menstrual cycle phase was determined by the last menstrual period, plasmatic progesterone level and endometrium biopsy. Seven lumps were extirpated in the proliferative phase and other 7 in the secretory period. The lysosomes were counted in the fibroadenoma and the normal breast epithelia. The statistical analysis of the findings showed that both fibroadenoma and normal breast epithelia present a number of lysosomes statistically higher in proliferative than in the secretory phase. It was also noted that the number of lysosomes in fibroadenomas is always greater than in the normal breast, independently of the phase of the menstrual cycle.

Morphological and ultrastructural aspects of the cyclical changes of human mammary gland during the menstrual cycle

Com a finalidade de avaliar se a glándula mamária humana sofre alteraçöes morfológicas e ultra-estruturais durante o ciclo menstrual, selecionaram-se 30 pacientes atendidas no período de janeiro de 1991 a setembro de 1992. As pacientes foram divididas, ao acaso, em dois grupos, isto é, 15 mulheres na fase proliferativa e igual número na fase secretora. Os espécimes de tecido mamário normal foram obtidos por ocasiäo da exérese de fibroadenomas. Na fase proliferativa, a glândula mamária encontra-se constituída por lóbulos pouco desenvolvidos e as figuras de mitose säo virtualmente ausentes. A maioria das células epiteliais apresenta núcleos pequenos, escuros, irregulares, ricos em heterocromatina. O estroma mamário apresenta comportamento semelhante; é compacto e denso e os fibrolastos säo desenvolvidos. Já durante a fase secretora, os lóbulos säo bem desenvolvidos e foi relativamente comum o encontro de mitoses. A grande maioria das células epiteliais, nesta fase apresenta núcleo claro, regular, volumoso, rico em eucromatina. Da mesma maneira, o estroma é constituído por fibroblastos com núcleos volumosos e ricos em eucromatina. Concluindo, podemos constatar alteraçöes marcantes na glândula mamária humana consoante a fase do ciclo menstrual

Morphological and ultrastructural aspects of the cyclical changes of human mammary gland during the menstrual cycle.

Analyzing the morphological and ultrastructural features of the mammary gland during the menstrual cycle, the authors observed that in the proliferative phase, the mammary gland consisted of few developed lobules and presented a relative absence of mitotic figures. In this phase, the great majority of the epithelial cells contained small, dark, irregular, rich heterochromatinic nuclei. The mammary stroma showed similar findings, presenting dense connective tissue with few enhanced fibroblasts. During the secretory phase, the lobules were well developed and mitotic figures were usually observed. In this phase, the great majority of the epithelial cells presented clear voluminous, rich euchromatinic nuclei; the stroma consisted of fibroblasts with voluminous nuclei full of euchromatin. There are evident modifications of the mammary tissue according to the phase of the menstrual cycle. The morphological and ultrastructural modifications indicate higher metabolical and mitotic activities during the secretory phase.