Prospecting of potentially probiotic lactic acid bacteria from bovine mammary ecosystem: imminent partners from bacteriotherapy against bovine mastitis / Prospección de bacterias ácido lácticas potencialmente probióticas del ecosistema mamario bovino: socios inminentes de la bacterioterapia contra la mastitis bovina

Mastitis is one of the most important causes of loss of cattle production, burdening producers due to the increased cost of milk production and decreased herd productivity. The development of alternative methods for the treatment and prevention of mastitis other than traditional chemical antibiotic therapy needs to be implemented to meet international pressures to reduce the use of these drugs and promote the elimination of multiresistant microbial strains from the environment. Treatment with probiotic bacteria or yeast strains offers a possible strategy for the control of mastitis. The objective of this work was to isolate, identify, and characterize lactic bacteria from milk and the intramammary duct of Gyr, Guzerat, Girolando 1/2, and Holstein cattle breeds from Brazil. Samples of 115 cows were taken, a total of 192 bacteria isolates belonging to 30 species were obtained, and 81 were selected to evaluate their probiotic potential in in vitro characterization tests. In general, bacteria isolated from the mammary gland have low autoaggregation, cell surface hydrophobicity, and co-aggregation with mastitis etiological bacteria Staphylococcus aureus and Escherichia coli. Also, they have biofilm assembly capacity, inability to produce exopolysaccharides, high production of H2O2, and strong antagonism against mastitis pathogens. Ten lactic bacteria isolates were used in co-culture with human MDA-MB-231 breast epithelial cells to assess their adhesion capacity and impairment of the S. aureus invasion. Our results, therefore, contribute to the future production of new prevention and treatment tools for bovine mastitis.(AU)

GAL3ST2 from mammary gland epithelial cells affects differentiation of 3T3-L1 preadipocytes

Introduction. In the mammary gland, the involution that occurs when lactation ends is an important period for cancer development. We have previously demonstrated stromal–epithelium interactions evaluating conditioned medium of adipose tissue on breast epithelial metalloproteases activity (Creydt et al., Clin Transl Oncol 15:124–131, 2013). Here, we evaluated the effects of conditioned medium of breast epithelial mammary cells on stromal cells. Materials and methods. Conditioned medium from normal murine mammary gland cell line (NMuMG) and conditioned medium proteins were obtained. Then, they were evaluated on modulation of adipocyte differentiation, using 3T3-L1 cell line. Results. We described, for the first time, that breast epithelial mammary cells could produce the enzyme galactose 3-O-sulfotransferase 2 (GAL3ST2). Importantly, GAL3ST2 is present in NMMuMG and two human breast cancer cell lines, and it is more strongly expressed in more metastatic tumors. When 3T3-L1 preadipocyte differentiation was triggered in the presence of conditioned medium from NMuMG or GAL3ST2, triglyceride accumulation was decreased by 40 % and C/EBPβ expression by 80 % in adipocytes. In addition, the expression of FABP4 (aP2), another marker of adipocyte differentiation, was inhibited by 40 % in GAL3ST2-treated cells. Conclusions. Taken together, these results suggest that GAL3ST2 would interfere with normal differentiation of 3T3-L1 preadipocytes; raising the possibility that it may affect normal differentiation of stromal preadipocytes and be a link to tumor metastatic capacity (AU)

No disponible

Expanding Sca-1(+) mammary stem cell in the presence of oestrogen and growth hormone

BACKGROUND: Sca-1 is controversial as a mammary stem cell marker in the literature, which may be due to the different isolation protocols and culture media used in different laboratories. The object of our study is to establish the Medium to promote the proliferation of mammary stem cell and explore the possibility of Sca-1 as mammary stem cell marker. METHODS: We used BM medium supplemented with different concentration of 17Β-oestradiol and GH to find out MaECM medium which promoted the proliferation of mouse mammary epithelial cells and inhibited the growth of fibroblasts. Flow cytometry was used to isolate Sca-1(+) and Sca-1(-) cell populations from cultured mammary epithelial cells. Mammary fat pad transplantation and Mammosphere- forming assay were done to confirm the stem cell potential of Sca-1(+) cells. Differentiating culture was used to detect the differentiation potential of Sca-1(+) cells. Real-time PCR was carried out to analyse the expression of mammary stem cell-related genes in Sca-1(+) cells. RESULTS: We first selected the medium suitable for mammary stem cell growth. Stem cell medium BM was used to culture mammary organoids, which generated many fibroblasts. We established MaECM medium supplemented with oestrogen and growth hormone (GH), in which oestrogen promoted mammary epithelial cell proliferation and inhibited fibroblast growth, and GH obviously enhanced the effect of oestrogen on mammary epithelial cell proliferation. Flow cytometry showed that 50% of cells were Sca-1(+) under the culture of MaECM medium. We confirmed that Sca-1(+) cells regenerated mammary outgrowths when transplanted in vivo, formed mammospheres in vitro and differentiated into luminal epithelial cells with milk-secreting function and myoepithelial cells under Matrigel culture. Furthermore, gene expression analysis by Real-time PCR revealed that Sca-1(+) cells expressed markedly higher levels of mammary stem cell-related genes in comparison to Sca-1(-) cells. CONCLUSION: Our research demonstrates that Sca-1(+) mammary stem cells can be more easily isolated when cultured in the presence of oestrogen and GH (AU)

Aislamiento de levaduras de glándulas mamarias con diferentes presentaciones de mastitis en el Altiplano Mexicano / Yeasts isolation from bovine mammary glands under different mastitis status in the Mexican High Plateu

Antecedentes. Las mastitis subclínicas y clínicas en bovinos por hongos se han incrementado principalmente por levaduras del género Candida. Objetivo. Conocer las levaduras presentes en la leche de glándulas mamarias de bovinos clínicamente sanos, y de aquellos con mastitis subclínica y clínica. Método. Se evaluó la presencia de levaduras en 1.095 muestras de leche de 342 glándulas mamarias sanas, 383 con mastitis subclínica y 370 con mastitis clínica, de los estados de Querétaro, Hidalgo, Puebla y la ciudad de México, Distrito Federal, que forman parte del Altiplano Mexicano. El estado sanitario de las glándulas mamarias se determinó por examen clínico y la prueba de California. La identificación de levaduras fue realizada por métodos morfológicos y bioquímicos. Resultados. Se identificaron 20 especies diferentes del género Candida a partir de 282 (25,75%) de las muestras de leche. Las especies encontradas con mayor frecuencia en los bovinos sanos y con mastitis clínica fueron Candida glabrata y Candida krusei. El grupo de las muestras con mastitis subclínica mostró una diversidad de especies de Candida, incluidas Candida zeylanoides, Candida norvegica, Candida viswanathii, Candida guilliermondii y Candida tropicalis. Candida albicans fue aislada solo en 11 (3,9%) de las muestras de mastitis clínica (6) y subclínica (5). Conclusiones. Estos resultados sugieren el posible papel de otras especies de Candida diferentes a C. albicans como causantes de mastitis micótica(AU)

Background. The mastitis subclinical and clinical in cows caused by fungi has been increased specially by yeast of the genus Candida. Objective. To identify what yeasts were present in milk samples obtained from mammary glands of healthy cows, and others suffering subclinical or clinical mastitis. Methods. From a total of 1,095 milk samples 342 were from mammary glands of healthy dairy cows, 383 with subclinical mastitis, and 370 with clinical mastitis, were taken, in the states of Querétaro, Hidalgo, Puebla and Mexico City (Distrito Federal) in the so called Mexican High Plateu. The clinical status of the mammary glands was determined by clinical examination and the California Mastitis Test. Yeasts identification was carried out by morphology and biochemical methods. Results. Twenty different species of Candida were identified out of 282 (25.75%) milk samples. The most frequently identified species in the healthy cows and cows with clinical mastitis groups were Candida glabrata and Candida krusei. On the other hand, samples from the subclinical mastitis group showed a diversity of Candida species, including Candida zeylanoides, Candida norvegica, Candida viswanathii, Candida guilliermondii, and Candida tropicalis. Candida albicans was isolated only in 11 (3.9%) samples from the clinical and subclinical mastitis groups. Conclusions. These results suggest the possible role that Candida species other than C. albicans may play in mycotic mastitis in cows(AU)

El citoesqueleto celular en la glándula mamaria y su aplicación diagnóstica / The cellular cytoskeleton in the mammary gland and its diagnostic application

Objetivos: Valorar la importancia de las modificaciones que aparecen en el citoesqueleto celular (en particular a nivel de unos filamentos intermedios denominados queratinas) a lo largo de los cambios que sufre la glándula mamaria, con especial referencia a la lactación. Su expresión será diferente y servirá como método de contraste. Diseño: 40 ratones distribuidos en cinco grupos según la etapa de estudio. El esquema experimental es común para todos ellos. Material y métodos: Análisis inmunohistoquímico de las muestras procedentes de las glándulas mamarias para evidenciar modificaciones a nivel de las citoqueratinas y comparación con otros estudios en diferentes modelos animales. Resultados: Cada citoqueratina presenta una expresión diferente según sea la estirpe celular y el periodo de lactación. Se diferencian mediante el uso de anticuerpos específicos marcados. Conclusiones: La expresión citoqueratínica varía de manera fisiológica y en células tumorales. La detección inmunohistoquímica de citoqueratinas puede servir como método de diagnóstico complementario (AU)

Objectives: To assess the importance of the changes that appear into the cytoskeleton (especially referred to some specific intermediate filaments called keratins) through different stages the mammary gland bears, focusing on the lactation. Their expression will be different in pathological conditions and this will be useful for contrasting other procedures. Design: 40 mice distributed into five groups according to the tage of study. The experimental project is the same for everyone. Material and methods: Immunohistochemical analysis from samples obtained from mammary glands in order to show changes into cytokeratin expression and comparison with other researches in different animal models. Results: The expression of every cytokeratin is different according to the cell lineage and the lactation period. Specific antibodies can reveal it. Conclusions: The cytokeratinic pattern changes in a physiological sense and also in malignant processes. Immunohistochemical detection of cytokeratins may be useful as complementary diagnosis (AU)

Evaluación del efecto del antiprogestágeno, aglepristone, sobre la regresión tumoral y sobrevida en perras con tumores mamarios estadio V / Assessment of aglepristone in dogs with mammary tumors stage V

Las metástasis pulmonares de tumores mamarios son generalmente la causa de muerte en las mascotas quepadecen una neoplasia de glándula mamaria. Esto marca la importancia en la búsqueda de nuevos tratamientos,enfocados a la modulación de blancos moleculares, con una acción predominante en la estabilización oregresión de la enfermedad metástasica. En este estudio se evaluaron 20 perras en estadio clínico V. Las perrasfueron distribuidas aleatoriamente en uno de los siguientes grupos. Aglespristone (Alizine ®, Virbac, Francia)(Agle, n = 10) o placebo (PLCB n = 10). El análisis de supervivencia por el método de Kaplan-Meier indicóque aglepristone no prolongo la mediana del tiempo de supervivencia general (Agle. 13 días vs. PLCB 14,5 días(P = 0,8). Este patrón de aplicación de aglepristone no fue efi caz en el tratamiento de metástasis de tumores demama caninos. Teniendo en cuenta los efectos positivos de los antagonistas de la progesterona en otras especiescon tumores en la glándula mamaria, no descartamos que otro esquema de tratamiento pudiera tener un efectobenefi cioso sobre la hembra canina portadora de metástasis de una neoplasia de glándula mamaria(AU)

Pulmonary metastasis of breast tumours is usually the cause of death in pets suffering a mammary glandneoplasm. This marks the importance of fi nding new treatments, focusing on the modulation of moleculartargets, with a predominant in the stabilization or regression of metastasis disease. In this study included 20dogs with mammary tumours in V clinical stage. The dogs were randomly placed in one of the following treatments. Aglespristone (Alizine ®, Virbac, France) (Agle, n = 10) or placebo (PLCB n = 10). Survivalanalysis by Kaplan-Meier method indicated that aglepristone not extended the median time of general survival(Agle. 13 days vs PLCB 14.5 days (P = 0.8). This application pattern in the aglepristone treatment in metastasisof canine breast tumours was not effective. Knowing the positive effects that some studies showed in mammarygland tumours treated with progesterone antagonist in other species; do not rule out that another treatmentschedule might have a benefi cial effect on the female dog(AU)