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Search Results: 1 - 10 of 10162 matches for " Elena Quaglino "
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Met Receptor Acts Uniquely for Survival and Morphogenesis of EGFR-Dependent Normal Mammary Epithelial and Cancer Cells
Paolo Accornero, Silvia Miretti, Francesca Bersani, Elena Quaglino, Eugenio Martignani, Mario Baratta
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0044982
Abstract: Mammary gland development and breast cancer growth require multiple factors both of endocrine and paracrine origin. We analyzed the roles of Epidermal Growth Factor Receptor (EGFR) and Hepatocyte Growth Factor Receptor (Met) in mammary epithelial cells and mammary tumor cells derived from a mutated-ErbB2 transgenic mice. By using highly specific tyrosine kinase inhibitors we found that MCF-10A and NMuMG mammary epithelial cell lines are totally dependent on EGFR activation for their growth and survival. Proliferation and 3D-morphogenesis assays showed that HGF had no role in maintaining mammary cell viability, but was the only cytokine able to rescue EGFR-inhibited mammary cells. Insulin-Like Growth Factor-I (IGF-I), basic-Fibroblast Growth Factor (b-FGF) and Neuregulin, which are well known mammary morphogenic factors, did not rescue proliferation or morphogenesis in these cell lines, following EGFR inhibition. Similarly, ErbB2-driven tumor cells are EGFR-dependent and also display HGF-mediated rescue. Western-blot analysis of the signaling pathways involved in rescue after EGFR inhibition indicated that concomitant ERK1/2 and AKT activation was exclusively driven by Met, but not by IGF-I or b-FGF. These results describe a unique role for EGFR and Met in mammary epithelial cells by showing that similar pathways can be used by tumorigenic cells to sustain growth and resist to EGFR-directed anti-tumorigenic drugs.
Chimeric DNA Vaccines against ErbB2+ Carcinomas: From Mice to Humans
Elena Quaglino,Federica Riccardo,Marco Macagno,Silvio Bandini,Rodica Cojoca,Elisabetta Ercole,Augusto Amici,Federica Cavallo
Cancers , 2011, DOI: 10.3390/cancers3033225
Abstract: DNA vaccination exploits a relatively simple and flexible technique to generate an immune response against microbial and tumor-associated antigens (TAAs). Its effectiveness is enhanced by the application of an electrical shock in the area of plasmid injection (electroporation). In our studies we exploited a sophisticated electroporation device approved for clinical use (Cliniporator, IGEA, Carpi, Italy). As the target antigen is an additional factor that dramatically modulates the efficacy of a vaccine, we selected ErbB2 receptor as a target since it is an ideal oncoantigen. It is overexpressed on the cell membrane by several carcinomas for which it plays an essential role in driving their progression. Most oncoantigens are self-tolerated molecules. To circumvent immune tolerance we generated two plasmids (RHuT and HuRT) coding for chimeric rat/human ErbB2 proteins. Their immunogenicity was compared in wild type mice naturally tolerant for mouse ErbB2, and in transgenic mice that are also tolerant for rat or human ErbB2. In several of these mice, RHuT and HuRT elicited a stronger anti-tumor response than plasmids coding for fully human or fully rat ErbB2. The ability of heterologous moiety to blunt immune tolerance could be exploited to elicit a significant immune response in patients. A clinical trial to delay the recurrence of ErbB2 + carcinomas of the oral cavity, oropharynx and hypopharynx is awaiting the approval of the Italian authorities.
Identification of Relevant Conformational Epitopes on the HER2 Oncoprotein by Using Large Fragment Phage Display (LFPD)
Federico Gabrielli, Roberto Salvi, Chiara Garulli, Cristina Kalogris, Serena Arima, Luca Tardella, Paolo Monaci, Serenella M. Pupa, Elda Tagliabue, Maura Montani, Elena Quaglino, Lorenzo Stramucci, Claudia Curcio, Cristina Marchini, Augusto Amici
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0058358
Abstract: We developed a new phage-display based approach, the Large Fragment Phage Display (LFPD), that can be used for mapping conformational epitopes on target molecules of immunological interest. LFPD uses a simplified and more effective phage-display approach in which only a limited set of larger fragments (about 100 aa in length) are expressed on the phage surface. Using the human HER2 oncoprotein as a target, we identified novel B-cell conformational epitopes. The same homologous epitopes were also detected in rat HER2 and all corresponded to the epitopes predicted by computational analysis (PEPITO software), showing that LFPD gives reproducible and accurate results. Interestingly, these newly identified HER2 epitopes seem to be crucial for an effective immune response against HER2-overexpressing breast cancers and might help discriminating between metastatic breast cancer and early breast cancer patients. Overall, the results obtained in this study demonstrated the utility of LFPD and its potential application to the detection of conformational epitopes on many other molecules of interest, as well as, the development of new and potentially more effective B-cell conformational epitopes based vaccines.
Marta Quaglino,José Pagura,Daniela Dianda,Evangelina Lupachini
SaberEs : Revista de Ciencias Económicas y Estadística , 2010,
Abstract: Los estudios de repetibilidad y reproducibilidad fueron dise ados con el propósito de analizar la bondad de los sistemas de medición, análisis cuya importancia radica en el hecho que un sistema inadecuado introducirá variabilidad adicional ocasionando que las mediciones no reflejen el verdadero comportamiento del proceso. El análisis se basa en la cuantificación de la variabilidad asociada al sistema de medición y su posterior comparación con la variabilidad total observada, siendo requerimiento fundamental para ello que resulte factible obtener mediciones repetidas de una misma unidad bajo las mismas condiciones experimentales, de lo contrario, la variabilidad en las mediciones estará confundida con la variabilidad propia de las partes medidas. Tal es el caso en que los ensayos de medición son “destructivos”, esto es, las unidades no son robustas frente al proceso de medición, o bien, las unidades no son temporalmente estables.En este trabajo se exponen diversas alternativas para el caso de estudios R&R con ensayos destructivos y una aplicación particular en un problema real sobre estimación de tiempos de producción en una empresa metalúrgica. El empleo de Modelos Lineales Generalizados permitió obtener estimaciones adecuadas de ciertas Componentes de Variancia, que advirtieron sobre características importantes a mejorar en el proceso de medición.
The Effect on Rat Thymocytes of the Simultaneous In Vivo Exposure to 50-Hz Electric and Magnetic Field and to Continuous Light
Daniela Quaglino,Miriam Capri,Luigi Zecca,Claudio Franceschi
The Scientific World Journal , 2004, DOI: 10.1100/tsw.2004.183
Hyaluronan uptake by adult human skin fibroblasts in vitro
MA Croce,F Boraldi,D Quaglino,R Tiozzo
European Journal of Histochemistry , 2003, DOI: 10.4081/808
Abstract: Low and high molecular weight hyaluronan (HA) was added to adult human fibroblasts grown in monolayer to assess its influence on CD44 expression, its internalisation and effect on cell growth. CD44 expression on the surface of in vitro fibroblasts was not modified by different concentrations of FCS, whereas it was sensitive to cell cycle, being higher in the growing than in the resting phase. Independently from molecular weight, upon addition of exogenous HA (from 0.1 up to 1 mg/mL) to fibroblasts in the growing phase, a slight but constant decrease of the expression of CD44 on the surface of fibroblasts was observed; moreover, HA induced a rearrangement of CD44 into patches in close relationship with the terminal regions of stress fibers, which became thicker and more rigid after a few hours from the addition of HA to the medium. Fluorescent HA, added to the culture medium, rapidly attached to the plasma membrane and in less than two minutes was observed within cells, partly in association with its receptor CD44. By the contemporary use of neutral red, which accumulates into functional lysosomes, the great majority of internalised HA was found within lysosomes. HA receptor RHAMM-IHABP was rather homogeneously localised within the cytoplasm of normal growing fibroblasts. Upon addition of HA, the RHAMM-IHABP distribution became discontinuous around the nucleus. Addition of HA to fibroblasts induced a significant inhibition of cell growth, which was dependent on HA concentration and irrespective of HA molecular weight, at least in the ranges tested. Results show that extra-cellular HA is rapidly taken up by human dermal fibroblasts together with its CD44 receptor, and transported mostly to the lysosomes. Both low and high molecular weight HA induced down-regulation of cell proliferation, which would seem to be mediated by HA catabolism.
Biocompatibility of Collagen Membranes Assessed by Culturing Human J111 Macrophage Cells
Claudia Gaetana Aruta,Maria Antonietta Croce,Daniela Quaglino,Deanna Guerra,Roberta Tiozzo
Materials , 2009, DOI: 10.3390/ma2030945
Abstract: We have carried out an in vitro study on the interactions of human macrophages (J111 cell line) with different scaffolds made of type I and II collagen, isolated from horse tendon and from horse articular and trachea cartilage, in order to assess growth properties and biocompatibility of these membranes. We have therefore evaluated cell adhesion and proliferation as well as cytokine production considered an indicator of macrophage activation. The inflammatory response is in fact one of the major causes of collagen destruction thus interfering with cell and tissue behaviour. Moreover, the morphology of cells, seeded on membranes selected for the best characteristics, was described. Results might be relevant for in vivo application such ad “tissue engineering” and/or specialized cells implants.
Mouse mammary tumors display Stat3 activation dependent on leukemia inhibitory factor signaling
Ana Quaglino, Carolina Schere-Levy, Leonardo Romorini, Roberto P Meiss, Edith C Kordon
Breast Cancer Research , 2007, DOI: 10.1186/bcr1777
Abstract: The studies were performed in different tumorigenic and non-tumorigenic mammary cells. The expression of LIF and LIF receptor was tested by RT-PCR analysis. In tumors, LIF and Stat3 proteins were analyzed by immunohistochemistry, whereas Stat3 and extracellular signal-regulated kinase (ERK)1/2 expression and phosphorylation were studied by Western blot analysis. A LIF-specific blocking antibody was used to determine whether this cytokine was responsible for Stat3 phosphorylation induced by conditioned medium. Specific pharmacological inhibitors (PD98059 and Stat3ip) that affect ERK1/2 and Stat3 activation were used to study their involvement in LIF-induced effects. To analyze cell survival, assays with crystal violet were performed.High levels of LIF expression and activated Stat3 were found in mammary tumors growing in vivo and in their primary cultures. We found a single mouse mammary tumor cell line, LM3, that showed low levels of activated Stat3. Incidentally, these cells also showed very little expression of LIF receptor. This suggested that autocrine/paracrine LIF would be responsible for Stat3 activation in mouse mammary tumors. This hypothesis was confirmed by the ability of conditioned medium of mammary tumor primary cultures to induce Stat3 phosphorylation, activity that was prevented by pretreatment with LIF-blocking antibody. Besides, we found that LIF increased tumor cell viability. Interestingly, blocking Stat3 activation enhanced this effect in mammary tumor cells.LIF is overexpressed in mouse mammary tumors, where it acts as the main Stat3 activator. Interestingly, the positive LIF effect on tumor cell viability is not dependent on Stat3 activation, which inhibits tumor cell survival as it does in normal mammary epithelium.The pleiotropic cytokine leukemia inhibitory factor (LIF) is a secreted 38 to 67 kDa glycoprotein first named for its ability to induce macrophage differentiation in the murine myeloid leukemic cell line M1 [1]. This factor has bee
Mechanical strain induces involution-associated events in mammary epithelial cells
Ana Quaglino, Marcelo Salierno, Jesica Pellegrotti, Natalia Rubinstein, Edith C Kordon
BMC Cell Biology , 2009, DOI: 10.1186/1471-2121-10-55
Abstract: We have designed and built a new device to analyze the biological consequences of applying mechanical stress to cells cultured on flexible silicone membranes. Subsequently, a geometrical model that predicted the percentage of radial strain applied to the elastic substrate was developed. By microscopic image analysis, the adjustment of these calculations to the actual strain exerted on the attached cells was verified. The studies described herein were all performed in the HC11 non-tumorigenic mammary epithelial cell line, which was originated from a pregnant BALB/c mouse. In these cells, as previously observed in other tissue types, mechanical stress induced ERK1/2 phosphorylation and c-Fos mRNA and protein expression. In addition, we found that mammary cell stretching triggered involution associated cellular events as Leukemia Inhibitory Factor (LIF) expression induction, STAT3 activation and AKT phosphorylation inhibition.Here, we show for the first time, that mechanical strain is able to induce weaning-associated events in cultured mammary epithelial cells. These results were obtained using a new practical and affordable device specifically designed for such a purpose. We believe that our results indicate the relevance of mechanical stress among the early post-lactation events that lead to mammary gland involution.Cells are able to act in response to multiple biochemical and biophysical stimuli, depending on their type and function. Particularly, it has been shown that mechanical forces trigger specific events in a variety of cell types under different physiological and pathological situations [1-5]. For example, it has been demonstrated that in solid tumors, cells are subjected to mechanical stress due to elevated interstitial pressure and perturbed vasculature [6]. The dramatic consequences of this strain were demonstrated when it was shown that extracellular matrix rigidity induced malignant phenotype in mammary epithelial cells [7].In the mouse mammary gland,
La Infección como Factor Pronóstico en Terapia Intensiva
Priscila Giavedoni,Daniel H. Daniel H. Bagilet,Claudio Settecase,Marta B. Quaglino
Medicrit : Revista de Medicina Crítica , 2008, DOI: 10.5413/mrmc.2008.53.103
Abstract: Objetivo. Estudiar la relación de la patología infecciosa como motivo de ingreso a Terapia Intensiva y la evolución a corto plazo. Método. Dise o: retrospectivo y observacional. Procedimiento: Se revisaron los pacientes internados entre el 01/01/1.996 y el 31/12/2.006. Para el análisis se crearon 6 grupos de gravedad según el escore APACHE II de ingreso. La influencia de la patología de ingreso, sexo y días de internación fue determinada con regresión logística múltiple. En cada grupo de APACHE II se analizó el riesgo de muerte de los pacientes que ingresaron por patología infecciosa. Resultados. Se analizaron 5.263 pacientes. Edad media 55.08 a os (r 11-98); sexo masculino 63%; promedio de internación 4.05 días (±5.44); APACHE II 11.3 (±8.27); mortalidad 22.4%; patología de ingreso: cardiovascular 31.50%, infecciosa 14.48%, neurológica 9,86%, respiratoria 8.09%, digestiva 7.30%, medio interno 3.55% y otras 25.22%. La patología infecciosa fue el único factor que fue significativo en todos los grupos de APACHE II. Cuando se analizaron en forma conjunta todos los pacientes y se incorporó al modelo de regresión logística el valor de APACHE II, resultaron significativos: patología cardiovascular, digestiva, infecciosa, neurológica, respiratoria y APACHE II. Siendo la patología infecciosa la de mayor peso estadístico. Conclusión. Nuestro estudio sugiere que la patología infecciosa es un factor de riesgo independiente de mortalidad. Por lo tanto si se pretende mejorar la sobrevida y disminuir los costos hospitalarios, estos enfermos deberían considerarse de alto riesgo. Palabras clave: Pronóstico; Infección; APACHE II. ............... Infections as Prognosis Factor in Critical Care Units ABSTRACT Objective. To study the prevalence of infectious diseases as a cause of critical care unit admission and its short-term evolution. Method. Design: Retrospective and observational study. Procedure: The patients admitted between January, 1st, 1,996 and December, 31st, 2,006 was reviewed. For the purpose of the analysis, there were 6 groups created according to the APACHE II score, to stratify severity at intensive-care unit admission. The influence of the illness at admission, gender and days at intensive care unit was determined through multiple logistic regression. In each APACHE II group, the mortality in patients admitted for infectious diseases were analyzed. Results. 5.263 patients were analyzed. Average age: 55.08 years (r 11-98); male gender: 63%; average days at intensive care unit: 4.05 days (±5.44); APACHE II 11.3 (±8.27); mortality: 22.4%; causes of admissio
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