Papers by Carmen Ramirez-Castillejo
Background: Breast cancer is the leading cause of death among females in developed countries. Alt... more Background: Breast cancer is the leading cause of death among females in developed countries. Although the implementation of screening tests and the development of new therapies has increased the probability of remission, relapse rates remain high. Numerous studies have indicated the connection between cancer initiating cells and slow cellular cycle cells, identified by their capacity to retain long labelling (LT+). Methods: In this study, we perform tumour cell culture dose-response assays in adherent and mamospheres cultures. We also probe our conclusion in Xenograft models. Cell culture and in vivo tumours were analysed by immunofluorescence, qPCR and flow cytometry assays showing how stem cell self-renewal modulating proteins, such as PEDF, can statistically significant modify the properties, cell-percentage expressing biomarkers, and carcinogenicity of cancer stem cells (by student t-test). Results: The PEDF signaling pathway could be a useful tool for controlling cancer stem c...
Biomedicines, May 6, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
International Journal of Molecular Sciences, Mar 14, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
International Journal of Molecular Sciences
Gold nanorods are the most commonly used nanoparticles in photothermal therapy for cancer treatme... more Gold nanorods are the most commonly used nanoparticles in photothermal therapy for cancer treatment due to their high efficiency in converting light into heat. This study aimed to investigate the efficacy of gold nanorods of different sizes (large and small) in eliminating two types of cancer cell: melanoma and glioblastoma cells. After establishing the optimal concentration of nanoparticles and determining the appropriate time and power of laser irradiation, photothermal therapy was applied to melanoma and glioblastoma cells, resulting in the highly efficient elimination of both cell types. The efficiency of the PTT was evaluated using several methods, including biochemical analysis, fluorescence microscopy, and flow cytometry. The dehydrogenase activity, as well as calcein-propidium iodide and Annexin V staining, were employed to determine the cell viability and the type of cell death triggered by the PTT. The melanoma cells exhibited greater resistance to photothermal therapy, bu...
Food Chemistry, 2021
A healthy life means a balance between physical activity and a diet rich in fruits and vegetables... more A healthy life means a balance between physical activity and a diet rich in fruits and vegetables, however, some plant-based foods can have certain adverse effects due to the presence of anti-nutritional factors, such as lectins, capable of binding molecules and preventing their normal assimilation. The level of lectins in Synsepalum dulcificum fruit was determined by hemagglutination assays in human blood, and its comparison with foods characterized as having high and low lectin content. The relative hemagglutinating activity of berries from Synsepalum dulcificum compared to our positive high lectin content food reference (Pinto bean) corresponds to 3.13-6.25%, representing safe levels for nutritional food.
Lecture Notes in Computer Science, 2014
Cell image processing and analysis is a crucial task in any health science laboratory. Cell count... more Cell image processing and analysis is a crucial task in any health science laboratory. Cell counting is a common task usually made by technicians with the support of a custom software tuned with experiment requirements. In this work we present a web services based platform focused on the cell counting problem. Using OpenCF, a web services development framework, we integrate in a single platform services oriented to image processing and classifying, cell counting based on a set of parameters, and data post-processing (plot generation, datasheets, etc.). A GUI added to the platform helps to launch jobs with image sets, and the execution of different tasks from a web service based client.
The FASEB Journal, 2015
We studied potential changes in the subventricular zone (SVZ) stem cell niche of the senescenceac... more We studied potential changes in the subventricular zone (SVZ) stem cell niche of the senescenceaccelerated mouse prone-8 (SAM-P8) aging model. Bromodeoxyuridine (BrdU) assays with longtime survival revealed a lower number of label-retaining stem cells in the SAM-P8 SVZ compared with the SAM-Resistant 1 (SAM-R1) control strain. We also found that in SAM-P8 niche signaling is attenuated and the stem cell pool is less responsive to the self-renewal niche factor pigmented epithelium-derived factor (PEDF). Protein analysis demonstrated stable amounts of the PEDF ligand in the SAM-P8 SVZ niche; however, SAM-P8 stem cells present a significant expression decrease of patatin-like phospholipase domain containing 2, a receptor for PEDF (PNPLA2-PEDF) receptor, but not of laminin receptor (LR), a receptor for PEDF (LR-PEDF) receptor. We observed changes in self-renewal related genes (hairy and enhancer of split 1 (Hes1), hairy and enhancer of split 1 (Hes5), Sox2] and report that although these genes are down-regulated in SAM-P8, differentiation genes (Pax6) are up-regulated and neurogenesis is increased. Finally, sheltering mammalian telomere complexes might be also involved given a down-regulation of telomeric repeat binding factor 1 (Terf1) expression was observed in SAM-P8 at young age periods. Differences between these 2 models, SAM-P8 and SAM-R1 controls, have been previously detected at more advanced ages. We now describe alterations in the PEDF signaling pathway and stem cell self-renewal at a very young age, which could be involved in the premature senescence observed in the SAM-P8 model.
Background: Breast cancer is the leading cause of death among females in developed countries. Alt... more Background: Breast cancer is the leading cause of death among females in developed countries. Although the implementation of screening tests and the development of new therapies has increased the probability of remission, relapse rates still remain high. Numerous studies have indicated the connection between cancer initiating cells and slow cellular cycle cells, identified by their capacity to retain long labelling (LT+). Methods: We have designed a transgenic protein consisting in the C-terminal part of this protein, which acts by blocking endogenous PEDF in culture cell assays. Present work is based in doses-response in vitro assays as well as flow cytometry analysis of surface markers and cell cycle kinetic study of the tumour initiating cells.Results: In this study we show that this type of cells is present not only in cancer cell lines but also in cancer cells from patients with metastatic and advanced stage tumours. We also present new assays showing how stem cell self-renewal...
Background: Breast cancer is the leading cause of death among females in developed countries. Alt... more Background: Breast cancer is the leading cause of death among females in developed countries. Although the implementation of screening tests and the development of new therapies has increased the probability of remission, relapse rates still remain high. Numerous studies have indicated the connection between cancer initiating cells and slow cellular cycle cells, identified by their capacity to retain long labelling (LT+). Methods: We have designed a transgenic protein consisting in the C-terminal part of this protein, which acts by blocking endogenous PEDF in culture cell assays. Present work is based in doses-response in vitro assays as well as flow cytometry analysis of surface markers and cell cycle kinetic study of the tumour initiating cells.Results: In this study we show that this type of cells is present not only in cancer cell lines but also in cancer cells from patients with metastatic and advanced stage tumours. We also present new assays showing how stem cell self-renewal...
Oncología (Barcelona), 2005
El gen mutado en el síndrome de Ataxia Telangiectasia está implicado en múltiples funciones celul... more El gen mutado en el síndrome de Ataxia Telangiectasia está implicado en múltiples funciones celulares que abarcan desde la respuesta al daño genotóxico causado por la radiación ionizante hasta la respuesta a insulina. Curiosamente la ruta de señalización mediada por Akt esta relacionada con los mismos estímulos. En el presente estudio demostramos como el gen mutado en Ataxia Telangiectasia (ATM) controla la fosforilación de Akt, en respuesta a estímulos como la radiación ionizante o la insulina. Estos datos han sido obtenidos usando modelos experimentales como células derivadas de enfermos de Ataxia o de ratones nulos para la expresión de ATM. Nuestro estudio propone nuevas explicaciones para entender la radio sensibilidad característica de estos enfermos y destaca el papel primordial de Akt en la respuesta a la radiación ionizante.
Journal of Biological Chemistry, 2008
This is an Open Access article under the CC BY license.
Journal of Analytical Oncology, 2013
We report on two patients, successfully treated by the combination therapy of gemcitabine and 24-... more We report on two patients, successfully treated by the combination therapy of gemcitabine and 24-h intravenous infusion of cisplatin, who were initially diagnosed with node-positive advanced urothelial cancer. Each patient had a very good clinical response and underwent curative radical surgery after gemcitabine/cisplatin chemotherapy. A microscopically detailed examination of surgically obtained specimens showed the complete disappearance of malignant cells in the two cases. As a pilot study, we have used the regimen of gemcitabine plus 24-h continuous infusion of cisplatin, instead of bolus injection, for the treatment of 20 patients with node-positive or metastatic urothelial cancer. The clinical response rate in this regimen was 75% (complete response 7/20; 35%, partial response 8/20; 40%). The median overall survival was 665 days. As for the adverse effects, the incidences of severe neutropenia and thrombocytopenia (grade 3-4) were 20% and 15%, which might be less toxic than co...
International Journal of Food Science & Technology
European Journal of Sport Science
The ontogeny of somatostatin immunoreactive interneurons in the cerebral cortex of the lizard Pod... more The ontogeny of somatostatin immunoreactive interneurons in the cerebral cortex of the lizard Podarcis hispanica has been studied in histological series of embryos, perinatal specimens, and adults. Somatostatin immunoreactive interneurons appear in the early stages of lizard cerebral cortex ontogeny, their number increases during embryonary development, reaches a peak in early postnatal life, and decreases in adult lizards. The first somatostatin immunoreactive somata in the lizard forebrain appeared on E36, and they were located in non cortical areas. Then, on E39 and later, somatostatin immunoreactive neurons were seen in the lizard cortex in a rostral-to-caudal spatial gradient, which parallels that of the normal histogenesis of the lizard cerebral cortex. On E39, labelled somata were seen in the medial and dorsal cortex inner plexiform layers; immunoreactive puncta and dendritic processes were detectable in the inner plexiform layer of the medial cortex. On E40, labelled neurons were observed in the inner plexiform layer of the lateral cortex; labelled processes were found in the inner plexiform layers (dorsomedial, dorsal, and lateral cortices) and the outer plexiform layers (medial and dorsomedial cortices). At hatching (P0), some somatostatin immunoreactive neurons populated the external plexiform layer of the dorsomedial cortex. On P28, groups of labelled neurons appeared in the cell layer of dorsal and lateral cortices, reaching the adult-mature pattern of somatostatin immunoreactivity in the lizard cerebral cortex, i.e., labelled somata and dendritic processes populating the inner plexiform layers in addition to an axonic labelled plexus in the outermost part of the outer plexiform layers. Immunoreactive somata and processes occupied all the cortical areas, but they were especially abundant in the dorsomedial cortex. Proliferating Cell Nuclear Antigen (PCNA) immunostaining in the same histological series revealed that the number of PCNA immunoreactive nuclei in the subjacent proliferative neuroepithelium followed an inverse-complementary evolution to somatostatin, suggesting some temporal relationship between somatostatin immunoreactive cells and neurogenesis in the lizard cerebral cortex.
The lizard medial cortex (lizard fascia dentata) is capable of neural regeneration after being le... more The lizard medial cortex (lizard fascia dentata) is capable of neural regeneration after being lesioned by the anti-metabolite 3-acetylpyridine (3AP). This study was aimed at detecting microglial behaviour during the medial cortex lesion-regeneration process using tomato lectin histochemistry to label microglia (both with light and electron microscopy) and proliferating cell nuclear antigen (PCNA) immunocytochemistry to label proliferating cells. As expected, 1-2 days post-injection lectin-labelled microglia cells could not be observed in the medial cortex plexiform layers, but later (7 days post-injection) abundant lectin-labelled microglia cells re-populated the regenerating medial cortex. Abundant PCNA-immunolabelled nuclei were detected both in the subjacent ependymal neuroepithelium (neuroblasts, maximum at 2 days post-injection) as well as in some parenchymal cells which were also lectin-labelled (microglia, maximum at 7-15 days post-injection). Re-invasive microglia were also detected in the vicinity of ventricular ependymal lining, blood vessels and meninges. The electron microscope demonstrated that these microglial cells participate in cell debris removal, especially of neural granular cell somata. Other cell types related to microglia (mast cells, peri-vascular cells and meningeal cells) were also present during the scavenging process. Significant numbers of microglial cells remained in close relationship with the ependymal proliferative areas, even in control non-lesioned animals. This is indirect evidence for the working hypothesis that microglia are not only implicated in cell debris removal, but also in the regulation of newly generated neuroblast incorporation onto the cortical areas. Whether they phagocytose immature neuroblasts or induce cell death in them or even prevent their migration onto the principal layer areas are likely possibilities that remain to be proven.
Radiation of experimental culture cells on plates with various wells can cause a risk of underdos... more Radiation of experimental culture cells on plates with various wells can cause a risk of underdosage as a result of the existence of multiple air–water interfaces. The objective of our study was to quantify this error in culture plates with multiple wells. Radiation conditions were simulated with the GAMOS code, based on the GEANT4 code, and this was compared with a simulation performed with PENELOPE and measured data. We observed a slight underdosage of ~4 % on the most superficial half of the culture medium. We believe that this underdosage does not have a significant effect on the dose received by culture cells deposited in a monolayer and adhered to the base of the wells. Keywords:Monte Carlo; plate irradiation; culture cells
Production and purification process of pigmented epithelium-derived retinal in a yeast system fac... more Production and purification process of pigmented epithelium-derived retinal in a yeast system factor. The present invention relates to a process for producing and purifying molecules PEDF and mutated versions and / or truncated said molecule in a yeast expression system. It also aims to protect the molecules themselves PEDF directly obtained and the use thereof in the preparation of pharmaceutical compositions. The method comprises the steps of obtaining the DNA of PEDF at least from cells pigment epithelium retina, obtain mutant versions and / or truncated clone constructions in an expression vector yeast, transform and select clones of yeast inoculate with the clones obtained, the growth medium of yeast induce PEDF full or truncated yeast factor, removing cells from the culture by centrifugation, change the culture medium in which the entire PEDF are present and / or truncated, purified and isolated.
En este trabajo se estudia la histogeneris y regeneracion del cortex medial ("facial dentada... more En este trabajo se estudia la histogeneris y regeneracion del cortex medial ("facial dentada") de Podarcis hispanica tras la lesion especifica con el neurotoxico 3AP. El estudio histologico de la histogenesis y regeneracion del cortex medial revela que ambos fenomenos comparten procesos similares de produccion(neurogenesis) migracion, incorparacion y maduracion de nuevas neuronas, tanto en el cortex medial embrionario como en el cortex medial regenerante. En este estudio se ha puesto a punto tecnicas de sintesis y deteccion de marcadores genicos especificos de tejido nervioso de lagartija que ayudan a entender ambos procesos. El microscopio electronico revela que la maduracion ultraestructural de las capas plexiformes externas del cortex medial es mas temprana que la de capas internas cercanas al neuroepitelio. Este patron de maduracion tambien ha sido confirmado con marcadores de migracion celular como la proteina NCAM en su estado polisialiazado(PSA-NCAM). Las aferencias...
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Papers by Carmen Ramirez-Castillejo