Pilar García-Palencia

Complutense University of Madrid, Pozuelo de Alarcón, Madrid, Spain

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Publications (25)87.14 Total impact

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    ABSTRACT: In Iberian pigs, a high conceptus loss occurs during the first 30 days of gestation. Although the exact causes for these losses have not been determined to date, the importance of blood vessel development during early pregnancy has been noted. The aim of this study was to analyze the messenger RNA (mRNA) and protein expression of VEGF-rs (vascular endothelial growth factor, the VEGFR1, and the VEGFR2 receptor system) and elucidate a possible relationship with the conceptus status (healthy or arrested) on gestational Days (gd) 22 and 32. Both mRNA and protein expression for VEGF-rs molecules were consistently expressed in conceptuses and endometrium during the pregnancy period analyzed. In endometrium, a significant increase in VEGF mRNA and VEGFR2 mRNA expression in healthy sites was observed as pregnancy advances (P < 0.001 and P < 0.05, respectively), whereas VEGFR1 mRNA expression was maintained at a constant level. Interestingly, a significantly elevated VEGFR2 mRNA expression (P < 0.05) was observed on gd 22 in endometrium from arrested conceptuses. Furthermore, VEGF mRNA and VEGFR1 mRNA expression in trophoblasts from healthy conceptuses decreased as pregnancy proceeded (P < 0.001). Arrested trophoblasts on gd 32 showed higher VEGFR2 mRNA expression than healthy conceptuses (P < 0.05). Although, in endometrium attachment sites, the pattern of VEGF-rs immunostaning was not affected by conceptus status, the immunoexpression of VEGF-rs in healthy attachment sites increased slightly but consistently as gestation proceeded. In arresting trophoblasts, VEGF and VEGFR2 staining decreased from gd 22 to 32. Moreover, the number of VEGF and VEGFR1-positive capillaries in the subepithelial vascular plexus of endometrium was related to the conceptus status, showing a moderate increase in healthy sites as pregnancy advances. In conclusion, it appears that VEGF-rs is expressed and related to vascular development in Iberian pigs between gd 22 and 32. The upregulated expression of VEGF mRNA and VEGFR2 mRNA in healthy uterine sites suggests a significant role for these angiogenic factors in early pregnancy. Published by Elsevier Inc.
    Theriogenology 10/2014; · 2.08 Impact Factor
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    ABSTRACT: Cellular senescence has been considered a novel target for cancer therapy. It has also been pointed out that p21cip1/waf1 and p27kip1 cyclin-dependent kinase inhibitors (CKIs) play a role in cellular senescence in some tumor types. Therefore, in order to address the possibility of a cooperative role between p21 and p27 proteins in senescence in vivo we analyzed cellular senescence in spontaneous glandular proliferative lesions (adrenal, thyroid and pituitary glands) in a double-KO mice model, using γH2AX, p53, p16, PTEN and Ki67 as senescence markers. The results obtained showed that p21p27 double-null mice had the lowest number of γH2AX positive cells in glandular hyperplasias and benign tumors. Also, in this group, Ki67 proliferation index correlated with a lower immunohistochemical expression of γH2AX and p53. The expression of p16 and PTEN do not seem to cause synergism of senescence in the benign lesions analyzed in p21p27 double-KO mice. These observations suggest an intrinsic cooperation between p21 and p27 CKIs in the activation of stress-induced cellular senescence and tumor progression in vivo, which would be a physiological mechanism to prevent tumor cell proliferation.
    Histology and histopathology 09/2013; · 2.28 Impact Factor
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    ABSTRACT: Cyclooxygenase-2 (COX-2) has been associated with cell growth regulation, tissue remodeling and carcinogenesis. Overexpression of COX-2 in hepatocytes constitutes an ideal condition to evaluate the role of prostaglandins (PGs) in liver pathogenesis. The effect of COX-2-dependent PGs in genetic hepatocarcinogenesis has been investigated in triple c-myc/transforming growth factor α (TGF-α) transgenic mice that express human COX-2 in hepatocytes on a B6CBAxCD1xB6DBA2 background. Analysis of the contribution of COX-2-dependent PGs to the development of hepatocarcinogenesis, evaluated in this model, suggested a minor role of COX-2-dependent prostaglandins to liver oncogenesis as indicated by liver histopathology, morphometric analysis and specific markers of tumor progression. This allows concluding that COX-2 is insufficient for modifying the hepatocarcinogenesis course mediated by c-myc/TGF-α.
    Prostaglandins & other lipid mediators 04/2013; · 2.42 Impact Factor
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    ABSTRACT: Cyclooxygenase-2 (COX-2) has been associated with cell growth regulation, tissue remodeling and carcinogenesis. Overexpression of COX-2 in hepatocytes constitutes an ideal condition to evaluate the role of prostaglandins (PGs) in liver pathogenesis. The effect of COX-2-dependent PGs in genetic hepatocarcinogenesis has been investigated in triple c-myc/transforming growth factor α (TGF-α) transgenic mice that express human COX-2 in hepatocytes on a B6CBAxCD1xB6DBA2 background. Analysis of the contribution of COX-2-dependent PGs to the development of hepatocarcinogenesis, evaluated in this model, suggested a minor role of COX-2-dependent prostaglandins to liver oncogenesis as indicated by liver histopathology, morphometric analysis and specific markers of tumor progression. This allows concluding that COX-2 is insufficient for modifying the hepatocarcinogenesis course mediated by c-myc/TGF-α.
    Prostaglandins & other lipid mediators 04/2013; · 2.42 Impact Factor
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    ABSTRACT: The present study compares two protocols for ovine estrus synchronization by assessing the caruncular angiogenic response to the establishment of pregnancy. The analysis consisted of the immunohistochemical evaluation of Vascular Endothelial Growth Factor (VEGF), Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1, CD31) and Von Willebrand Factor (vWF) in ovine caruncular stroma. A flock of thirty-eight adult ewes was divided in two groups and synchronized with either progestagens (Group P) or prostaglandin analogues (Group PG). Immunohistochemistry was performed in uterine samples obtained from pregnant ewes (P, n=15; PG, n=13) on days 15 post coitus (pc), 17pc and 21pc (day 0 =day of estrus). Each factor was assessed by total vascular density (TVD, total positive blood vessels/mm2), capillary vascular density (CVD, positive blood capillaries/mm2) and arteriolar vascular density (AVD, positive arterioles/mm2). Group P demonstrated higher VEGF-CVD (P=0.045) when compared to prostaglandin treated animals. Vascular CD31-expression decreased on days 15pc and 21pc (TVD, P=0.007 and CVD, P=0.014) in both groups. vWF analysis did not show significant differences between groups or days of study. These results demonstrate a different influence of progestagen-based and prostaglandin analogues-based synchronization treatments over VEGF vascular expression during caruncular development taking place in response to pregnancy establishment. In addition, observations pointed out in this study support the involvement of CD31 in the angiogenic stimulus that occurs during early maternal placentation in the ewe.
    Histology and histopathology 03/2013; 28(3):373-83. · 2.28 Impact Factor
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    ABSTRACT: The aim of this work was to compare PR, ERα and OTR uterine expression between days 9 and 21 of pregnancy in ewes whose estrus had been synchronized with two different protocols. Sixty-four adult Manchega ewes were synchronized with either conventional progestagens (P) or prostaglandin analogues (PG), and mated. Uterine samples were obtained from pregnant animals (group P, n=24; group PG, n=25) on days 9 post coitus (pc), 13pc, 15pc, 17pc and 21pc. Immunohistochemical detection of progesterone receptor (PR), estrogen receptor-α (ERα) and oxytocin receptor (OTR) was assessed in different uterine cell compartments including luminal and glandular epithelium, stroma and myometrium. Interaction day × treatment was obtained when assessing PR expression in the caruncular stroma (P=0.027) and myometrium (P=0.000), as well as for ERα in the superficial stroma (P=0.05). Significant "day post coitus" effect was found regarding to PR (P<0.01, with the exception of the superficial stroma, deep stroma and myometrium), ERα (P<0.01), and OTR (P<0.05, except in the deep compartments). No significant "treatment" effect was found for PR, ERα or OTR protein immunoexpression. This study supports the implication of PR, ERα and OTR within days 9-21 of the ovine pregnancy. Moreover, different expression pattern of PR and ERα proteins has been found between treatments in various compartments studied. Collectively, these results indicate that PR, ERα and OTR expression during early pregnancy is similar between ewes treated with either progestagens or prostaglandin analogues-based protocols for estrus synchronization.
    Animal reproduction science 06/2012; 133(1-2):93-100. · 1.56 Impact Factor
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    ABSTRACT: Microsporidiosis of adult honeybees caused by Nosema apis and Nosema ceranae is a common worldwide disease with negative impacts on colony strength and productivity. Few options are available to control the disease at present. The role of the queen in bee population renewal and the replacement of bee losses due to Nosema infection is vital to maintain colony homeostasis. Younger queens have a greater egg laying potential and they produce a greater proportion of uninfected newly eclosed bees to compensate for adult bee losses; hence, a field study was performed to determine the effect of induced queen replacement on Nosema infection in honey bee colonies, focusing on colony strength and honey production. In addition, the impact of long-term Nosema infection of a colony on the ovaries and ventriculus of the queen was evaluated. Queen replacement resulted in a remarkable decrease in the rates of Nosema infection, comparable with that induced by fumagillin treatment. However, detrimental effects on the overall colony state were observed due to the combined effects of stressors such as the queenless condition, lack of brood and high infection rates. The ovaries and ventriculi of queens in infected colonies revealed no signs of Nosema infection and there were no lesions in ovarioles or epithelial ventricular cells.
    Environmental Microbiology 11/2011; 14(4):845-59. · 6.24 Impact Factor
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    ABSTRACT: The cell cycle inhibitors p21(Waf1/Cip1) and p27(Kip1) are frequently downregulated in many human cancers, and correlate with a worse prognosis. We show here that combined deficiency in p21 and p27 proteins in mice is linked to more aggressive spontaneous tumorigenesis, resulting in a decreased lifespan. The most common tumors developed in p21p27 double-null mice were endocrine, with a higher incidence of pituitary adenomas, pheochromocytomas and thyroid adenomas. The combined absence of p21 and p27 proteins delays the incidence of radiation-induced thymic lymphomas with a higher apoptotic rate, measured by active caspase-3 and cleaved PARP-1 immunoexpresion. These results provide experimental evidence for a cooperation of both cyclin-dependent kinase inhibitors in tumorigenesis in mice.
    Laboratory Investigation 08/2011; 91(11):1634-42. · 3.96 Impact Factor
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    ABSTRACT: The objective of this study was to investigate differences on the endometrial immunoexpression of type I IFN receptor subunit 1 (IFNAR1) and oxytocin receptor (OTR) during the time of maternal recognition of pregnancy in sheep, when oestrus is synchronized with either prostaglandin analogues (group PG) or conventional progestagens (group P). Plasma progesterone was measured from day 0 to 21 post-coitus (pc) (day 0 = day of oestrus). Immunohistochemistry was performed in samples of uterine horns from pregnant sheep on days 9pc, 13pc, 15pc, 17pc and 21pc to locate IFNAR1 and OTR expression in different endometrial compartments. Mean levels of plasma progesterone were different between treatments, obtaining higher levels in the PG group than in the P group (p < 0.05). Comparing days of pregnancy, IFNAR1 protein expression was different in the luminal epithelium (LE) (p < 0.05), while OTR was different in the LE and in the superficial glandular epithelium (SG) (p < 0.05). Temporal variation on the expression of both proteins from day 9pc to 21pc has been evidenced. IFNAR1 and OTR expression did not show significant differences between treatments. However, the response observed in the endometrium was highly inconsistent when prostaglandin analogues were used. Therefore, the protocol based on prostaglandin analogues still needs to be optimized before being considered as a better alternative to progestagens for oestrous synchronization in sheep.
    Reproduction in Domestic Animals 07/2011; 47(2):274-80. · 1.39 Impact Factor
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    ABSTRACT: The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri-implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen-treated group showed smaller corpora lutea than cloprostenol-treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen-treated group, which even showed a decrease between days 11 and 13. In progestagen-treated sheep, VEGF expression in granulosa-derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.
    Reproduction in Domestic Animals 06/2011; 46(3):481-8. · 1.39 Impact Factor
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    ABSTRACT: The aim of the current study was to determine possible differences in ovarian and pituitary features explaining lower fertility rates in sheep with oestrus induced with intravaginal progestagens or prostaglandin analogues (group FGA and PGF, n=8 in both) when compared to a control group (group C, n=8). The growth profiles and the mean individual sizes of preovulatory follicles were similar between groups; however, the number of preovulatory follicles per ewe and, consequently, the number of ovulations were higher in groups FGA and PGF (2.3±0.3 and 2.0±0.1, respectively) than in group C (1.4±0.1, P<0.05). However, plasma oestradiol concentrations were similar between groups suggesting a defective function in some preovulatory follicles of groups FGA and PGF. In group FGA, the basal LH levels during the follicular phase were lower (0.21±0.0 ng/mL, P<0.005) than in groups C (0.41±0.1 ng/mL) and PGF (0.55±0.1 ng/mL); the onset of preovulatory discharge being later (21.0±2.3h vs. 12.8±1.5 in C and 14.5±1.5 in PGF; P<0.05 for both). Finally, luteal activity was also found to be affected in group FGA; the rate of progesterone secretion per total luteal tissue was lower (range: 0.46-0.65 ng/mL/cm(2)) than in ewes treated with cloprostenol (2.1-3.3 ng/mL/cm(2)) and control sheep (2.0-3.4 ng/mL/cm(2)).
    Animal reproduction science 04/2011; 126(1-2):61-9. · 1.56 Impact Factor
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    ABSTRACT: Cyclooxygenase-2 (COX-2) has been associated with cell growth regulation, tissue remodeling, and carcinogenesis. Ectopic expression of COX-2 in hepatocytes constitutes a nonphysiological condition ideal for evaluating the role of prostaglandins (PGs) in liver pathogenesis. The effect of COX-2-dependent PGs in chronic liver disease, hepatitis, fibrosis, and chemical hepatocarcinogenesis, has been investigated in transgenic (Tg) mice that express human COX-2 in hepatocytes and in Tg hepatic human cell lines. We have used three different complementary approaches: i) diethylnitrosamine (DEN)-induced chemical hepatocarcinogenesis in COX-2 Tg mice, ii) DEN/phenobarbital treatment of human COX-2 Tg hepatocyte-like cells, and iii) COX-2 Tg hepatocyte-like cells implants in nude mice. The data suggest that PGs produced by COX-2 in hepatocytes promoted mild hepatitis in 60-week-old mice, as assessed by histological examination, but failed to contribute to the development of liver fibrogenesis after methionine- and choline-deficient diet treatment. Moreover, liver injury, collagen content, and hepatic stellate cell activation were equally severe in wild-type and COX-2 Tg mice. The contribution of COX-2-dependent PGs to the development of DEN-induced hepatocarcinogenesis was evaluated in Tg mice, Tg hepatocyte-like cells, and nude mice and the analysis revealed that COX-2 expression favors the development of preneoplastic foci without affecting malignant transformation. Endogenous COX-2 expression in wild-type mice is a late event in the development of hepatocellular carcinoma.
    American Journal Of Pathology 03/2011; 178(3):1361-73. · 4.60 Impact Factor
  • Journal of Comparative Pathology. 11/2010; 143(4):335.
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    ABSTRACT: In cerebral malaria, the most severe complication of malaria, both neurotransmission mechanisms and energy metabolism are affected. To understand how metabolic changes modify neurotransmission, we examined P2 receptor expression in a murine model of cerebral malaria. Quantitative polymerase chain reaction experiments revealed that parasite deposition was greatest in the cerebellum, compared with other areas of the brain, suggesting a correlation between brain parasitemia and loss of control of movement. Infected mice showed modified patterns of expression of P2 receptor subtype messenger RNA (mRNA), depending on both the specific purinergic receptor and the cerebral region analyzed. Immunohistochemical studies indicated altered levels of protein expression by these receptors in infected brains and, in some cases, a pattern of expression different from that noted in control mice. These differences in both the amount of mRNA and the protein distribution of P2 receptors observed in the different brain sites in infected mice suggest an important role for P2 receptors in either provoking cerebral damage or conferring neuroprotection.
    The Journal of Infectious Diseases 10/2009; 200(8):1279-88. · 5.85 Impact Factor
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    ABSTRACT: To determine, in a sheep model, the effect of a short-term progestative treatment on growth dynamics and functionality of induced corpora lutea. Observational, model study. Public university. Sixty adult female sheep. Synchronization and induction of ovulation with progestogens and prostaglandin analogues; ovarian ultrasonography, blood sampling, and ovariectomy. Determination of pituitary function and morphologic characteristics, expression of luteinizing hormone (LH) receptors, and progesterone secretion of corpora lutea. The use of progestative pretreatments for assisted conception affect the growth patterns, the expression of LH receptors, and the progesterone secretion of induced corpora lutea. The current study indicates, in a sheep model, the existence of deleterious effects from progestogens on functionality of induced corpora lutea.
    Fertility and sterility 06/2009; 93(4):1308-15. · 3.97 Impact Factor
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    ABSTRACT: The biological cycle of Nosema spp. in honeybees depends on temperature. When expressed as total spore counts per day after infection, the biotic potentials of Nosema apis and N. ceranae at 33 degrees C were similar, but a higher proportion of immature stages of N. ceranae than of N. apis were seen. At 25 and 37 degrees C, the biotic potential of N. ceranae was higher than that of N. apis. The better adaptation of N. ceranae to complete its endogenous cycle at different temperatures clearly supports the observation of the different epidemiological patterns.
    Applied and Environmental Microbiology 03/2009; 75(8):2554-7. · 3.95 Impact Factor
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    ABSTRACT: The importance of transmission factor identification is of great epidemiological significance. The bee-eater (Merops apiaster) is a widely distributed insectivorous bird, locally abundant mainly in arid and semi-arid areas of southern Europe, northern Africa and western Asia but recently has been seen breeding in central Europe and Great Britain. Bee-eaters predominantly eat insects, especially bees, wasps and hornets. On the other hand, Nosema ceranae is a Microsporidia recently described as a parasite in Apis mellifera honeybees in Europe. Due to the short time since its description scarce epidemiological data are available. In this study we investigate the role of the regurgitated pellets of the European bee-eater as fomites of infective spores of N. ceranae. Spore detection in regurgitated pellets of M. apiaster is described [phase-contrast microscopy (PCM) and polymerase chain reaction (PCR) methods]. Eighteen days after collection N. ceranae spores still remain viable and their infectivity is shown after artificial infection of Nosema-free 8-day-old adult bees. The epidemiological consequences of the presence of Nosema spores in this fomites are discussed.
    Environmental Microbiology 06/2008; 10(5):1374-9. · 6.24 Impact Factor
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    ABSTRACT: Nosema ceranae is a Microsporidia recently described as a parasite in Apis mellifera honeybees in Europe. Due to the short time since its description, no epidemiological data are available. In this study, spore detection in both pollen baskets and pollen collected from commercial traps is described (PCM, TEM and PCR methods). Spore infectivity is shown after artificial infection of Nosema-free adult bees. The epidemiological consequences of the presence of Nosema spores in corbicular pollen require more study and must be considered in beekeeping practices.
    Journal of Invertebrate Pathology 02/2008; 97(1):76-8. · 2.67 Impact Factor
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    ABSTRACT: In this report, an experimental infection of Apis mellifera by Nosema ceranae, a newly reported microsporidian in this host is described. Nosema free honeybees were inoculated with 125,000 N. ceranae spores, isolated from heavily infected bees. The parasite species was identified by amplification and sequencing the SSUrRNA gene of the administered spores. Three replicate cages of 20 honeybees each were prepared, along with one control cage (n=20) supplied with sugar syrup only. The infection rate was 100% at the dosage administered. The presence of Nosema inside ventricular cells was confirmed in the samples using ultrathin sectioning and transmission electron microscopy. By day 3 p.i. a few cells (4.4%+/-1.2) were observed to be parasitized, whereas by 6 days p.i. more than half of the counted cells (66.4%+/-6) showed different parasite stages, this value increasing on day 7 p.i. (81.5%+/-14.8). Only one control bee died on day 7 p.i. In the infected groups, mortality was not observed until day 6 p.i. (66.7%+/-5.6). Total mortality on day 7 p.i. was 94.1% in the three infected replicates and by day 8 p.i. no infected bee was alive. After the infection, the parasites invaded both the tip of folds and the basal cells of the epithelium and the autoinfective capacity of the spores seemed to spread the infection rapidly between epithelial cells. On day 3 p.i., mature spores could be seen inside host cell tissue implying that the developmental cycle had been completed. The large number of parasitized cells, even the regenerative ones, the presence of autoinfective spores and the high mortality rate demonstrate that N. ceranae is highly pathogenic to Apis mellifera. Possible relation with bee depopulation syndrome is discussed by authors.
    Journal of Invertebrate Pathology 04/2007; 94(3):211-7. · 2.67 Impact Factor
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    ABSTRACT: The objective of this study was to investigate differences in the expression of estrogen receptor-alpha (ERalpha), progesterone receptor (PR) and the proliferative indexes (Ki-67), in the uterus and oviduct of sheep with estrus synchronized either by prostaglandin analogues (Group PA, n=27) or by treatment with progestagens (Group P, n=29) on days 4 and 7 (day 0=estrus), when the embryos were collected. Immunohistochemical methods were used to quantify ERalpha, PR and Ki-67 in six superficial and deep compartments in the uterus and oviduct. The expression of ERalpha was significantly (P<0.01) lower in progestagen treated ewes than in prostaglandin analogues treated group in the luminal epithelium, superficial glands and superficial stroma in the uterus on day 4. The expression of PR was significantly lower in progesterone treated ewes than in the PA Group in the superficial gland (P<0.05) in both days studied. The lowest expression of PR was observed in the luminal caruncular epithelium and superficial glands in both treatments, obtaining the lowest levels on day 4 (P<0.05). There were significant differences between days 4 and 7 in the Ki-67 immunostaining in the luminal epithelium (P<0.01) and superficial glands (P<0.05). A higher cell proliferation was observed in the uterine epithelium (P<0.05) on day 4 in the animals treated with progestagens. Results indicate that sheep with synchronization of estrus with progestagens showed a reduction of ERalpha and PR protein expression in most of oviductal and uterine cells.
    Animal Reproduction Science 01/2007; 97(1-2):25-35. · 1.90 Impact Factor

Publication Stats

504 Citations
87.14 Total Impact Points

Institutions

  • 2004–2013
    • Complutense University of Madrid
      • • Departamento de Medicina y Cirugía Animal
      • • Facultad de Veterinaria
      • • Departamento de Sanidad Animal
      Pozuelo de Alarcón, Madrid, Spain
  • 2011
    • Austral University of Chile
      • Instituto de Ciencia Animal
      Puerto Montt, Region de Los Lagos, Chile
    • Facultad de Medicina
      Madrid, Madrid, Spain
  • 2009–2011
    • Spanish National Research Council
      • Department of Stellar Physics
      Madrid, Madrid, Spain