Activation of p53 by Nutlin-3a Induces Apoptosis and Cellular Senescence in Human Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor in adults. Despite concerted efforts to improve current therapies and develop novel clinical approaches, patient survival remains poor. As such, increasing attention has focused on developing new therapeutic strategies that specifically target the apoptotic pathway in order to improve treatment responses. Recently, nutlins, small-molecule antagonists of MDM2, have been developed to inhibit p53-MDM2 interaction and activate p53 signaling in cancer cells. Glioma cell lines and primary cultured glioblastoma cells were treated with nutlin-3a. Nutlin-3a induced p53-dependent G1- and G2-M cell cycle arrest and apoptosis in glioma cell lines with normal TP53 status. In addition, nutlin-arrested glioma cells show morphological features of senescence and persistent induction of p21 protein. Furthermore, senescence induced by nutlin-3a might be depending on mTOR pathway activity. In wild-type TP53 primary cultured cells, exposure to nutlin-3a resulted in variable degrees of apoptosis as well as cellular features of senescence. Nutlin-3a-induced apoptosis and senescence were firmly dependent on the presence of functional p53, as revealed by the fact that glioblastoma cells with knockdown p53 with specific siRNA, or cells with mutated or functionally impaired p53 pathway, were completely insensitive to the drug. Finally, we also found that nutlin-3a increased response of glioma cells to radiation therapy. The results provide a basis for the rational use of MDM2 antagonists as a novel treatment option for glioblastoma patients

Hazte un cell-fie

[EN] The objective of this project is to develop inclusive scientific workshops which combine promoting "scientific literacy" in primary schools and addressing the training needs of university students through the Service-Learning methodology. The project involved the participation of 50 students of Pharmacy (1st year) and Primary Education (3rd year) at San Jorge University, along with 6 primary school teachers and 75 primary schoolchildren (2nd year) from various schools in Zaragoza. The project was divided into four phases. The first one focused on the search of specific information by all students involved and the development of learning materials to organize all the content related to cells. The second phase took the form of a cooperative seminar for students of both degree programmes. During the seminar, Education students received specific training (which allowed them to clarify and expand their knowledge about cell biology) and taught Pharmacy students how to adapt scientific language to a non-specialized audience (didactic transposition). The third phase was dedicated to creating informative materials (videotutorial recording, construction of cell models and design of a microscopy workshop for children). As a fourth phase of the project, a series of practical workshops were held with different groups of primary schoolchildren, using the materials developed in the previous phase. The activity was very highly rated by both Pharmacy and Education students whereas the workshops had a very positive impact on schoolchildren. In this respect, 98.5% of the children liked the activity and 93% indicated that they had learned things they did not know about cells.[ES] Este proyecto tiene como objetivo desarrollar talleres científicos inclusivos destinados a promover la “alfabetización científica” en centros de Educación Primaria a la vez que se satisfacen las necesidades formativas de estudiantes universitarios a través de la metodología de Aprendizaje-Servicio. En el proyecto han participado 50 estudiantes de 1º del grado en Farmacia y de 3º del grado en Educación Primaria de la Universidad San Jorge, junto con 6 maestros y 75 estudiantes de 2º de Educación Primaria en varios colegios de Zaragoza. El proyecto se estructuró en cuatro fases. La primera se concretó en la búsqueda de información específica por parte de los estudiantes de ambos grados y el desarrollo de materiales de aprendizaje para organizar toda los contenidos relativos a las células. La segunda fase se materializó en un seminario cooperativo entre discentes de ambos grados en el cual los estudiantes de educación recibieron formación específica (que les permitió aclarar y ampliar sus conocimientos sobre biología celular) a la vez que enseñaron a los estudiantes de farmacia a adaptar el lenguaje científico para un público no especializado (transposición didáctica). La tercera fase se enfocó en la creación de materiales divulgativos (grabación de videotutoriales, construcción de maquetas celulares y diseño de un taller de microscopía para niños). Como cuarta fase del proyecto, se llevaron a cabo una serie de talleres prácticos con diversos grupos de estudiantes de 2º de Educación Primaria, utilizando los materiales desarrollados en la fase anterior.Tanto los estudiantes del grado en Farmacia como los estudiantes del Grado en Educación valoraron de forma muy positiva la actividad. Por otro lado, los talleres realizados tuvieron un impacto muy positivo en los escolares así un 98,5% de los mismos indicó que la actividad les había gustado mucho y un 93% que habían aprendido cosas que no sabían sobre las células.Gómez Rincón, C.; Langa, E.; Verón, JJ.; Uriel, M.; Acebes, D.; Woźniak, M.; Terrado, EM. (2019). Hazte un cell-fie. En IN-RED 2019. V Congreso de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 353-363. https://doi.org/10.4995/INRED2019.2019.10352OCS35336

The nuclear receptor LXRα controls the functional specialization of splenic macrophages.

Macrophages are professional phagocytic cells that orchestrate innate immune responses and have considerable phenotypic diversity at different anatomical locations. However, the mechanisms that control the heterogeneity of tissue macrophages are not well characterized. Here we found that the nuclear receptor LXRα was essential for the differentiation of macrophages in the marginal zone (MZ) of the spleen. LXR-deficient mice were defective in the generation of MZ and metallophilic macrophages, which resulted in abnormal responses to blood-borne antigens. Myeloid-specific expression of LXRα or adoptive transfer of wild-type monocytes restored the MZ microenvironment in LXRα-deficient mice. Our results demonstrate that signaling via LXRα in myeloid cells is crucial for the generation of splenic MZ macrophages and identify an unprecedented role for a nuclear receptor in the generation of specialized macrophage subsets

Neutrophils instruct homeostatic and pathological states in naive tissues

Immune protection relies on the capacity of neutrophils to infiltrate challenged tissues. Naive tissues, in contrast, are believed to remain free of these cells and protected from their toxic cargo. Here, we show that neutrophils are endowed with the capacity to infiltrate multiple tissues in the steady-state, a process that follows tissue-specific dynamics. By focusing in two particular tissues, the intestine and the lungs, we find that neutrophils infiltrating the intestine are engulfed by resident macrophages, resulting in repression of Il23 transcription, reduced G-CSF in plasma, and reinforced activity of distant bone marrow niches. In contrast, diurnal accumulation of neutrophils within the pulmonary vasculature influenced circadian transcription in the lungs. Neutrophil-influenced transcripts in this organ were associated with carcinogenesis and migration. Consistently, we found that neutrophils dictated the diurnal patterns of lung invasion by melanoma cells. Homeostatic infiltration of tissues unveils a facet of neutrophil biology that supports organ function, but can also instigate pathological states.S

A Neutrophil Timer Coordinates Immune Defense and Vascular Protection

Neutrophils eliminate pathogens efficiently but can inflict severe damage to the host if they over-activate within blood vessels. It is unclear how immunity solves the dilemma of mounting an efficient anti-microbial defense while preserving vascular health. Here, we identify a neutrophil-intrinsic program that enabled both. The gene Bmal1 regulated expression of the chemokine CXCL2 to induce chemokine receptor CXCR2-dependent diurnal changes in the transcriptional and migratory properties of circulating neutrophils. These diurnal alterations, referred to as neutrophil aging, were antagonized by CXCR4 (C-X-C chemokine receptor type 4) and regulated the outer topology of neutrophils to favor homeostatic egress from blood vessels at night, resulting in boosted anti-microbial activity in tissues. Mice engineered for constitutive neutrophil aging became resistant to infection, but the persistence of intravascular aged neutrophils predisposed them to thrombo-inflammation and death. Thus, diurnal compartmentalization of neutrophils, driven by an internal timer, coordinates immune defense and vascular protection.We thank all members of the Hidalgo Lab for discussion and insightful comments; J.M. Ligos, R. Nieto, and M. Viton for help with sorting and cytometric analyses; I. Ortega and E. Santos for animal husbandry; D. Rico, M.J. Gomez, C. Torroja, and F. Sanchez-Cabo for insightful comments and help with transcriptomic analyses; V. Labrador, E. Arza, A.M. Santos, and the Microscopy Unit of the CNIC for help with microscopy; S. Aznar-Benitah, U. Albrecht, Q.-J. Meng, B. Staels, and H. Duez for the generous gift of mice; J.A. Enriquez and J. Avila for scientific insights; and J.M. Garcia and A. Diez de la Cortina for art. This study was supported by Intramural grants from A* STAR to L.G.N., BES-2013-065550 to J.M.A., BES-2010-032828 to M.C.-A, and JCI-2012-14147 to L.A.W (all from Ministerio de Economia, Industria y Competitividad; MEIC). Additional MEIC grants were SAF2014-61993-EXP to C.L.-R.; SAF2015-68632-R to M.A.M. and SAF-2013-42920R and SAF2016-79040Rto D.S. D.S. also received 635122-PROCROP H2020 from the European Commission and ERC CoG 725091 from the European Research Council (ERC). ERC AdG 692511 PROVASC from the ERC and SFB1123-A1 from the Deutsche Forschungsgemeinschaft were given to C.W.; MHA VD1.2/81Z1600212 from the German Center for Cardiovascular Research (DZHK) was given to C.W. and O.S.; SFB1123-A6 was given to O.S.; SFB914-B08 was given to O.S. and C.W.; and INST 211/604-2, ZA 428/12-1, and ZA 428/13-1 were given to A.Z. This study was also supported by PI12/00494 from Fondo de Investigaciones Sanitarias (FIS) to C.M.; PI13/01979, Cardiovascular Network grant RD 12/0042/0054, and CIBERCV to B.I.; SAF2015-65607-R, SAF2013-49662-EXP, and PCIN-2014-103 from MEIC; and co-funding by Fondo Europeo de Desarrollo Regional (FEDER) to A.H. The CNIC is supported by the MEIC and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (MEIC award SEV-2015-0505).S

Phagocytosis imprints heterogeneity in tissue-resident macrophages

Tissue-resident macrophages display varying phenotypic and functional properties that are largely specified by their local environment. One of these functions, phagocytosis, mediates the natural disposal of billions of cells, but its mechanisms and consequences within living tissues are poorly defined. Using a parabiosis-based strategy, we identified and isolated macrophages from multiple tissues as they phagocytosed blood-borne cellular material. Phagocytosis was circadianally regulated and mediated by distinct repertoires of receptors, opsonins, and transcription factors in macrophages from each tissue. Although the tissue of residence defined the core signature of macrophages, phagocytosis imprinted a distinct antiinflammatory profile. Phagocytic macrophages expressed CD206, displayed blunted expression of Il1b, and supported tissue homeostasis. Thus, phagocytosis is a source of macrophage heterogeneity that acts together with tissue-derived factors to preserve homeostasis

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Sympatric guanacos and livestock share water resources in drylands of Argentina

There is growing concern about the effect of livestock on wild ungulate populations, particularly in arid ecosystems, where waterholes are an extremely scarce resource, around which animals tend to gather, primarily in the dry season. This situation is worrying in South American deserts, where guanaco is the native species that often shares trophic and water resources with livestock from local communities, even inside protected areas. We assess through general linear modeling (GLM) the use of waterholes by guanaco and two introduced species, free-ranging cattle and feral donkeys, during the summer-wet and winter-dry seasons, in an arid, water-limited region in northwestern Argentina. Waterholes were more intensively used in the dry than the wet season by all three herbivores. However, introduced ungulates did not use all of the waterholes, whereas guanaco used them all with equal intensity, which points to an apparent absence of interference probably due to the low density of the introduced species. Nevertheless these results could mask negative effects regarding the risk of parasite transmission, the under-use of one of the waterholes, and the risk related to increasing livestock density in a near future. Therefore, it would be advisable to make long-term monitoring to prevent potentially negative effects on guanacos.Fil: Acebes, Pablo. Universidad Autónoma de Madrid; EspañaFil: Malo, Juan E.. Universidad Autónoma de Madrid; EspañaFil: Ovejero Aguilar, Ramiro Jose Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Traba, Juan. Universidad Autónoma de Madrid; Españ

Density and habitat use at different spatial scales of a guanaco population (Lama guanicoe) in the Monte desert of Argentina

The first density estimates of a peripheral guanaco population and its habitat use at different spatial scales are presented for a protected area of Monte desert, Argentina. Transects were surveyed in the wet and dry seasons of 2005. All guanaco herds observed during systematic surveys using roads and tracks were GPS located and their habitat use was identified. Herd size differed significantly between the dry and wet seasons. Population densities differed between wet (0.10-0.12 individuals/km2) and dry seasons (0.60-0.75 individuals/km2). The population estimates ranged from 75 individuals (dry season) to 388 individuals (wet season). Guanacos showed differential habitat use, the first determinant being abiotic factors, such as topography, soil characteristics or microclimate conditions, animals being detected in rougher rocky substrata in the dry season and in open flat terrain in the wet season, followed by a mesoscale selection defined by plant communities. At the latter scale, guanaco preferentially used mixed creosote bushland and saltbush more intensively during the wet season, and open scrub and columnar-cactus slopes in the dry season. The estimated population of this protected area was small but its population density was within the range of other populations and was relatively high for this dry and unproductive area.Fil: Acebes, Pablo. Universidad Autónoma de Madrid; EspañaFil: Traba, Juan. Universidad Autónoma de Madrid; EspañaFil: Malo, Juan E.. Universidad Autónoma de Madrid; EspañaFil: Ovejero Aguilar, Ramiro Jose Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Provincia de Mendoza. Instituto Argentino de Investigaciones de las Zonas Áridas. Universidad Nacional de Cuyo. Instituto Argentino de Investigaciones de las Zonas Áridas; ArgentinaFil: Borghi, Carlos Eduardo. Universidad Nacional de San Juan. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto y Museo de Ciencias Naturales; Argentin