Developmental profiles of SUMOylation pathway proteins in rat cerebrum and cerebellum

<div><p>Protein SUMOylation regulates multiple processes involved in the differentiation and maturation of cells and tissues during development. Despite this, relatively little is known about the spatial and temporal regulation of proteins that mediate SUMOylation and deSUMOylation in the CNS. Here we monitor the expression of key SUMO pathway proteins and levels of substrate protein SUMOylation in the forebrain and cerebellum of Wistar rats during development. Overall, the SUMOylation machinery is more highly-expressed at E18 and decreases thereafter, as previously described. All of the proteins investigated are less abundant in adult than in embryonic brain. Furthermore, we show for first time that the profiles differ between cerebellum and cerebrum, indicating differential regional regulation of some of the proteins analysed. These data provide further basic observation that may open a new perspective of research about the role of SUMOylation in the development of different brain regions.</p></div

Laboratory-based spectral data acquisition of roof materials

Roof characteristics such as material type and their properties information are essential to integrating urban agriculture (UA), rainwater harvesting systems (RWHS), and energy systems on roofs. Roof materials can be identified from their spectral signatures. However, this identification requires a priori knowledge of the materials’ spectral characteristics. The main perspective of this work is the future use of spectral data for roof classification. A common practice in mapping materials is the use of spectral libraries. In this regard, this work describes a novel framework for laboratory-based spectral data acquisition. The reflectance data of common, recently introduced (plastics and metals), and representative roof materials from the Mediterranean region were obtained. Data acquisition was conducted in a laboratory under controlled conditions using a high-spatial-resolution (HSR) sensor, which is usually used for airborne surveys. Large variations in the spectral reflectance data were observed due to the composition of the roof material. Flat spectral signatures were found for fibre cement, concrete, gravels and some metals, especially from the near-infrared (NIR) spectral region. Colour and surface finish greatly influence the visible (VIS) spectral range. It was confirmed that the view angle did not modify the spectral shapes. A collection of 39 spectral data of roof materials (ceramics, concrete, fibre cement, metals, plastics, paints, stone, and wood) were compiled into a spectral library that is available online.This work is part of the Fertilecity II project supported by the Spanish Ministry of Economy and Competitiveness (CTM2016-75772-C3-3-R and CTM2016-75772-C3-1-R, AEI/FEDER, UE); from the Spanish Ministry of Science, Innovation and Universities; through the María de Maeztu program for Units of Excellence (MDM-2015-0552). Authors want to thank to the University of Guadalajara (Mexico) for awarding a research scholarship to Perla Zambrano-Prado, and to the reviewers for their valuable comments.Peer ReviewedPostprint (author's final draft

Herpes Simplex Virus type 1 inhibits autophagy in glial cells but requires ATG5 for the success of viral replication

Herpes Simplex Virus type 1 (HSV-1) 1 is a neurotropic virus that has been associated with neurodegenerative disorders. The dysregulation of autophagy by HSV-1 has been proposed as a potential cause of neurodegeneration. While studies have extensively tackled the interaction between autophagy and HSV-1 in neurons, research in glial cells is currently limited. Our studies demonstrate that HSV-1 inhibits, but not completely blocks, the formation of autophagosomes in human oligodendroglioma- and astrocytoma- derived cell lines. These findings have been confirmed in murine oligodendrocyte precursor cells (OPCs). Finally, this study investigates the impact of autophagy on HSV-1 infection in glial cells. While the lack of basal autophagy in LC3B knockout glial cells does not have a significant effect on viral infection, cells without the autophagy-related protein ATG5 exhibit reduced viral production. The absence of ATG5 leads to a decrease in the transcription and replication of viral genes, as well as a delay in the initial stages of the formation of HSV-1 replication compartments. These findings indicate that while autophagy may not play a significant role in antiviral defense in glial cells, HSV-1 may be inhibiting autophagy to exploit non-canonical functions of certain components of the autophagic machinery, such as ATG5, to benefit its lifecycleWe would want to thank Verónica Dominguez (Transgenesis Core Facility, Centro Nacional de Biotecnología, CNB-CSIC, Madrid, Spain) all the help in both the design and development of ATG5 knockout cell lines. We also want to thank Laura Tabero and María Gabriela Atencia (Genomics and NGS Facility, Centro de Biología Molecular Severo Ochoa, CBMSO-CSIC, Madrid, Spain) for all the training received on RT-qPCR. PID2019-109858RB-I00 and PID2022-143110OB-I00 (funded by MCIN/AEI/10.13039/501100011033 FEDER/EU) to FC. BF-G was predoctoral researcher contracted under IND2018/BMD-9751 (Programa de Doctorados Industriales de la Comunidad de Madrid, Spain), to FC. FJ-P was hired under grant CIVP19A5917 from Fundación Ramón Areces (Spain) to F

Functional Heterogeneity of Mouse and Human Brain OPCs: Relevance for Preclinical Studies in Multiple Sclerosis.

Besides giving rise to oligodendrocytes (the only myelin-forming cell in the Central Nervous System (CNS) in physiological conditions), Oligodendrocyte Precursor Cells (OPCs) are responsible for spontaneous remyelination after a demyelinating lesion. They are present along the mouse and human CNS, both during development and in adulthood, yet how OPC physiological behavior is modified throughout life is not fully understood. The activity of adult human OPCs is still particularly unexplored. Significantly, most of the molecules involved in OPC-mediated remyelination are also involved in their development, a phenomenon that may be clinically relevant. In the present article, we have compared the intrinsic properties of OPCs isolated from the cerebral cortex of neonatal, postnatal and adult mice, as well as those recovered from neurosurgical adult human cerebral cortex tissue. By analyzing intact OPCs for the first time with 1H High Resolution Magic Angle Spinning Nuclear Magnetic Resonance (1H HR-MAS NMR) spectroscopy, we show that these cells behave distinctly and that they have different metabolic patterns in function for their stage of maturity. Moreover, their response to Fibroblast Growth Gactor-2 (FGF-2) and anosmin-1 (two molecules that have known effects on OPC biology during development and that are overexpressed in individuals with Multiple Sclerosis (MS)) differs in relation to their developmental stage and in the function of the species. Our data reveal that the behavior of adult human and mouse OPCs differs in a very dynamic way that should be very relevant when testing drugs and for the proper design of effective pharmacological and/or cell therapies for MS.post-print753 K

Vuelo con sensores remotos hiperspectrales para clasificación de cubiertas

Informe del Projecte FERTILECITYSe describe el proceso de captación de las imágenes hiperespectrales en los rangos VisNIR (400-1000 nm) y LWIR (8,5-11 μm) en el área de interés (AOI) que incluye la una parte del Valles Occidental (Badia, Barbera del Valles, Cerdanyola del Valles, Sabadell y Santa Perpetua de la Mogoda) y el proceso posterior de cálculo de radiancia, separación de temperatura y emisividad (LWIR), corrección atmosférica y clasificación de tipo de cubiertas por sus características espectrales

Herpes Simplex Virus type 1 inhibits autophagy in glial cells but requires ATG5 for the success of viral replication

Herpes Simplex Virus type 1 (HSV-1) 1 is a neurotropic virus that has been associated with neurodegenerative disorders. The dysregulation of autophagy by HSV-1 has been proposed as a potential cause of neurodegeneration. While studies have extensively tackled the interaction between autophagy and HSV-1 in neurons, research in glial cells is currently limited. Our studies demonstrate that HSV-1 inhibits, but not completely blocks, the formation of autophagosomes in human oligodendroglioma- and astrocytoma- derived cell lines. These findings have been confirmed in murine oligodendrocyte precursor cells (OPCs). Finally, this study investigates the impact of autophagy on HSV-1 infection in glial cells. While the lack of basal autophagy in LC3B knockout glial cells does not have a significant effect on viral infection, cells without the autophagy-related protein ATG5 exhibit reduced viral production. The absence of ATG5 leads to a decrease in the transcription and replication of viral genes, as well as a delay in the initial stages of the formation of HSV-1 replication compartments. These findings indicate that while autophagy may not play a significant role in antiviral defense in glial cells, HSV-1 may be inhibiting autophagy to exploit non-canonical functions of certain components of the autophagic machinery, such as ATG5, to benefit its lifecycle

Crystal structures of taxane-tubulin complexes: Implications for the mechanism of microtubule stabilization by Taxol

30 p.-9 fig.-1 tab.Paclitaxel (Taxol®) is a first-line chemotherapeutic drug that promotes the curved-to-straight conformational transition of tubulin, an activation step that is necessary for microtubule formation. Crystallization of Taxol bound to tubulin has been long elusive. We found that baccatin III, the core structure of paclitaxel which lacks the C13 side chain, readily co-crystallizes with curved tubulin. Tailor-made taxanes with alternative side chains also co-crystallized, allowing us to investigate their binding modes. Interestingly, these Taxol derived compounds lost their microtubule stabilizing activity and cytotoxicity but kept their full microtubule binding affinity, and all induced lattice expansion upon binding. Additional nuclear magnetic resonance studies propose that Taxol binds to a small fraction of straight tubulin present in solution. Our results suggest a mode of action of Taxol, where the core structure is responsible for the interacting energy while the bulky hydrophobic C13 side chain enables binding selectively to straight tubulin and promotes stabilization.N

Study of exclusive two-photon production of W+W- in pp collisions at √s=7 TeV and constraints on anomalous quartic gauge couplings

A search for exclusive or quasi-exclusive W+W- production by photon-photon interactions, pp -> p(*)W(+)W(-)p(*), at \/s = 7 TeV is reported using data collected by the CMS detector with an integrated luminosity of 5.05 fb(-1). Events are selected by requiring a mu(+/-)mu(-/+) vertex with no additional associated charged tracks and dilepton transverse momentum p(T)(mu(+/-)mu(-/+)) > 30 GeV. Two events passing all selection requirements are observed in the data, compared to a standard model expectation of 2.2 +/- 0.4 signal events with 0.84 +/- 0.15 background. The tail of the dilepton p(T) distribution is studied for deviations from the standard model. No events are observed with p(T) > 100 GeV. Model-independent upper limits are computed and compared to predictions involving anomalous quartic gauge couplings. The limits on the parameters a(0,c)(W)/A(2) with a dipole form factor and an energy cutoff A(cutoff) = 500 GeV are of the order of 10(-4)

Identification of pyrrolopyrimidine derivative PP-13 as a novel microtubule-destabilizing agent with promising anticancer properties

14 p.-6 fig-.1 tab. Gilson, Pauline et al.Despite the emergence of targeted therapies and immunotherapy, chemotherapy remains the gold-standard for the treatment of most patients with solid malignancies. Spindle poisons that interfere with microtubule dynamics are commonly used in chemotherapy drug combinations. However, their troublesome side effects and the emergence of chemoresistance highlight the need for identifying alternative agents. We performed a high throughput cell-based screening and selected a pyrrolopyrimidine molecule (named PP-13). In the present study, we evaluated its anticancer properties in vitro and in vivo. We showed that PP-13 exerted cytotoxic effects on various cancer cells, including those resistant to current targeted therapies and chemotherapies. PP-13 induced a transient mitotic blockade by interfering with both mitotic spindle organization and microtubule dynamics and finally led to mitotic slippage, aneuploidy and direct apoptotic death. PP-13 was identified as a microtubule-targeting agent that binds directly to the colchicine site in β-tubulin. Interestingly, PP-13 overcame the multidrug-resistant cancer cell phenotype and significantly reduced tumour growth and metastatic invasiveness without any noticeable toxicity for the chicken embryo in vivo. Overall, PP-13 appears to be a novel synthetic microtubule inhibitor with interesting anticancer properties and could be further investigated as a potent alternative for the management of malignancies including chemoresistant ones.This work was supported by the Institut Curie, the CNRS, the INSERM, and by grants from La Fondation de France, La Ligue contre le Cancer (comité de l’Isère), Cancéropôle CLARA (Lyon Auvergne Rhône-Alpes),and BFU2016–75319-R (AEI/FEDER, UE) from Ministerio de Economia y Competitividad (JFD).Peer reviewe

SUMOylation of Argonaute-2 regulates RNA interference activity

AbstractPost-translational modification of substrate proteins by small ubiquitin-like modifier (SUMO) regulates a vast array of cellular processes. SUMOylation occurs through three sequential enzymatic steps termed E1, E2 and E3. Substrate selection can be determined through interactions between the target protein and the SUMO E2 conjugating enzyme Ubc9 and specificity can be enhanced by substrate interactions with E3 ligase enzymes. We used the putative substrate recognition (PINIT) domain from the SUMO E3 PIAS3 as bait to identify potential SUMO substrates. One protein identified was Argonaute-2 (Ago2), which mediates RNA-induced gene silencing through binding small RNAs and promoting degradation of complimentary target mRNAs. We show that Ago2 can be SUMOylated in mammalian cells by both SUMO1 and SUMO2. SUMOylation occurs primarily at K402, and mutation of the SUMO consensus site surrounding this lysine reduces Ago2-mediated siRNA-induced silencing in a luciferase-based reporter assay. These results identify SUMOylation as a potential regulator of Ago2 activity and open new avenues for research into the mechanisms underlying the regulation of RNA-induced gene silencing