Role of p75 Neurotrophin Receptor in the Neurotoxicity by β-amyloid Peptides and Synergistic Effect of Inflammatory Cytokines

The neurodegenerative changes in Alzheimer's disease (AD) are elicited by the accumulation of β-amyloid peptides (Aβ), which damage neurons either directly by interacting with components of the cell surface to trigger cell death signaling or indirectly by activating astrocytes and microglia to produce inflammatory mediators. It has been recently proposed that the p75 neurotrophin receptor (p75NTR) is responsible for neuronal damage by interacting with Aβ. By using neuroblastoma cell clones lacking the expression of all neurotrophin receptors or engineered to express full-length or various truncated forms of p75NTR, we could show that p75NTR is involved in the direct signaling of cell death by Aβ via the function of its death domain. This signaling leads to the activation of caspases-8 and -3, the production of reactive oxygen intermediates and the induction of an oxidative stress. We also found that the direct and indirect (inflammatory) mechanisms of neuronal damage by Aβ could act synergistically. In fact, TNF-α and IL-1β, cytokines produced by Aβ-activated microglia, could potentiate the neurotoxic action of Aβ mediated by p75NTR signaling. Together, our results indicate that neurons expressing p75NTR, mostly if expressing also proinflammatory cytokine receptors, might be preferential targets of the cytotoxic action of Aβ in AD

Nitric Oxide Protects Neuroblastoma Cells from Apoptosis Induced by Serum Deprivation through cAMP-response Element-binding Protein (CREB) Activation

The transcription factor cAMP-response element-binding protein (CREB) mediates survival in many cells, including neurons. Recently, death of cerebellar granule neurons due to nitric oxide (NO) deprivation was shown to be accompanied by down-regulation of CREB activity (1). We now provide evidence that overproduction of endogenous NO or supplementation with exogenous NO renders SK-N-BE human neuroblastoma cells more resistant to apoptosis induced by serum deprivation. Parental cells underwent apoptosis after 24 h of serum deprivation, an outcome largely absent in clones overexpressing human neuronal nitric oxide synthase (nNOS). This protective effect was reversed by the inhibition of NOS itself or soluble guanylyl cyclase, pointing at cGMP as an intermediate effector of NO-mediated rescue. A slow-releasing NO donor protected parental cells to a significant extent, thus confirming the survival effect of NO. The impaired viability of serum-deprived parental cells was accompanied by a strong decrease of CREB phosphorylation and transcriptional activity, effects significantly attenuated in nNOS-overexpressing clones. To confirm the role of CREB in survival, the ectopic expression of CREB and/or protein kinase A largely counteracted serum deprivation-induced cell death of SK-N-BE cells, whereas transfection with a CREB negative mutant was ineffective. These experiments indicate that CREB activity is an important step for NO-mediated survival in neuronal cells

The C-Terminal Domain of CENP-C Displays Multiple and Critical Functions for Mammalian Centromere Formation

CENP-C is a fundamental component of functional centromeres. The elucidation of its structure-function relationship with centromeric DNA and other kinetochore proteins is critical to the understanding of centromere assembly. CENP-C carries two regions, the central and the C-terminal domains, both of which are important for the ability of CENP-C to associate with the centromeric DNA. However, while the central region is largely divergent in CENP-C homologues, the C-terminal moiety contains two regions that are highly conserved from yeast to humans, named Mif2p homology domains (blocks II and III). The activity of these two domains in human CENP-C is not well defined. In this study we performed a functional dissection of C-terminal CENP-C region analyzing the role of single Mif2p homology domains through in vivo and in vitro assays. By immunofluorescence and Chromatin immunoprecipitation assay (ChIP) we were able to elucidate the ability of the Mif2p homology domain II to target centromere and contact alpha satellite DNA. We also investigate the interactions with other conserved inner kinetochore proteins by means of coimmunoprecipitation and bimolecular fluorescence complementation on cell nuclei. We found that the C-terminal region of CENP-C (Mif2p homology domain III) displays multiple activities ranging from the ability to form higher order structures like homo-dimers and homo-oligomers, to mediate interaction with CENP-A and histone H3. Overall, our findings support a model in which the Mif2p homology domains of CENP-C, by virtue of their ability to establish multiple contacts with DNA and centromere proteins, play a critical role in the structuring of kinethocore chromatin

The p75NTR-induced Apoptotic Program Develops through a Ceramide-Caspase Pathway Negatively Regulated by Nitric Oxide

SK-N-BE neuroblastoma cell clones transfected with p75(NTR) and lacking Trk neurotrophin receptors, previously reported to undergo extensive spontaneous apoptosis and to be protected by nerve growth factor (NGF) (Bunone, G., Mariotti, A., Compagni, A., Morandi, E., and Della Valle, G. (1997) Oncogene 14, 1463-1470), are shown to exhibit (i) increased levels of the pro-apoptotic lipid metabolite ceramide and (ii) high activity of caspases, the proteases of the cell death cascade. In the p75(NTR)-expressing cells, these parameters were partially normalized by prolonged NGF treatment, which, in addition, decreased apoptosis, similar to caspase blockers. Conversely, exogenous ceramide increased caspase activity and apoptosis in both wild-type and p75(NTR)-expressing cells. A new p75(NTR)-expressing clone characterized by low spontaneous apoptosis exhibited high endogenous ceramide and low caspase levels. A marked difference between the apoptotic and resistant clones concerned the very low and high activities of nitric-oxide (NO) synthase, respectively. Protection from apoptosis by NO was confirmed by results with the NO donor S-nitrosoacetylpenicillamine and the NO-trapping agent hemoglobin. We conclude that the p75(NTR) receptor, while free of NGF, triggers a cascade leading to apoptosis; the cascade includes generation of ceramide and increased caspase activity; and the protective role of NO occurs at step(s) in between the latter events

MITS: the Multi-Imaging Transient Spectrograph for SOXS

The Son Of X-Shooter (SOXS) is a medium resolution spectrograph R~4500 proposed for the ESO 3.6 m NTT. We present the optical design of the UV-VIS arm of SOXS which employs high efficiency ion-etched gratings used in first order (m=1) as the main dispersers. The spectral band is split into four channels which are directed to individual gratings, and imaged simultaneously by a single three-element catadioptric camera. The expected throughput of our design is >60% including contingency. The SOXS collaboration expects first light in early 2021. This paper is one of several papers presented in these proceedings describing the full SOXS instrument

Optical design of the SOXS spectrograph for ESO NTT

An overview of the optical design for the SOXS spectrograph is presented. SOXS (Son Of X-Shooter) is the new wideband, medium resolution (R>4500) spectrograph for the ESO 3.58m NTT telescope expected to start observations in 2021 at La Silla. The spectroscopic capabilities of SOXS are assured by two different arms. The UV-VIS (350-850 nm) arm is based on a novel concept that adopts the use of 4 ion-etched high efficiency transmission gratings. The NIR (800- 2000 nm) arm adopts the '4C' design (Collimator Correction of Camera Chromatism) successfully applied in X-Shooter. Other optical sub-systems are the imaging Acquisition Camera, the Calibration Unit and a pre-slit Common Path. We describe the optical design of the five sub-systems and report their performance in terms of spectral format, throughput and optical quality. This work is part of a series of contributions describing the SOXS design and properties as it is about to face the Final Design Review.Comment: 9 pages, 9 figures, published in SPIE Proceedings 1070

The VIS detector system of SOXS

SOXS will be a unique spectroscopic facility for the ESO NTT telescope able to cover the optical and NIR bands thanks to two different arms: the UV-VIS (350-850 nm), and the NIR (800-1800 nm). In this article, we describe the design of the visible camera cryostat and the architecture of the acquisition system. The UV-VIS detector system is based on a e2v CCD 44-82, a custom detector head coupled with the ESO continuous ow cryostats (CFC) cooling system and the NGC CCD controller developed by ESO. This paper outlines the status of the system and describes the design of the different parts that made up the UV-VIS arm and is accompanied by a series of contributions describing the SOXS design solutions.Comment: 9 pages, 13 figures, to be published in SPIE Proceedings 1070

The Acquisition Camera System for SOXS at NTT

SOXS (Son of X-Shooter) will be the new medium resolution (R$\sim$4500 for a 1 arcsec slit), high-efficiency, wide band spectrograph for the ESO-NTT telescope on La Silla. It will be able to cover simultaneously optical and NIR bands (350-2000nm) using two different arms and a pre-slit Common Path feeding system. SOXS will provide an unique facility to follow up any kind of transient event with the best possible response time in addition to high efficiency and availability. Furthermore, a Calibration Unit and an Acquisition Camera System with all the necessary relay optics will be connected to the Common Path sub-system. The Acquisition Camera, working in optical regime, will be primarily focused on target acquisition and secondary guiding, but will also provide an imaging mode for scientific photometry. In this work we give an overview of the Acquisition Camera System for SOXS with all the different functionalities. The optical and mechanical design of the system are also presented together with the preliminary performances in terms of optical quality, throughput, magnitude limits and photometric properties.Comment: 9 pages, 7 figures, SPIE conferenc

The Highly Energetic Expansion of SN2010bh Associated with GRB 100316D

We present the spectroscopic and photometric evolution of the nearby (z = 0.059) spectroscopically confirmed type Ic supernova, SN 2010bh, associated with the soft, long-duration gamma-ray burst (X-ray flash) GRB 100316D. Intensive follow-up observations of SN 2010bh were performed at the ESO Very Large Telescope (VLT) using the X-shooter and FORS2 instruments. Owing to the detailed temporal coverage and the extended wavelength range (3000--24800 A), we obtained an unprecedentedly rich spectral sequence among the hypernovae, making SN 2010bh one of the best studied representatives of this SN class. We find that SN 2010bh has a more rapid rise to maximum brightness (8.0 +/- 1.0 rest-frame days) and a fainter absolute peak luminosity (L_bol~3e42 erg/s) than previously observed SN events associated with GRBs. Our estimate of the ejected (56)Ni mass is 0.12 +/- 0.02 Msun. From the broad spectral features we measure expansion velocities up to 47,000 km/s, higher than those of SNe 1998bw (GRB 980425) and 2006aj (GRB 060218). Helium absorption lines He I lambda5876 and He I 1.083 microm, blueshifted by ~20,000--30,000 km/s and ~28,000--38,000 km/s, respectively, may be present in the optical spectra. However, the lack of coverage of the He I 2.058 microm line prevents us from confirming such identifications. The nebular spectrum, taken at ~186 days after the explosion, shows a broad but faint [O I] emission at 6340 A. The light-curve shape and photospheric expansion velocities of SN 2010bh suggest that we witnessed a highly energetic explosion with a small ejected mass (E_k ~ 1e52 erg and M_ej ~ 3 Msun). The observed properties of SN 2010bh further extend the heterogeneity of the class of GRB supernovae.Comment: 37 pages and 12 figures (one-column pre-print format), accepted for publication in Ap