Epstein-Barr virus nuclear antigen 3A protein regulates CDKN2B transcription via interaction with MIZ-1

The Epstein-Barr virus (EBV) nuclear antigen 3 family of protein is critical for the EBV-induced primary B-cell growth transformation process. Using a yeast two-hybrid screen we identified 22 novel cellular partners of the EBNA3s. Most importantly, among the newly identified partners, five are known to play direct and important roles in transcriptional regulation. Of these, the Myc-interacting zinc finger protein-1 (MIZ-1) is a transcription factor initially characterized as a binding partner of MYC. MIZ-1 activates the transcription of a number of target genes including the cell cycle inhibitor CDKN2B. Focusing on the EBNA3A/MIZ-1 interaction we demonstrate that binding occurs in EBV-infected cells expressing both proteins at endogenous physiological levels and that in the presence of EBNA3A, a significant fraction of MIZ-1 translocates from the cytoplasm to the nucleus. Moreover, we show that a trimeric complex composed of a MIZ-1 recognition DNA element, MIZ-1 and EBNA3A can be formed, and that interaction of MIZ-1 with nucleophosmin (NPM), one of its coactivator, is prevented by EBNA3A. Finally, we show that, in the presence of EBNA3A, expression of the MIZ-1 target gene, CDKN2B, is downregulated and repressive H3K27 marks are established on its promoter region suggesting that EBNA3A directly counteracts the growth inhibitory action of MIZ-1

Rubbing induced reversible fluorescence switching in thiophene-based organic semiconductor films by mechanical amorphisation

Here, we applied rubbing on thiophene-basedorganic semiconductor thin films to induce a reversible mechanical amorphisation. Amorphisation is associated with fluorescence switching, which is regulated by the polymorphic nature of the film. Thermal annealing of rubbed films produces an opposite effect with respect to rubbing, inducing film crystallization. Notably, thermal crystallisation starts at a low temperature but generates the polymorph stable at a high temperature in the bulk. The mechanism of mechanical transformation is explained considering the mechanical properties of the material and demonstrated through combined X-ray diffraction, atomic force microscopy and photoluminescence at confocal microscopy. This journal i

Dyads of G-Quadruplex Ligands Triggering DNA Damage Response and Tumour Cell Growth Inhibition at Subnanomolar Concentration

Naphthalene diimide (NDI) dyads exhibiting a different substitution pattern and linker length have been synthesised and evaluated as G-quadruplex (G4) ligands, by investigating their cytotoxicity in selected cell lines. The dyads with the long C7 linker exhibit extremely low IC50 values, below 10\u2005nm, on different cancer cell lines. Contrary, the dyads with the shorter C4 linker were much less effective, with IC values increasing up to 1\u2005\u3bcm. Among the three dyads with the longest linker, small differences in the IC50 values emerge, suggesting that the linker length plays a more important role than the substitution pattern. We have further shown that the dyads are able to induce cellular DNA damage response, which is not limited to the telomeric regions and is likely the origin of their cytotoxicity. Both absorption titration and dynamic light scattering of the most cytotoxic dyads in the presence of hTel22 highlight their ability to induce effective G4 aggregation, acting as non-covalent cross-linking agents

Temporal Tracking of Microglia Activation in Neurodegeneration at Single-Cell Resolution

Microglia, the tissue-resident macrophages in the brain, are damage sensors that react to nearly any perturbation, including neurodegenerative diseases such as Alzheimer's disease (AD). Here, using single-cell RNA sequencing, we determined the transcriptome of more than 1,600 individual microglia cells isolated from the hippocampus of a mouse model of severe neurodegeneration with AD-like phenotypes and of control mice at multiple time points during progression of neurodegeneration. In this neurodegeneration model, we discovered two molecularly distinct reactive microglia phenotypes that are typified by modules of co-regulated type I and type II interferon response genes, respectively. Furthermore, our work identified previously unobserved heterogeneity in the response of microglia to neurodegeneration, discovered disease stage-specific microglia cell states, revealed the trajectory of cellular reprogramming of microglia in response to neurodegeneration, and uncovered the underlying transcriptional programs. Mathys et al. use single-cell RNA sequencing to determine the phenotypic heterogeneity of microglia during the progression of neurodegeneration. They identify multiple disease stage-specific cell states, including two molecularly distinct reactive microglia phenotypes that are typified by modules of co-regulated type I and type II interferon response genes, respectively.National Institutes of Health (U.S.) (Grant RF1 AG054321

The structure of the Shiga toxin 2a A-subunit dictates the interactions of the toxin with blood components

Hemolytic uremic syndrome (eHUS) is a severe complication of human infections with Shiga toxins (Stxs)-producing Escherichia coli. A key step in the pathogenesis of eHUS is the interaction of Stxs with blood components before the targeting of renal endothelial cells. Here, we show that a single proteolytic cleavage in the Stx2a A-subunit, resulting into two fragments (A1 and A2) linked by a disulfide bridge (cleaved Stx2a), dictates different binding abilities. Uncleaved Stx2a was confirmed to bind to human neutrophils and to trigger leukocyte/platelet aggregate formation, whereas cleaved Stx2a was ineffective. Conversely, binding of complement factor H was confirmed for cleaved Stx2a and not for uncleaved Stx2a. It is worth noting that uncleaved and cleaved Stx2a showed no differences in cytotoxicity for Vero cells or Raji cells, structural conformation, and contaminating endotoxin. These results have been obtained by comparing two Stx2a batches, purified in different laboratories by using different protocols, termed Stx2a(cl; cleaved toxin, Innsbruck) and Stx2a(uncl; uncleaved toxin, Bologna). Stx2a(uncl) behaved as Stx2a(cl) after mild trypsin treatment. In this light, previous controversial results obtained with purified Stx2a has to be critically re-evaluated; furthermore, characterisation of the structure of circulating Stx2a is mandatory to understand eHUS-pathogenesis and to develop therapeutic approaches

Surgical management of giant pituitary neuroendocrine tumors: Meta-analysis and consensus statement on behalf of the EANS skull base section.

The optimal surgical treatment for giant pituitary neuroendocrine tumors(GPitNETs) is debated. The aim of this paper is to optimize the surgical management of these patients and to provide a consensus statement on behalf of the EANS Skull Base Section. We constituted a task force belonging to the EANS skull base committee to define some principles for the management of GPitNETs. A systematic review was performed according to PRISMA guidelines to perform a meta-analysis on surgical series of GPitNETs. Weighted summary rates were obtained for the pooled extent of resection and according to the surgical technique. These data were discussed to obtain recommendations after evaluation of the selected articles and discussion among the experts. 20articles were included in our meta-analysis, for a total of 1263 patients. The endoscopic endonasal technique was used in 40.3% of cases, the microscopic endonasal approach in 34% of cases, transcranial approaches in 18.7% and combined approaches in 7% of cases. No difference in terms of gross total resection (GTR) rate was observed among the different techniques. Pooled GTR rate was 36.6%, while a near total resection (NTR) was possible in 45.2% of cases. Cavernous sinus invasion was associated with a lower GTR rate (OR: 0.061). After surgery, 35% of patients had endocrinological improvement and 75.6% had visual improvement. Recurrent tumors were reported in 10% of cases. After formal discussion in the working group, we recommend the treatment of G-PitNETs tumors with a more complex and multilobular structure in tertiary care centers. The endoscopic endonasal approach is the first option of treatment and extended approaches should be planned according to extension, morphology and consistency of the lesion. Transcranial approaches play a role in selected cases, with a multicompartmental morphology, subarachnoid invasion and extension lateral to the internal carotid artery and in the management of residual tumor apoplexy

A Hetero-Bifunctional Spacer for the Smart Engineering of Carbon-Based Nanostructures

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Efforts have been made in recent years to develop novel functionalisation protocols aimed at imparting multimodality and improved properties to complex carbon-based nanostructures. The incorporation of cleavable bonds to the nanomaterial surface for the controlled release (or exchange) of specific molecules under appropriate chemical and biological settings is relatively unexplored. The design and synthesis of a hetero-bifunctional linker joining a "cleavable" disulfide moiety for the covalent anchoring of a wide range of thiol end-capped (bio)molecules and a "clickable" terminal acetylene group is described. The strategy is based on the well-established copper-mediated acetylene-azide coupling reaction between the acetylene linker and single-walled carbon nanotubes decorated with phenylazido pendant arms. As a result, easily "post-derivatisable" and traceable nanostructured platforms containing a linking group potentially available for a wide range of biological probes are prepared and completely characterised. Building on solid foundations: A hetero-bifunctional linker joining a "cleavable" disulfide moiety and a "clickable" terminal acetylene group was synthesized and used to decorate carbon nanotubes (CNTs). When used in combination with other selected terminal acetylene molecules, the linker can impart multimodality through a controlled click reaction to give carbon nanohybrids (see figure)

A Hetero-Bifunctional Spacer for the Smart Engineering of Carbon-Based Nanostructures

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. The cover picture shows a multifunctional platform based on carbon nanotubes, where a modular organic spacer acts as the anchoring site for controlled covalent functionalization of the surface. The combination of fluorescent dyes and post-derivatizable disulfide pendant arms capable of reacting with thiol end-capped (bio)molecules, generates optically traceable (bio)conjugates. The release of a pyridinic dye allows for a precise estimation of the functionalization loading through simple UV/Vis measurements. Details are given in the Full Paper by Giuliano Giambastiani etal. (DOI: 10.1002/cplu.201402391)

Quantitative description of temperature induced self-aggregation thermograms determined by differential scanning calorimetry

A novel thermodynamic approach for the description of differential scanning calorimetry (DSC) experiments on self-aggregating systems is derived and presented. The method is based on a mass action model where temperature dependence of aggregation numbers is considered. The validity of the model was confirmed by describing the aggregation behavior of poly(ethylene oxide)-poly(propylene oxide) block copolymers, which are well-known to exhibit a strong temperature dependence. The quantitative description of the thermograms could be performed without any discrepancy between calorimetric and van 't Hoff enthalpies, and moreover, the aggregation numbers obtained from the best fit of the DSC experiments are in good agreement with those obtained by light scattering experiments corroborating the assumptions done in the derivation of the new model

ALMA-IMF IX: Catalog and Physical Properties of 315 SiO Outflow Candidates in 15 Massive Protoclusters

We present a catalog of 315 protostellar outflow candidates detected in SiO J=5-4 in the ALMA-IMF Large Program, observed with ~2000 au spatial resolution, 0.339 km/s velocity resolution, and 2-12 mJy/beam (0.18-0.8 K) sensitivity. We find median outflow masses, momenta, and kinetic energies of ~0.3 M$_{\odot}$, 4 M$_{\odot}$ km/s, and 10$^{45}$ erg, respectively. Median outflow lifetimes are 6,000 years, yielding median mass, momentum, and energy rates of $\dot{M}$ = 10$^{-4.4}$ M$_{\odot}$ yr$^{-1}$, $\dot{P}$ = 10$^{-3.2}$ M$_{\odot}$ km/s yr$^{-1}$, and $\dot{E}$ = 1 L$_{\odot}$. We analyze these outflow properties in the aggregate in each field. We find correlations between field-aggregated SiO outflow properties and total mass in cores (~3$-$5$\sigma$), and no correlations above 3$\sigma$ with clump mass, clump luminosity, or clump luminosity-to-mass ratio. We perform a linear regression analysis and find that the correlation between field-aggregated outflow mass and total clump mass - which has been previously described in the literature - may actually be mediated by the relationship between outflow mass and total mass in cores. We also find that the most massive SiO outflow in each field is typically responsible for only 15-30% of the total outflow mass (60% upper limit). Our data agree well with the established mechanical force-bolometric luminosity relationship in the literature, and our data extend this relationship up to L $\geq$ 10$^6$ L$_{\odot}$ and $\dot{P}$ $\geq$ 1 M$_{\odot}$ km/s yr$^{-1}$. Our lack of correlation with clump L/M is inconsistent with models of protocluster formation in which all protostars start forming at the same time.Comment: 46 pages, 14 figures, 10 tables. This publication has an associated Zenodo entry, which can be found here: https://zenodo.org/records/835059