Physical interaction between MYCN oncogene and polycomb repressive complex 2 (PRC2) in neuroblastoma: Functional and therapeutic implications

This article is made available through the Brunel Open Access Publishing Fund. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.CLU (clusterin) is a tumor suppressor gene that we have previously shown to be negatively modulated by the MYCN proto-oncogene, but the mechanism of repression was unclear. Here, we show that MYCN inhibits the expression of CLU by direct interaction with the non-canonical E box sequence CACGCG in the 5′-flanking region. Binding of MYCN to the CLU gene induces bivalent epigenetic marks and recruitment of repressive proteins such as histone deacetylases and Polycomb members. MYCN physically binds in vitro and in vivo to EZH2, a component of the Polycomb repressive complex 2, required to repress CLU. Notably, EZH2 interacts with the Myc box domain 3, a segment of MYC known to be essential for its transforming effects. The expression of CLU can be restored in MYCN-amplified cells by epigenetic drugs with therapeutic results. Importantly, the anticancer effects of the drugs are ablated if CLU expression is blunted by RNA interference. Our study implies that MYC tumorigenesis can be effectively antagonized by epigenetic drugs that interfere with the recruitment of chromatin modifiers at repressive E boxes of tumor suppressor genes such as CLU.SPARKS, The Neuroblastoma Society, a Wellcome Trust grant (to A. S.), and the Italian Association for Cancer Research

Open Source evaluation of kilometric indexes of abundance.

Kilometric Abundance Index (KAI) is a common measure used in wildlife studies because it allows a straightforward comparison of species abundance in different sites or at different times. KAI expresses the ratio of the total number of individuals (or of signs of presence) observed along a transect by the total transect length covered at each site. v.transect.kia is a new tool for GRASS GIS, developed for automating the evaluation of KAI, reducing the risk of manual errors especially when handling large datasets. It can also split the transects according to one environmental variable (typically habitat type) and evaluate true 3D transect length. It calculates KAI using a point map of sightings and saves the results in the attribute table, the output can be displayed in any GIS or used for further statistical analysis. The tool has been tested on field data from Northern Italy for mountain hare (Lepus timidus), allowing a first wide-area estimate

Polymerogenic neuroserpin causes mitochondrial alterations and activates NFκB but not the UPR in a neuronal model of neurodegeneration FENIB

The neurodegenerative condition FENIB (familiar encephalopathy with neuroserpin inclusion bodies) is caused by heterozygous expression of polymerogenic mutant neuroserpin (NS), with polymer deposition within the endoplasmic reticulum (ER) of neurons. We generated transgenic neural progenitor cells (NPCs) from mouse fetal cerebral cortex stably expressing either the control protein GFP or human wild type, polymerogenic G392E or truncated (delta) NS. This cellular model makes it possible to study the toxicity of polymerogenic NS in the appropriated cell type by in vitro differentiation to neurons. Our previous work showed that expression of G392E NS in differentiated NPCs induced an adaptive response through the upregulation of several genes involved in the defence against oxidative stress, and that pharmacological reduction of the antioxidant defences by drug treatments rendered G392E NS neurons more susceptible to apoptosis than control neurons. In this study, we assessed mitochondrial distribution and found a higher percentage of perinuclear localisation in G392E NS neurons, particularly in those containing polymers, a phenotype that was enhanced by glutathione chelation and rescued by antioxidant molecules. Mitochondrial membrane potential and contact sites between mitochondria and the ER were reduced in neurons expressing the G392E mutation. These alterations were associated with a pattern of ER stress that involved the ER overload response but not the unfolded protein response. Our results suggest that intracellular accumulation of NS polymers affects the interaction between the ER and mitochondria, causing mitochondrial alterations that contribute to the neuronal degeneration seen in FENIB patients

Regional Precuneus Cortical Hyperexcitability in Alzheimer's Disease Patients

Objective: Neuronal excitation/inhibition (E/I) imbalance is a potential cause of neuronal network malfunctioning in Alzheimer's disease (AD), contributing to cognitive dysfunction. Here, we used a novel approach combining transcranial magnetic stimulation (TMS) and electroencephalography (EEG) to probe cortical excitability in different brain areas known to be directly involved in AD pathology. Methods: We performed TMS-EEG recordings targeting the left dorsolateral prefrontal cortex (l-DLPFC), the left posterior parietal cortex (l-PPC), and the precuneus (PC) in a large sample of patients with mild-to-moderate AD (n = 65) that were compared with a group of age-matched healthy controls (n = 21). Results: We found that patients with AD are characterized by a regional cortical hyperexcitability in the PC and, to some extent, in the frontal lobe, as measured by TMS-evoked potentials. Notably, cortical excitability assessed over the l-PPC was comparable between the 2 groups. Furthermore, we found that the individual level of PC excitability was associated with the level of cognitive impairment, as measured with Mini-Mental State Examination, and with corticospinal fluid levels of Aβ42 . Interpretation: Our data provide novel evidence that precuneus cortical hyperexcitability is a key feature of synaptic dysfunction in patients with AD. The current results point to the combined approach of TMS and EEG as a novel promising technique to measure hyperexcitability in patients with AD. This index could represent a useful biomarker to stage disease severity and evaluate response to novel therapies. ANN NEUROL 2022

Cortico-cortical stimulation and robot-assisted therapy (CCS and RAT) for upper limb recovery after stroke: study protocol for a randomised controlled trial

Background: Since birth, during the exploration of the environment to interact with objects, we exploit both the motor and sensory components of the upper limb (UL). This ability to integrate sensory and motor information is often compromised following a stroke. However, to date, rehabilitation protocols are focused primarily on recovery of motor function through physical therapies. Therefore, we have planned a clinical trial to investigate the effect on functionality of UL after a sensorimotor transcranial stimulation (real vs sham) in add-on to robot-assisted therapy in the stroke population. Methods: A randomised double-blind controlled trial design involving 32 patients with a single chronic stroke (onset > 180 days) was planned. Each patient will undergo 15 consecutive sessions (5 days for 3 weeks) of paired associative stimulation (PAS) coupled with UL robot-assisted therapy. PAS stimulation will be administered using a bifocal transcranial magnetic stimulator (TMS) on the posterior-parietal cortex and the primary motor area (real or sham) of the lesioned hemisphere. Clinical, kinematics and neurophysiological changes will be evaluated at the end of protocol and at 1-month follow-up and compared with baseline. The Fugl-Meyer assessment scale will be the primary outcome. Secondly, kinematic variables will be recorded during the box-and-block test and reaching tasks using video analysis and inertial sensors. Single pulse TMS and electroencephalography will be used to investigate the changes in local cortical reactivity and in the interconnected areas. Discussion: The presented trial shall evaluate with a multimodal approach the effects of sensorimotor network stimulation applied before a robot-assisted therapy training on functional recovery of the upper extremity after stroke. The combination of neuromodulation and robot-assisted therapy can promote an increase of cortical plasticity of sensorimotor areas followed by a clinical benefit in the motor function of the upper limb. Trial registration: ClinicalTrials.gov NCT05478434. Registered on 28 Jul 2022

COVID-eVax, an electroporated DNA vaccine candidate encoding the SARS-CoV-2 RBD, elicits protective responses in animal models

The COVID-19 pandemic caused by SARS-CoV-2 has made the development of safe and effective vaccines a critical priority. To date, four vaccines have been approved by European and American authorities for preventing COVID-19, but the development of additional vaccine platforms with improved supply and logistics profiles remains a pressing need. Here we report the preclinical evaluation of a novel COVID-19 vaccine candidate based on the electroporation of engineered, synthetic cDNA encoding a viral antigen in the skeletal muscle. We constructed a set of prototype DNA vaccines expressing various forms of the SARS-CoV-2 spike (S) protein and assessed their immunogenicity in animal models. Among them, COVID-eVax—a DNA plasmid encoding a secreted monomeric form of SARS-CoV-2 S protein receptor-binding domain (RBD)—induced the most potent anti-SARS-CoV-2 neutralizing antibody responses (including against the current most common variants of concern) and a robust T cell response. Upon challenge with SARS-CoV-2, immunized K18-hACE2 transgenic mice showed reduced weight loss, improved pulmonary function, and lower viral replication in the lungs and brain. COVID-eVax conferred significant protection to ferrets upon SARS-CoV-2 challenge. In summary, this study identifies COVID-eVax as an ideal COVID-19 vaccine candidate suitable for clinical development. Accordingly, a combined phase I-II trial has recently started

Certainties and uncertainties affecting the postdoctoral phase of the scientific career in Argentina

En la etapa posdoctoral varios desafíos e interrogantes determinan nuestra consolidación como investigadores. Sobre la base de reuniones con ocho investigadores formados, identifcamos algunas certezas e incertidumbres de esta etapa. También destacamos herramientas para lograr buenos antecedentes académicos y poder acceder a la Carrera de Investigador Científco (CIC) del Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). La principal certeza fue que hay que publicar: es fundamental para la trasmisión del conocimiento a la comunidad y es el aspecto más valorado por el CONICET. Sin embargo, publicar implica mucho más que redactar un manuscrito. Es fundamental la contribución conceptual y la originalidad de nuestro trabajo. Identifcamos la incertidumbre que nos genera cómo administrar el tiempo para lograr varias publicaciones de alto impacto y realizar otras actividades como formar recursos humanos, hacer docencia o pedir subsidios o estadías en el exterior, durante los siete años de becas que normalmente disponemos (cinco para doctorarnos y dos de posdoctorado). Otras incertidumbres son el cambio constante y la exigencia creciente de requisitos para ingresar a la CIC. Además, la poca estabilidad de los recursos para investigar y la necesidad de buscar ideas originales son incertidumbres importantes en esta etapa. Quienes quieran desarrollar una carrera científica en la Argentina tendrán que delinear una estrategia. Proponemos perfles de becarios posdoctorales 'exitosos' a fn de encontrar alternativas efcientes para acceder a una posición y un buen desarrollo de la carrera de científco en el CONICET. Para generar un debate enriquecedor, valoraremos su opinión en una encuesta que diseñamos especialmente (goo.gl/forms/k185nQG8ABxjL4bo2, abierta hasta 31 de marzo de 2019). La encuesta está dirigida a becarios doctorales, posdoctorales e investigadores con un cargo permanente, y nos permitirá abrir el abanico de perfles y opiniones. Este debate servirá de guía a jóvenes que quieran desempeñarse exitosamente como científcos en nuestro país.In the postdoctoral stage several challenges and questions define our development and consolidation as researchers. Based on the guidance of trained researchers, we identified various certainties and uncertainties during this phase. We also highlight several tools to achieve a good academic performance and obtain a position as researchers of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). The main certainty was that publishing is a 'must': it is essential to transmit the knowledge to the community and it is the most valued aspect by the CONICET. However, a publication involves much more than writing a manuscript. Detecting the conceptual contribution and the originality of our work is fundamental. One of the uncertainties identified was producing several high-impact publications, while performing other activities (e.g., human resources training, teaching, grant applications and stays abroad) during the seven years that fellowships usually last (five for Ph.D. and two for postdoctoral fellowships). Other uncertainties are the constantly changing rules and the increasing requisites to meet CONICET conditions for researchers. Besides, the scarce stability of resources to research and the need to develop original ideas are particularly uncertain at this stage. Those wanting to develop a scientific career in Argentina must outline a strategy. We propose ‘successful’ postdoctoral fellows’ profiles to find pathways towards a permanent position at the CONICET scientific career, with good development prospects. As a way to open a fruitful debate, your survey feedback will be highly valued. Said survey is available at (goo.gl/forms/k185nQG8ABxjL4bo2, until March 31st, 2019), addressed to doctoral students, postdoctoral fellows and researchers with permanent positions. This will allow opening the range of profiles and opinions. This debate may be a guide for those young researchers willing to perform successfully as Scientists within our country

Functional connectivity studies in migraine: What have we learned?

Background: Resting-state functional connectivity (FC) MRI has widely been used to understand migraine pathophysiology and to identify an imaging marker of the disorder. Here, we review what we have learned from FC studies. Methods: We performed a literature search on the PubMed website for original articles reporting data obtained from conventional resting-state FC recording in migraine patients compared with healthy controls or during and outside of migraine attacks in the same patients. Results: We found 219 articles and included 28 in this review after screening for inclusion and exclusion criteria. Twenty-five studies compared migraine patients with healthy controls, whereas three studies investigated migraine patients during and outside of attacks. In the studies of interictal migraine more alterations of more than 20 FC networks (including amygdala, caudate nucleus, central executive, cerebellum, cuneus, dorsal attention network, default mode, executive control, fronto-parietal, hypothalamus, insula, neostriatum, nucleus accumbens, occipital lobe, periaqueductal grey, prefrontal cortex, salience, somatosensory cortex I, thalamus and visual) were reported. We found a poor level of reproducibility and no migraine specific pattern across these studies. Conclusion: Based on the findings in the present review, it seems very difficult to extract knowledge of migraine pathophysiology or to identify a biomarker of migraine. There is an unmet need of guidelines for resting-state FC studies in migraine, which promote the use of homogenous terminology, public availability of protocol and the a priori hypothesis in line with for instance randomized clinical trial guidelines

Gli scavi dell’Università degli Studi di Napoli “L’Orientale” nell’abitato greco-romano di Cuma (2007-2013)

Sintesi degli scavi nell'abitato greco-romano di Cuma, anni 2007-201