The molecular chaperone Hsp90 is a component of the cap-binding complex and interacts with the translational repressor Cup during Drosophila oogenesis

In metazoa, the spatio-temporal translation of diverse mRNAs is essential to guarantee proper oocyte maturation and early embryogenesis. The eukaryotic translation initiation factor 4E (eIF4E), which binds the 5′ cap structure of eukaryotic mRNAs, associates with either stimulatory or inhibitory factors to modulate protein synthesis. In order to identify novel factors that might act at the translational level during Drosophila oogenesis, we have undertaken a functional proteomic approach and isolated the product of the Hsp83 gene, the evolutionarily conserved chaperone Hsp90, as a specific component of the cap-binding complex. Here we report that Hsp90 interacts in vitro with the translational repressor Cup. In addition, we show that Hsp83 and cup interact genetically, since lowering Hsp90 activity enhances the oogenesis alterations linked to diverse cup mutant alleles. Hsp90 and Cup co-localize in the cytoplasm of the developing germ-line cells within the germarium, thus suggesting a common function from the earliest stages of oogenesis. Taken together, our data start elucidating the role of Hsp90 during Drosophila female germ-line development and strengthen the idea that Cup has multiple essential functions during egg chamber development

Treatment with FRAX486 rescues neurobehavioral and metabolic alterations in a female mouse model of CDKL5 deficiency disorder

Introduction: CDKL5 deficiency disorder (CDD) is a rare neurodevelopmental condition, primarily affecting girls for which no cure currently exists. Neuronal morphogenesis and plasticity impairments as well as metabolic dysfunctions occur in CDD patients. The present study explored the potential therapeutic value for CDD of FRAX486, a brain-penetrant molecule that was reported to selectively inhibit group I p21-activated kinases (PAKs), serine/threonine kinases critically involved in the regulation of neuronal morphology and glucose homeostasis.Methods: The effects of treatment with FRAX486 on CDD-related alterations were assessed in vitro (100 nM for 48h) on primary hippocampal cultures from Cdkl5-knockout male mice (Cdkl5-KO) and in vivo (20 mg/Kg, s.c. for 5 days) on Cdkl5-KO heterozygous females (Cdkl5-Het).Results: The in vitro treatment with FRAX486 completely rescued the abnormal neuronal maturation and the number of PSD95-positive puncta in Cdkl5-KO mouse neurons. In vivo, FRAX486 normalized the general health status, the hyperactive profile and the fear learning defects of fully symptomatic Cdkl5-Het mice. Systemically, FRAX486 treatment normalized the levels of reactive oxidizing species in the whole blood and the fasting-induced hypoglycemia displayed by CdklS-Het mice. In the hippocampus of Cdkl5-Het mice, treatment with FRAX486 rescued spine maturation and PSD95 expression and restored the abnormal PAKs phosphorylation at sites which are critical for their activation (P-PAK-Ser144/141/139) or for the control cytoskeleton remodeling (P-PAK1-Thr212).Conclusions: Present results provide evidence that PAKs may represent innovative therapeutic targets for CDD

Communities’ Development in CVEs and Sustaining Functions of On-line Tutorship

Abstract. The impact of Collaborative Virtual Environments (CVEs) on communities ’ development is a topic still rather unexplored. All the interaction processes mediated by these technological environments are characterised, from both psychosocial and discursive points of view, by absolutely new practices and by the absence of some communication features, typical of face-to face. The “Community of Practice ” model, allows us to analyse the growth of virtual communities along the construction of a shared repertoire, the involvement in a common enterprise and the sharing of a mutual engagement. Communities of practice use technology in social and material contexts, sharing meanings about the technology uses and peculiarities through a continuous negotiating process. The analysis is aimed at describing how a community of practice is established in a virtual world where both textual and graphical interactions are allowed. Conversational and quantitative results show: a) how the community of practice evolves over the time, b) the specificity of the interactions mediated by the virtual environment; c) the role of different participants in the management of discourse in interaction, with a detailed analysis of the on-line tutorship impact on the development of the community

Inhibition of p21-activated kinase rescues disrupted phenotype in a mouse model of CDKL5 deficiency disorder

Introduction: CDKL5 Deficiency Disorder (CDD) is a rare neurodevelopmental condition characterized by severe behavioral and physiological symptoms. About 90 % of patients diagnosed with CDD are females. CDD is caused by mutations in the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene. This gene encodes for the homonym serine/threonine kinase, a protein involved in several neurodevelopmental processes, including regulation of spine morphogenesis and synapse formation. To date, no cure is available for this disorder. Mice carrying mutations in the CDKL5 gene have been generated, and represent good models for this disorder as they recapitulate both the behavioral and morphological phenotype disrupted in CDD patients [1], [2]. We recently demonstrated that a CDD mouse model present brain overactivation of group I p21-activated kinases (PAKs), a family of serine/threonine kinases that are critically involved in the regulation of ultrastructural neuronal morphology and activity-dependent actin dynamics [3]. Prolonged activation of group I PAKs affects cognitive function and social behavior in mice and has been associated with several neurologic disorders [4]. Furthermore, recent studies demonstrate that pharmacological inhibition of group I PAKs rescues neurobehavioral defects in preclinical models of Fragile X and schizophrenia [5]. Aim: The present study evaluated whether overactivation of group I PAKs might contribute to the behavioral phenotype observed in CDD, and whether pharmacological inhibition of these kinases might rescue CDD-related behavioral alterations in female mice, the gender most frequently affected by CDD. To this aim, we used Frax486 (supplied by DBA Italia), a brain penetrant PAK inhibitor with an excellent selectivity over group I PAKs. Methods: Cdkl5 heterozygous (Het) female mice were injected daily with FRAX486 (20 mg/kg, s.c. injections, for 5 days) or saline at an advanced stage of the disease (1 year of age). Wild-type (wt) littermates treated with saline were used as controls. A battery of behavioral tests was carried out to evaluate genotype differences and treatment efficacy. Data were analyzed with one way ANOVA. Post-hoc comparisons were performed using the Tukey's HSD. When relevant, repeated measures ANOVA was applied. Results: A 5-day long treatment with FRAX486 at an advanced stage of the disease normalized the general health status (p<0.001 after post hoc comparison on the experimental group: F(1,36)=6.414; p=0.0001) and the hyperactive profile of Cdkl5-Het mice (p<0.01 after post hoc comparison on the experimental group: F(1,35)= 6.414; p= 0.004). It also increased the defective preference for the novel social stimulus of CDD mice in the Three-chamber social test, thus restoring wt-like levels of social novelty (p<0.05 after post hoc comparison on the experimental group: F(1,34)= 3.796; p= 0.032). Conclusion: Present results provide evidence that a 5-day long treatment with FRAX486 rescues relevant behavioral alterations in a CDD female mouse model, suggesting that group I PAK inhibitors may represent an innovative therapeutic strategy for this disorder

Giuseppe Riva Carlo Galimberti (Eds.)

The impact of Collaborative Virtual Environments (CVEs) on communities' development is a topic still rather unexplored. All the interaction processes mediated by these technological environments are characterised, from both psychosocial and discursive points of view, by absolutely new practices and by the absence of some communication features, typical of face-to face. The "Community of Practice" model, allows us to analyse the growth of virtual communities along the construction of a shared repertoire, the involvement in a common enterprise and the sharing of a mutual engagement. Communities of practice use technology in social and material contexts, sharing meanings about the technology uses and peculiarities through a continuous negotiating process

Laboratori di Collaborative Knowledge: sperimentazioni itineranti per il Recupero e la Manutenzione dell’ambiente costruito

La diffusione della cultura del recupero e della manutenzione sul territorio costituisce oggi una strategia per la conservazione e la cura dell’ambiente costruito, fondata su obiettivi di miglioramento nella gestione delle risorse e di supporto dei soggetti coinvolti (PINTO, TALAMO, 2016). La sostenibilità degli interventi dovrebbe essere fondata sulla capacità di promuovere sviluppo locale beneficiando delle qualità di molti edifici abbandonati o in cattivo stato di conservazione, del loro valore culturale, della loro ubicazione in aree urbane centrali o in zone turistiche. Il recupero e la manutenzione costituiscono strategie di azione sostenibili poiché considerano il patrimonio architettonico come risorsa. La conservazione e la fruizione dell’ambiente costruito è dunque una questione economica, oltre che culturale, da perseguire attraverso nuovi modelli di gestione in grado di ottimizzare le risorse finanziarie disponibili e rafforzare i legami esistenti tra patrimonio culturale e persone (PINTO, DE MEDICI E CECERE, 2016)

Laboratori di Collaborative Knowledge: sperimentazioni itineranti per il Recupero e la Manutenzione dell’ambiente costruito

The architectural recovery design, which has always been qualified by a wide involvement of experts in the information-decision process, is characterized by recent experiences in the promotion of collaborative knowledge management tools, based on the cooperation between Researchers, Entrepreneurship, Public Administrations, NGOs and Citizens. These tools start from a critical reformulation of territorial intervention strategies, in line with the new roles of the local community towards objectives of resilience and sustainability. The paper presents the Collaborative Knowledge Laboratories experience promoted by the SITdA Cluster Recovery and Maintenance of the Built Environment, with a specific reference to the so-called slow territories. The Laboratories are intended as ‘itinerant’ experiences, aimed at re-empowering the population and stakeholders to collaborate as local communities, interacting with the public administrations in adopting management and adaptive settlement governance forms, with a co-planning and co-creation concept