Eastern European parents’ experiences of parenting a child with SEN in England

Parenting a child with Special Educational Needs (SEN) presents numerous challenges for families. For immigrant parents, these challenges can be particularly difficult to overcome when faced with structural, cultural and linguistic barriers. This qualitative study explored the lived experiences of eight Eastern European immigrants parenting a child with SEN in England. Semi-structured interviews were conducted, and a data-driven thematic analysis of a series of interviews was carried out. The study identified two key themes: (a) embarking on an unpredicted journey and (b) navigating through challenges. The analyses highlight discrepancies in partnership working between parents and educators and shortcomings in advice that professionals provided to these parents, potentially placing pupils and their families at a disadvantage. The implications for educational psychologists (EPs) and other professionals working with Eastern European parents raising a child with SEN are also discussed

The Parkinson-related E193K LRRK2 variant impacts neuronal vesicles dynamics through perturbed protein interactions

The Leucine-Rich Repeat Kinase 2 (LRRK2) is a complex protein, expressed in neurons and implicated in Parkinson disease (PD). LRRK2 contains a dual enzymatic activity and several structural domains that constitute a versatile platform for multiple protein interactions at the synapses. In this study, we characterize the functional role of the N-terminal Armadillo repeats domain of LRRK2 and the impact on synaptic vesicle (SV) dynamics of a novel variant, E193K, harboured within this domain and identified in an Italian family affected by PD. Using a genetically encoded sensor of recycling, synaptopHluorine, and total internal reflection fluorescence microscopy, we visualized SV trafficking in the N2A neuroblastoma cells expressing the wild type LRRK2 protein, a mutant lacking the Armadillo domain (\u394N LRRK2) or the E193K variant. We found that expression of the \u394N construct increased the frequency and the amplitude of spontaneous synaptic events. A similar phenotype was detected in the presence of the E193K variant, suggesting that this mutation behaves as a loss-of-function mutation. A domain-based pulldown approach demonstrated that the LRRK2 N-terminus binds to cytoskeletal (\u3b2-actin and \u3b1-tubulin) and SV (synapsin I) proteins and the E193K substitution alters strength and quality of LRRK2 interactions. The results support a role of the Armadillo domain in interaction with synaptic proteins and suggest that the E193K mutation affects LRRK2 function via perturbation of its physiological network of interactors, resulting in impaired vesicular trafficking. These findings may have important implications for understanding the role of LRRK2 at the synapses and the pathophysiological mechanism for LRRK2-linked disease

Mechanotransduction in human and mouse beta cell lines: reliable models to characterize novel signaling pathways controlling beta cell fate

Background and aims: Attempts to influence \u3b2-cell differentiation by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the fact that profound details of mechanosensing/transduction complexity remain elusive. We recently demonstrated that human islets of Langerhans sense the ECM nanotopography and activate a mechanotransductive pathway, which is essential for preserving long-term \u3b2-cell differentiation and function in vitro. However, human islets of Langerhans are extremely heterogeneous and their availability for research purpose is limited. Therefore, aim of the proposed research was to investigate whether mouse and human \u3b2-cell lines might sense changes innthe ECM topography and might be used as a simplified model to dissect the molecular pathways involved in mechanotransduction. Materials and methods: We used supersonic cluster beam deposition to fabricate nanostructured substrates characterized by a quantitatively controllable ECM-like nanoroughness. Mouse \u3b2TC3 and human 1.1B4 cells were seeded on these substrates and after five days in culture, the activation of the mechanotransductive pathway was verified by means of morphological (super-resolution fluorescence microscopy), functional and proteomic techniques. Results: Quantitative immunofluorescence studies demonstrated that the cell-nanotopography interaction affects the focal adhesion structures (smaller vinculin clusters), the organization of the actin cytoskeleton (shorter actin fiber) and the nuclear architecture. Functional studies revealed that nanostructured surfaces improve the \u3b2-cell mitochondrial activity and increase the glucose-stimulated Ca2+currents and insulin release. Label-free shotgun proteomics broadly confirmed the morphological and functional studies and showed the upregulation of a number of mechanosensors and transcription factors involved in \u3b2-cell differentiation in cells grown on nanostructured substrates compared to those grown on flat standard control surfaces. Conclusion: Our data reveal that mouse and human \u3b2-cell lines sense changes in extracellular mechanical forces and activate a mechanotransductive pathway. The findings from this study will be useful to clarify the link between mechanotransduction and cell fate and to successfully engineer scaffolds in order to have functional beta cells

Proteomic Analysis Reveals a Mitochondrial Remodeling of βTC3 Cells in Response to Nanotopography

Recently, using cluster-assembled zirconia substrates with tailored roughness produced by supersonic cluster beam deposition, we demonstrated that \u3b2 cells can sense nanoscale features of the substrate and can translate these stimuli into a mechanotransductive pathway capable of preserveing \u3b2-cell differentiation and function in vitro in long-term cultures of human islets. Using the same proteomic approach, we now focused on the mitochondrial fraction of \u3b2TC3 cells grown on the same zirconia substrates and characterized the morphological and proteomic modifications induced by the nanostructure. The results suggest that, in \u3b2TC3 cells, mitochondria are perturbed by the nanotopography and activate a program involving metabolism modification and modulation of their interplay with other organelles. Data were confirmed in INS1E, a different \u3b2-cell model. The change induced by the nanostructure can be pro-survival and prime mitochondria for a metabolic switch to match the new cell needs

DECIPHERING THE ROLE OF THE SERINE THREONINE KINASE LRRK2 IN THE CENTRAL NERVOUS SYSTEM AND IN PERIPHERAL TISSUES: IMPLICATION FOR PARKINSON\ubfS DISEASE TREATMENT

Leucine-rich repeat kinase 2 (LRRK2) encodes a large and complex protein which is widely expressed in brain and peripheral organs. Mutations in LRRK2 are a major cause of inherited and sporadic Parkinson\u2019s disease (PD), an age-dependent neurodegenerative disorder characterized by neuronal damage in multiple brain regions and consequent motor defects. Studies performed in the Central Nervous System (CNS) support a role of the protein in different functions, but despite extensive studies, the pathophysiological role of LRRK2 still remains enigmatic. LRRK2 encompasses two enzymatic domains (a kinase and GTPase domain) and multiple protein-protein interaction modules that can scaffold several protein complexes. Therefore, defining the LRRK2 interactome and understanding how its composition is modulated can be useful to identify its function. Since LRRK2 is notably involved in vesicular trafficking, the first aim for the proposed study was to investigate the role of the N-terminal domain of LRRK2 as a scaffold and its functional impact on synaptic vesicle (SV) dynamics in the N2a neuroblastoma cell line. High resolution fluorescence microscopy techniques coupled to genetically encoded sensors of SV fusion and recycling, demonstrated that the N-terminal domain is crucial for LRRK2 targeting to membrane structures and for its interaction with proteins involved in the control of SV mobilization and recycling. Our data confirm LRRK2 as a component of the presynaptic protein network, coordinating both storage and mobilization of SV through its N-terminal domain. Mutations in this domain (PD-linked E193K variant studied in this work) or modulation of its phosphorylation (in particular at serine 935) affects LRRK2 function via perturbation of its physiological network of interactors, thus resulting in impaired SV dynamics (Chapter I). In addition to being expressed in the CNS, LRRK2 can be detected in a variety of peripheral organs, including the endocrine pancreas, thus leading to the suggestion that the protein may be implicated in other functions. Therefore, in the second part of the project we focused on the endocrine pancreas and explored the possible involvement of LRRK2 in the control of insulin release and glucose homeostasis, using both cellular and animal models. Through pharmacological, molecular, and functional approaches we demonstrated that LRRK2 and its kinase activity control the glucose-stimulated insulin secretion, by modulating the trafficking of insulin granules. Expression of PD-associated LRRK2 mutants, characterized by enhanced kinase-activity, increased basal insulin release, and decreased the glucose-stimulated secretion, thus affecting the animal\u2019s metabolic profile. These findings identify a new role of LRRK2 in the control of glucose homeostasis and support the idea that LRRK2 may contribute to PD development and/or progression also indirectly through modulation of glucose homeostasis and cellular energetics. Not surprisingly, drugs used in type 2 diabetes and intended to restore the glucose homeostasis are among the most promising treatments currently being considered as a possible new therapy for PD (Chapter II). In the complex, our data indicate LRRK2 as a master regulator of secretory vesicles trafficking in both neuronal and endocrine cells of the pancreas and suggest the protein as important therapeutic target in multiple diseases

Malattie virali dell’apparato respiratorio

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Malattie virali dell’apparato respiratorio

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Eastern European parents' experiences of parenting a child with SEN in England

Parenting a child with Special Educational Needs (SEN) presents numerous challenges for families. For immigrant parents, these challenges can be particularly difficult to overcome when faced with structural, cultural and linguistic barriers. This qualitative study explored the lived experiences of 8 Eastern European immigrants parenting a child with SEN in England. Semi-structured interviews were conducted, and a data-driven thematic analysis of a series of interviews was carried out. The study identified two key themes: (a) embarking on an unpredicted journey and (b) navigating through challenges. The analyses highlight discrepancies in partnership working between parents and educators and shortcomings in advice that professionals provided to these parents, potentially placing pupils and their families at a disadvantage. The implications for educational psychologists (EP) and other professionals working with Eastern European parents raising a child with SEN are also discussed

Eastern European parents' experiences of parenting a child with SEN in England

Parenting a child with Special Educational Needs (SEN) presents numerous challenges for families. For immigrant parents, these challenges can be particularly difficult to overcome when faced with structural, cultural and linguistic barriers. This qualitative study explored the lived experiences of 8 Eastern European immigrants parenting a child with SEN in England. Semi-structured interviews were conducted, and a data-driven thematic analysis of a series of interviews was carried out. The study identified two key themes: (a) embarking on an unpredicted journey and (b) navigating through challenges. The analyses highlight discrepancies in partnership working between parents and educators and shortcomings in advice that professionals provided to these parents, potentially placing pupils and their families at a disadvantage. The implications for educational psychologists (EP) and other professionals working with Eastern European parents raising a child with SEN are also discussed