Papers by Alessia Pascale
Pharmacological Research, 1999
Calcium plays a pivotal role in mediating many important biological functions. The intracellular ... more Calcium plays a pivotal role in mediating many important biological functions. The intracellular calcium concentration is tightly regulated by a variety of systems and mechanisms. Calcium is sequestered by various organelles such as mitochondria andror endoplasmic reticulum and extruded across the plasma membrane by energy-dependent transport systems. Different Ca 2q -binding proteins are also involved in these processes. Alterations in calcium homeostasis might be critically implicated in brain aging and in the neuropathology Ž . of Alzheimer's disease AD . In fact, one of the postulated mechanisms of -amyloid toxicity seems to involve a Ca 2q dysregulation accompanied with enhanced vulnerability to excitotoxic stimuli. Although brain characteristic lesionsᎏplaques and tanglesᎏconstitute the hallmarks of AD, accumulated evidence suggests the systemic feature of this disease. Therefore peripheral cell lines may represent a useful approach to explore the cellular pathophysiology of AD, including calcium alterations and associated phenomena.
International Journal of Molecular Sciences, Dec 21, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Acta scientific microbiology, Aug 1, 2022
Amyloid-β peptide (Aβ) represents the main component of amyloid plaques in Alzheimer's disease (A... more Amyloid-β peptide (Aβ) represents the main component of amyloid plaques in Alzheimer's disease (AD); Aβ belongs to the group of antimicrobial peptides (AMPs) small peptides that kill pathogens through their antimicrobial activity and also have affinity for bacterial lipopolysaccharide (LPS). If amyloid is part of the antimicrobial mechanism of Aβ, fibrillar material would also be expected to accumulate as long as the innate immune system, correctly or incorrectly perceives an infection. Repeated reactivations of the chronic latent infection are constantly producing new Aß peptide, this situation lasts for a long time in the decades preceding the manifestation of AD, progressively leading to neurodegeneration and neuroinflammation. Aim of this work was to evaluate the concomitant synergizing action of Aβ 1-42 and LPS in human SH-SY5Y cells; AMPs and LPS have an amphipatic structure that is able to form heterogeneous micelles, in this way LPS acts as a fibrillogenesis promoter, Furthermore, depending on peptide concentration, the action of Aβ as AMP can be bacteriostatic or bactericidal.
A low-frequency electromagnetic (LF-EMF) exposure scheme induces autophagy activation to counteract in vitro Abeta-amyloid neurotoxicity
HuR/ELAVL1 expression in the human cataractous lens
Acta Ophthalmologica, Sep 14, 2016
PurposeCataract is a common age‐related ocular disease having as major determinants oxidative str... more PurposeCataract is a common age‐related ocular disease having as major determinants oxidative stress, accumulation of protein aggregates and inflammation. It has been previously reported that the expression of the RNA‐binding HuR/ELAVL1 (Embryonic Lethal Abnormal Vision‐Like 1) protein, a master regulator of many cellular functions including cell stress response, is altered in various ocular disorders. Here we develop a method to assess, by means of several techniques, the expression of HuR/ELAVL1 in human lens.MethodsHuman anterior lens capsules (aLCs) were collected from patients undergoing uneventful cataract surgery for mixed nuclear and cortical cataracts after informed consent was obtained. Control aLCs were obtained from non‐cataractous lenses derived from human cadaver eyes removed within 24 h of death. The samples collected were snap frozen for further processing. Quantitative real‐time PCR, Western blotting and ELISA assay were used to determine the expression and distribution of HuR/ELAVL1 in the lenses.ResultsHuR/ELAVL1 is expressed at both mRNA and protein level in the epithelial cells and aLC from age‐related cataract patients and healthy subjects.ConclusionsThis is the first study documenting the expression of HuR/ELAVL1 in human lens. Considering the importance of oxidative stress in the development of cataract, further experiments will allow to determine whether HuR/ELAVL1, which exerts in other ocular pathologies a post‐transcriptional control of stress response genes, may play a role in cataract pathogenesis.
Chemistry, Aug 26, 2022
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Evidence for novel cell defense mechanisms sustained by dimethyl fumarate in multiple sclerosis patients: the HuR/SOD2 cascade
Multiple sclerosis and related disorders, Dec 1, 2022
Chiral 2‐phenyl‐3‐hydroxypropyl esters as PKC‐alpha modulators: HPLC enantioseparation, NMR absolute configuration assignment, and molecular docking studies
Chirality, Dec 28, 2021
Protein kinase C (PKC) isoforms play a pivotal role in the regulation of numerous cellular functi... more Protein kinase C (PKC) isoforms play a pivotal role in the regulation of numerous cellular functions, making them extensively studied and highly attractive drug targets. In our previous work, we identified in racemate 1–2, based on the 2‐benzyl‐3‐hydroxypropyl ester scaffold, two new potent and promising PKCα and PKCδ ligands, targeting the C1 domain of these two kinases. Herein, we report the resolution of the racemates by enantioselective semi‐preparative HPLC. The attribution of the absolute configuration (AC) of homochirals 1 was performed by NMR, via methoxy‐α‐trifluoromethyl‐α‐phenylacetic acid derivatization (MTPA or Mosher's acid). Moreover, the match between the experimental and predicted electronic circular dichroism (ECD) spectra confirmed the assigned AC. These results proved that Mosher's esters can be properly exploited for the determination of the AC also for chiral primary alcohols. Lastly, homochiral 1 and 2 were assessed for binding affinity and functional activity against PKCα. No significative differences in the Ki of the enantiopure compounds was observed, thus suggesting that chirality does not seem to play a significant role in targeting PKC C1 domain. These results are in accordance with the molecular docking studies performed using a new homology model for the human PKCαC1B domain.
Tailored coating of gold nanostars: rational approach to prototype of theranostic device based on SERS and photothermal effects at ultralow irradiance
Nanotechnology, Apr 9, 2018
The last decade has come across an increasing demand for theranostic biocompatible nanodevices po... more The last decade has come across an increasing demand for theranostic biocompatible nanodevices possessing the double ability of diagnosis and therapy. In this work, we report the design, synthesis and step-by-step characterization of rationally coated gold nanostars (GNSs) for the SERS imaging and photothermal therapy of HeLa cancer cells. The nanodevices were realized by synthesizing GNSs with a seed growth approach, coating them with a controlled mixture of thiols composed of a Raman reporter and a polyethylene glycol with a terminal amino group, and then reacting these amino groups with folic acid (FA), in order to impart selectivity towards cancer cells which overexpress folate receptors on their membranes. After a complete characterization, we demonstrate that these FA-functionalized GNSs (FA-GNSs) are able to bind selectively to the membranes of HeLa cells, acting as SERS tags and allowing SERS imaging. Moreover, we demonstrate that once bound to HeLa cell membranes, FA-GNSs exhibit photothermal effect which can be exploited to kill the same cells in vitro using laser irradiation in the NIR at a very low and safe irradiance. We thus demonstrate that the FA-GNSs designed following the described approach are an efficient prototype of theranostic nanodevices.
Autophagy stimulus affects different kinase pathways and promotes HuR protein activation and SQSTM1/p62 protein synthesis in ARPE-19 cells
Acta Ophthalmologica, Sep 23, 2015
PurposeAge‐related macular degeneration (AMD) pathogenesis is characterized by protein degradatio... more PurposeAge‐related macular degeneration (AMD) pathogenesis is characterized by protein degradation impairment in retinal pigment epithelial (RPE) cells. We previously found that the expression of autophagy receptor SQSTM1/p62 is positively regulated by the RNA‐binding HuR/ELAV protein. We investigated the effects of AICAR (autophagy inducer, 5‐aminoimidazole‐4‐ carboxamide‐1‐β‐D‐ribofuranoside) and MG‐132 (proteasome inhibitor) co‐treatment on HuR activation, p62 expression, and the kinases potentially involved.MethodsARPE‐19 cells were treated with MG‐132 (1 µM) and/or AICAR (2 mM) for increasing times (up to 2 h) and subjected to cell fractionation. SQSTM1/p62 mRNA and protein levels were measured by qRT‐PCR and Western blotting, respectively. HuR protein levels and its phosphorylated status were evaluated by Western blotting. The effects of puromycin (1 mM, protein synthesis inhibitor) and various kinase inhibitors were also tested.ResultsAICAR+MG‐132 co‐treatment for 2 h induces HuR protein up‐regulation, its cytoplasmic translocation and phosphorylation, as well as increased expression of p62 protein, being the latter one blunted by puromycin. AICAR+MG‐132 co‐treatment affects various kinases with differential outcomes.ConclusionsAICAR+MG‐132 co‐treatment leads to HuR activation and p62 protein translation. Different protein kinase pathways are likely involved in these events.
The brain-gut-microbiota interplay in depression: A key to design innovative therapeutic approaches
Pharmacological Research, Jun 1, 2023
Differential modulation of protein kinase C isoforms in NG108-15 cell differentiation
Pharmacological Research, 1995
International Journal of Molecular Sciences, Apr 2, 2023
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Tunable coating of gold nanostars: tailoring robust SERS labels for cell imaging
Nanotechnology, May 20, 2016
Surface modification of noble metal nanoparticles with mixed molecular monolayers is one of the m... more Surface modification of noble metal nanoparticles with mixed molecular monolayers is one of the most powerful tools in nanotechnology, and is used to impart and tune new complex surface properties. In imaging techniques based on surface enhanced Raman spectroscopy (SERS), precise and controllable surface modifications are needed to carefully design reproducible, robust and adjustable SERS nanoprobes. We report here the attainment of SERS labels based on gold nanostars (GNSs) coated with a mixed monolayer composed of a poly ethylene glycol (PEG) thiol (neutral or negatively charged) that ensure stability in biological environments, and of a signalling unit 7-Mercapto-4-methylcoumarin as a Raman reporter molecule. The composition of the coating mixture is precisely controlled using an original method, allowing the modulation of the SERS intensity and ensuring overall nanoprobe stability. The further addition of a positively charged layer of poly (allylamine hydrocloride) on the surface of negatively charged SERS labels does not change the SERS response, but it promotes the penetration of GNSs in SH-SY5Y neuroblastoma cells. As an example of an application of such an approach, we demonstrate here the internalization of these new labels by means of visualization of cell morphology obtained with SERS mapping.
Journal of Alzheimer's Disease, Oct 29, 2019
It is now more than two decades since amyloid- (A), the proteolytic product of the amyloid- pr... more It is now more than two decades since amyloid- (A), the proteolytic product of the amyloid- protein precursor (APP), was first demonstrated to be a normal and soluble product of neuronal metabolism. To date, despite a growing body of evidence suggests its regulatory role on synaptic function, the exact cellular and molecular pathways involved in A-driven synaptic effects remain elusive. This review provides an overview of the mounting evidence showing A-mediated effects on presynaptic functions and neurotransmitter release from axon terminals, focusing on its interaction with synaptic vesicle cycle. Indeed, A peptides have been found to interact with key presynaptic scaffold proteins and kinases affecting the consequential steps of the synaptic vesicle dynamics (e.g., synaptic vesicles exocytosis, endocytosis, and trafficking). Defects in the fine-tuning of synaptic vesicle cycle by A and deregulation of key molecules and kinases, which orchestrate synaptic vesicle availability, may alter synaptic homeostasis, possibly contributing to synaptic loss and cognitive decline. Elucidating the presynaptic mechanisms by which A regulate synaptic transmission is fundamental for a deeper comprehension of the biology of presynaptic terminals as well as of A-driven early synaptic defects occurring in prodromal stage of AD. Moreover, a better understating of A involvement in cellular signal pathways may allow to set up more effective therapeutic interventions by detecting relevant molecular mechanisms, whose imbalance might ultimately lead to synaptic impairment in AD.
PKCβII/HuR/VEGF Cascade in Experimental Diabetic Retinopathy
Investigative Ophthalmology & Visual Science, Apr 28, 2009
Memoria e aumento dell'espressione di HuD
Activation of Elav-Like Rna Binding Proteins by Learning
Elav-Like Proteins And Protein Kinase C: A New Cascade For Memory Trace Formation And Alzheimer's Disease?
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Papers by Alessia Pascale