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Abstract Background Amyloid-β42 (Aβ42) aggregation consists of a complex chain of nucleation events producing soluble oligomeric intermediates, which are considered the major neurotoxic agents in Alzheimer’s disease (AD). Cerebral lesions in the brain of AD patients start to develop 20 years before symptom onset; however, no preventive strategies, effective treatments, or specific and sensitive diagnostic tests to identify people with early-stage AD are currently available. In addition, the isolation and characterisation of neurotoxic Aβ42 oligomers are particularly difficult because of their transient and heterogeneous nature. To overcome this challenge, a rationally designed method generated a single-domain antibody (sdAb), named DesAb-O, targeting Aβ42 oligomers. Methods We investigated the ability of DesAb-O to selectively detect preformed Aβ42 oligomers both in vitro and in cultured neuronal cells, by using dot-blot, ELISA immunoassay and super-resolution STED microscopy, and to counteract the toxicity induced by the oligomers, monitoring their interaction with neuronal membrane and the resulting mitochondrial impairment. We then applied this approach to CSF samples (CSFs) from AD patients as compared to age-matched control subjects. Results DesAb-O was found to selectively detect synthetic Aβ42 oligomers both in vitro and in cultured cells, and to neutralise their associated neuronal dysfunction. DesAb-O can also identify Aβ42 oligomers present in the CSFs of AD patients with respect to healthy individuals, and completely prevent cell dysfunction induced by the administration of CSFs to neuronal cells. Conclusions Taken together, our data indicate a promising method for the improvement of an early diagnosis of AD and for the generation of novel therapeutic approaches based on sdAbs for the treatment of AD and other devastating neurodegenerative conditions.

AbstractIntroduction: Several neurodegenerative conditions are associated with a common histopathology within neurons of the central nervous system, consisting of the deposition of cytoplasmic inclusions of TAR DNA-binding protein 43 (TDP-43). Such inclusions have variably been described as morphologically and molecularly ordered aggregates having amyloid properties, as filaments without the cross-β-structure and dye binding specific for amyloid, or as amorphous aggregates with no defined structure and fibrillar morphology.Aims and Methods: Here we have expressed human full-length TDP-43 in neuroblastoma x spinal cord 34 (NSC-34) cells to investigate the morphological, structural, and tinctorial properties of TDP-43 inclusions in situ. We have used last-generation amyloid diagnostic probes able to cross the cell membrane and detect amyloid in the cytoplasm and have adopted Raman and Fourier transform infrared microspectroscopies to study in situ the secondary structure of the TDP-43 protein in the inclusions. We have then used transmission electron microscopy to study the morphology of the TDP-43 inclusions.Results: The results show the absence of amyloid dye binding, the lack of an enrichment of cross-β structure in the inclusions, and of a fibrillar texture in the round inclusions. The aggregates formed in vitro from the purified protein under conditions in which it is initially native also lack all these characteristics, ruling out a clear amyloid-like signature.Conclusions: These findings indicate a low propensity of TDP-43 to form amyloid fibrils and even non-amyloid filaments, under conditions in which the protein is initially native and undergoes its typical nucleus-to-cell mislocalization. It cannot be excluded that filaments emerge on the long time scale from such inclusions, but the high propensity of the protein to form initially other types of inclusions appear to be an essential characteristic of TDP-43 proteinopathies.KEY MESSAGESCytoplasmic inclusions of TDP-43 formed in NSC-34 cells do not stain with amyloid-diagnostic dyes, are not enriched with cross-β structure, and do not show a fibrillar morphology.TDP-43 assemblies formed in vitro from pure TDP-43 do not have any hallmarks of amyloid.

The misfolding and aggregation of α-synuclein is the general hallmark of a group of devastating neurodegenerative pathologies referred to as synucleinopathies, such as Parkinson’s disease, dementia with Lewy bodies, and multiple system atrophy. In such conditions, a range of different misfolded aggregates, including oligomers, protofibrils, and fibrils, are present both in neurons and glial cells. Growing experimental evidence supports the proposition that soluble oligomeric assemblies, formed during the early phases of the aggregation process, are the major culprits of neuronal toxicity; at the same time, fibrillar conformers appear to be the most efficient at propagating among interconnected neurons, thus contributing to the spreading of α-synuclein pathology. Moreover, α-synuclein fibrils have been recently reported to release soluble and highly toxic oligomeric species, responsible for an immediate dysfunction in the recipient neurons. In this review, we discuss the current knowledge about the plethora of mechanisms of cellular dysfunction caused by α-synuclein oligomers and fibrils, both contributing to neurodegeneration in synucleinopathies.

Summary: High cholesterol levels are a risk factor for the development of Alzheimer’s disease. Experiments investigating the influence of cholesterol on the proteolytic processing of the amyloid precursor protein (APP) by the β-secretase Bace1 and on their proximity in cells have led to conflicting results. By using a fluorescence bioassay coupled with flow cytometry we found a direct correlation between the increase in membrane cholesterol amount and the degree of APP shedding in living human neuroblastoma cells. Analogue results were obtained for cells overexpressing an APP mutant that cannot be processed by α-secretase, highlighting the major influence of cholesterol enrichment on the cleavage of APP carried out by Bace1. By contrast, the cholesterol content was not correlated with changes in membrane dynamics of APP and Bace1 analyzed with single molecule tracking, indicating that the effect of cholesterol enrichment on APP processing by Bace1 is uncoupled from changes in their lateral diffusion.

Cancer’s impact on sexuality has been well documented across all types of cancers, both in patients with active cancers and long-term survivors. As we make progresses in extending survival for cancer patients, we must face the challenge of providing a satisfactory quality of life, on which sexual disfunction impacts profoundly. Although some treatment-related sexual adverse effects are short lived, many chronic cancer patients and survivors face long-term physical and psychological sequelae. Partners are also affected by a cancer diagnosis, with both negative and positive consequences for the relationship. Patients with sexual apparatus and non-sexual apparatus cancer are affected by sexual dysfunction, therefore each group should be evaluated for support, nevertheless considering the peculiarities of each cancer treatment and side effects, the person individuality and his cultural and religious background. Although sexual dysfunction has been studied quite extensively and a range of intervention strategies that can help patients cope with treatment-related sexual problems are now available, many patients do not feel that they are receiving adequate support from healthcare providers. By taking a holistic approach through appropriate screening, communication, medical therapy, surgical prosthesis and psychological support for cancer patients and their partners, sexual dysfunction and the accompanying distress can be signi cantly alleviated. The healthcare barriers to such interventions perceived by healthcare providers (primary care physicians, oncologists, other specialized healthcare professionals) are mainly a lack of formal training, access to appropriate referrals, the lack of standards of care, and assuming that non-sexual related cancer and elderly people sexual needs are minimal.

The cerebrospinal fluid (CSF) of 19 subjects who received a clinical diagnosis of Alzheimer’s disease (AD) as well as of 5 pathological controls was collected and analyzed by Raman spectroscopy (RS). We investigated whether the raw and preprocessed Raman spectra could be used to distinguish AD from controls. First, we applied standard Machine Learning (ML) methods obtaining unsatisfactory results. Then, we applied ML to a set of topological descriptors extracted from raw spectra, achieving a very good classification accuracy (>87%). Although our results are preliminary, they indicate that RS and topological analysis may provide an effective combination to confirm or disprove a clinical diagnosis of AD. The next steps include enlarging the dataset of CSF samples to validate the proposed method better and, possibly, to investigate whether topological data analysis could support the characterization of AD subtypes.

The self-assembly of α-synuclein (αS) is a pathological feature of Parkinson’s disease. The αS species responsible for neuronal damage are not well characterized. Here, the authors show that αS fibrils release soluble prefibrillar oligomeric species responsible for neurotoxicity in vitro.

Safety data regarding BNT162b2 in cancer patients (CPs) are scarce. Herein we report the side effects (SEs), the adverse events (AEs), and the patient-reported outcomes (PROs) following BNT162b2 administration in CPs treated at the San Luigi Gonzaga University Hospital. All CPs who agreed to participate in our vaccination campaign received BNT162b2 and were included in the descriptive analysis. An anonymous questionnaire investigating the occurrence of SEs/AEs and PROs was administered to the study population 21 days after the first dose. Pearson’s chi-squared test was used to estimate the risk of experiencing SEs/AEs according to selected variables. A total of 997 patients were included in the study: 62.0% had stage IV cancer, and 68.8% were receiving an active treatment, of whom 15.9% were receiving immunotherapy. SEs/AEs were recorded in 37.1% of cases after the first dose and in 48.5% of cases after the second dose. The most common SEs were muscle pain/local rash (27.9% and 28%, after the first and second dose, respectively). Patients older than 70 years showed lower risk of SEs/AEs, while women showed a higher risk. Before receiving the vaccine, 18.2% of patients felt fearful and/or insecure about the vaccination. After the first dose, 57.5% of patients changed their feelings positively. Our data support the short-term safety of BNT162b2 in CPs, regardless of disease stage and concurrent treatments. Overall, the vaccination showed a positive impact on quality of life.

The aberrant aggregation of proteins is a key molecular event in the development and progression of a wide range of neurodegenerative disorders. We have shown previously that squalamine and trodusquemine, two natural products in the aminosterol class, can modulate the aggregation of the amyloid-β peptide (Aβ) and of α-synuclein (αS), which are associated with Alzheimer’s and Parkinson’s diseases. In this work, we expand our previous analyses to two squalamine derivatives, des-squalamine and α-squalamine, obtaining further insights into the mechanism by which aminosterols modulate Aβ and αS aggregation. We then characterize the ability of these small molecules to alter the physicochemical properties of stabilized oligomeric species in vitro and to suppress the toxicity of these aggregates to varying degrees toward human neuroblastoma cells. We found that, despite the fact that these aminosterols exert opposing effects on Aβ and αS aggregation under the conditions that we tested, the modifications that they induced to the toxicity of oligomers were similar. Our results indicate that the suppression of toxicity is mediated by the displacement of toxic oligomeric species from cellular membranes by the aminosterols. This study, thus, provides evidence that aminosterols could be rationally optimized in drug discovery programs to target oligomer toxicity in Alzheimer’s and Parkinson’s diseases.

Transient oligomeric species of the amyloid-β peptide (Aβ42) have been identified as key pathogenic agents in Alzheimer’s disease. Here the authors find that the aminosterol trodusquemine enhances Aβ42 aggregation and suppresses Aβ42-induced toxicity by displacing oligomers from cell membranes.

α-Synuclein (αS) is an intrinsically disordered and highly dynamic protein involved in dopamine release at presynaptic terminals. The abnormal aggregation of αS as mature fibrils into intraneuronal inclusion bodies is directly linked to Parkinson’s disease. Increasing experimental evidence suggests that soluble oligomers formed early during the aggregation process are the most cytotoxic forms of αS. This study investigated the uptake by neuronal cells of pathologically relevant αS oligomers and fibrils exploiting a range of conformation-sensitive antibodies, and the super-resolution stimulated emission depletion (STED) microscopy. We found that prefibrillar oligomers promptly penetrate neuronal membranes, thus resulting in cell dysfunction. By contrast, fibril docking to the phospholipid bilayer is accompanied by αS conformational changes with a progressive release of A11-reactive oligomers, which can enter into the neurons and trigger cell impairment. Our data provide important evidence on the role of αS fibrils as a source of harmful oligomers, which resemble the intermediate conformers formed de novo during aggregation, underling the dynamic and reversible nature of protein aggregates responsible for α-synucleinopathies.

Accumulation of ubiquitin-positive, tau- and α-synuclein-negative intracellular inclusions of TDP-43 in the central nervous system represents the major hallmark correlated to amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin-positive inclusions. Such inclusions have variably been described as amorphous aggregates or more structured deposits having an amyloid structure. Following the observations that bacterial inclusion bodies generally consist of amyloid aggregates, we have overexpressed full-length TDP-43 and C-terminal TDP-43 in E. coli, purified the resulting full-length and C-terminal TDP-43 containing inclusion bodies (FL and Ct TDP-43 IBs) and subjected them to biophysical analyses to assess their structure/morphology. We show that both FL and Ct TDP-43 aggregates contained in the bacterial IBs do not bind amyloid dyes such as thioflavin T and Congo red, possess a disordered secondary structure, as inferred using circular dichroism and infrared spectroscopies, and are susceptible to proteinase K digestion, thus possessing none of the hallmarks for amyloid. Moreover, atomic force microscopy revealed an irregular structure for both types of TDP-43 IBs and confirmed the absence of amyloid-like species after proteinase K treatment. Cell biology experiments showed that FL TDP-43 IBs were able to impair the viability of cultured neuroblastoma cells when added to their extracellular medium and, more markedly, when transfected into their cytosol, where they are at least in part ubiquitinated and phosphorylated. These data reveal an inherently high propensity of TDP-43 to form amorphous aggregates, which possess, however, an inherently high ability to cause cell dysfunction. This indicates that a gain of toxic function caused by TDP-43 deposits is effective in TDP-43 pathologies, in addition to possible loss of function mechanisms originating from the cellular mistrafficking of the protein.

The Manifesto serves as a unifying framework for academics, degree courses, departments, schools and universities seeking to institutionalise service-learning into their curriculum. It has been developed by the Italian Universities Network for Service-Learning (UNiSL). It represents the initial outcome of a comprehensive analysis of research findings and documented experiences from Italy and abroad. This document introduces the Italian definition of service-learning in higher education, accompanied by a glossary consisting of nine key terms, each followed by an explanatory description. Its primary objective is to comprehensively understand the approaches, theoretical models and perspectives that characterise the Network. Il Manifesto della Rete delle Università italiane per il Service-Learning (UNiSL). Il Manifesto intende offrire una piattaforma comune per docenti, Corsi di Laurea, Dipartimenti, Scuole o Atenei che intendano istituzionalizzare il service-learning all’interno del proprio curriculum. Il documento è stato elaborato dalla Rete delle UNiversità Italiane per il Service-Learning (UNiSL) e rappresenta un primo risultato raggiunto grazie al confronto sugli esiti delle ricerche condotte e sulle esperienze documentate in Italia e all’estero. Nel documento si propone la definizione italiana di service-learning in contesto universitario e un glossario con nove parole chiave, ognuna accompagnata da una spiegazione, con l’obiettivo di comprendere in modo esaustivo gli approcci, i modelli teorici e le prospettive che caratterizzano la Rete.

According to the most recent provisions of the European Union, public buildings should play an exemplary role in sustainable development, adopting accelerated renovation rates aimed at improving their energy performance. Within this category, university buildings are a case study of great interest to experiment with new approaches for energy refurbishment and sustain-able management of architectural assets. The research presents a work-flow that originates from easily available input data, to reach the definition of a multi-scale spatial database, founded on the synergy between GIS (Geographic Information System) and BIM (Building Information Mod-eling) and defined according to standard and shared data models. Tools of this kind are crucial for promoting efficient information management building assets, by organizing data into navigable three-dimensional mod-els. In addition to the clear benefits associated with structured archiving, the provision of a relational database makes it possible to capitalize on the already available knowledge and to activate decision support tools for comparative assessment of transformation scenarios. In particular, the use of the cost-optimal methodology is proposed: it is a multi-criteria assess-ment aimed at identifying a set of optimal energy refurbishment solutions concerning energy consumption and management costs. The paper pres-ents the methodological framework and examines its application at differ-ent scales, from the case of the University of Pavia real estate asset to the application to a single building complex.

Empiric rheology is considered a useful tool for assessing the technological quality of wheat. Over the decades, several tests have been adapted from common to durum wheat, and new approaches have been proposed to meet the needs of the players of the durum wheat value chain. Breeders are looking for reliable methods to test the functional quality of wheat lines at early stages, where there are limited amounts of sample; millers need fast and reliable methods for checking wheat quality right at the point of the receiving station; and pasta-makers are looking for suitable methods to predict end product quality. This review provides an overview of the strengths and weaknesses of the rheological tests currently used to evaluate the quality of durum wheat semolina, with the emphasis on Europe. Moreover, the relationships among the parameters obtained from different rheological approaches are extrapolated from the literature and integrated with the data obtained from 74 samples of durum wheat semolina. Although numerous efforts have been made to propose rapid and reliable tests for semolina characterization, the ideal test has yet to be proposed, indicating that researchers and pasta companies need to focus on perfecting the way to assess the quality of durum wheat and pasta.

Worldwide, organizations and small and medium-sized enterprises have already disruptively changed in many ways their physiological inner mechanisms, because of information and communication technologies (ICT) revolution. Nevertheless, the still ongoing COVID-19 worldwide emergency definitely promoted a wide adoption of teleworking modalities for many people around the world, making it more relevant than before to understand the real impact of virtual environments (VEs) on teamwork dynamics. From a psychological point of view, a critical question about teleworking modalities is how the social and cognitive dynamics of collaborative facilitation and collaborative inhibition would affect teamwork within VEs. This study analyzed the impact of a virtual environment (VE) on the recall of individuals and members of nominal and collaborative groups. The research assessed costs and benefits for collaborative retrieval by testing the effect of experimental conditions, stimulus materials, group size, experimental conditions order, anxiety state, personality traits, gender group composition and social interactions. A total of 144 participants were engaged in a virtual Deese-Roediger-McDermott (DRM) classical paradigm, which involved remembering word lists across two successive sessions, in one of four protocols: I-individual/nominal, I I -nominal/individual, I I I -nominal/collaborative, I V -collaborative/nominal. Results suggested, in general, a reduced collaborative inhibition effect in the collaborative condition than the nominal and individual condition. A combined effect between experimental condition and difficulty of the task appears to explain the presence of collaborative inhibition or facilitation. Nominal groups appeared to enhance the collaborative groups’ performance when virtual nominal groups come before collaborative groups. Variables such as personality traits, gender and social interactions may have a contribution to collaborative retrieval. In conclusion, this study indicated how VEs could maintain those peculiar social dynamics characterizing the participants’ engagement in a task, both working together and individually, and could affect their intrinsic motivation as well as performances. These results could be exploited in order to design brand new and evidenced-based practices, to improve teleworking procedures and workers well-being, as well as teleworking teamwork effectiveness.

A colored and fiber-rich fraction from the debranning of purple wheat was incorporated at 25% into semolina- and flour-based pasta produced on a pilot-plant scale, with the aim of increasing anthocyanin and total phenolic content with respect to pasta obtained from whole pigmented grains. The debranning fraction impaired the formation of disulfide-stabilized protein networks in semolina-based systems. Recovery of phenolics was impaired by the pasta making process, and cooking decreased the phenolic content in both enriched samples. Cooking-related losses in anthocyanins and total phenolics were similar, but anthocyanins in the cooked semolina-based pasta were around 20% of what was expected from the formulation. HPLC (High Performance Liquid Chromatography) profiling of phenolics was carried out on extracts from either type of enriched pasta both before and after cooking and indicate possible preferential retention of specific compounds in each type of enriched pasta. Extracts from cooked samples of either enriched pasta were tested as inhibitors of enzymes involved in glucose metabolism and uptake, as well as for their capacity of suppressing the response to inflammatory stimuli. Results of both biological tests indicate that the phenolics in extracts from both cooked pasta samples had inhibitory capacities higher than extracts of the original debranning fraction at identical concentrations of total bioactives.

The aim of control the economic, social and environmental aspects connected to the construction of a building imposes a systematic approach for which it is necessary to make test models aimed to a coordinate analysis of different and independent performance issues. BIM technology, referring to interoperable informative models, offers a significant operative basis to achieve this necessity. In most of the cases, informative models concentrate on a product-based digital models collection built in a virtual space, more than on the simulation of their relational behaviors. This relation, instead, is the most important aspect of modelling because it marks and characterizes the interactions that can define the building as a system. This study presents the use of standard classification systems as tools for both the activation and validation of an integrated performance-based building process. By referring categories and types of the informative model to the codes of a technological and performance-based classification system, it is possible to link and coordinate functional units and their elements with the indications required by the AEC standards. In this way, progressing with an incremental logic, it is possible to achieve the management of the requirements of the whole building and the monitoring of the fulfilment of design objectives and specific normative guidelines. The informative model, indeed, offers through the whole building process, the possibility to list the expected performance requirements defined at the design level for the singles technical elements. Once filled in those values and given the model interoperability to analysis platforms, we can gather the results and publish them by using computational design algorithm in parameters of functional units with the aim to monitor the relation with the expressed specifications. In that way, the model assumes the role of decisional basis for the building process because it allows to gather the technical specifications of simple components and functional units and in the same time compare them to the performance requirements of building products and to all the system composing the building, both active or passive. This research is proposed as a hypothesis for the extension of the project informative content, with the aim to support the decision-making skills related to information management to ensure the success on projects. The work has been developed during a research program in partnership between University of Padova and F&M Ingegneria S.p.A., an Italian company operating on engineering and architecture. In the first phase of work the study focused on the literature analysis dealing with development and spread of standard classification codes, with more attention on OmniClass and its implementation in BIM platforms. In the second stage, it concentrated on the implementation of the standard classification system and the performance specifications on the BIM platforms used for project design and management. The result has been achieved through the use of Computational design and data integration solutions, developed with the help of visual programming language (VPL). The same solutions have been used to fill in the performances of the whole system that has been simulated with FEA software and to validate the choices in relation to the proposed aims and to the normative prescriptions by comparing requirements and performances. The project has made possible to activate a real integrate performance-based design process thanks to the realization of the map previously mentioned and to the fact that BIM technology can define the relationship between technical elements and determined spaces and, at the same time, to monitor functions and requirements on that space.