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Background: The differential diagnosis of frontotemporal dementia (FTD) is still a challenging task due to its symptomatic overlap with other neurological diseases and the lack of biofluid-based biomarkers. Objective: To investigate the diagnostic potential of a combination of novel biomarkers in cerebrospinal fluid (CSF) and blood. Methods: We included 135 patients from the Center for Memory Disturbances, University of Perugia, with the diagnoses FTD (n = 37), mild cognitive impairment due to Alzheimer’s disease (MCI-AD, n = 47), Lewy body dementia (PDD/DLB, n = 22), and cognitively unimpaired patients as controls (OND, n = 29). Biomarker levels of neuronal pentraxin-2 (NPTX2), neuronal pentraxin receptor, neurofilament light (NfL) and glial fibrillary acidic protein (GFAP) were measured in CSF, as well as NfL and GFAP in serum. We assessed biomarker differences by analysis of covariance and generalized linear models (GLM). We performed receiver operating characteristics analyses and Spearman correlation to determine biomarker associations. Results: CSF NPTX2 and serum GFAP levels varied most between diagnostic groups. The combination of CSF NPTX2, serum NfL and serum GFAP differentiated FTD from the other groups with good accuracy (FTD versus MCI-AD: area under the curve (AUC) [95% CI] = 0.89 [0.81–0.96]; FTD versus PDD/DLB: AUC = 0.82 [0.71–0.93]; FTD versus OND: AUC = 0.80 [0.70–0.91]). CSF NPTX2 and serum GFAP correlated positively only in PDD/DLB (ρ= 0.56, p

Background and Objectives.Patients with Lewy body disease (LBD) often show a co-occurring Alzheimer’s disease (AD) pathology. Cerebrospinal fluid (CSF) biomarkers allow the detectionin vivoof AD-related pathological hallmarks included in the AT(N) classification system. Here, we aimed to investigate whether CSF biomarkers of synaptic and neuro-axonal damage are correlated with the presence of AD co-pathology in LBD and can be useful to differentiate LBD patients with different AT(N) profiles.Methods.We retrospectively measured CSF levels of AD core biomarkers (Aβ42/40 ratio, p-tau, t-tau) and of synaptic (β-synuclein, α-synuclein, SNAP-25, neurogranin) and neuro-axonal proteins (NfL) in 28 cognitively unimpaired participants with non-degenerative neurological conditions and 161 participants with a diagnosis of either LBD or AD (at both mild cognitive impairment, AD-MCI, and dementia stages, AD-dem). We compared CSF biomarker levels in clinical and AT(N)-based subgroups.Results.CSF β-synuclein, α-synuclein, SNAP-25, neurogranin and NfL levels did not differ between LBD (n = 101, age = 67.2 ± 7.8 years, 27.7% females) and controls (age = 64.8 ± 8.6 years, 39.3% females) and were increased in AD (AD-MCI n = 30, AD-dem n = 30, age = 72.3 ± 6.0 years, 63.3% females) compared to both groups (p < 0.001 for all comparisons). In LBD, we found increased levels of synaptic and neuro-axonal degeneration biomarkers in patients with A+T+ (LBD/A+T+) than with A-T- profiles (LBD/A-T-) (p < 0.01 for all), and β-synuclein showed the highest discriminative accuracy between the two groups (AUC = 0.938, 95%CI = 0.884 - 0.991). CSF β-synuclein (p = 0.0021), α-synuclein (p = 0.0099) and SNAP-25 concentrations (p = 0.013) were also higher in LBD/A+T+ than in LBD/A+T- cases, which had synaptic biomarkers levels within the normal range. CSF α-synuclein was significantly decreased only in LBD patients with T- profiles compared to controls (p = 0.0448). Moreover, LBD/A+T+ and AD cases did not differ in any biomarker level.Discussion.LBD/A+T+ and AD cases showed significantly increased CSF levels of synaptic and neuro-axonal biomarkers compared to LBD/A-T- and control subjects. LBD patients with AD co-pathology might, thus, experience similar degrees of synaptic dysfunction than pure AD cases.Classification of evidence.This study provides Class II evidence that CSF levels of β-synuclein, α-synuclein, SNAP-25, neurogranin and NfL are higher in patients with AD than in patients with LBD.

Objectives The core cerebrospinal fluid (CSF) biomarkers; total tau (tTau), phospho-tau (pTau), amyloid β 1-42 (Aβ 1-42), and the Aβ 1-42/Aβ 1-40 ratio have transformed Alzheimer’s disease (AD) research and are today increasingly used in clinical routine laboratories as diagnostic tools. Fully automated immunoassay instruments with ready-to-use assay kits and calibrators has simplified their analysis and improved reproducibility of measurements. We evaluated the analytical performance of the fully automated immunoassay instrument LUMIPULSE G (Fujirebio) for measurement of the four core AD CSF biomarkers and determined cutpoints for AD diagnosis. Methods Comparison of the LUMIPULSE G assays was performed with the established INNOTEST ELISAs (Fujirebio) for hTau Ag, pTau 181, β-amyloid 1-42, and with V-PLEX Plus Aβ Peptide Panel 1 (6E10) (Meso Scale Discovery) for Aβ 1-42/Aβ 1-40, as well as with a LC-MS reference method for Aβ 1-42. Intra- and inter-laboratory reproducibility was evaluated for all assays. Clinical cutpoints for Aβ 1-42, tTau, and pTau was determined by analysis of three cohorts of clinically diagnosed patients, comprising 651 CSF samples. For the Aβ 1-42/Aβ 1-40 ratio, the cutpoint was determined by mixture model analysis of 2,782 CSF samples. Results The LUMIPULSE G assays showed strong correlation to all other immunoassays (r>0.93 for all assays). The repeatability (intra-laboratory) CVs ranged between 2.0 and 5.6%, with the highest variation observed for β-amyloid 1-40. The reproducibility (inter-laboratory) CVs ranged between 2.1 and 6.5%, with the highest variation observed for β-amyloid 1-42. The clinical cutpoints for AD were determined to be 409 ng/L for total tau, 50.2 ng/L for pTau 181, 526 ng/L for β-amyloid 1-42, and 0.072 for the Aβ 1-42/Aβ 1-40 ratio. Conclusions Our results suggest that the LUMIPULSE G assays for the CSF AD biomarkers are fit for purpose in clinical laboratory practice. Further, they corroborate earlier presented reference limits for the biomarkers.

The aggregation of amyloid-β 42 (Aβ42) is directly related to the pathogenesis of Alzheimer's disease. Here, we have investigated the early stages of the aggregation process, during which most of the cytotoxic species are formed. Aβ42 aggregation kinetics, characterized by the quantification of Aβ42 monomer consumption, were tracked by real-time solution NMR spectroscopy (RT-NMR) allowing the impact that low-molecular-weight (LMW) inhibitors and modulators exert on the aggregation process to be analysed. Distinct differences in the Aβ42 kinetic profiles were apparent and were further investigated kinetically and structurally by using thioflavin T (ThT) and transmission electron microscopy (TEM), respectively. LMW inhibitors were shown to have a differential impact on early-state aggregation. Insight provided here could direct future therapeutic design based on kinetic profiling of the process of fibril formation.

Background: Aggregation of α-synuclein (α-syn) is a prominent feature of Parkinson’s disease (PD) and other synucleinopathies. In these diseases, the extracellular spreading of misfolded α-syn significantly contributes to the cell-to-cell propagation of the α-syn misfolding pathology in a prion-like fashion. Therefore, extracellular α-syn aggregates are considered primary targets both for diagnostics and for novel disease modifying therapies. Currently, α-syn seed amplification assays (SAAs) using cerebrospinal fluid (CSF) represent the most promising diagnostic tools for synucleinopathies. However, CSF itself contains several compounds that can modulate the aggregation of α-syn in a patient-dependent manner, potentially sabotaging unoptimized α-syn SAAs and preventing seed quantification. Methods: In this study, we characterized the inhibitory effect of CSF on in vitro α-syn aggregation by means of CSF fractionation, mass spectrometry, dot-blot, Western blot, transmission electron microscopy, solution nuclear magnetic resonance spectroscopy, a highly accurate and standardized diagnostic SAA, and different in vitro aggregation conditions to evaluate spontaneous aggregation of α-syn. Results: We found the high-molecular weight fraction of CSF (>100,000 Da) to be highly inhibitory and identified lipoproteins to be the main drivers of this effect. We evaluated direct interaction between lipoprotein and α-syn and observed lipoprotein-α-syn complexes by transmission electron microscopy. Direct interaction between lipoproteins and monomeric α-syn was not detected by solution nuclear magnetic resonance spectroscopy, suggesting interaction between lipoproteins and oligomeric/proto-fibrillary α-syn intermediates instead. Lastly, we observed significantly slower amplification of α-syn seeds in PD CSF when lipoproteins were added to the reaction mix of a highly accurate diagnostic SAA. Conclusions: Our results describe a novel interaction between lipoproteins and α-syn aggregates that inhibits the formation of α-syn fibrils and could have relevant biological and translational implications. Indeed, the donor-specific inhibition of CSF on α-syn aggregation explains the lack of quantitative results so far obtained by the analysis of SAA-derived kinetic parameters. Furthermore, our data show that apolipoproteins are the main inhibitory components of CSF, suggesting that lipoprotein concentration measurements could be incorporated into data analysis models to eliminate the confounding effects of CSF milieu on α-syn quantification efforts.

Objectives Cerebrospinal fluid α-synuclein (CSF α-syn) represents a possible biomarker in Parkinson’s disease (PD) diagnosis. CSF blood contamination can introduce a bias in α-syn measurement. To date, CSF samples with a red blood cells (RBC) count >50 RBC × 106/L or haemoglobin (Hb) concentration >200 μg/L are excluded from biomarker studies. However, investigations for defining reliable cut-off values are missing. Methods We evaluated the effect of blood contamination on CSF α-syn measurement by a systematic approach in a cohort of 42 patients with different neurological conditions who underwent lumbar puncture (LP) for diagnostic reasons. CSF samples were spiked with whole blood and serially diluted to 800, 400, 200, 100, 75, 50, 25, 5, 0 RBC × 106/L. CSF α-syn and Hb levels were measured by ELISA. Results In neat CSF, the average concentration of α-syn was 1,936 ± 636 ng/L. This value increased gradually in spiked CSF samples, up to 4,817 ± 1,456 ng/L (+149% α-syn variation) in samples with 800 RBC × 106/L. We established different cut-offs for discriminating samples with α-syn level above 5, 10, and 20% variation, corresponding to a Hb (RBC) concentration of 1,569 μg/L (37 RBC × 106/L), 2,082 μg/L (62 RBC × 106/L), and 3,118 μg/L (87 RBC × 106/L), respectively. Conclusions Our data show the high impact of CSF blood contamination on CSF α-syn levels, highlighting the measurement of Hb concentration as mandatory when assessing CSF α-syn. The thresholds we calculated are useful to classify CSF samples for blood contamination, considering as reliable only those showing a Hb concentration

INFN is developing at LASA laboratory (Milano, Italy) the prototypes of five high order corrector magnets, from skew quadrupole to dodecapole, which will equip the high-luminosity interaction regions of the High Luminosity-LHC (HL-LHC). These magnets are based on a superferric design, which allows a relatively simple, modular and easy to construct magnet. This activity takes place within the framework of a collaboration agreement between CERN and INFN. Four prototypes, from sextupole to dodecapole, have been built and tested starting in 2016. We present here the last prototype of the high order correctors, to be installed in LHC, the skew quadrupole: magnetic and mechanical design are discussed together with quench protection. We report also on the overall experience gained during construction aiming toward the series production. The power test of the quadrupole, including the training, the qualification and the quench behavior in operational conditions are also described.

We present a processing method, based on the multivariate curve resolution approach (MCR), to denoise 2D solid-state NMR spectra, yielding a substantial S/N ratio increase while preserving the lineshapes and relative signal intensities. These spectral features are particularly important in the quantification of silicon species, where sensitivity is limited by the low natural abundance of the 29Si nuclei and by the dilution of the intrinsic protons of silica, but can be of interest also when dealing with other intermediate-to-low receptivity nuclei. This method also offers the possibility of coprocessing multiple 2D spectra that have the signals at the same frequencies but with different intensities (e.g.: as a result of a variation in the mixing time). The processing can be carried out on the time-domain data, thus preserving the possibility of applying further processing to the data. As a demonstration, we have applied Cadzow denoising on the MCR-processed FIDs, achieving a further increase in the S/N ratio and more effective denoising also on the transients at longer indirect evolution times. We have applied the combined denoising on a set of experimental data from a lysozyme-silica composite.

Parkinson disease (PD) is the second most common neurodegenerative disease, and the most common synucleinopathy, as alpha-synuclein (α-syn), a prion-like protein, plays an important pathophysiologic role in its onset and progression. Although neuropathologic changes begin many years before the onset of motor manifestations, diagnosis still relies on the identification of the motor symptoms, which hinders to formulate an early diagnosis. Because α-syn misfolding and aggregation precede clinical manifestations, the possibility to identify these phenomena in patients with PD would allow us to recognize the disease at the earliest, premotor phases, as a consequence of the transition from a clinical to a molecular diagnosis. Seed amplification assays (SAAs) are a group of techniques that currently support the diagnosis of prion subacute encephalopathies, namely Creutzfeldt-Jakob disease. These techniques enable the detection of minimal amounts of prions in CSF and other matrices of affected patients. Recently, SAAs have been successfully applied to detect misfolded alpha-synuclein (α-syn) in CSF, olfactory mucosa, submandibular gland biopsies, skin, and saliva of patients with Parkinson disease (PD) and other synucleinopathies. In these categories, they can differentiate PD and dementia with Lewy bodies (DLBs) from control subjects, even in the prodromal stages of the disease. In differential diagnosis, SAAs satisfactorily differentiated PD, DLB, and multiple system atrophy (MSA) from nonsynucleinopathy parkinsonisms. The kinetic analysis of the SAA fluorescence profiles allowed the identification of synucleinopathy-dependent α-syn fibrils conformations, commonly referred to as strains, which have demonstrated diagnostic potential in differentiating among synucleinopathies, especially between Lewy body diseases (LBDs) (PD and DLB) and MSA. In front of these highly promising data, which make the α-syn seeding activity detected by SAAs as the most promising diagnostic biomarker for synucleinopathies, there are still preanalytical and analytical issues, mostly related to the assay standardization, which need to be solved. In this review, we discuss the key findings supporting the clinical application of α-syn SAAs to identify PD and other synucleinopathies, the unmet needs, and future perspectives.

Background: Phosphatidylethanolamine binding protein 1 (PEBP1) is a multifunctional protein, mainly known for its specific binding of phosphatidylethanolamine and the ability to suppress the Raf1-MAPK pathway. Its potential role as an Alzheimer’s disease (AD) biomarker has been proposed in several studies. However, evaluation of its discriminative value in clinical cohorts is missing. Objective: We aimed to develop a new immunoassay for the measurement of PEBP1 in cerebrospinal fluid (CSF) and assess the possible role of this protein as AD biomarker. Methods: We developed a sandwich enzyme-linked immunosorbent assay (ELISA) for detection of PEBP1 in CSF and performed a technical and a clinical validation on two well-characterized cohorts. The first cohort included 14 mild cognitive impairment due to AD (MCI-AD) and 11 other neurological diseases (OND) patients. The second, larger cohort, included 25 MCI-AD, 29 AD dementia (AD-dem), and 21 OND patients. Results: PEBP1 is highly sensitive to pre-analytical conditions, especially to prolonged storage at room temperature or 4°C. Analysis of the first cohort showed a trend of an increase of PEBP1 level in MCI-AD patients versus OND subjects. Analysis of the second cohort did not show significant differences among diagnostic groups. Weak, positive correlation was found between CSF PEBP1 and t-tau, p-tau, and Aβ40 in the AD-dem group. Conclusion: A novel ELISA for the detection of PEBP1 in CSF was developed. Further research is needed to assess the potential of PEBP1 in AD diagnostics. The observed dependence of the PEBP1 signal on operating procedures encourages its potential application as CSF quality control.

Introductionβ-synuclein (β-syn) is a presynaptic protein, whose cerebrospinal fluid (CSF) levels are increased in patients with Alzheimer’s diseases (AD) showing mild cognitive impairment (MCI) and dementia (dem). Here, we aimed to investigate CSF β-syn in subjects at different AD stages, including preclinical AD (pre-AD), and to compare its behaviour with another synaptic biomarker, α-synuclein (α-syn), and two biomarkers of neuro-axonal damage, namely neurofilament light chain protein (NfL) and total tau protein (t-tau).MethodsWe measured β-syn, α-syn, t-tau and NfL in CSF of 75 patients with AD (pre-AD n=17, MCI-AD n=28, dem-AD n=30) and 35 controls (subjective memory complaints, SMC-Ctrl n=13, non-degenerative neurological disorders, Dis-Ctrl n=22).ResultsCSF β-syn, α-syn, t-tau were significantly elevated in pre-AD patients compared with controls (pDiscussionCSF β-syn increases in the whole AD continuum since the preclinical stage and represents a promising biomarker of synaptic damage in AD.

The synuclein family includes three neuronal proteins, named α-synuclein, β-synuclein, and γ-synuclein, that have peculiar structural features. α-synuclein is largely known for being a key protein in the pathophysiology of Parkinson's disease (PD) and other synucleinopathies, namely, dementia with Lewy bodies and multisystem atrophy. The role of β-synuclein and γ-synuclein is less well understood in terms of physiological functions and potential contribution to human diseases. α-synuclein has been investigated extensively in both cerebrospinal fluid (CSF) and blood as a potential biomarker for synucleinopathies. Recently, great attention has been also paid to β-synuclein, whose CSF and blood levels seem to reflect synaptic damage and neurodegeneration independent of the presence of synucleinopathy. In this review, we aim to provide an overview on the pathophysiological roles of the synucleins. Because γ-synuclein has been poorly investigated in the field of synucleinopathy and its pathophysiological roles are far from being clear, we focus on the interactions between α-synuclein and β-synuclein in PD. We also discuss the role of α-synuclein and β-synuclein as potential biomarkers to improve the diagnostic characterization of synucleinopathies, thus highlighting their potential application in clinical trials for disease-modifying therapies. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Background: The prion-like misfolding and aggregation of α-synuclein (α-syn) is involved in the pathophysiology of Parkinson's disease and other synucleinopathies. Seed amplification assays (SAAs) are biophysical tools that take advantage on the peculiar properties of prion proteins by amplifying small amounts of aggregates in biological fluids at the expense of recombinant monomeric protein added in solution. SAAs have emerged as the most promising tools for the diagnosis of synucleinopathies in vivo. However, the diagnostic outcome of SAAs depends on the aggregation kinetics of α-syn, which in turn is influenced by several experimental variables. Methods: In our work, we analysed the impact on SAAs of some of the most critical experimental factors by considering models that describe the aggregation kinetics of α-syn. Results: We started our analysis by making simulations to understand which kinetic models could explain the aggregation kinetics of α-syn during incubation/shaking cycles. Subsequently, under shaking/incubation cycles similar to the ones commonly used in SAAs, we tested the influence of some analytical variables such as monomer concentration, presence/absence of glass beads, pH, addition of human cerebrospinal fluid, and use of detergents on α-syn aggregation. Conclusions: Our investigation highlighted how optimization and standardization of experimental procedures for α-syn SAAs is of utmost relevance for the ultimate goal of applying these assays in clinical routine. Although these aspects have been evaluated with specific SAA protocols, most of the experimental variables considered influenced very general aggregation mechanisms of α-syn, thus making most of the results obtained from our analyses extendable to other protocols.

The EuroCirCol project is part of the Future Circular Collider (FCC) study under the European Union leadership in the framework of a H2020 project. In particular, the Italian Institute for Nuclear Physics, in collaboration with CERN and other European laboratories, is developing the design of a cos theta ${{\rm Nb}_{3}{\rm Sn}}$ dipole magnet which will be part of the Conceptual Design Report of the FCC studies in 2019. The magnet, with an aperture diameter of 50 mm and a bore field of 16 T, will be able to bend the beams at final energies and within collider size constraints. Here we present an update of the electromagnetic design of the ${{\rm Nb}_{3}{\rm Sn}}$ cos theta dipole in double-aperture configuration, the 2-D mechanical analysis, and also the 3-D coil-ends study.

The High Luminosity LHC (HL-LHC) is a program to upgrade the performance of the collider in order to achieve, in the ATLAS and CMS collision regions, instantaneous luminosities a factor of five larger than the actual nominal value. To obtain this result, all the magnets in the collision region will have to be substituted, together with other important devices. In the framework of this program, the Italian National Institute for Nuclear Physics (INFN-LASA laboratory) is entrusted to develop five corrector magnets (skew quadrupole, sextupole, octupole, decapole, and dodecapole), which in the magnet chain will have to correct the magnetic imperfections of the main magnets in the regions (low-beta quadrupoles Q1/Q3 and dipole D1/D2). These corrector magnets have been designed with relatively simple, robust, and easy to construct concepts, using superferric configuration. The first prototypes, a sextupole, an octupole, and a decapole, have been tested in 2016 and 2017. We present here the prototype of dodecapole. We report magnetic and mechanical designs and the experience during construction. The qualification, the cold test, and the quench behavior in operational conditions are also described.

SerpinA1 (α1-antitrypsin) is a soluble glycoprotein, the cerebrospinal fluid (CSF) isoforms of which showed disease-specific changes in neurodegenerative disorders that are still unexplored in Alz-heimer’s disease (AD). By means of capillary isoelectric focusing immunoassay, we investigated six serpinA1 isoforms in CSF samples of controls (n = 29), AD-MCI (n = 29), AD-dem (n = 26) and Lewy body disease (LBD, n = 59) patients and correlated the findings with CSF AD core biomarkers (Aβ42/40 ratio, p-tau, t-tau). Four CSF serpinA1 isoforms were differently expressed in AD patients compared to controls and LBD patients, especially isoforms 2 and 4. AD-specific changes were found since the MCI stage and significantly correlated with decreased Aβ42/40 (p < 0.05) and in-creased p-tau and t-tau levels in CSF (p < 0.001). Analysis of serpinA1 isoform provided good di-agnostic accuracy in discriminating AD patients versus controls (AUC = 0.80) and versus LBD patients (AUC = 0.92), with best results in patients in the dementia stage (AUC = 0.97). SerpinA1 isoform expression is altered in AD patients, suggesting a common, albeit disease-specific, in-volvement of serpinA1 in most neurodegenerative disorders.

Amyloid-β (Aβ) 1-40 and 1-42 peptides are key mediators of synaptic and cognitive dysfunction in Alzheimer’s disease (AD). Whereas in AD, Aβ is found to act as a pro-epileptogenic factor even before plaque formation, amyloid pathology has been detected among patients with epilepsy with increased risk of developing AD. Among Aβ aggregated species, soluble oligomers are suggested to be responsible for most of Aβ’s toxic effects. Aβ oligomers exert extracellular and intracellular toxicity through different mechanisms, including interaction with membrane receptors and the formation of ion-permeable channels in cellular membranes. These damages, linked to an unbalance between excitatory and inhibitory neurotransmission, often result in neuronal hyperexcitability and neural circuit dysfunction, which in turn increase Aβ deposition and facilitate neurodegeneration, resulting in an Aβ-driven vicious loop. In this review, we summarize the most representative literature on the effects that oligomeric Aβ induces on synaptic dysfunction and network disorganization.

INFN is developing at the LASA lab (Milano, Italy) the High Order (HO) corrector magnets for the High Luminosity-LHC (HL-LHC) project, which will equip the interaction regions. Five prototypes, from skew quadrupole to dodecapole, have been designed and tested at LASA. All the HO correctors are based on a superferric design, which allows a relatively simple, modular, and easy to construct magnet. This modularity has been exploited for an engineering change request. The tradeoff between safe handling, force management during cooldown, powering and protection of the magnet is explained in detail. The design of the coils and the mechanical assembly have been improved to increase the efficiency of the series production. Most of the optimizations are aimed to guarantee both a good integration in the LHC framework (i.e., radiation hardness, easiness of installation, safe operation over years), and compatibility with the series production (i.e., standardization of procedures and components for different magnets, reduction of the number of components, standardization of the quality control systems and acceptance tests). All of the produced magnets will be tested at LASA and then delivered to CERN. The design of the test-bed and the optimization of cryogenic and testing procedures is also described in this paper. Following the completion of the prototyping phase, we report in detail the solutions chosen for the HO correctors and we give a perspective of the series construction in industry and acceptance tests at LASA.

Cerebrospinal fluid (CSF) contains different types of extracellular vesicles (EVs) with undisclosed biomarker potential for neurodegenerative diseases. The aims of the present study were: i) to compare the proteome EVs isolated using different ultracentrifugation speed ii) to preliminary explore the EVs proteome in a common neurodegenerative disorder, Alzheimer’s disease (AD) compared to neurological controls. CSF samples from control subjects and AD patients were pooled separately (15 mL) and subjected to ultracentrifugation (UC) at different speeds (20,000g and 100,000g) to isolate separate EV fractions (P20 and P100). The proteome was analysed using high-resolution mass spectrometry (LC-MS/MS) and comparisons were made using bioinformatic analysis. EVs isolated at 100,000g (P100) had a proteome consistent with vesicles secreted via an ESCRT-dependent mechanism, being highly enriched in alix (PDCD6IP), syntenin-1 (SDCBP) and TSG101. EVs isolated at 20,000g were substantially different, showing enrichment in cytoskeletal and cell adhesion molecules. The pools from patients diagnosed with AD showed a distinct protein profile of CSF EVs, with increased levels of ADAM10, SPON1, CH3IL1 and MDK in the P100 fraction. CSF EV offer a new potential biosource of protein markers for AD detection and a complementary framework to the analysis of whole biological fluids for biomarker discovery.

Background: Conspicuous concentric ring artifacts in phantom reconstructions triggers retuning SPECT systems. These evaluations are visual, not quantitative. Our study was undertaken to determine the degree to which observers agree about SPECT concentric ring artifacts, and to test whether quantitative texture analysis metrics correspond to significant artifacts.Methods: Test data were acquired as part of quarterly quality assurance using standardized SPECT phantoms containing solid spheres, solid rods and volumes of uniform activity concentration loaded with 99mTc. Forty SPECT studies were identified as having concentric ring artifacts or were acquired to assess whether artifacts were resolved following camera retuning after obtaining an unacceptably non-uniform result. Transaxial reconstructions were reviewed independently by two medical physicists who graded severity of artifacts on a 5-point scale. Counts were tabulated in volumes of interest created in uniform phantom sections, from which were computed 72 radiomics image texture analysis metrics. Radial contrast (RContrast) derived from the radial profile of summed slices transformed into polar coordinates and radial noise-to-signal (RNSR) also calculated. Results: Artifacts were considered sufficiently severe to require camera retuning in 10 rods sections, 17 sphere sections, and 16 uniform sections. In uniform sections, there was “good agreement” for inter-observer and intra-rater assessments (κ = 0.66, Fisher exact p < 0.0001 and κ = 0.61, Fisher exact p = 0.001, respectively). While 3 radiomics image analysis features agreed significantly (p = 0.001) with visual detection of significant artifacts in uniform sections, the parameters most strongly associated with severe artifacts were RContrast > 4.75% and RNSR > 2.7%, for which ROC AUC accuracy = 88%±5%, sensitivity = 83%, specificity = 83%, p < 0.0001. Accuracy was 76%-78% for the 3 radiomics features, with significantly lower specificity (48%-61%, p < 0.05) than RContrast and RNSR. Increasing magnitude of RContrast and RNSR correlated significantly with increasingly severe artifact scores (ρ = 0.71-0.72, p < 0.0001). Conclusion: There was good agreement among physicists as to the presence of circular ring artifacts in uniform sections of SPECT quality assurance scans, with artifacts accurately detected by radial contrast and noise-to-signal ratio measurements.

INFN is developing at LASA Laboratory (Milano, Italy) the prototypes of five corrector magnets, from skew quadrupole to dodecapole, which will equip the high-luminosity interaction regions (IRs) of the High Luminosity-LHC (HL-LHC). These magnets are based on a superferric design, to allow a relatively simple, modular, and easy to construct magnet. This program takes place within the framework of a collaboration agreement between CERN and INFN. In this paper, we present an overview of the activity, from the design, to the construction and test at the operation condition.

INFN is developing at LASA lab (INFN Milano, Italy) the prototypes of five corrector magnets, from skew quadrupole to dodecapole, which will equip the high-luminosity interaction regions of the High Luminosity-LHC. These magnets are based on a superferric design, which allows a relatively simple, modular, and easy to construct magnet. This activity takes place within the framework of a collaboration agreement between CERN and INFN. The first prototype, a sextupole, has been assembled in early 2016. We present here the results of the cold test, including the training performed at 4.2 K and the qualification at 2.2 K. We report also on the overall experience gained during its construction and test. An important aspect is represented by the analysis of the manufacturing tolerances of the mechanical pieces and the alignment accuracy achieved. These results will serve as a basis for the design of the next magnets, the octupole, and the decapole. Other important results concern the cold performance of the coil-to-coil electrical joints (especially critical in a magnet with a large number of coils), dynamic inductance measurements and quench studies.

Neuropathological investigations report that in synucleinopathies with dementia, namely Parkinson&rsquo, s disease (PD) with dementia (PDD) and dementia with Lewy bodies (DLB), the histopathological hallmarks of Alzheimer&rsquo, s Disease (AD), in particular amyloid plaques, are frequently observed. In this study, we investigated the cerebrospinal fluid (CSF) AD biomarkers in different clinical phenotypes of synucleinopathies. CSF A&beta, 42/A&beta, 40 ratio, phosphorylated tau and total tau were measured as markers of amyloidosis (A), tauopathy (T) and neurodegeneration (N) respectively, in 98 PD (48 with mild cognitive impairment, PD-MCI, 50 cognitively unimpaired, PD-nMCI), 14 PDD and 15 DLB patients, and 48 neurological controls (CTRL). In our study, CSF AD biomarkers did not significantly differ between CTRL, PD-MCI and PD-nMCI patients. In PD-nMCI and PD-MCI groups, A-/T-/N- profile was the most represented. Prevalence of A+ was similar in PD-nMCI and PD-MCI (10% and 13%, respectively), being higher in PDD (64%) and in DLB (73%). DLB showed the lowest values of A&beta, 40 ratio. Higher total tau at baseline predicted a worse neuropsychological outcome after one year in PD-MCI. A+/T+, i.e., AD-like CSF profile, was most frequent in the DLB group (40% vs. 29% in PDD).

Neuroinflammatory and neurodegenerative diseases are characterized by the interplay of a number of molecular pathways that can be assessed through biofluids, especially cerebrospinal fluid and blood. Accordingly, the definition and classification of these disorders will move from clinical and pathological to biological criteria. The consequences of this biomarker-based diagnostic and prognostic approach are highly relevant to the field of drug development. Indeed, in view of the availability of disease-modifying drugs, fluid biomarkers offer a unique opportunity for improving the quality and applicability of results from clinical trials. Herein, we discuss the benefits of using fluid biomarkers for patient stratification, target engagement, and outcome assessment, as well as the most recent developments in neuroinflammatory and neurodegenerative diseases.

Background Cerebrospinal fluid (CSF) amyloid-beta (Aβ) 42/40 ratio, threonine-181-phosphorylated-tau (p-tau), and total-tau (t-tau) represent core biomarkers of Alzheimer disease (AD). The recent availability of automated platforms has represented a significant achievement for reducing the pre-analytical variability of these determinations in clinical setting. With respect to classical manual ELISAs, these platforms give us also the possibility to measure any single sample and to get the result within approximately 30 min. So far, reference values have been calculated from measurements obtained in frozen samples. In this work, we wanted to check if the values obtained in fresh CSF samples differ from those obtained in frozen samples, since this issue is mandatory in routine diagnostic work. Methods Fifty-eight consecutive CSF samples have been analyzed immediately after lumbar puncture and after 1-month deep freezing (− 80 °C). As an automated platform, we used Lumipulse G600-II (Fujirebio Inc.). Both the fresh and the frozen aliquots were analyzed in their storage tubes. Results In fresh samples, a mean increase of Aβ40 (6%), Aβ42 (2%), p-tau (2%), and t-tau (4%) was observed as compared to frozen samples, whereas a slight decrease was observed for Aβ42/Aβ40 ratio (4%), due to the higher deviation of Aβ40 in fresh samples compared to Aβ42. These differences are significant for Aβ40, Aβ42/Aβ40 ratio, p-tau, and t-tau. Nevertheless, the Aβ42/Aβ40 ratio showed a lower variability (smaller standard deviation of relative differences) with respect to Aβ42. With respect to the AD profile according to the A/T/(N) criteria for AD diagnosis, no significant changes in classification were observed when comparing results obtained in fresh vs frozen samples. Conclusions Small but significant differences have been found for Aβ40, Aβ42/Aβ40 ratio, p-tau, and t-tau in fresh vs frozen samples. Importantly, these differences did not imply a modification in the A/T/(N) classification system. In order to know if different cutoffs for fresh and frozen samples are required, larger, multi-center investigations are needed.

The Italian Institute for Nuclear Physics (INFN), in collaboration with CERN, is going to build the short model in Nb $_3$ Sn of the main bending dipole for the hadron-hadron Future Circular Collider (hh-FCC). The magnet will be developed on the basis of the baseline design presented in the FCC Conceptual Design Report (CDR) in the end of 2018. In particular, it will be based on cosine-theta design, with an internal aperture diameter of 50 mm and a Bladder & Key configuration for the mechanics. The main purpose of the model construction is to demonstrate the feasibility of a magnet dipole with field quality characteristic suitable for a collider and magnetic field above the LHC frontier.The mechanical structure, which is a critical aspect of the magnet design, especially for the brittleness of the Nb $_3$ Sn cables, will have to demonstrate the effectiveness to reach the highest performance achievable in terms of bore magnetic field. Here we present both the electromagnetic and mechanical design study of the model.