Characterization of protein complexes associated with TRP channels in the context of nociception

The transient receptor potential A1 (TRPA1) channel is essential for vertebrate pain. TRPA1 plays a fundamental role as a primary detector of noxious stimuli of physical and chemical nature, and is critically involved in different pain states. Even though TRPA1 activation modalities have been studied extensively, the network of protein interactions regulating TRPA1 (the so-called TRPA1 interactome) is only poorly understood. Considering the crucial role of TRPA1 in pain signaling, it is mandatory to shed light on the elusive molecular machinery regulating TRPA1 channels in sensory neurons. This project was therefore aimed at getting insights into the mechanisms of TRPA1 regulation by identifying TRPA1-protein complexes and characterizing their biological meaning in the context of nociception. A mass spectrometry-based proteomics approach led to the discovery of the physical association of Annexin A2 (AnxA2) with native TRPA1 in mouse sensory neurons. AnxA2 is enriched in a subpopulation of sensory neurons and coexpressed with TRPA1. Furthermore, we observed an increase of TRPA1 membrane levels in cultured sensory neurons from AnxA2-deficient mice. Functional studies suggest that AnxA2 limits the availability and consequently the activity of TRPA1 channels at the plasma membrane of sensory neurons. Moreover, our in vitro observations were reflected in enhanced nocifensive responses of AnxA2 knock-out mice specifically upon TRPA1 activation in vivo. TRPA1 channels have been shown to contribute to hypersensitivity to cold stimuli during inflammation. We therefore investigated the possible impact of AnxA2 on TRPA1 function also in this context employing the well-established complete Freund´s adjuvant (CFA) model of persistent inflammation in mice. AnxA2-deficient mice showed enhanced hypersensitivity on a cold plate upon CFA injection, whereas the hypersensitivity to heat and mechanical stimulation were not affected. In conclusion, we characterized AnxA2 as a novel TRPA1-associated protein which specifically regulates TRPA1 channels in vitro and in vivo. These findings pave the way for a more thorough investigation of the dynamic changes in TRPA1-associated protein complexes in different conditions. In this context we submitted mice to the established Complete Freund’s Adjuvant (CFA)-model of inflammatory pain and isolated TRPA1-protein complexes from sensory neurons. Samples were submitted to state-of-the-art quantitative mass spectrometry analysis to compare complexes between CFA and control mice and therefore identify dynamic changes in TRPA1 interactome. This work revealed a dramatic alteration of TRPA1-associated protein complexes in the context of inflammatory pain, and the functional characterization of selected candidates that will follow this study might uncover new players in the development and maintenance of inflammatory pain. In a separate set of experiments we focused our attention on a protein called 4-Nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1 (NIPSNAP1), which function is largely still unknown. NIPSNAP1 showed to be expressed in nociceptive neurons of mouse dorsal root ganglia and functional studies suggested that it can modulate TRPA1 expression and/or function upon overexpression. A role of NIPSNAP1 in nociceptive transmission has been recently uncovered by the identification of its physical association with Nocistatin, a neuropeptide involved in pain signaling. In this context we investigated Nocistatin and found that it enhances TRPA1-mediated cellular calcium responses in DRG neurons. Mechanistically, this effect does not seem to be mediated by a modulation of TRPA1 plasma membrane expression, and NIPSNAP1 knock-down did not affect it. Future studies will contribute to characterize the underlying mechanisms and also the potential relevance for TRPA1-mediated nociception in vivo

Effects of bus-based disruptive business models with limited capacity on rail monopolies: Social welfare implications

Long distance passenger transport markets are facing important changes as new entrants, e-Platform based bus services retailer (PBSR) operators, are challenging the railway incumbents applying judo economic strategies. Traditionally, European policymakers tended to favour railway services over road services in the long-haul markets, often leading the rail operators in monopolistic-alike positions. Recently, several countries deregulated their national intercity bus markets, gradually introducing intermodal competition in the sector. The competition led to important improvements in service quality, but it also had negative impacts on rail operators’ profitability, especially after PBSR operators started to work, due to their disruptive business model based on aggregative online platforms and production externalization. PBSR companies (e.g. Flixbus, BlaBlaBus) are characterized by high flexibility and low production costs, which use as advantage against the incumbents. The rail operators are instead characterized by high indivisibility, high production costs and, usually, big sizes. Losses in either revenues or market shares could easily force them into reducing services quantity or even exit the market. Our paper aims to analyse these new competitive relations in the intercity intermodal market, focusing on resulting impacts on market shares, demand satisfaction and social welfare. Since the bus operators present limited capacity due to technical feasibility (e.g. minimum headway) and the need to limit road congestion (to preserve service quality), the mobility right fulfilment is put in jeopardy. We modelled the competitive relations through game theory, excluding high speed rail from the perimeter to preserve service comparability. Profit levels and optimal social welfare are then studied through simulations. Results confirm that for increasing PBSR production capacity, railway operators tend to have fewer profits or be forced to leave the market, resulting in unsatisfied demand. Furthermore, from a social point of view, the rail monopoly seems to be, under specific circumstances, preferred to a duopoly

Determinants of the incidence of non-academic staff in European and US HEIs

In this article, we contribute to the scant literature covering quantitative studies on the determinants of the non-academic staff incidence in higher education institutions by analysing how the proportion of non-academic staff is related to key features such as size, prestige, year of foundation and financial structure of universities. We apply nonlinear regression analysis to compare HEIs across Europe and the USA, taking into account time and cross-country heterogeneity of the two balanced panel datasets concerning European and American universities over a period of 6 years (2011–2016 for Europe and 2012–2017 for the USA). Evidence suggests that in both Europe and the USA, public and larger (if sufficiently large) as well as more research-oriented units are characterised by a higher proportion of non-academic staff. In Europe, we observe an inverted U-shaped effect of the share of non-personnel expenditure and the foundation year on the proportion of non-academic staff, while the proportion of non-academic staff decreases with the share of core and third-party funding. For the USA, we obtain similar findings except that the share of core funding and third-party funding is characterised by a U-shaped effect, and the impact of the share of non-personnel expenditure has no empirical effect on the proportion of non-academic staff. Additionally, we discover that some factors that contribute to the proportion of non-academic staff may constitute indicators of performance, suggesting the need for further research to extend our knowledge on the complex issue of the role played by non-academic staff in university performance

Strategic formation and welfare effects of airline-high speed rail agreements

Policy makers encourage airline-high speed rail (HSR) cooperation to promote intermodal passenger transport. We study the strategic formation of airline-HSR partnerships (depending on sunk costs and firms’ bargaining power) and their effects on consumer surplus and social welfare. We assume that airline-HSR agreements serve to offer a bundle of domestic HSR and international air services. In a capacity purchase (CP) agreement, the airline buys train seats to sell the bundle, whereas in a joint venture (JV) agreement firms create a distinct business unit. We find that both agreements increase traffic in the network, and thereby may not reduce congestion at hub airports. We provide antitrust authorities with a simple two-tier test for the CP agreement to improve consumer surplus. Contrary to airline-HSR mergers, the JV agreement benefits consumers independent of hub congestion and mode substitution. Simulation results show that, in case of cooperation, public agencies should prefer firms to create a JV, unless the related sunk costs are far greater than the costs of the CP agreement

Who is at Risk of Parkinson Disease? Refining the Preclinical Phase of GBA1 and LRRK2 Variant Carriers: a Clinical, Biochemical, and Imaging Approach

Purpose of Review: Genetic variants in GBA1 and LRRK2 genes are the commonest genetic risk factor for Parkinson disease (PD); however, the preclinical profile of GBA1 and LRRK2 variant carriers who will develop PD is unclear. This review aims to highlight the more sensitive markers that can stratify PD risk in non-manifesting GBA1 and LRRK2 variant carriers. Recent Findings: Several case–control and a few longitudinal studies evaluated clinical, biochemical, and neuroimaging markers within cohorts of non-manifesting carriers of GBA1 and LRRK2 variants. Summary: Despite similar levels of penetrance of PD in GBA1 and LRRK2 variant carriers (10–30%), these individuals have distinct preclinical profiles. GBA1 variant carriers at higher risk of PD can present with prodromal symptoms suggestive of PD (hyposmia), display increased α-synuclein levels in peripheral blood mononuclear cells, and show dopamine transporter abnormalities. LRRK2 variant carriers at higher risk of PD might show subtle motor abnormalities, but no prodromal symptoms, higher exposure to some environmental factors (non-steroid anti-inflammatory drugs), and peripheral inflammatory profile. This information will help clinicians tailor appropriate screening tests and counseling and facilitate researchers in the development of predictive markers, disease-modifying treatments, and selection of healthy individuals who might benefit from preventive interventions

A proxy cost model for tramway services

In this paper, we build a proxy cost model for tramway services. we estimate separately: (i) transport services production costs; (ii) infrastructure costs; (iii) maintenance costs; (iv) administrative and general costs and (v) the cost of capital. we apply the proposed methodology to estimate the standard cost of italian tramway services. detailed data about costs, technical and environmental characteristics were collected by means of questionnaires sent to italian companies providing 100% of tramway services in 2012. we perform a simulation study in order to highlight the marginal impact of efficiency gains obtained by manipulating cost-driving variables both under the control of the operators (trains and drivers productivity) and of the local authority who assigns the service (number of train revenue kilometers (trK) assigned within the service contract, average fleet age). the simulations show how the local authority should allocate extra resources if it wants to increase the quality-quantity mix of tramway services. our results might help the decision-maker to define the maximum economic compensation (auction base) in competitive tendering procedures or a benchmark for the bargaining with the local monopolist

The remote assessment of parkinsonism supporting ongoing development of interventions in Gaucher disease

Mutations in GBA which are causative of Gaucher disease in their biallelic form, are the most common genetic risk factor for Parkinson's disease (PD). The diagnosis of PD relies upon clinically defined motor features which appear after irreversible neurodegeneration. Prodromal symptoms of PD may provide a means to predict latent pathology, years before the onset of motor features. Previous work has reported prodromal features of PD in GBA mutation carriers, however this has been insufficiently sensitive to identify those that will develop PD. The Remote Assessment of Parkinsonism Supporting Ongoing Development of Interventions in Gaucher Disease (RAPSODI GD) study assesses a large cohort of GBA mutation carriers, to aid development of procedures for earlier diagnosis of PD

STANDARD COST OF ITALIAN METRO SERVICES: THE INFLUENCE OF AUTOMATISM, WHEELS TECHNOLOGY AND CAPACITY

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Airway stenting in a child with spondyloepiphyseal dysplasia congenita: 13-Year survival.

We describe the case of a boy with spondyloepiphyseal dysplasia congenita. At birth, he experienced severe respiratory distress necessitating tracheotomy. Endoscopy done because mechanical ventilation failed to resolve desaturations disclosed severe tracheo-bronchomalacia. A Polyflex silicone stent was placed in the trachea (replaced by Y-Dumon stent) and 2 Palmaz metallic stents in the mainstem bronchi (overlapped with 2 Jomed stents 5 years later). Airway stenting guaranteed a suitable respiratory status and allowed a child who was expected to die at birth, to reach 13.5 years old in good conditions