Muscle Proteomic Profile before and after Enzyme Replacement Therapy in Late-Onset Pompe Disease

Mutations in the acidic alpha-glucosidase (GAA) coding gene cause Pompe disease. Late-onset Pompe disease (LOPD) is characterized by progressive proximal and axial muscle weakness and atrophy, causing respiratory failure. Enzyme replacement therapy (ERT), based on recombinant human GAA infusions, is the only available treatment; however, the efficacy of ERT is variable. Here we address the question whether proteins at variance in LOPD muscle of patients before and after 1 year of ERT, compared withhealthy age-matched subjects (CTR), reveal a specific signature. Proteins extracted from skeletal muscle of LOPD patients and CTR were analyzed by combining gel based (two-dimensional difference gel electrophoresis) and label-free (liquid chromatography-mass spectrometry) proteomic approaches, and ingenuity pathway analysis. Upstream regulators targeting autophagy and lysosomal tethering were assessed by immunoblotting. 178 proteins were changed in abundance in LOPD patients, 47 of them recovered normal level after ERT. Defects in oxidative metabolism, muscle contractile protein regulation, cytoskeletal rearrangement, and membrane reorganization persisted. Metabolic changes, ER stress and UPR (unfolded protein response) contribute to muscle proteostasis dysregulation with active membrane remodeling (high levels of LC3BII/LC3BI) and accumulation of p62, suggesting imbalance in the autophagic process. Active lysosome biogenesis characterizes both LOPD PRE and POST, unparalleled by molecules involved in lysosome tethering (VAMP8, SNAP29, STX17, and GORASP2) and BNIP3. In conclusion this study reveals a specific signature that suggests ERT prolongation and molecular targets to ameliorate patient’s outcome.     

Indoor air quality in schools of a highly polluted south Mediterranean area

This study aimed at surveying lower secondary schools in southern Italy, in a highly polluted area. A community close to an industrial area and three villages in rural areas was investigated. Indoor temperature, relative humidity (RH), gaseous pollutants (CO₂ and NO₂), selected biological pollutants in indoor dust, and the indoor/outdoor mass concentration and elemental composition of PM_2.5 were ascertained. Temperature and RH were within, or close to, the comfort range, while CO₂ frequently exceeded the threshold of 1000 ppm, indicating inadequate air exchange rate. In all the classrooms, median NO₂ levels were above the WHO threshold value. Dermatophagoides p. allergen concentration was below the sensitizing threshold, while high endotoxin levels were detected in the classrooms, suggesting schools may produce significant risks of endotoxin exposure. Concentration and solubility of PM_2.5 elements were used to identify the sources of indoor particles. Indoor concentration of most elements was higher than outdoors. Resuspension was responsible for the indoor increase in soil components. For elements from industrial emission (Cd, Co, Ni, Pb, Sb, Tl, V), the indoor concentration depended on penetration from the outside. For these elements, differences in rural vs industrial concentrations were found, suggesting industrial sources may influence indoor air quality nearby schools.     

A Petri nets semantics for data flow networks

This work presents a truly concurrent operational semantics for nondeterministic data flow networks. We introduce a model, the df-process, which is a notion similar to that of non-sequential process for a Petri net: a df-process is defined as a mapping from an occurrence net K to a data flow net N, such that the places and the transitions of K are mapped onto the channels and the nodes of N. A df-process contains, by means of some labelling of the places, information on the value and the order in which data flow through the channels during a computation. Df-processes for a data flow network are characterized in an abstract way by a set of properties and in general a df-process corresponds to a set of computations of the network. We give a way to build the df-process corresponding to a computation incrementally at each event occurrence: a main result of the paper is that the incremental construction yields exactly the same set of df-processes as abstractly defined. We also show that df-processes are compositional. The model is intended to be used by distributed systems designers: it contains enough information to be a guideline for the designer and to be a base to develop dynamic checking tools. We outline how df-processes can be used in the design phase of a system.Research supported in part by Progetto Coordinato CNR ANATRA     

Dexamethasone abolishes the activation by nerve growth factor of protein kinase N: effects of nerve growth factor and dexamethasone on protein kinase N

Post-operative therapeutic rehabilitation in ligamentous-capsular injuries has a great importance and for the final result it is as the very operation. We begin it from making the patient realize that the good final result can be obtained only with patient, persistence and discipline. Early therapeutic rehabilitation after surgical treatment of ligamentous-capsular injuries is possible only when the ligament was reconstructed in a motor stable way. Painless, dosed, passive motion exercises with a limited range of movements did on a mechanical splint TELOS have a great importance for the final results.     

Characterization of an ecto-phosphorylated protein of cultured cerebellar granule neurons

Previous work identified the phosphorylation by extracellular ATP of an endogenous 45-kDa protein substrate and established the presence of ecto-protein kinase activity associated with cultured cerebellar granule neurons (Volonté et al.: J Neurochem 63:2028-2037, 1994). In this work, we characterize such ecto-phosphorylated 45-kDa protein substrate and its association with the cellular membrane. The total radioactive content of the 45-kDa protein is stable for the first 15 min after phosphorylation, and decreases about 70% in 30 min and 90% in approximately 2 hr. Rinsing the cells after the phosphorylating reaction causes a 50% removal of the incorporated radioactivity. Glycosidic residues are present on the 45-kDa ecto-protein, which is held in position on the cellular membrane through a specific glycosyl-phosphatidylinositol anchor. The extracellular incorporation of phosphate on the 45-kDa protein is not modulated by agents interfering with cytoskeleton stability, such as colchicine and taxol, or by gangliosides. The extracellular phosphorylation occurs mostly on serine residues, since the phosphate ester linkage is unstable at high pH and only antibodies raised against phosphoserine are capable of recognizing the 45-kDa ecto-protein.     

A dirichlet process elaboration diagnostic for binomial goodness of fit

Useful model checking tools can be constructed by measuring the distance between a prior distribution that concentrates most of its mass around a model of interest, and the resulting posterior distribution. In this paper we use this approach to construct a diagnostic measure for detecting lack of fit in discrete data, with special focus on binomial data. We begin by constructing a suitable probability model “around” the model of interest, via a Dirichlet Process elaboration. We derive the resulting diagnostic and show that, approximately, it is the sum of two terms: the first is the logarithm of the Bayes factor and the second is proportional to the Pearson chi-square statistics. We give details of a simulation algorithm for computing the diagnostic and illustrate its use in an application to biomedical data.     

Efficiency,effectiveness, and impacts assessment in the rail transport sector: a state‐of‐the‐art critical analysis of current research

This paper aims at being a comprehensive reference for stakeholders, policy makers, and scholars interested in analyzing the problem of efficiency, effectiveness, and impacts of rail transport systems in a sound empirical way, paying specific attention to passenger transport services. The paper combines different analytical frameworks (engineering, economics, impacts), systematic review techniques, and advanced mappings. Framing economic efficiency studies into a transport planning perspective permits to move from efficiency to effectiveness issues. In addition, including impacts offers a critical discussion of the existing empirical studies, relating them to the main methodological approaches used. This analysis can be useful for those interested in developing new techniques for the evaluation of this sector. The critical analysis developed in this paper provides a catalog of inputs, outputs, external factors, possible impacts to account for, data, and approaches, which allows us to identify areas in which new methodological developments, new approaches, are needed to address the relevant societal challenges of the rail transport sector.     

Verifying data secure flow in AUTOSAR models

This paper presents an approach for enhancing the design phase of AUTOSAR models when security annotations are required. The approach is based on information flow analysis and abstract interpretation. The analysis evaluates the correctness of the model by assessing if the flow of data is secure with respect to causal data dependencies within the model. To find these dependencies an exhaustive search through the model would be required. Abstract interpretation is used as a trade-off between the precision and complexity of the analysis. The approach also provides annotated models without oversizing the set of annotations.     

SRAM-Based FPGA Systems for Safety-Critical Applications：A Survey on Design Standards and Proposed Methodologies

As the ASIC design cost becomes affordable only for very large-scale productions, the FPGA technology is currently becoming the leading technology for those applications that require a small-scale prod...