TBPH/TDP-43 modulates translation of Drosophila futsch mRNA through an UG-rich sequence within its 5'UTR.

Abstract Nuclear factor TDP-43 is an evolutionarily conserved multifunctional RNA-binding protein associated with frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). In recent years, Drosophila models of ALS based on TDP-43 knockdown/overexpression have allowed to find several connections with disease. Among these, we have previously described that silencing the expression of its fly ortholog (TBPH) can alter the expression of the neuronal microtubule-associated protein Futsch leading to alterations of neuromuscular junction (NMJ) organization. In particular, TBPH knocked out flies displayed a significant reduction of Futsch protein levels, although minimal variation in the futsch mRNA content was observed. These conclusions were recently validated in an independent study. Together, these observations strongly support the hypothesis that TBPH might regulate the translation of futsch mRNA. However, the mechanism of TBPH interference in futsch mRNA translation is still unknown. In this work, we use EMSA experiments coupled with RNA-protein co-immunprecipitations and luciferase assays to show that TBPH interacts with a stretch of UG within the 5′UTR of futsch mRNA and translation is positively modulated by this binding. Most importantly, this function is also conserved in human TDP-43. This result can therefore represent the first step in elucidating the relationship between TDP-43, protein translation, and eventual disease onset or progression. This article is part of a Special Issue entitled SI:RNA Metabolism in Disease

Drosophila Answers to TDP-43 Proteinopathies

Initially implicated in the pathogenesis of CFTR and HIV-1 transcription, nuclear factor TDP-43 was subsequently found to be involved in the origin and development of several neurodegenerative diseases. In 2006, in fact, it was reported for the first time the cytoplasmic accumulation of TDP-43 in ubiquitin-positive inclusions of ALS and FTLD patients, suggesting the presence of a shared underlying mechanism for these diseases. Today, different animal models of TDP-43 proteinopathies are available in rodents, nematodes, fishes, and flies. Although these models recapitulate several of the pathological features found in patients, the mechanisms underpinning the progressive neuronal loss observed in TDP-43 proteinopathies remain to be characterized. Compared to other models, Drosophila are appealing because they combine the presence of a sophisticated brain with the possibility to investigate quickly and massively phenotypic genetic modifiers as well as possible therapeutic strategies. At present, the development of TDP-43-related Drosophila models has further strengthened the hypothesis that both TDP-43 “loss-of-function” and “gain-of-function” mechanisms can contribute to disease. The aim of this paper is to describe and compare the results obtained in a series of transgenic and knockout flies, along with the information they have generated, towards a better understanding of the mechanisms underlying TDP-43 proteinopathies

Climate change and water abstraction impacts on the long-term variability of water levels in Lake Bracciano (Central Italy): A Random Forest approach

Abstract Study Region Lake Bracciano has been historically used as a strategic water reservoir for the city of Rome (Italy) since ancient times. However, following the severe water crisis of 2017, water abstraction has been completely stopped. Study Focus The relative impact of the various drivers of change (climatological and management) on fluctuations in lake water level is not yet clear. To quantify this impact, we applied the Random Forest (RF) machine learning approach, taking advantage of a century of observations. New Hydrological Insights for the Region Since the late 1990s the monthly variation in lake water levels has doubled, as has variation in monthly abstraction. Increased variation in annual cumulated precipitation and a rise in mean air temperature have also been observed. The RF machine learning approach made it possible to confirm the marginal role of temperature, the increasing role of abstraction during the last two decades (from 24 % to 39 %), and the key role played by the increased precipitation variability. These results highlight the notable prediction and inference capabilities of RF in a complex and partially unknown hydrological context. We conclude by discussing the limits of this approach, which are mainly associated with its capacity to generates scenarios compared to physical based models

Indirect monitoring and early detection of faults in trains motors

This paper investigates the ability of temperature sensors installed in the traction core of trains to early detect incipient faults. For instance, the breaking of a bearing is known to be critical as it may cause an increase of the temperature in the motor compartment, that in turn may eventually lead to a winding fault in the induction motor. The technique proposed in this contribution is characterised by extreme generality, since most frequent incipient faults lead to temperature increase that, if properly analyzed, can be a tool for preventive maintenance. In particular, the measured data, provided by the main Italian railway company, are processed by two different methodologies which are characterized by positive, yet different, performances. The results show that preventive maintenance with the proposed approach is feasibl

Oxy-turbine for Power Plant with CO2Capture

The IEA Greenhouse Gas R&D (IEAGHG) programme contracted Amec Foster Wheeler to perform a study providing an evaluation of the performance and costs of a number of oxy-turbine plants for utility scale power generation with CO2capture. The main outcomes of the detailed technical and economical modelling of the most promising oxy-turbine cycles is presented in this paper, including sensitivity analyses on main technical and financial parameters. Each cycle configuration and optimization is developed jointly with the main cycle developers, i.e. Clean Energy Systems, Graz University of Technology and NET Power. The modelling of the gas turbine, including efficiency and blade cooling requirement, have been performed using a calculation code developed by Politecnico di Milano

Chronological requirements of TDP-43 function in synaptic organization and locomotive control.

Abstract Alterations in TDP-43 are commonly found in patients suffering from amyotrophic lateral sclerosis (ALS) and the genetic suppression of the conserved homologue in Drosophila (TBPH) provokes alterations in the functional organization of motoneuron synaptic terminals, resulting in locomotive defects and reduced life span. To gain more insight into this pathological process, it is of fundamental importance to establish when during the fly life cycle the lack of TBPH affects motoneuron activity and whether this is a reversible phenomenon. To achieve this, we conditionally expressed the endogenous protein in TBPH minus Drosophila neurons and found that TBPH is a short lived protein permanently required for Drosophila motility and synaptic assembly through the direct modulation of vesicular proteins, such as Syntaxin 1A, indicating that synaptic transmission defects are early pathological consequences of TBPH dysfunction in vivo. Importantly, TBPH late induction is able to recover synaptogenesis and locomotion in adult flies revealing an unexpected late-stage functional and structural neuronal plasticity. These observations suggest that late therapeutic approaches based on TDP-43 functionality may also be successful for the human pathology

Depletion of TDP 43 overrides the need for exonic and intronic splicing enhancers in the human apoA-II gene

Exon 3 of the human apolipoprotein A-II (apoA-II) gene is efficiently included in the mRNA although its acceptor site is significantly weak because of a peculiar (GU)(16) tract instead of a canonical polypyrimidine tract within the intron 2/exon 3 junction. Our previous studies demonstrated that the SR proteins ASF/SF2 and SC35 bind specifically an exonic splicing enhancer (ESE) within exon 3 and promote exon 3 splicing. In the present study, we show that the ESE is necessary only in the proper context. In addition, we have characterized two novel sequences in the flanking introns that modulate apoA-II exon 3 splicing. There is a G-rich element in intron 2 that interacts with hnRNPH1 and inhibits exon 3 splicing. The second is a purine rich region in intron 3 that binds SRp40 and SRp55 and promotes exon 3 inclusion in mRNA. We have also found that the (GU) repeats in the apoA-II context bind the splicing factor TDP-43 and interfere with exon 3 definition. Significantly, blocking of TDP-43 expression by small interfering RNA overrides the need for all the other cis-acting elements making exon 3 inclusion constitutive even in the presence of disrupted exonic and intronic enhancers. Altogether, our results suggest that exonic and intronic enhancers have evolved to balance the negative effects of the two silencers located in intron 2 and hence rescue the constitutive exon 3 inclusion in apoA-II mRNA

An Open Architecture for Signal Monitoring and Recording Based on SDR and Docker Containers: A GNSS Use Case

Signal monitoring and recording station architectures based on software-defined radio (SDR) have been proposed and implemented since several years. However, the large amount of data to be transferred, stored, and managed when high sampling frequency and high quantization depth are required, poses a limit to the performance, mostly because of the data losses during the data transfer between the front-end and the storage unit. To overcome these limitations, thus allowing a reliable, high-fidelity recording of the signals as required by some applications, a novel architecture named SMART (Signal Monitoring, Analysis and Recording Tool) based on the implementation of Docker containers directly on a Network Attached Storage (NAS) unit is presented. Such paradigms allow for a fully open-source system being more affordable and flexible than previous prototypes. The proposed architecture reduces computational complexity, increases efficiency, and provides a compact, cost-effective system that is easy to move and deploy. As a case study, this architecture is implemented to monitor Radio-Frequency Interferences (RFI) on Global Navigation Satellite System (GNSS) L1/E1 and L5/E5 bands. The sample results show the benefits of a stable, long-term capture at a high sampling frequency to characterize the RFIs spectral signature effectively

Major hnRNP proteins act as general TDP-43 functional modifiers both in Drosophila and human neuronal cells

Nuclear factor TDP-43 is known to play an important role in several neurodegenerative pathologies. In general, TDP-43 is an abundant protein within the eukaryotic nucleus that binds to many coding and non-coding RNAs and influence their processing. Using Drosophila, we have performed a functional screening to establish the ability of major hnRNP proteins to affect TDP-43 overexpression/depletion phenotypes. Interestingly, we observed that lowering hnRNP and TDP-43 expression has a generally harmful effect on flies locomotor abilities. In parallel, our study has also identified a distinct set of hnRNPs that is capable of powerfully rescuing TDP-43 toxicity in the fly eye (Hrb27c, CG42458, Glo and Syp). Most importantly, removing the human orthologs of Hrb27c (DAZAP1) in human neuronal cell lines can correct several pre-mRNA splicing events altered by TDP-43 depletion. Moreover, using RNA sequencing analysis we show that DAZAP1 and TDP-43 can co-regulate an extensive number of biological processes and molecular functions potentially important for the neuron/motor neuron pathophysiology. Our results suggest that changes in hnRNP expression levels can significantly modulate TDP-43 functions and affect pathological outcomes

Peripheral Biomarkers in Manifest and Premanifest Huntington’s Disease

Huntington's disease (HD) is characterized by clinical motor impairment (e.g., involuntary movements, poor coordination, parkinsonism), cognitive deficits, and psychiatric symptoms. An inhered expansion of the CAG triplet in the huntingtin gene causing a pathogenic gain-of-function of the mutant huntingtin (mHTT) protein has been identified. In this review, we focus on known biomarkers (e.g., mHTT, neurofilament light chains) and on new biofluid biomarkers that can be quantified in plasma or peripheral blood mononuclear cells from mHTT carriers. Circulating biomarkers may fill current unmet needs in HD management: better stratification of patients amenable to etiologic treatment; the initiation of preventive treatment in premanifest HD; and the identification of peripheral pathogenic central nervous system cascades