Molecular Dissection of TDP-43 as a Leading Cause of ALS/FTLD

TAR DNA binding protein 43 (TDP-43) is a DNA/RNA binding protein involved in pivotal cellular functions, especially in RNA metabolism. Hyperphosphorylated and ubiquitinated TDP-43-positive neuronal cytoplasmic inclusions are identified in the brain and spinal cord in most cases of amyotrophic lateral sclerosis (ALS) and a substantial proportion of frontotemporal lobar degeneration (FTLD) cases. TDP-43 dysfunctions and cytoplasmic aggregation seem to be the central pathogenicity in ALS and FTLD. Therefore, unraveling both the physiological and pathological mechanisms of TDP-43 may enable the exploration of novel therapeutic strategies. This review highlights the current understanding of TDP-43 biology and pathology, describing the cellular processes involved in the pathogeneses of ALS and FTLD, such as post-translational modifications, RNA metabolism, liquid–liquid phase separation, proteolysis, and the potential prion-like propagation propensity of the TDP-43 inclusions.     

Overview of the potent cyanobacterial neurotoxin β-methylamino-L-alanine (BMAA) and its analytical determination

Blue-green algae are responsible for the production of different types of toxins which can be neurotoxic, hepatotoxic, cytotoxic and dermatotoxic and that can affect both aquatic and terrestrial life. Since its discovery the neurotoxin β-methylamino-L-alanine (BMAA) has been a cause for concern, being associated with the neurodegenerative disease amyotrophic lateral sclerosis/Parkinsonism–dementia complex (ALS/PDC). The initial focus was on Guam where it was observed that a high number of people were affected by the ALS/PDC complex. Subsequently, researchers were surprised to find levels of BMAA in post mortem brains from Canadian patients who also suffered from ALS/PDC. Recent research demonstrates that BMAA has been found at different levels in the aquatic food web in the brackish waters of the Baltic Sea. There is emerging evidence to suggest that sand-borne algae from Qatar can also contain BMAA. Furthermore, there is now concern because BMAA has been found not only in warmer regions of the world but also in temperate regions like Europe. The aim of this review is to focus on the methods of extraction and analysis of the neurotoxic non-protein amino acid BMAA. We also consider the neurotoxicity, aetiology, and diverse sources and routes of exposure to BMAA. In recent years, different methods have been developed for the analysis of BMAA. Some of these use HPLC-FD, UPLC-UV, UPLC-MS and LC-MS/MS using samples that have been derivatised or underivatised. To date the LC-MS/MS approach is the most widely used analytical technique as it is the most selective and sensitive method for BMAA determination.     

Integral analysis of large LOCA beyond design basis accident scenario in RBMK-1500

The integral analysis of severe accident scenario for RBMK-1500 was performed using combined approach with RELAP5, RELAP/SCDAPSIM, ASTEC and COCOSYS codes. The performed analysis covered response of the reactor core, the reactor cooling system and the confinement. There were performed several analyses: the first analysis assumed that operators take no action or their actions are not successful to provide the coolant injection to the reactor core; the other analyses were performed to investigate the accident management measures to restore the core cooling at different temperatures of the reactor core. The results of performed analyses showed that the operators have ∼5 h before the ruptures of fuel claddings occur and ∼8 h before the onset of exothermic steam-zirconium reaction. The coolant injection to the reactor core should be restored as soon as possible in order to prevent high hydrogen concentrations in the confinement and significant release of the fission products to the environment.     

Extended HALS algorithm for nonnegative Tucker decomposition and its applications for multiway analysis and classification

Analysis of high dimensional data in modern applications, such as neuroscience, text mining, spectral analysis, chemometrices naturally requires tensor decomposition methods. The Tucker decompositions allow us to extract hidden factors (component matrices) with different dimension in each mode, and investigate interactions among various modalities. The alternating least squares (ALS) algorithms have been confirmed effective and efficient in most of tensor decompositions, especially Tucker with orthogonality constraints. However, for nonnegative Tucker decomposition (NTD), standard ALS algorithms suffer from unstable convergence properties, demand high computational cost for large scale problems due to matrix inverse, and often return suboptimal solutions. Moreover they are quite sensitive with respect to noise, and can be relatively slow in the special case when data are nearly collinear. In this paper, we propose a new algorithm for nonnegative Tucker decomposition based on constrained minimization of a set of local cost functions and hierarchical alternating least squares (HALS). The developed NTD-HALS algorithm sequentially updates components, hence avoids matrix inverse, and is suitable for large-scale problems. The proposed algorithm is also regularized with additional constraint terms such as sparseness, orthogonality, smoothness, and especially discriminant. Extensive experiments confirm the validity and higher performance of the developed algorithm in comparison with other existing algorithms.     

MPEG-4音频压缩技术的新进展--ALS

在语音信号处理的许多应用中,采用了无损音频压缩的方法.回顾了MPEG-4音频技术的现状,MPEG-4 ALS(Audio Lossless Coding)标准化过程以及编解码技术.详细介绍了编码器中的线性预测、量化、熵编码等模块,展望了MPEG-4 ALS的应用前景.     

Comparing airborne laser scanning-imagery fusion methods based on geometric accuracy in forested areas

In this study, different methods for fusing airborne laser scanning (ALS) and multispectral imagery were compared. The improvement observed when computing the position of individual ALS returns in the original unrectified scenes (back-projecting ALS) was contrasted against the geometric errors observed in many orthorectified images. Results showed that back-projecting the ALS is the most accurate technique for fusing high-resolution datasets. Potential sources of systematic errors were studied for all techniques, finding the nadir angle as the most influencing one. The capacity of back-projecting ALS and true orthorectification for reducing the tree leaning problem found in traditional orthophotos was evaluated. Modelled tree lean correlated with geometric errors even while individual factors alone did not. The technique used for true orthorectification solved tree leaning near nadir, but it was affected by the variance of the kriging model, resulting in a low overall accuracy. In view of the results observed in back-projected ALS, the spatial resolution of the imagery was regarded as its main source of uncertainty. The convenience of adding a correction of the effect of the atmospheric refraction and the Earth's curvature was also discussed. Even though the magnitude of those corrections was low, they succeeded in avoiding nadir angle-dependent systematic errors.     

Impaired 26S Proteasome Assembly Precedes Neuronal Loss in Mutant UBQLN2 Rats

Proteasomal dysfunction is known to be associated with amyotrophic lateral sclerosis and frontotemporal degeneration (ALS/FTD). Our previous reports have shown that a mutant form of ubiquilin-2 (UBQLN2) linked to ALS/FTD leads to neurodegeneration accompanied by accumulations of the proteasome subunit Rpt1 in transgenic rats, but the precise pathogenic mechanisms of how this mutation impairs the proteasome remains to be elucidated. Here, we reveal that this UBQLN2 mutation in rats disrupted the proteasome integrity prior to neurodegeneration, that it dissociated the 26S proteasome in vitro, and that its depletion did not affect 26S proteasome assembly. During both disease progression and in an age-dependent manner, we found that proteasome subunits were translocated to the nucleus, including both of the 20S core particles (PSMA1 and PSMB7) and the 19S regulatory particles (Rpt1 and Rpn1), suggesting that defective proteasome function may result from the proteasome-subunit mislocalization. Taken together, the present data demonstrate that impaired proteasome assembly is an early event in the pathogenesis of UBQLN2-associated neurodegeneration in mutant UBQLN2 rats.     

Mechanisms of TDP-43 Proteinopathy Onset and Propagation

TDP-43 is an RNA-binding protein that has been robustly linked to the pathogenesis of a number of neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal dementia. While mutations in the TARDBP gene that codes for the protein have been identified as causing disease in a small subset of patients, TDP-43 proteinopathy is present in the majority of cases regardless of mutation status. This raises key questions regarding the mechanisms by which TDP-43 proteinopathy arises and spreads throughout the central nervous system. Numerous studies have explored the role of a variety of cellular functions on the disease process, and nucleocytoplasmic transport, protein homeostasis, RNA interactions and cellular stress have all risen to the forefront as possible contributors to the initiation of TDP-43 pathogenesis. There is also a small but growing body of evidence suggesting that aggregation-prone TDP-43 can recruit physiological TDP-43, and be transmitted intercellularly, providing a mechanism whereby small-scale proteinopathy spreads from cell to cell, reflecting the spread of clinical symptoms observed in patients. This review will discuss the potential role of the aforementioned cellular functions in TDP-43 pathogenesis, and explore how aberrant pathology may spread, and result in a feed-forward cascade effect, leading to robust TDP-43 proteinopathy and disease.     

New Strategies to Overcome Present CRISPR/Cas9 Limitations in Apple and Pear: Efficient Dechimerization and Base Editing

Despite recent progress, the application of CRISPR/Cas9 in perennial plants still has many obstacles to overcome. Our previous results with CRISPR/Cas9 in apple and pear indicated the frequent production of phenotypic and genotypic chimeras, after editing of the phytoene desaturase (PDS) gene conferring albino phenotype. Therefore, our first objective was to determine if adding an adventitious regeneration step from leaves of the primary transgenic plants (T0) would allow a reduction in chimerism. Among hundreds of adventitious buds regenerated from a variegated T0 line, 89% were homogeneous albino. Furthermore, the analysis of the target zone sequences of twelve of these regenerated lines (RT0 for “regenerated T0” lines) indicated that 99% of the RT0 alleles were predicted to produce a truncated target protein and that 67% of RT0 plants had less heterogeneous editing profiles than the T0. Base editors are CRISPR/Cas9-derived new genome-editing tools that allow precise nucleotide substitutions without double-stranded breaks. Hence, our second goal was to demonstrate the feasibility of CRISPR/Cas9 base editing in apple and pear using two easily scorable genes: acetolactate synthase—ALS (conferring resistance to chlorsulfuron) and PDS. The two guide RNAs under MdU3 and MdU6 promoters were coupled into a cytidine base editor harboring a cytidine deaminase fused to a nickase Cas9. Using this vector; we induced C-to-T DNA substitutions in the target genes; leading to discrete variation in the amino-acid sequence and generating new alleles. By co-editing ALS and PDS genes; we successfully obtained chlorsulfuron resistant and albino lines in pear. Overall; our work indicates that a regeneration step can efficiently reduce the initial chimerism and could be coupled with the application of base editing to create accurate genome edits in perennial plants.